Abstracts of CSCC Conference 2016

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 P101 Analysis of Point of Care (POC) glucose test utilization patterns for hospital inpatients using centralized POC data management software Julie Shaw, The Ottawa Hospital, The University of Ottawa and EORLA Introduction: Point of care (POC) glucose monitoring is widely used in hospitals to facilitate rapid treatment of fluctuating blood glucose concentrations in patients. The Ottawa Hospital (TOH) is a 1117-bed academic health centre, located in Eastern Ontario, which includes the University of Ottawa Heart Institute. TOH employs use of the Roche Inform II glucose meter in the POC environment. Approximately 350 meters are in-use across three campuses and all meters are connected centrally to Roche Cobas IT 1000 data management software. Inpatient POC glucose results are automatically transmitted to the electronic patient record from Cobas IT 1000 via the laboratory information system. Methods: All inpatient POC glucose results were mined in Cobas IT 1000 from a one week period, September 23-29, 2015, to analyze utilization patterns. Results: Approximately 8000 POC glucose tests were performed during the one-week period examined across all inpatient units, including the emergency departments. A summary of POCT glucose utilization for the highest volume clinical areas is shown in Table 1. Discussion: POCT device connectivity to centralized data management software allows for data mining to examine POCT utilization patterns. These data can be shared with the clinical areas to help educate and identify inappropriate test utilization.

P102 Calculation of pediatric reference intervals for electrolytes on the Ortho Vitros 5.1 FS and Siemens Vista 1500 platforms Julie Shaw, The Ottawa Hospital, The University of Ottawa and EORLA Christopher McCudden, The Ottawa Hospital, EORLA, University of Ottawa Nathalie Lepage, Children’s Hospital of Eastern Ontario, Ottawa Khosrow Adeli, Clinical Biochemistry, The Hospital for Sick Children, Toronto Introduction: Clinicians rely upon accurate reference intervals for interpretation of laboratory test results. Reference interval establishment studies are difficult, time consuming and expensive, particularly for pediatrics. As such, accurate pediatric reference intervals are lacking. CALIPER is a Canadian national initiative aimed at calculating pediatric reference intervals for an extensive number of analytes. Here, pediatric reference intervals were calculated for serum sodium, potassium and chloride measured on the Ortho Vitros 5.1 FS and Siemens Vista 1500 chemistry platforms. Methods: Healthy children (n=207) were recruited as part of the CALIPER study. Written, informed consent and a health questionnaire were obtained from each participant and a blood sample collected. Sodium, potassium and chloride were measured by the Ortho Vitros 5.1 FS and Siemens Vista 1500 methods. Hemolysis was checked by HIL indice measurement

on the Vista 1500. Reference intervals were calculated according to CLSI guidelines (C28-A3), although the number of samples per partition was sometimes lower than 120. Results: Calculated pediatric reference intervals and confidence intervals are shown in Table 1. Discussion: None of the electrolytes studied showed gender-related differences. Sodium did not show any agedifferences. Potassium and chloride concentrations were found to be higher in children less than 1 year of age.

P103 A quality perspective on disclosure of medical errors Jawahar (Jay) Kalra, University of Saskatchewan Ashish Kopargaonkar, University of Saskatchewan Objective: The quality of healthcare is an emerging concern worldwide. Disclosure of an adverse event is an important element in managing the consequences of a medical error. We have previously reported a non-punitive, “no-fault” model for reporting medical and clinical errors. The objective of this study was to review and compare the disclosure policies recommended by various provincial colleges of physicians and surgeons in Canada. Methods: We reviewed and compared various error disclosure initiatives that are in practice in Canada to analyze the progress made in this area. A letter was sent to the Registrars of the College of Physician and Surgeons of all provinces in Canada, requesting their medical error/ adverse event disclosure policies. Results: Designing of an error disclosure policy requires integration of various aspects including bioethics, physicianpatient communication, quality of care, and team-based care delivery. The data from 13 College of physicians and surgeons of Canada was received, collated and analyzed. Many of these disclosure policies are in practice or they are mandated in the code of ethics. These Canadian provincial initiatives, though similar in content, remain isolated because of their nonmandatory nature and absence of federal or provincial laws on disclosure. Conclusions: The complexities of medical error disclosure to patients present ideal opportunities for medical educators to probe how learners are balancing the ethical complexities involved in error disclosure with other related fields. We suggest that a uniform policy centered on addressing errors in a non-punitive manner and respecting the patient’s right to an honest disclosure be a standard of care. P104 Role of advanced glycation end products (AGEs) and its receptor in the pathogenesis and complication of hyperthyroidism in Graves’ disease Mabood Qureshi, Department of Pathology and Laboratory Medicine, Royal University Hospital Background: Oxidative stress has been implicated in the pathogenesis of hyperthyroidism and its complications. Interaction of advanced glycation end products (AGEs) with its cell receptor RAGE (receptor for AGEs), generates reactive

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 oxygen species (ROS). Circulating soluble receptor for AGEs (sRAGE) competes with RAGE for binding with AGEs and reduces generation of ROS. Objectives: It is hypothesized that low levels of sRAGE and high levels of AGEs would increase the generation of ROS culminating in development of hyperthyroidism and its complication. The objective is to investigate if serum levels of sRAGE are lower, and the levels of AGEs and AGEs/sRAGE ratio are higher in hyperthyroidism associated with Graves’ disease compared to control subjects. Methods: The study population comprised of 23 patients of hyperthyroidism associated with Graves’ disease and 20 control subjects. Blood samples were collected for the measurement of serum sRAGE and AGEs using commercially available enzyme linked immune assay (ELISA) kits. Results: The Serum levels of AGEs in patients were significantly higher compared to controls (694481.3 ± 39903.4 vs 476700 ± 96549.8 pg/mL). The serum levels of sRAGE were lower in patients compared to controls but the values were not significantly different from each other (1295.8 ±128.6 vs 1418.04 ± 86.62 pg/mL). The AGEs/sRAGE ratio were significantly greater in patients with Graves’ disease as compared to control ( 719.9 ± 97.53 vs 340.10 ± 70.08). Conclusions: In conclusion, high levels of serum AGEs and AGEs/sRAGE ratio may be a risk marker for hyperthyroidism and its complications in patients with Graves’ disease. P105 Continuous and partitioned reference intervals for antistreptolysin on the Siemens BNII nephelometer Christopher McCudden, The Ottawa Hospital, EORLA, University of Ottawa Julie Shaw, The Ottawa Hospital, The University of Ottawa and EORLA Nathalie Lepage, Children’s Hospital of Eastern Ontario, Ottawa Khosrow Adeli, Clinical Biochemistry, The Hospital for Sick Children Objectives: Anti-streptolysin O antibodies (ASO) are measured to detect past streptococcal infection, where exposure can cause serious, but avoidable complications including rheumatic fever, scarlet fever, and glomerulonephritis. The objective of this study was to establish pediatric reference intervals for ASO on the Siemens BNII System. Design and Methods: A total of 221 CALIPER initiative samples were included in the study. ASO was measured on the Siemens BNII (Ref method K7022) according to the manufacturer’s directions. Non-parametric partitioned reference intervals were calculated according to CLSI guidelines (C28-A3). In addition, continuous reference intervals were determined using a quadratic (2nd degree polynomial) quantile regression on Box-Cox power-transformed values; a 2nd degree polynomial was determined as the best (lowest AIC) of several different fits. For consistency with literature, the 90th percentile upper limit was selected for partitioned and continuous intervals. Results: ASO values increased with age. There was a significant, but small ( Conclusions: Partitioned and continuous reference intervals were largely consistent with published estimates. These cutoffs will be useful for assessing ASO values in a pediatric population on the Siemens BNII.

P106 Robustness of the Reichert Unistat Bilirubinometer for analysis of hemolyzed samples from neonates Christopher McCudden, The Ottawa Hospital, EORLA, University of Ottawa Michelle Warr, EORLA Kathy Fleming Objectives: Bilirubin is routinely measured in neonates to avoid the irreversible effects of kernicterus. Grossly hemolyzed are routinely rejected under the assumption that bilirubin cannot be accurately measured in hemolyzed samples. The objective of this study was to determine the effect of hemolysis on bilirubin measurement in neonates. Design/Methods: Data included 2-years of results (n=70) where there were hemolyzed and non-hemolyzed samples measured within 6hrs of each other. Bilirubin results were compared by Passing-Bablock linear regression and difference plots. Bilirubin results were also compared using the Bhutani nomogram to determine if hemolysis affected the hyperbilirubinemia risk-category. Results: Gross hemolysis resulted in a mean negative bias of -5.2 umol/L (95% CI: -30.7 to 20.3 umol/L). Based on the Bhutani nomogram, 1/70 samples would have been classified as Low-intermediate instead of High-intermediate risk and 3/70 would have been classified as Low instead of Lowintermediate risk; 5/70 samples would have been classified as Low-intermediate instead of Low risk. Conclusions: Collectively the data support that neonatal bilirubin may be reported from grossly hemolyzed samples measured using the Reichert Unistat Bilirubinometer. This practice has been adopted at TOH where results are reported with comments describing the effects of hemolysis. Implementation of this approach has decreased the number of blood re-collections in neonates.

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 P107 Development and validation of a LC-MS/MS method for salicylate analysis Lufang Yang, Cathy Rawluk, Gordon Ball, Gordon Hoag, Department of Laboratory Medicine, Pathology and Medical Genetics, Island Health, Victoria BC Objective: To develop a clinically robust, rapid and reliable LC-MS/MS method for salicylate analysis. Design and Methods: Salicylate was extracted with 1:10 (v:v) methanol containing the internal standard (IS) 6methoxysalicylic acid. The supernatant was diluted with water and analyzed on a Shimadzu Prominence HPLC and AB SCIEX 4000 QTRAP mass spectrometer with electrospray ionization in negative polarity. The transitions used for the detection of salicylate and the IS were 137 > 93 and 167 >123, respectively. The salicylate was separated using gradient elution at 30 °C with a 0.5ml/min flow rate. This method was evaluated for precision, accuracy, linearity, limit of quantification, ion suppression, and interference, followed by correlation to the Trinder-based DRI® salicylate assay from Thermo Scientific using patient and Bio-Rad EQA samples. Results: Total run time was two minutes. Imprecision in two levels of QC was less than 7%. Linearity was within the range 0.06 –5.79 mmol/L. The limit of quantification was 0.05 mmol/L. No ion suppression effects were observed. LCMS/MS results correlated well with DRI (R² > 0.95), but showed that the DRI produced higher values for salicylate levels by 7.5% for the 12 EQA samples, by 23% for the 20 patient samples with normal indices at level > 1.0 mmol/L. The salicylate levels of 30 samples with abnormal lipemic or hemolysis indices were undetectable in LC-MS/MS but had the range 0.5 – 4.2 mmol/L in DRI. Conclusion: This simple, highly selective, and sensitive LC-MS/MS method is an alternative method for salicylate levels assessment for clinical specimens with abnormal indices. P108 Evaluation of the Accuracy of Friedewald-Estimation LDL Cholesterol at Clinically Relevant Very Low Levels on Architect C16000 Amy Lou, IWK Health Centre, Halifax, NS Manal elnenaei, IWK Health Centre, Halifax, NS Bassam Nassar, IWK Health Centre, Halifax, NS Objectives: Guidelines recommend target levels of LDLC of < 2.0 mmol/L for patients who are at an intermediate- or high risk for cardiovascular disease. Moreover, patients on lipid-lowering agents, such as PCSK9 inhibitors may be expected to have LDL-C levels even lower than 1.5 mmol/L. Questions have arisen about the accuracy of the Friedwald formula in calculating LDL-C at such low values . Our goal was therefore to evaluate the accuracy of Friedewald-estimated LDL-C (LDLf-C), compared with directly-measured LDL-C (LDLd-C) at very low LDL-C . Design and Methods: Autorules were employed to filter results of LDLf-C Results: Results of LDLf-C did not show significant differences from those of LDLd-C with the mean difference of 0.015 mmol/L (P= 0.46). The Passing-Bablok fit was Y=0.97x+0.09, indicating a positive bias of 3% from LDLf-C at LDL-d level of 1.5 mmol/L and 6% bias at LDL-d level of 1 mmol/L. Thirty samples were identified with LDL-d Conclusion: LDLf-C shows slight overestimation of LDLC when compared to LDLd-C for LDL-C. P109 Analytical evaluation of the Roche automated electrochemiluminescence immunoassay for cyclosporine and tacrolimus

Michael Knauer, Department of Laboratory Medicine and Pathobiology, University of Toronto Ivan Blasutig, University Health Network, Toronto David Colantonio, The Hospital for Sick Kids, Toronto Vathany Kulasingam, University Health Network, Toronto Objectives: To evaluate the analytical performance of the semi-automated Roche Elecsys immunoassays for cyclosporine (CSA) and tacrolimus (TAC) on the Roche e411 platform. Design and Methods: Precision was evaluated using manufacturer and BioRad Whole Blood Immunosuppressant controls. Linearity across the measuring range was assessed using CAP Linearity Survey material. Method comparison studies comparing Roche e411 with Abbott Architect and LCMS/MS was performed, along with analytical sensitivity and lotto-lot assessment. Results: Precision ranged from 3.4 to 8.0% for CSA and 4.1 to 9.9% for TAC. Linearity was verified from 48.0 μg/L to 960.9 μg/L for CSA and from 1.4 μg/L to 27.1 μg/L for TAC. The functional sensitivity was determined to be 44 μg/L for CSA and 0.7 μg/L for TAC (CV ≤ 20%). Deming regression analysis of the Abbott Architect method comparison (n = 102) yielded slopes of 0.92 (95%CI: 0.89-0.95) for CSA and 0.92 (95%CI: 0.88-0.97) for TAC. Deming regression analysis of the LC–MS/MS method comparison (n = 20) yielded slopes of 1.33 (95%CI: 1.17-1.50) for CSA and 0.92 (95%CI: 0.84-1.01) for TAC. Lot-to-Lot comparison (n = 20) yielded slopes of 0.998 (95%CI: 0.97-1.03) for CSA and 0.97 (95%CI: 0.94-1.01) for TAC. Conclusions: The Roche Elecsys CSA and TAC assays have acceptable precision, linearity, and functional sensitivity and are comparable to Abbott Architect and LC–MS/MS methods. P110 Anti-nuclear antibody testing by indirect immunofluorescence and a multiplexed immunoassay: Clinical performance of discordant results against chart review by a Rheumatologist Ivan Blasutig, University Health Network, Toronto, ON Kevin Gorsky Irvin Bromberg, Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, ON Barry Hoffman, Mount Sinai Hospital, Toronto, ON Shikha Mittoo, Mount Sinai Hospital, Toronto, ON Amanda Steiman Background: Indirect immunofluorescence (IFA) is considered the gold standard for detecting anti-nuclear antibodies (ANA) and many clinicians and laboratorians are wary of screening with more convenient non-IFA immunoassays. Objective and Study Design: The aim of the study was to assess the clinical performance of IFA and Bio-Rad ® BioPlex 2200 ANA assay in samples submitted by a Rheumatology service. Results from specimens tested st simultaneously by both IFA and BioPlex from May 1 2011 to th April 30 2012 were analyzed for concordance. Patients with discordant results were subjected to an objective chart review by a single Rheumatologist to identify the presence of connective tissue disease (CTD; subdivided into those meeting classification criteria and those with incomplete–but clinically suggestive–presentations). Results: A total of 1,206 samples were tested by both methods and 920 samples were concordant (76.3%; 654 + + IFA /BioPlex and 266 IFA /BioPlex ). Of the 286 samples with discordant results, 230 were from patients visiting a Rheumatologist and were subjected to chart review. The BioPlex was clinically concordant in 100 (43.5%) of these:

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 +

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39/160 (24.4%) of the IFA /BioPlex results and 61/70 (87.1%) + of the IFA /BioPlex results. For these 230 samples, the sensitivity and specificity at the time of the discordant result was 34% and 81% for the BioPlex and 67% and 19% for IFA compared to the clinician’s overall impression. Conclusions: IFA and BioPlex assays disagreed in less than one quarter of the samples tested by both methods. In the patients with discordant result, the two assays demonstrated about equal clinical performance. P111 Marked Influence of Body Mass Index (BMI) on biochemical markers in the CALIPER cohort of healthy children and adolescents Victoria Higgins, CALIPER Program, Divisions of Clinical Biochemistry and Hematopathology, Pediatric Laboratory Medicine, The Hospital for Sick Children, University of Toronto Michelle Niewesteeg Kian Gordanifar, SickKids Hospital, Toronto Rathena Maheshan, SickKids Research Institute, Toronto Jobin Simon, SickKids Hospital, Toronto Kimia Khoee, SickKids Hospital, Toronto Mankhun Chan, The Hospital for Sick Children, Toronto Khosrow Adeli, Clinical Biochemistry, The Hospital for Sick Children, Toronto Objectives: Reference intervals, essential to accurately interpret laboratory tests, are severely lacking in pediatrics, potentially causing misdiagnosis. The CALIPER project established the first age- and sex-specific Canadian pediatric reference interval database (www.caliperproject.ca) to address this gap. Body mass index (BMI) is another key covariate that may significantly affect analyte levels. Here we determine the effect of BMI on several biochemical markers in a healthy pediatric population. Design and Methods: Biochemical markers were measured in the healthy CALIPER cohort (n=681-998) and levels compared between normal weight (NW), overweight (OW) and obese (OB) children using one-way ANOVA and Bonferroni’s Post-Hoc test. Independent Sample T-Test compared NW and OW/OB. Results: OW/OB adolescent males had higher ALT and ferritin, and lower HDL-C than NW. Triglycerides, apoB, and CRP were elevated in OW/OB adolescents, although more pronounced in males. Vitamin B12, C3, and C4 were elevated in OW/OB adolescents compared to NW. OW/OB children had higher triglycerides and C3 than NW. Conclusions: Increased triglycerides and apoB, and decreased HDL-C in OW/OB adolescents suggest lipid abnormalities manifest prior to developing insulin resistance. Elevated inflammatory proteins, C3, C4, and CRP suggest chronic low-grade inflammation in OW/OB children/adolescents. Lower vitamin B12 in OW/OB adolescents could result in hyperhomocysteinemia and increased CVD risk. Thus, these biomarker levels could be monitored in OW/OB children/adolescents to identify early risk factors for T2D and CVD. Therefore, reference intervals for these markers should be partitioned by BMI or OW/OB subjects should be excluded if increased levels predict disease development. P112 Development and validation of a spectrophotometric porphyrin screening method using statistics sensitive nonlinear iterative peak-clipping. Matthew P.A. Henderson, Children's Hospital of Eastern Ontario, Ottawa Objectives: The objective of this study was automation and standardization of data analysis required for urine and

fecal porphyrin screening using automated background subtraction and classification of results. Design and Methods: Statistics sensitive non-linear iterative peak-clipping (SNIP) was used to automate porphyrin spectra background subtraction prior to quantitation (Ryan et al., 1988). Optimization of the SNIP algorithm window size and number of iterations was performed via simulation. The results of SNIP based quantitation were compared to three-point drop line back-ground subtraction in which base line and maximum points are selected manually (Allen and Rieman, 1953). An application was written in ANSI Common Lisp to automate data analysis, classification and report generation. Results: SNIP based porphyrin screening was validated according to CLSI EP-15. The precision of the field (fecal CV = 8.2, urine CV = 8.6) and test (fecal CV = 6.1 , urine CV = 5.69) methods are comparable, however automated background subtraction removes inter-operator subjectivity when analyzing patient samples. Linear regression equations from the urine (y=0.81x-3.2) and fecal (y=0.78x+1.08) method comparison studies were used to transfer screening thresholds. Conclusions: Automation of data analysis, classification and report generation for urine and fecal porphyrin spectrophotometric screening removes subjectivity and improves productivity.

P114 Multi-cohort evaluation of Extractable Nuclear Antigen (ENA) and Anti-Nuclear Antibody (ANA) testing methods for autoimmune disease diagnosis Alex Chin, Calgary Laboratory Services Dennis Orton, Calgary Laboratory Services Marvin J. Fritzler, Mitogen Advanced Diagnostics Laboratory, Cumming School of Medicine Haiyan Hou, Mitogen Advanced Diagnostics Laboratory, Cumming School of Medicine Ivan Blasutig, University Health Network Objectives: Laboratory investigations of autoimmune disease routinely include anti-nuclear antibody (ANA) and extractable nuclear antigen (ENA) autoantibody analyses. This study compared results from three facilities utilizing distinct methods for measuring ANA and ENA to characterize differences between the assay platforms. Design and Methods: 100 patient serum samples were analyzed for ANA by indirect immunofluorescence (IFA) by two methods (Euroimmun and Inova) at different sites, and by multiplex bead-based assays (BioPlex and TheraDiag/FIDIS) at three different sites. Results were interpreted as positive/negative (concordance) or by staining pattern for ANA. ENA results were interpreted both quantitatively (between BioPlex instruments) and qualitatively (between BioPlex and TheraDiag/FIDIS). Results: Comparison of the ANA screening results from the BioPlex instruments employing 44 of the 100 patient samples showed good concordance (95.5%) and quantitative 2 correlation (slopes ranging 0.82 – 0.98, R values between 0.840 – 0.998 for ENA antigens). Conversely, BioPlex and IFA (Euroimmun) methods showed relatively poor concordance at 61.0 %. Despite this, IFA methods showed good agreement for staining pattern (80.8 %). ENA antigens compared between BioPlex and TheraDiag/FIDIS demonstrated good, although varied, concordance for the various analytes (81 – 97 %). The differences between BioPlex and Theradiag/FIDIS were attributed to differences in autoantigen preparations. Conclusions: This study demonstrated some differences between platforms for ENA and ANA immunoassays. BioPlex instruments were consistent between sites; however IFA and ENA methods from different yielded dissimilar results, attributed to a lack of standardization and autoantigen

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 preparations between vendors. Overall, laboratory results should be interpreted in the context of the clinical picture. P115 Setting a reference interval for aldosterone-renin ratio using direct renin concentration Alex Chin, Calgary Laboratory Services Dennis Orton, Calgary Laboratory Services Alexander A. Leung, Division of Endocrinology and Metabolism, Department of Medicine, Cumming School of Medicine, University of Calgary Gregory Kline, Division of Endocrinology and Metabolism, Department of Medicine, Cumming School of Medicine, University of Calgary S.M. Hossein Sadrzadeh, Calgary Laboratory Services Objectives: Methods used to derive the aldosterone– renin ratio (ARR), the standard screening test for primary aldosteronism (PA), generally employ plasma renin activity (PRA) or direct renin concentration (DRC). This study characterized DRC-ARR ranges appropriate for evaluation of PA using PRA-ARR as a benchmark. Design and Methods: Data was obtained from Calgary Laboratory Services (CLS) from January, 2010 to December, 2015. CLS converted to DRC from PRA in February, 2014, allowing access to data for both methods within a single population. Indirect comparison of the performance of DRCbased ARR and PRA thresholds was assessed using ROC curve analysis analyses using PRA-ARR values as the gold standard. Renin levels from each method were also evaluated for classification of “low renin”. Results: Results from 5864 patients were obtained (associated with 6074 PRA-ARRs and 1405 DRC-ARRs). The published PRA-ARR threshold of >550 pmol/L/ng/mL/h showed a prevalence of PA of 37.2%. Within the elevated PRA-ARR th patients, “low renin” was defined as the 95 percentile of the renin values obtained (25 pmol/LmIU/L the prevalence was 38.9% and “low renin” was Conclusions: A DRC-ARR threshold of >25 pmol/L/mIU/L performed similarly to the conventional PRA-ARR threshold of >550 pmol/L/ng/mL/h for PA. Additionally, DRC levels P116 CALIPER pediatric reference intervals for Ortho Vitros 5600 immunoassays Michelle Niewesteeg Victoria Higgins, CALIPER Program, Divisions of Clinical Biochemistry and Hematopathology, Pediatric Laboratory Medicine, The Hospital for Sick Children, University of Toronto Khosrow Adeli, Clinical Biochemistry, The Hospital for Sick Children, Toronto Mankhun Chan, The Hospital for Sick Children, Toronto Objectives: Accurate reference intervals based on a healthy pediatric population are essential for test result interpretation, since growth and development can significantly influence biomarker concentrations. The CALIPER (Canadian Laboratory Initiative on Pediatric Reference Intervals) project, a national research initiative, has made considerable strides to address the gaps in pediatric reference intervals. This current study expands the CALIPER database by establishing covariate-stratified reference intervals for Ortho Vitros 5600 immunoassays. Design and Methods: Healthy children and adolescents recruited for the CALIPER study completed a health questionnaire and provided a blood sample. We measured 28 biomarkers using the Ortho Vitros 5600 Immunoassay System, utilizing approximately 300 samples per assay. Analyte concentrations were visually inspected and statistically relevant age/sex- partitions were determined. After removing outliers,

age- and sex-specific reference intervals with corresponding 90% confidence intervals were calculated using CSLI C28-A3 guidelines. Results: AFP, prolactin, rubella IgG, and beta-hCG were fairly consistent over the pediatric age range, also requiring no further sex partitioning. Testosterone, progesterone, LH, FSH, ferritin, estradiol, and CEA all showed sex differences, the majority of which were seen after pubertal age. Conclusions: The complex expression profiles of 28 immunoassays were observed, allowing calculation of age- and sex-specific reference intervals for the Ortho Vitros 5600 Immunoassays. This will enable accurate diagnosis and laboratory assessment of children monitored by this immunoassays on the Ortho platform in health care institutions across Canada and worldwide. These reference intervals should be validated for each laboratory’s local pediatric population and specific analyzer as per CLSI guidelines. P117 Immunosuppressive drug monitoring in renal transplantation: comparison between Elecsys® immunoassays and high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Robert Robitaille, Service de biochimie, Hôpital MaisonneuveRosemont, CIUSSS Est-de-l’Île-de-Montréal Marie-Ève Gingras, Service de biochimie, Hôpital Maisonneuve-Rosemont, CIUSSS Est-de-l’Île-de-Montréal Chantal Cossette, Service de biochimie, Hôpital MaisonneuveRosemont, CIUSSS Est-de-l’Île-de-Montréal ®

Objectives: To compare the Elecsys immunoassays (Roche Diagnostics, Laval, QC) to the LC-MS/MS methods routinely used in a reference laboratory for immunosuppressive drug monitoring after kidney transplant. Design and Methods: The within-run CV (5 replicates twice a day) and between run CV (5 replicates twice a day for 5 days) were evaluated using three different levels of control material (PreciControl ISD, Roche Diagnostics, Laval, QC). Residual whole blood samples from 60 and 57 patients undergoing tacrolimus and cyclosporine therapy, respectively, ® were used to compare the Elecsys immunoassay methods to LC–MS/MS. Results: Within-run imprecision calculated for the tacrolimus immunoassay was 10.25% at 2.5 µg/L, 6.99% at 10 µg/L and 4.30% at 18 µg/L and the between run imprecision was 12.55% at 2.5 µg/L, 6.82% at 10 µg/L and 4.52% at 18 µg/L. For the cyclosporine immunoassay, within-run imprecision was 11.90% at 100 µg/L, 6.58% at 350 µg/L and 4.82% at 1250 µg/L and between run imprecision was 13.87% at 100 µg/L, 7.91% at 350 µg/L and 4.92% at 1250 µg/L. Passing-Bablok regression analysis of method comparison to LC–MS/MS yielded a slope of 1.17 (95%CI: 1.10,1.26), intercept of -0.22 (95%CI: ‑ 0.86,0.16) and R2 = 0.9168 for tacrolimus (range 1.8-16.1 µg/L) and a slope of 1.17 (95%CI: 1.12,1.23), intercept of ‑ 5.20 (95%CI: -34.94,11.54) and R2 = 0.9673 for cyclosporine (range 44-1173 µg/L). Conclusions: The proportional bias observed can be managed by reference value changes. The Elecsys® immunoassays meet the requirements for monitoring immunosuppressant concentrations in current regimens used for kidney transplantation. ® Keywords: Elecsys , tacrolimus, cyclosporine, kidney, transplantation P118 Natural killer cell activity (NKA) in vitro diagnostic device (IVDD) – a fast and simple method that brings NKA testing to the clinic Mathieu Provençal, Hôpital Maisonneuve-Rosemont Plante Geneviève, Hôpital Maisonneuve-Rosemont

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Background and Objectives. For several decades, patients with different types of cancers have been shown to have lower NKA compared to controls. To date, NKA has been 51 measured with the Research Use Only Cr or immunofluorescence-based cytotoxicity assays, requiring PBMC isolation. Herein, a novel, simple IVDD is described (NK ® Vue ; ATGen Canada). It uses standard blood collection tubes containing a proprietary formulation engineered to activate NK cells in whole blood. NKA is determined by the amount of interferon-gamma released into the plasma by ELISA after incubation. Assay reproducibility, sample stability, and precision of the quantitative ELISA test were evaluated. Methods. Blood was drawn from healthy donors, NK cells were activated, and NKA determined using NK Vue as per the manufacturer’s recommendations. Results. Plasma samples were stable at least 6 months at -80°C (p = 0.98) and interferon-gamma determination was reproducible (between-run precision of 6%). NKA and NKA per NK cell values were consistent for same-patient samples taken on three different days. Data from 40 healthy adults provided a potential reference range for NKA. Conclusions. The NK Vue tubes are an easy and efficient way to activate NK cells. Samples obtained with this IVDD were stable for a minium of 6 months frozen, and the provided ELISA assay offers a highly reproducible method. A preliminary NKA reference range in the normal Canadian population was obtained and could prove useful in patient assessment. Additional studies are required to optimize the ELISA part of the test on different automated platforms. P119 Pre-analytical storage conditions may affect Abbott ARCHITECT high-sensitivity cardiac troponin I results Terence Agbor, McMaster University Lorna Clarke, Hamilton Regional Laboratory Medicine Program Peter Kavsak, Juravinski Hospital & Cancer Centre, Core Lab Objectives: Laboratories receive requests, infrequently, to measure cardiac troponin (cTn) on previously stored patient samples to help determine myocardial injury onset in hospitalized patients. Lithium-heparin or K2EDTA plasma are often the sample types used for cTn testing. Our study objective was to investigate the effects of anticoagulants, the duration of sample storage, the presence/absence of cells and centrifugation on Abbott Diagnostics’ high-sensitivity cTnI (hscTnI) assay. Design and methods: After clinical testing, 10 random patients’ samples collected at the same time with sufficient volume for both K2EDTA and lithium-heparin plasma were selected and de-identified for this study. All samples were treated similarly before baseline hs-cTnI measurements. o Samples were then stored at 4 C on-cells or off-cells (plasma aliquoted into separate tubes) with hs-cTnI measured on day1&3 after refrigeration, and again on day3 after recentrifugation. Results: Two of the lithium-heparin plasma samples yielded significantly higher hs-cTnI results. The first sample; stored on-cells, hs-cTnI concentration increased from 1ng/L to 2066ng/L on day3 (by comparison, the aliquot hs-cTnI concentration was only 2ng/L on day3). The second lithiumheparin sample hs-cTnI concentration increased from 21ng/L to 491ng/L (stored on-cells) and from 20ng/L to 116ng/L (aliquot) after only 1day storage. Re-centrifugation of all samples yielded hs-cTnI concentrations equivalent to baseline. This effect was not observed in K2EDTA plasma. Conclusions: Pre-analytical variables including anticoagulant, duration and type of storage, presence of cells, and centrifugation may affect hs-cTnI results with K2EDTA plasma possibly being the preferred matrix over lithium-heparin

plasma. Further studies are required to fully characterize the effect of these pre-analytical variables. P120 Hyperammonemia resulting from inadequate specimen centrifugation and plasma contamination with platelets Lawrence de Koning, Calgary Laboratory Services Dennis Orton, Calgary Laboratory Services Jessica Gifford, Calgary Laboratory Services Isolde Seiden-Long, Calgary Laboratory Services Aneal Khan, Alberta Children's Hospital Background: A community physician contacted the laboratory after observing hyperammonemia (>47 µmol/L) in several asymptomatic patients. Specimens exhibited slight turbidity (elevated L-index), however this was below the threshold for interference. No other laboratory-related errors were detected. The laboratory was later contacted by the metabolics service after several admissions for hyperammonemia, including one patient who was refractory to treatment. Further review revealed a laboratory centrifuge was set at 1200 RPM (~300 × g) instead of 1200 × g. Methods: The centrifuge setting was immediately corrected. To evaluate whether this was the proximal cause, EDTA-whole blood was collected from 10 healthy volunteers and separated at 1200 RPM and 1200 × g. Plasma was tested for complete blood count (CBC), L-index and ammonia. Root cause analysis was applied to determine why the centrifuge settings were changed. Results: Patient ammonia concentrations normalized after the centrifuge change. Plasma separated at 1200 RPM had on average 2.9X higher ammonia, 11X higher platelets, 9 and a 10X higher L-index (71 µmol/L, 261 10 /L, 240) than 9 tubes spun at 1200 x g (25 µmol/L, 24 10 /L, 24). Ammonia 2 was strongly correlated with platelet count (R = 0.97). The root causes were identified as (1) a centrifuge speed change for bone-marrow processing, (2) a difficult to interpret centrifuge display, and (3) a lack of procedural clarity on centrifuge monitoring. Conclusions: We describe a case of inadequate centrifugation leading to plasma platelet contamination, and falsely-elevated ammonia concentrations. Adequate removal of platelets is critical for the validity of ammonia analysis. P121 Out of alignment? Optimization of thyroid test utilization through analysis of physician ordering patterns Jasmine Gill, University of Alberta Vilte E Barakauskas, Pathology and Laboratory Medicine, BC Children's Hospital Dylan Thomas, DynaLIFEDx Karina Rodriguez-Capote, DynaLIFEDx and the University of Alberta Trefor Higgins, DynaLIFEDx Amanda VanSpronsen, Laboratory Medicine and Pathology, University of Alberta Oksana Babenko , University of Alberta Roberta Martindale, University of Alberta Mathew Estey, DynaLIFEdx and the University of alberta Objectives: Thyroid-stimulating hormone (TSH) is the first-line test recommended for screening and monitoring thyroid abnormalities. At DynaLIFEDx laboratories, there are two TSH test codes. TSHB is a reflexive algorithm for initial investigation, where free T4 (FT4) and free T3 (FT3) are measured if necessary. TSHBO will test TSH only. Additionally, FT4 and FT3 can be ordered individually. The main aim of this study was to characterize thyroid test utilization in Northern Alberta and determine whether it aligns with the Alberta

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Toward Optimized Practice (TOP) guidelines. The validity of current TSH reflex cut-offs was also assessed. Design and Methods: In this retrospective study, thyroid test results from January 2014 to January 2015 were analyzed. Each pattern of thyroid test combinations was designated as appropriate or inappropriate according to TOP guidelines. Frequencies of each pattern were calculated. Receiver operating characteristic (ROC) curves were generated using MedCalc to determine optimal cut-offs for reflexing to FT4. Results: Of 759,822 specimens analyzed, 11.2% of all thyroid test utilization was found to be inappropriate. TSHBO was ordered for 381,917 (50.3%) specimens with 39,552 (10.4%) as inappropriate. TSHB was ordered for 370,719 (48.8%) with 41,951 (11.3%) as inappropriate. For 7186 specimens, FT4 and/or FT3 were ordered without TSH. FT4 and FT3 ordered with TSH accounted for 58% of all inappropriate orders. Conclusions: Despite well-defined clinical practice guidelines, inappropriate use of thyroid tests is still occurring. Future work will focus on elucidating the source of inappropriate requests. The reflex cut-offs currently in use were validated by ROC analysis. P122 The conundrum of evaluating lot change over multiple lots of reagent Waleed Alomaim, McMaster University Joceline Turner, Hamilton Regional Laboratory Medicine Program Patricia Hudecki, Hamilton Regional Laboratory Medicine Program Maria Howell, Hamilton Regional Laboratory Medicine Program Andrew Don-Wauchope, Hamilton Regional Laboratory Medicine Program Objectives: Reagent Lot-Lot drift should be monitored to minimise the opportunity for systematic bias. Most laboratories will have lot-to-lot change protocols that record the change from one lot to the next lot. There are suggestions of how to evaluate lot change in context of multiple previous lots. We investigated the proposed methods with a series of lot changes for our 25OH vitamin D (vitD) assay. Methods: We used five techniques to compare the lots of vitD reagents between December 2012 to January 2016. 1: Percentage bias and linear regression between consecutive lots (patient samples). 2: t-Test of data from Deming regression between accumulating lots (patient samples). 3: ANOVA on patient reported results for each lot of reagent. 4: Confidence interval (CI) calculations around the proportion of all patient results in each diagnostic category. 5: Mean and 95% CI of Quality Control(QC) data. Results: Twelve lots were evaluated in the time period. By assessment of linear regression one lot should be rejected. WDR assessment demonstrated cumulative change in the last two lots p one lot had significant change. QC data analysis confirmed this finding. Conclusion: Different reagent lot change evaluation methods provide discrepant information. Use of QC material may provide the best option for evaluating vitD lot drift retrospectively. P123 Prenatal carrier screening for thalassemia and hemoglobinopathy in Northern Alberta Tara Dixon, University of Alberta Mathew Estey, DynaLIFEdx and the University of alberta Don Zhang, DynaLIFEDx Dylan Thomas, DynaLIFEDx Trefor Higgins, DynaLIFEDx

Karina Rodriguez-Capote, DynaLIFEDx and the University of Alberta Introduction: Prenatal carrier screening for thalassemia and hemoglobinopathy is recommended by the Society of Obstetricians and Gynaecologists of Canada (SOGC) to provide prenatal diagnosis in couples with significant risk of an affected pregnancy. The aim of this study was to determine the incidence of thalassemia and hemoglobinopathy diagnosed with prenatal carrier screening in Northern Alberta. Methods: Retrospective analysis of structural Hb variants and thalassemias reported between February 1, 2013 to December 31, 2014 in patients of Edmonton-based obstetricians. Investigations included a CBC, ferritin, and Hb fractionation by HPLC (BioRad Variant II). Hb electrophoresis at alkaline and acid pH (Sebia Hydrasys) was conducted on specimens with an abnormal peak on HPLC. Detection of common α-thalassemia mutations and HbH disease was performed by GAP-PCR analysis (Calgary Laboratory Services). Results were extracted from the Laboratory Information System; statistical analysis was performed using MedCalc® version 14.12.0. Results: Of 1960 prenatal carrier screens ordered by an Edmonton-based obstetrician group, five cases of beta thalassemia minor and ten cases of alpha thalassemia minor were diagnosed, along with twenty-three heterozygous structural hemoglobin variants (fourteen HbS, four HbE, and five HbC). One case of homozygous HbS, and five heterozygous structural hemoglobin variants had been previously reported (four HbS trait; one HbE trait). Conclusions: The incidence of thalassemia/hemoglobinopathy diagnosed in this study was 2.3%, similar to that described for the general North American population. This suggests that non-selective screening is common in Northern Alberta; education on appropriate patient selection may be warranted. P124 Evaluation of the Roche fructosamine method on the Beckman AU400 Joceline Turner, Hamilton Regional Laboratory Medicine Program Sheri Campbell, Hamilton Regional Laboratory Medicine Program Kelly Lambert, Hamilton Regional Laboratory Medicine Program Janet Sundin, Hamilton Regional Laboratory Medicine Program Andrew Don-Wauchope, Hamilton Regional Laboratory Medicine Program Objectives: The purpose of this evaluation was to determine if the fructosamine method from Roche Diagnostics could be run on the Beckman AU400. Method: The parameters for the assay were reviewed and adjustments made for the AU400 with advice from Beckman technical support. A full method validation was planned that included simple precision, complex precision over a period of 20 days, sensitivity was determined using the limit of blank and linearity was evaluated. Patient comparison was done with another Laboratory using the same Roche method on a Roche instrument. For accuracy the RIQAS External Quality Assessment (EQA) was used. Verification of reference interval was performed using 20 non-diabetic patient samples. All data was analyzed with EP-Evaluator. Results: Modifications were made to the package insert parameters to make allowance for lower volumes and shorter read times for the AU400. Simple precision: Lyphochek 2 Mean=246.9 µmol/L, SD=1.2, CV=0.5%; Precinorm Mean=283.0 µmol/L, SD=3.0, CV=1.0%; Precipath Mean=521.1 µmol/L, SD=2.6, CV=0.5%. Complex precision: Precinorm Mean=280.9 µmol/L, SD=8.8, CV=3.1% and

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Precipath Mean=528.1 µmol/L, SD=24.0, CV=4.5%. Results of the limit of blank testing the manufacturer’s claim of 10 µmol/L passed. We performed a patient comparison of 73 samples. Passing-Bablok regression y=1.03 + 0.8. EQA results were acceptable. The assay is linear from 0.0 to 951.7 µmol/L. Verification of reference range was acceptable. Conclusion: The Roche fructosamine method can be run on the Beckman AU400 with adjustments to the wavelength, reaction time, sample and reagent volumes. Reference interval and bias will be carefully monitored to confirm ongoing accuracy. P125 Acquired hemoglobinopathy and thalassemia Tara Dixon, University of Alberta Mathew Estey, DynaLIFEdx and the University of Alberta Joy Adekanmbi, University of Alberta Trefor Higgins, DynaLIFEDx Dylan Thomas, DynaLIFEDx Karina Rodriguez-Capote, DynaLIFEDx and the University of Alberta Objectives: Hemoglobinopathy or thalassemia usually results from inherited defects in globin chain production. We report three cases of acquired hemoglobin defects. Methods: EDTA-anti-coagulated blood samples were analyzed by high performance liquid chromatography (HPLC) using the ß thalassemia program on the Bio-Rad VARIANT II, followed by electrophoresis at alkaline and acid pH (Sebia Hydrasys Electrophoresis System). A complete blood count (CBC) and ferritin level were included as part of the hemoglobinopathy/thalassemia investigation. Results/Case Reports: Case A: Transfusion-acquired hemoglobinopathy in a paediatric male who was found to have a hemoglobinopathy following erythrocytopheresis. Case B: Bone marrow transplant-acquired thalassemia in an adult patient who developed thalassemic indices and an elevated HbA2% following bone marrow transplantation for acute myeloid leukemia. Case C: Pre-analytical error identified in an adult female with an abnormal HbA1c chromatogram. Reflexive testing indicated the presence of homozygous HbS. A review of her medical record revealed previously normal HbA1c results, and no history of sickle cell disease. Conclusions: When an unexpected hemoglobinopathy or thalassemia is identified, resolution may require a thorough review of the clinical record along with a high degree of suspicion. Interestingly, donor testing does not include investigation for hemoglobin defects which likely have minimal significance beyond the resultant investigation when identified unexpectedly. P126 Effects of an intervention on test utilization of serum protein electrophoresis Albert Tsui, University of Alberta Karina Rodriguez-Capote, DynaLIFEDx and the University of Alberta Alison Hunt, University of Alberta Mathew Estey, DynaLIFEdx and the University of alberta Dylan Thomas, DynaLIFEDx Irwindeep Sandhu, University of Alberta Trefor Higgins, DynaLIFEDx Objectives: Serum protein electrophoresis (SPE) is used to diagnose and monitor monoclonal gammopathies. In northern Alberta, SPE is performed using gel electrophoresis, which is labor intensive and time consuming. Inappropriate utilization of SPE results in increase burden to the laboratory.

The purpose of this study is to review SPE utilization within the context of the current medical guidelines and demonstrate the effects of intervention to optimize SPE utilization. Design/Methods: A retrospective analysis of the SPEs was performed at DynaLIFEDX in 2014 and 2015. Patient age, gender, interpretative results and ordering physician was extracted from the Laboratory Information System. Criteria to assess test utilization appropriateness include age and frequency. In July 2015, two family physicians with high SPE orders were contacted by an oncologist to reduce SPE orders. Results: In 2014, a total of 40930 SPE tests were performed in 32868 patients (48% male). There was 566 SPEs performed in patients < 19 years old, all of which were negative for M-protein. A list of 20 physicians who ordered the most SPE in patients <19 years old were identified and were primarily family physicians. For monitoring the disease, 19% of all repeat SPE was ordered within the one-month period as suggested by the guideline, with most of these orders from oncologists. Intervention resulted in a 90% drop in monthly SPE orders from the two family physicians. Conclusion: By identifying the potential key areas of SPE over-utilization, it will help develop strategies to optimize laboratory test utilization. Keywords: Monoclonal protein, serum protein electrophoresis, test utilization P127 Comparison of serum protein electrophoresis and immunofixation electrophoresis in the assessment of complete response in multiple myeloma Alison Hunt, University of Alberta Albert Tsui, University of Alberta Mathew Estey, DynaLIFEdx and the University of alberta Dylan Thomas, DynaLIFEDx Trefor Higgins, DynaLIFEDx Irwindeep Sandhu, University of Alberta Karina Rodriguez-Capote, DynaLIFEDx and the University of Alberta Objectives: A monoclonal protein (M-protein) is identified by serum protein electrophoresis (SPE) followed by immunofixation electrophoresis (IFE) in the diagnosis and monitoring of multiple myeloma (MM). According to the Alberta Myeloma guidelines, a complete response (CR) requires two consecutive negative IFE results. Recent technological advances suggested that SPE is as sensitive as IFE for Mprotein detection. The purpose of this study is to compare the performance of SPE and IFE in the assessment of CR. Design and Methods: Patient sera with M-protein (IgG, IgA or IgM >1g/L) were serially diluted with a normal serum pool. SPE was performed on the normal, neat, and diluted specimens. The dilution at which the M-protein was absent was scored by five independent observers. IFE was performed on the dilutions suggested by the observers. Results: For samples with IgG-kappa and IgG-lambda Mproteins in the gamma region, there was an excellent agreement between SPE and IFE results. The theoretical mean concentration at which M-protein was absent in both SPE and IFE was at 0.17 g/L. However, for M-proteins located in the alpha and beta regions, the M-protein was still detectable by IFE at the dilution where M-protein was judged absent in SPE. Conclusion: SPE was as sensitive and reliable as IFE in assessing CR for M-proteins located in the gamma region. The result of this study may suggest reassessing the need for repeated IFE in the context of CR in select patients with MM. Keywords: Monoclonal protein, serum protein electrophoresis, immunofixation electrophoresis, multiple myeloma P128

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Clinically significant differences in anion gap results among different chemistry platforms Albert Tsui, University of Alberta George Cembrowski Karina Rodriguez-Capote, DynaLIFEDx and the University of Alberta Rob Sharman, Alberta Health Services EMS Community Care Program Anna Füzéry, Alberta Health Services Objectives: An increased anion gap (AG) suggests the presence of unmeasured anions and often leads to further laboratory investigations. A study from 1998 indicated that AG results vary widely among chemistry platforms, suggesting the use of a single reference range is inappropriate. Our present study evaluated whether this variation still exists twenty years later. Design and Methods: Retrospective patient data from November 2014 to October 2015 was analyzed for differences in electrolytes and AG results for Beckman Coulter DxC, Siemens Advia, and Ortho-Clinical Diagnostics Vitros platforms. In a prospective study, electrolytes and AG were determined on 21 plasma samples obtained from healthy volunteers. A combination of central lab (Beckman Coulter DxC, Ortho Clinical Diagnostics Vitros) and point-of-care (Alere EPOC) platforms were used. Data was expressed as mean ± standard deviation. Results: Retrospective data analysis included a total of 997,423 tests. Average AG were 8.3 ± 2.9 mmol/L, 9.4 ± 2.6 mmol/L, and 11.4 ± 3.6 mmol/L for the DxC, Advia, and Vitros, respectively. A positive bias for sodium and negative bias for chloride was observed for Vitros in both the retrospective and prospective studies. However, the Alere EPOC showed a negative bias for sodium and positive bias for chloride, resulting in significantly lower AG (3.6 ± 1.2 mmol/L). Conclusions: Our results confirm the continued existence of clinically significant differences in AG results amongst different chemistry platforms. Health care systems using a variety of platforms need to consider these differences when establishing their reference ranges. Keywords: Anion gap, instrument differences, reference range P129 A quality assurance investigation of syringe dependent hemolysis in blood gas specimens Nick Baniak, Royal University Hospital, University of Saskatchewan Martha Lyon, Saskatoon Health Region Objectives - To determine the effect of syringe type on hemolysis rate, a source of pre-analytical error, in venous and arterial blood gases. Design and Methods – Leftover clinical blood gas specimens were collected for 50 days at Royal University Hospital, allowing 100 samples/syringe type to be collected from two syringe types (Westmed): arterial and venous. After the clinically required blood gas analysis, syringes were centrifuged and assessed for hemolysis (visual and spectrophotometric H-index). Hemolysis rates were compared between the syringe types. Percentages of hypokalemia (< 3.5 mmol/L), normokalemia (3.5 – 5.1 mmol/L) and hyperkalemia (> 5.1 mmol/L) in hemolyzed specimens for both syringe types were assessed. Specimens with an H-index of ≥ 60 were considered hemolyzed. Results - Overall hemolysis rates were similar between the syringe types (venous 14% vs arterial 13%). The arterial syringe was used 95% of the time to collect arterial specimens, while venous syringes were used to collect venous specimens in 40% of cases. The hemolysis rates for venous versus arterial

specimens collected into venous syringes were 7.5% and 18.3%, respectively. Hemolysis rates for arterial syringe use were 12.6% (arterial specimen) and 25% (venous specimen). The percentage of hemolyzed specimens with hypokalemia were 23.1% (arterial), 28.6% (venous), normokalemia 76.9% (arterial), 50% (venous) and hyperkalemia were 0% (arterial), 21.4% (venous). Conclusions – Higher rates of hemolysis were detected with inappropriate collection of arterial or venous specimens into their designated syringes. Improper collection may diminish actual hypokalemia, making samples appear inappropriately normal. Keywords - Hemolysis, pre-analytic error, venous/arterial blood gases P130 Method validation for Tin analysis of [18F]FDOPA by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) Lusia Sepiashvili, Mayo Clinic, Rochester, MN Melissa Maras, Mayo Clinic, Rochester, MN Michael Mbughuni, Mayo Clinic, Rochester, MN David Murray, Mayo Clinic, Rochester, MN Paul Jannetto, Mayo Clinic, Rochester, MN Objective: Tin (Sn) is the major chemical impurity present 18F during the production of [ ]FDOPA, a diagnostic research Positron Emission Tomography contrast agent administered to Mayo Clinic patients as an Investigational New Drug. The total allowable Sn content in the final drug formulation is 1.8 μg/kg. Excessive exposure to Sn can lead to anemia, hepatic, renal, or neurological problems. The objective of this study was to 18F validate a method to quantify Sn in [ ]FDOPA to facilitate compliance with Sn limits. Design and Methods: Standard and control solutions ranging from 100 to 50,000 ng/mL Sn were evaluated over multiple runs to assess specimen stability, analytical sensitivity, precision, accuracy, analytical specificity, and reportable range. Replicate limits were 30 ng/mL or 10%, whichever is greater. Sn analysis was performed on the PerkinElmer NexION 350 D ICP-MS instrument in Kinetic Energy Discrimination (KED) mode using helium for signal suppression. The Hamilton MicroLab 500 Digital Diluter was used to dispense and dilute specimens. Results: Specimen stability at ambient temperature was demonstrated for up to 28 days (CV<3%). The limit of detection 2 (LOD) was >0.999, y=0.98x-10.418) from 100-50,000 ng/mL. Carryover could be mitigated through diluter rinsing and additional auto sampler washout. Lastly, the method demonstrated acceptable accuracy and recovery. Conclusion: Method validation for Sn analysis by ICP-MS was successful. Keywords: Tin, ICP-MS, assay validation P131 Validation of high sensitivity, conventional and POC Troponin I for use within a regional laboratory program Ronald A Booth, University of Ottawa, The Ottawa Hospital & EORLA Matthew P.A. Henderson, Children's Hospital of Eastern Ontario Nathalie Lepage, Children’s Hospital of Eastern Ontario Christopher McCudden, The Ottawa Hospital, EORLA, University of Ottawa Sherry L. Perkins, The Ottawa Hospital Julie Shaw, The Ottawa Hospital, The University of Ottawa and EORLA Michele Benoit, Eastern Ontario Regional Laboratory Association

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Objective: Laboratory services are provided to 14 community and 1 multi-site academic hospital by EORLA, a single not for profit laboratory services provider. Three TnI assays are offered, Abbott iSTAT at small hospitals and backup for large community hospitals, Abbott Architect at large community hospitals and Siemens Vista at the tertiary-care hospital. With the change from contemporary to high-sensitivity TnI by Abbott, we undertook a regional project to implement hsTnI at the large community hospitals along with test-specific 3-hour acute coronary syndrome biomarker protocols (not discussed here). Methods: Prior to implementation, we performed a full validation of the Abbott Architect hsTnI and analytical correlation with Architect contemporary, iSTAT and Vista TnI assays. Results: Assay precision profiles were calculated, hsTnI performed the best; hsTnI =2.1% to 5.2% for mean values of 3.9ng/L to 20,373ng/L, Vista=7.7% to 2.2% from 18ng/L to 20,000ng/L, and iSTAT =13.5% to 6.2% from 50ng/L to 1810ng/L. Using all data points, Abbott hsTnI correlated surprisingly well with both iSTAT and Vista TnI assays (hsTnI vs Vista, y = 0.9537x + 370.52 R²=0.8432; hsTnI vs iSTAT, y=0.7419x + 47.317 R²=0.90246). hsTnI below 500 ng/L showed a small negative bias with contemporary laboratorybased assays (Abbott= -26ng/L, Vista= -31ng/L) and a minor th positive bias with iSTAT (+8ng/L). Agreement around the 99 percentile was good, 89% for contemporary Architect TnI, 86% for Vista and 81% for iSTAT, with iSTAT being the least sensitive assay. Conclusions: The Abbott hsTnI is a high-precision assay that correlates surprisingly well with other TnI assays. P133 Regional Implementation of Emergency Department 3-hour Biomarker Algorithms for POC, Conventional and HighSensitivity Troponin Ronald A Booth, University of Ottawa, The Ottawa Hospital & EORLA Matthew P.A. Henderson, Children's Hospital of Eastern Ontario Nathalie Lepage, Children’s Hospital of Eastern Ontario, Ottawa Christopher McCudden, The Ottawa Hospital, EORLA, University of Ottawa Sherry L. Perkins, The Ottawa Hospital Julie Shaw, The Ottawa Hospital, The University of Ottawa and EORLA Objective: The Champlain Local Health Integration Network (LHIN) consists of 14 community hospitals and 1 multisite academic health-science centre. Three different troponin assays are offered within the LHIN, Abbott iSTAT at small hospitals, Abbott Architect high-sensitivity (hsTnI) at large community hospitals and Siemens Vista at the academic health-science centre. Recent guidelines have advocated use of serial testing for assessment of acute coronary syndrome. We undertook a LHIN-wide project to develop and implement test-specific 3-hour troponin biomarker algorithms. Methods: Correlation between all troponin assays were performed including short and long-term precision studies. International recommendations for serial troponin testing were reviewed. 3-hour diagnostic algorithms were developed for each assay based on published reference change values, assay precision and guideline recommendations. Numerous educational sessions and tools were provided for ED physicians, residents and nurses. Results: Critical change criteria for all assays were identified. Negative and positive predictive values were assigned where available. When possible, laboratory information systems were programmed to report presentation,

3h and troponin delta values to encourage use of the algorithm. Conclusion: Initial response to implementation by regional emergency departments has been positive. Impact on ED wait times and diagnostic accuracy will be performed after a 3-6 month trial period. P134 Assay of plasma ammonia: whether delay in centrifugation of drawn blood, duration of storage and degree of hemolysis is acceptable as judged by quality goals. Angela W.S. Fung, Department of Laboratory Medicine and Pathobiology, University of Toronto Dorothy Truong, Department of Laboratory Medicine and Pathobiology, University of Toronto Nicole White-Al Habeeb, Department of Laboratory Medicine and Pathobiology Barry Hoffman, Mount Sinai Hospital Objectives: Plasma ammonia methods stipulate minimal delay from blood draw to centrifugation to assay, transport on ice to attenuate metabolism and no hemolysis in order to minimize artifactual increases in measured levels. We investigated how much we could depart from such fastidious pre-analytical handling by varying the time to centrifugation and assay, the temperature at which the blood/plasma was stored, and the degree of (syringe-induced) hemolysis and compare the resulting incremental increase to available allowable limits of error. Methods: Lithium heparin plasma was assayed for ammonia by the Roche Modular glutamate dehydrogenase method (normal a16%), local clinical opinion (±20 µmol/L) and RCPA/AACB Chemical Pathology QAP (±5 µmol/L 50 µmol/L). Results: Blood incubated 0, 15 (ice), 30 (ice) and 30 (RT) minutes prior to centrifugation and testing gave ammonia measurements of 20, 21, 24 and 29 µmol/L, respectively. Incubation of plasma for a further 30 minutes at RT increased levels Conclusions: Quality goals are met when the delay to assay is less than 30 minutes after drawing the blood and hemolysis is ≤0.5 g/L. Keywords: ammonia, pre-analytical error, quality goals P135 Tackling the interference problem for estradiol analysis by LC-MS/MS using differential ion mobility spectrometry Michael Jarvis, Sciex, Toronto Evelyn McClure, Sciex, Toronto Objectives: Clinical researchers familiar with the analysis of estradiol by LC-MS/MS will be acutely aware of the urgent selectivity challenge posed by the presence of ubiquitous, lowlevel, non-specific chromatographic interferences that frequently mask the presence of the target estradiol peak. In this work, we have employed differential ion mobility spectrometry (DMS) to enhance the selectivity of the analysis of estradiol. The DMS cell filters out interferences prior to detection by MS/MS, ensuring that isobaric components do not obfuscate the analysis. Design and Methods: LC separation was performed using the SCIEX ExionLC™ 20AD system, and employing a Phenomenex C18 (100x2.1mm, 2.6um) column, with 50uL injection. MS/MS detection was accomplished using the SCIEX + + Triple Quad™ 6500 system, equipped with SelexION® ion mobility technology. Calibration standards were prepared in double-charcoal stripped serum. Sample preparation consisted of a liquid-liquid extraction using MTBE. Results: Using conventional LC-MS/MS, a large number of interfering chromatographic features were observed, which have the potential to mask the presence of the estradiol

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 analyte. Upon re-analysis by LC-DMS-MS/MS, all interference peaks were removed, enabling quantitation of estradiol at <1 pg/mL. This level of sensitivity is necessary for measurements of estradiol in certain populations such as males, children, and post-menopausal females. Conclusions: The use of ion mobility separation, combined with MS/MS detection, effectively removed uncharacterized interferences that obfuscate the analysis of estradiol using conventional LC-MS/MS methods. This method provided analytical sensitivity that has traditionally only been achieved by derivatizing the target estradiol analyte. P136 Drug monitoring of methotrexate in breast milk: understanding transport from cells to humans Sarah Delaney, Hospital for Sick Children, Toronto Shinya Ito, The Hospital for Sick Kids David Colantonio, The Hospital for Sick Kids Objectives: Methotrexate (MTX) is routinely used to treat inflammatory conditions common in women of childbearing age. Many women taking MTX are concerned with the possibility for drug excretion into breast milk and the risk of exposure to the infant. Study objectives are to: 1) Determine the role of drug transporting proteins in excreting MTX into milk, and 2) determine the pharmacokinetic profile of MTX in milk to examine risk of infant exposure and toxicity. Methods: RT-PCR was carried out to measure mRNA expression of drug transporting proteins in mammary gland tissue. MTX uptake in Breast Cancer Resistance Protein (Bcrp)-transfected HEK293 cells was determined. MTX and was measured in the milk and plasma of human, wild-type (FVB) mice, and Bcrp knockout mice using a validated LCMS/MS assay. Results: mRNA expression of Folr1 and Abcg2 (Bcrp) were up-regulated in lactating mammary glands. Intracellular accumulation of MTX was significantly higher in the presence of a Bcrp inhibitor. Bcrp knockout mice had lower excretion of MTX into breast milk compared to wild-type mice. MTX was excreted into human milk the low nanomolar range across the dosing interval. Discussion: Our data suggests that Bcrp plays a role in transporting MTX into milk. The concentration of MTX excreted into breast milk following low-dose MTX therapy is minimal and may not pose a risk to the nursing infant. Therapeutic drug monitoring and drug transporter data from this study provide a comprehensive view of MTX excretion and will help guide decision making for drug use during lactation. Keywords: Drug Transporters, Bcrp, Drug Excretion, Lactation, Methotrexate P137 A validated LC-MS/MS method for quantitative analysis of methotrexate and 7-hydroxymethotrexate in human and mouse milk Sarah Delaney, Hospital for Sick Children, Toronto Shinya Ito, The Hospital for Sick Kids David Colantonio, The Hospital for Sick Kids Objectives: Methotrexate (MTX) is used to treat inflammatory diseases that commonly occur in women during reproductive years. MTX is contraindicated during breastfeeding; however, only two cases have been published measuring MTX in milk, both with high limits of detection and no measurement of MTX metabolites. The objective of this study was to develop a sensitive LC-MS/MS method to measure the kinetics of low-dose MTX in milk to assess risk of infant exposure and potential for toxicity. Design and Methods: A liquid-liquid extraction procedure using a mixture of organic solvents was used to prepare milk

samples. MTX and metabolite 7-hydroxymethotrexate (7OHMTX) were measured using LC-MS/MS. Performance of the method was assessed. Results: Calibration curves for both MTX and 7OH-MTX followed a regression in the linear range of 0.0016-0.05 μmol/L. The LoQ was 0.0002 μmol/L for MTX and 0.0008 μmol/L for 7OH-MTX. Drug dilution recovery and carry-over was 96% and 0.12%, respectively for MTX; and 91% and 0.11%, respectively for 7OH-MTX. Within-day precision CVs were: < 7% for MTX and < 9 %for 7-OH MTX; and between-day precision CVs were: < 13% for MTX and < 17% for 7OH-MTX across three levels of QC. Conclusions: We have developed a sensitive LCMS/MS method to measure MTX and 7OH-MTX in breast milk with higher sensitivity than the methods published in the literature. The results from this study examining the kinetics of low-dose MTX in milk will help clarify infant risk assessment and help guide decision making for MTX use during breastfeeding. Keywords: LC-MS/MS, Methotrexate, Breast Milk P138 Strict reference values for post-vasectomy semen analyzes obtained by flow cytometry allow to bypass motility analysis and to extend the time-to-analysis window Lyne Massicotte, Nasci Biologie Médicale inc Mathieu Boilard, Nasci Biologie Médicale inc Objectives : Twelve weeks after vasectomy, sterility has to be confirmed by a spermogram that shows : 1- Spermatozoa (spz) concentration inferior to 100 000/ml and 2- absence of motility. The motility criteria forces the laboratory to complete the analysis within 2h otherwise, motility could be lost which would lead to false negatives. Viability analysis using cytometry must be completed within 5h for the same reason. The goal of this study was to extend the time-to-analysis window. Design and methods: Spz concentration and motility were evaluated in parallel by phase contrast microscopy (n=137). For cytometry, spermatozoa were double-labeled with Hoechst (spz detection) and DiOC6 (viability assessment). Concentration and motility scores obtained by microscopy were compared to total spz (tspz) and viable spz (vspz) concentration scores (TCONC and VCONC) obtained by cytometry. Reference values for TCONC and VCONC were determined according to Clinical Laboratory Standard Institute guidelines using StatisPro-CLSI software. Results: After outlier data were removed, 97.5% of TCONC scores were below 8480 tspz/ml and 97.5 % of VCONC scores were below 1600 vspz/ml. No pregnancy ever occurred when VCONC<1600 vspz/ml but it happened at least once when TCONC<8480 tspz/ml. Therefore, VCONC<1600 vspz/ml was chosen as the threshold to confirm sterility. In this study, 99% of samples with a TCONC<5000 tspz/ml had a VCONC<1600 vspz/ml. Conclusions: Thus, TCONC <5000 samples can be considered sterile in cases where time-to-analysis exceeds 2h or 5h. Microscopy is likely to generate false negative results since no spz were observed in 57% of samples that were above the sterility threshold. Keywords: Semen, vasectomy, spermatozoa. P139 Evaluating microtainer neonatal whole blood minimum volume requirements Jessica Gifford, Calgary Laboratory Services Amy Thommasen, Calgary Laboratory Services Isolde Seiden-Long, Calgary Laboratory Services Objectives: While microtainers can potentially reduce the amount of blood drawn from neonates, short draws can

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 potentially lead to high rates of recollects. To avoid this, we have re-evaluated our minimum volume requirements for the new wider bore design of BD Microtainers®. Design/Methods: We examined two questions: First, when transferring separated serum using a micro-pipette, transfer pipette, or pouring, what volumes can we recover from the new BD Microtainer® serum separator tubes (SSTs) with different initial volumes of whole blood with higher neonatal hematocrits (0.61±0.02 L/L)? Second, how many chemistry tests can be performed on the recovered serum using a Roche Cobas 6000 analyzer? Results: The micro-pipette is the most effective at transferring serum with a mean ± avg deviation recovery of 68% ± 3% of total theoretical serum volume, but at volumes of 300 µL and 400 µL, pouring is nearly as effective (67% ± 1.5%). Whole blood volumes of 300 µL yielded enough serum to measure serum indices plus four tests. For the whole blood volumes below 300 µL, there was not enough serum to measure serum indices plus one chemistry test. In addition, we found that a significant volume of serum is lost per test run (10.1 ± 0.4 µL/test) as part of the sampling process on this analyzer type. Conclusions: For neonates, a minimum volume of 300 µL should be drawn into the BD Microtainers® and pouring or using a micro-pipette can transfer enough serum/plasma for 3 4 general chemistry tests. Our findings will prevent redraws in neonates. P140 A liquid chromatography-tandem mass spectrometry method for the detection of beta-carotene in serum Jessica Gifford, Calgary Laboratory Services Joshua Buse, Calgary Laboratory Services Jessica Boyd, Calgary Laboratory Services S.M. Hossein Sadrzadeh, Calgary Laboratory Services Objectives: Beta-carotene is a member of the carotenoid family and it is routinely measured in serum to assess malabsorption conditions, carotenodema, and liver disease. Many current techniques to measure beta-carotene suffer from poor analyte sensitivity, specificity and accuracy. To address these issues, we are developing a liquid-chromatographyelectrospray ionization-tandem mass spectrometry (LC-ESIMS/MS) method for measuring beta-carotene in serum. Methods: Using an Agilent 6460 triple quadrupole mass spectrometer, our LC-ESI-MS/MS method characterizes betacarotene through separation on a C30 column followed by multiple reaction monitoring in positive mode. An isocratic mobile phase of 75:20:5 ethanol: methanol: tetrahydrofuran is run at 1.4 mL/min over a total run time of 15 minutes. Standard solutions of beta-carotene were prepared in ethyl acetate and this solvent was used to perform liquid:liquid extractions of beta-carotene out of patient serum samples. Results: We can achieve chromatographic isolation of beta-carotene in standard solutions (retention time of 10.02 minutes). The method is linear between 0.05 and 5.0 μmol/L for beta-carotene. Stability is a major issue for accurate measurement of beta-carotene. The addition of 0.1 % butylated hydroxytoluene (BHT) greatly improved the stability of this analyte for up to 7 days, especially at more ambient temperatures (22 % vs 100%, recovery without BHT vs with BHT, respectively). Conclusion: We developed an accurate and cost effective method to measure beta-carotene in human serum. More work is in progress to fully evaluate the method and simultaneously measure other members of the carotenoid family and it will be presented at the meeting. P141

Sample preparation methodology for the analysis of steroid hormones by LCMS John Vukovic, Waters Limited Dominic Foley, Lisa Calton, Robert Wardle, Waters Corporation Tandem mass spectrometry provides a way to selectively detect steroids and the LC dimension provides for separation of isobaric species. However, good sample preparation is the key in providing optimal LC-MS/MS analytical sensitivity. To this end, different sample preparation techniques were evaluated. Standard solutions (2 ng/ ml, Sigma Aldrich, USA) of Testosterone, Androstenedione, 17-OHP, Cortisol, DHT and DHEAS were spiked into pooled plasma (Seralab UK). Techniques evaluated; 1) Protein precipitation (PPt); 2) LiquidLiquid Extraction (LLE); 3) PPt with Solid Phase Extraction ® ® Oasis MAX and Oasis Prime HLB (two different protocols; a) Methanol elution & b) Acetonitrile/Methanol elution (85/15, v/v)). ® LC-MS/MS analysis performed on ACQUITY UPLC I-Class ® using a 2.1x50mm ACQUITY HSS T3 UPLC column with VanGuard™ T3 pre-column, a water/methanol/ammonium ® acetate/formic acid gradient and MRM detection on a Xevo TQ-S tandem MS. Simultaneous Precursor Scanning of m/z 184 identified phospholipids. ® Elution protocol b with Oasis Prime HLB provided a 97% reduction in extract phospholipid content relative to PPt. Use of this protocol results in a concomitant increase in sensitivity reflected in the Mean S/N values obtained from each extraction method (n=3) ranked 1–5 (1 indicating highest S/N, 5 the lowest S/N) for each steroid, as seen below.

P142 Detection of non-innoculated fecal immunochemical test kits Jessica Boyd, Calgary Laboratory Services Dennis Orton, Calgary Laboratory Services S.M. Hossein Sadrzadeh, Calgary Laboratory Services Objectives: Fecal immunochemical testing (FIT) is commonly used for colorectal cancer screening. Recently, we have identified a number of non-innoculated FIT kits being received by our lab, which poses a patient safety risk as the instrument gives a negative result for non-innoculated kits. Most manufacturers include a window on the kit to check for inoculation. However, this is not a practical solution for labs that runs several hundred FITs a day. Here we report our investigation into detection of uninnoculated FIT kits. Design and Methods: In Alberta, FIT is performed using the OC-Sensor Diana (Polymedco) with a screening cutoff of 75 ng/mL. Kits are distributed provincially through laboratory services in each health zone. Testing is centralized in Edmonton and Calgary. FIT data was obtained from our laboratory information system (Cerner Millennium) as per institutional data request policies. Results: A result cutoff of 15 ng/mL was derived from the analysis of 20 non-inoculated FIT kits. However, review of the previous 14 months of FIT results indicated that over 70% of

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 tests performed had a result ≤ 15 ng/mL, making the cutoff non-viable. Administrative controls, such as requiring patients to log their FIT kit in with accession staff was successful in reducing the number of non-innoculated FIT kits arriving at the laboratory from 2/week to one every six months. The manufacturer was also contacted and have redesigned the FIT kit labels to include a break away seal. Conclusion: Labs running FIT should evaluate their local process to ensure that non-innoculated FIT kits are detected prior to analysis.

test) after; and abnormal HTC was 50/430 and 42/334 (p=0.69), respectively. The frequency of abnormal MMA and HTC results did not significantly change in cases where B12 results were low (B12<138pmol/L). Conclusions: The intervention reduced B12 requests by >75%, but did not result in improved selection of patients at risk for B12 deficiency based on MMA or HTC. Keywords: Utilization Management, vitamin B12, Methylmalonic Acid, Holotranscobalamin

P143 Evaluation of a point of care strip for urine fentanyl screening Jessica Boyd, Calgary Laboratory Services Anita Klavins S.M. Hossein Sadrzadeh, Calgary Laboratory Services

P145 Investigation of posture specific reference intervals for plasma metanephrine and normetanephrine Jessica Boyd, Calgary Laboratory Services Gregory Kline, Division of Endocrinology and Metabolism, Department of Medicine, Cumming School of Medicine, University of Calgary S.M. Hossein Sadrzadeh, Calgary Laboratory Services

Objectives: Fentanyl is a powerful opioid that is estimated to be 100X more potent than morphine and 40X more potent than heroin. In Alberta in 2015, over 250 deaths were associated with fentanyl. Few immunoassay fentanyl screening kits are available. This leaves mass spectrometry as the first line test, which is only available in specialized labs and has a longer turnaround time. Here we present the evaluation of a point of care fentanyl strip for use in urine drug screening. Design and Methods: The point of care strip (Innovacon) is a qualitative lateral flow immunoassay calibrated against norfentanyl. The cutoff is 20 ng/mL and results are manually read after 15 minutes. The strip performance was compared to our quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) opioid confirmation assay that detects both fentanyl and norfentanyl (lower limit of quantitation = 10 ng/mL). Results: 50 samples were analyzed using the fentanyl strip and LC-MS/MS. The fentanyl and norfentanyl concentrations in the patient specimens ranged from Conclusion: The point of care fentanyl stip compares well with our LC-MS/MS opioid confirmation assay P144 Effect of utilization management rules on ordering patterns for serum vitamin B12 levels. Edward Randell Wael Demian, Eastern Health Authority Georgina M. Randell, Eastern Health Authority David Parry, Eastern Health Authority Christopher S. Kovacs, Faculty of Medicine, Memorial University Brendan Barrett, Faculty of Medicine, Memorial University Sergei LIkhodi, Faculty of Medicine, Memorial University Objectives: A vitamin B12 (B12) test utilization intervention was developed to curtail overuse and to improve patient selection toward higher risk cases. The intervention involved a laboratory operated algorithm whereby all low and low normal B12 results <221pmol/L were reflex tested for methylmalonic acid (MMA), and restricting testing to situations where the reason for the B12 test was provided. This study evaluated the effectiveness of this intervention to reduce B12 tests and improve patient selection. Design and Methods: B12 tests were monitored for 6 months prior to the intervention and during 3 months afterward. In order to assess for improved patient selection relative to B12 status, serum MMA and holotranscobalamin (HTC) were also examined in 2 anonymized cohorts with B12 results 0.40µmol/L and HTC Results: The intervention reduced B12 requests from about 9300/month to 2120/month; and reduced the total 2 number of B12 deficiency cases from 135 to 30/month (B12

Objectives: The Endocrine Society pheochromocytoma guidelines recommend supine collection of plasma metanephrines (PM) to reduce false positive results. However, most laboratories do not require supine collection or provide a posture specific reference interval. We investigated the effect of collection posture on the distribution of our patient population. Design and Methods: Our lab’s PM patient preparation requirements are fasting and abstaining from smoking for 4 hrs prior to phlebotomy. Specimens (EDTA Plasma) are spun, aliquoted and frozen as soon as possible. The majority are collected in the seated position with other bloodwork. The exception are specimens collected at our local endocrine unit, which follows the same preparation as above, except PM collection occurs after the patient has been supine for 30 min. All PM results between May 2010 and September 2015 were analyzed and the distribution of results compared to the current reference intervals for metanephrine (<0.5 nmol/L) and normetanephrine (<0.9 nmol/L). Results: 5452 PM results from 5068 patients were obtained. Specimens collected at the endocrinology unit consisted of 313 specimens on 269 patients and was analyzed separately. No difference in distribution was observed for metanephrine between the two groups. However, the distribution of normetanephrine collected in the seated position had a large tail that extended over 0.9 nmol/L, resulting in 14% of all patients between 0.9 and 1.8 nmol/L. This tailing was not seen when collected supine. Conclusion: Our normetanephrine reference range may not be appropriate for seated collections, resulting in an increased number of false positive results. Further investigation and possible reference interval revision may be necessary. P146 Comparison of cobalt and chromium results from two Canadian laboratories Jessica Boyd, Calgary Laboratory Services James Powell S.M. Hossein Sadrzadeh, Calgary Laboratory Services Objectives: Patients with metal on metal (MoM) joint prosthetics commonly have increased concentrations of cobalt and chromium in their blood, which may be monitored as an indicator of implant failure and/or to assess for possible toxicity. At present, there is little consensus on the preferred specimen type for testing (serum vs whole blood). Recently, our lab changed trace metals referral testing from Lab A (cobalt and chromium in whole blood) to Lab B (cobalt in whole blood and chromium in serum). Here we present the comparison of cobalt and chromium results from these two labs.

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Design and Methods: Lab A and B both use inductively coupled plasma-mass spectrometry for trace element analysis. Specimens were collected and sent to Lab B as per normal practice. An aliquot of the whole blood tube was also sent to Lab A. Results were collated and analyzed using our in-house patient comparison spreadsheet. Results: Specimens were collected from 68 patients with a MoM joint replacement. Comparison of whole blood chromium (Lab A) to serum chromium (Lab B) showed a clear positive bias towards Lab B (R2=0.9944; y=1.41x-5.05). Percent bias ranged from -30% to +60%. Whole blood cobalt comparisons displayed a bias towards Lab A (R2=0.9969, y=0.79x+2.16). With cobalt concentrations below 100 mmol/L, the percent bias ranged from +10% to -30%. Above 100 ng/mL, the percent bias was -20%. This was of interest as better agreement was expected when using the same specimen type. Conclusion: Cobalt and chromium measurements can differ substantially between labs, even when using the same specimen type. P147 Functional genetic variants in the vitamin D pathway: phenotypes in an inner-city pediatric population Lei Fu, Sunnybrook Health Sciences Centre and University of Toronto Betty Y. L. Wong, Sunnybrook Health Sciences Centre Rashida Williams, University of Toronto Bonnie Lee, University of Toronto Thomas O. Carpenter David E.C. Cole, Sunnybrook Health Sciences Centre and University of Toronto Objectives: In a large cohort of healthy infants and toddlers 6 to 38 months of age, we have been exploring the potential role of genetic variation in predisposition to vitamin D insufficiency. The genes encoding the vitamin D binding protein (GC) and the 25-hydroxyvitamin D 24-hydroxylase (CYP24A1) harbor uncommon mutations of uncertain effect. This study was undertaken to look for biochemically recognizable effects of the mutations on vitamin D metabolism, in our at-risk population. Methods: Genotyping for the GC-A246del (c.737739delCTG, rs.529043158) and CYP24A1 mutations (c.686A>G, rs.111622401) were performed using denaturing high performance liquid chromatography and single nucleotide extension assay, respectively, followed by Sanger sequencing confirmation. Vitamin D metabolites and vitamin D binding protein (DBP) were measured by established methods. Results: Genotype Groups Metabolites

Wild-Type GC (A246del) CYP24A1 n = 728 n = 19 n = 10

25(OH)D (nM) 65 ± 16 55 ± 18 1,25(OH)2 D (pM) 157 ± 58 148 ± 48 24,25(OH)2 D (nM) 4.9 ± 2.0 2.9 ± 2.1* DBP (mg/L) 197 ± 29 174 ± 19* † Free 25(OH)D (pM) 24 ± 6 21 ± 6 † *Differs from Wild-Type (p Calculated

73 ± 13* 174 ± 70 5.0 ± 1.8 191 ± 29 27 ± 5

Subjects with the GC-A246del had significantly lower DBP (p=0.034), and 24,25(OH)2 D (p=0.029), but 1,25(OH)2 D and free 25(OH)D were normal. In subjects with the CYP24A1 mutation, only the 25(OH)D was significantly higher (p=0.031). Conclusions: The GC-246delA and the CYP24A1 variants have small but measurable effects on the vitamin D pathway. It seems unlikely that they will be clinically relevant in isolation, but they may be members of the large pool of infrequent mutations that contribute to overall genetic predisposition.

Key Words: Functional genetic variants, vitamin D binding protein, 25-hydroxyvitamin D 24-hydroxylase, vitamin D metabolites, pediatric population P148 Utilization of renal calculi analyses in Calgary S.M. Hossein Sadrzadeh, Calgary Laboratory Services Dennis Orton, Calgary Laboratory Services Ellen Burgess David Ward, University of Calgary Objectives: Analyses of kidney stone composition in patients with chronic stone formation assists in evaluating etiology of stones to evaluate treatment. As a quality improvement initiative and to improve utilization of renal stone testing, an assessment of renal calculi analyses was conducted. Design and Methods: Calgary Laboratory Services (CLS) reports kidney stone composition as determined by infrared spectroscopy. Data was analyzed from January, 2010 to October, 2015 using the majority composition (component > 49%) of the stone and test utilization looking at the number of analyses per patient and by composition of retested stones. Results: Data for 12,903 kidney stones were analyzed from 10,846 patients; 1,463 (13.5%) patients had more than one stone analysis performed, totalling 2,057 (15.9%) tests. 899 (61.4%) patients had 1,101 stones analyzed (53.5%) within 1 year of their first stone, while 1,349 patients (92.2%) had 1,806 (87.8%) repeat analyses performed within 3 years. Composition showed 78.5% of all of the stones were calcium oxalate, 10.8% calcium phosphate, 9.6% urate, 0.68% struvite, and 0.2% cystine. In all, the primary component changed in 201 (12.4%) cases, 112 of which changed from calcium oxalate to calcium phosphate or vice versa, thus only 89 (5.5% of patients with repeats) patients had stones change from calcium to other. Conclusions: Analysis of renal calculi in Calgary showed that the majority of stones contained calcium (phosphate or oxalate) or urate. Repeat stone analyses accounted for 15.9% of the total workload and rarely (5.5%) changed composition, thus indicating repeat testing is not generally required. P149 Pediatric reference intervals for 1,25-dihydroxyvitamin D in the CALIPER Cohort of healthy children and adolescents Dorothy Truong, Department of Laboratory Medicine and Pathobiology, University of Toronto Angela W.S. Fung, Department of Laboratory Medicine and Pathobiology, University of Toronto Nicole White-Al Habeeb, Department of Laboratory Medicine and Pathobiology Victoria Higgins, CALIPER Program, Divisions of Clinical Biochemistry and Hematopathology, Pediatric Laboratory Medicine, The Hospital for Sick Children, University of Toronto Mankhun Chan Barry Hoffman, Mount Sinai Hospital Khosrow Adeli, Clinical Biochemistry, The Hospital for Sick Children Objectives: 1,25 dihyroxyvitamin D (1,25(OH)2 D), the biologically active metabolite of vitamin D, is essential to childhood growth and development, yet age and gender stratified reference intervals do not exist. Traditional 1,25(OH)2 D assays require complex manual preparation, however Diasorin has developed a fully automated in vitro chemiluminescent immunoassay (CLIA) to measure 1,25(OH)2 D, requiring no sample pretreatment or preparation. In alignment with CALIPER [Canadian Laboratory Initiative for Pediatric Reference Intervals], we aimed to establish age- and sex-specific reference intervals.

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Design and Methods: 405 blood samples were collected from apparently healthy children and adolescents aged 0-18y. Those aged 1-18y were from the CALIPER cohort, while those aged 0-1y were Mount Sinai Hospital outpatient samples. 1,25(OH)2 D levels were measured using Diasorin Liaison XL, an in vitro non-competitive three step sandwich CLIA. Statistical analysis was performed using R software, in accordance with CLSI C28-A3 guidelines. Age- and sexspecific reference intervals with corresponding 90% confidence intervals were calculated. Results: There was a significant age-dependent decline in 1,25(OH)2 D levels over the first few years of life requiring data partitioning and calculation of reference values for three age groups: 0-2 D levels in our study group (p=0.364 based on the Mann Whitney U-Test) and sex-partitioning was not necessary. Conclusions: This study provides novel and robust pediatric reference intervals for the 1,25(OH)2 D Diasorin Liaison assay and will improve the accuracy of pediatric test result interpretation for 1,25(OH)2D. Keywords: Reference Intervals, Pediatrics, 1,25 Vitamin D P150 Lymphocyte toxicity assay demonstrates the immunogenetic component of clopidogrel hypersensitivity Manuela Neuman, In Vitro Drug Safety and Biotechnology Background: Clopidogrel (Plavix) is an antiplatelet agent for patients with acute coronary syndrome or undergoing stent placement. Hypersensitivity reaction (HSR) after clopidogrel initiation manifesting as rash and organ involvement affects up to 3% of treated patients. The HSR diagnosis is clinical only. Lymphocyte-toxicity-assay (LTA) measures mitochondrial succinate dehydrogenase activity and is the only validated blood test for HSR. Objective: To characterize the immunogenetic mechanism of clopidogrel-HSR using LTA for the diagnosis in affected individuals. Methods: Blood and skin samples from 32 patients with a confirmed diagnosis of clopidogrel-HSR were analyzed during active HSR and after a minimum of 12 months post clopidogrel discontinuation. Control patients were 32 individuals taking the drug. Tumor necrosis factor (TNF)-alpha, IL-8 and caspase assay were performed in all patients. Results: The hematological screen identified a significant increase in neutrophils, significant decrease in lymphocytes, with no change in eosinophils. The immunohistochemistry of affected areas showed a predominance of CD4+, CD1+ cells with few CD8+ and CD 68+ cells and absence of caspase staining. The mean percentage-LTA was significantly higher for clopidogrel HSR compared to control patients (20±6 vs. 7±4, p=0.0004). The TNF-alpha levels (116±49 vs. 29±9 pg/mL) and IL-8 (92±22 vs 18± 5 pg/mL) were significantly higher (p16% showed a sensitivity of 86% and a specificity of 100% for the diagnosis of clopidogrel. Conclusion: Elevated LTA confirms clinical diagnosis of Clopidogrel-HSR. P151 Human herpes virus-6B can be preset in POEMS syndrome. Manuela Neuman, In Vitro Drug Safety and Biotechnology Susan Goodwin, McMaster Hospital Steven Baker, McMaster Hospital John Neary, McMaster Hospital Hasan Ghaffar, St. Michael Hospital George Moussa, Mount Sinai Hospital Tony Mazzulli, Mount Sinai Hospital

Background: POEMS' syndrome is characterized by Polyneuropathy, Organomegaly, Endocrinopathy, Monoclonal protein, Skin changes. Objectives: 1-to demonstrate that diagnosis and the efficacy of the therapeutic intervention is monitored by quantitative levels of serum vascular endothelial growth factor (VEGF); 2- overproduction of pro-inflammatory cytokines is a characteristic of POEMS; 3-to show that human herpes virus 6 (HHV6) can be present in POEMS Methods: We followed 15 patients with POEM with levels of VEGF between 200 and 7500 pg/mL. Four of the individuals presented human herpes virus (HHV)6. Serum levels of cytokines and chemokines were compared between the patients with 4-POEMS+HHV6, 20 DILI (drug induced liver injury), healthy controls and 11-POEM-without HHV6 (graph). We quantified (ELISA pg/mL) the levels of VEGF, Interferon (IFN-g), Tumor Necrosis Factor (TNF-a), Regulated-uponActivation Normal-T-cell-Expressed and presumably-Secreted; (RANTES), and Nuclear factor kappa-B (NFkB). Results: In POEMS-individuals, VEGF levels were significantly (p<0.001) elevated versus control or DILI. HHV-6POEM, TNF levels were significantly higher (p<0.001) vs. POEM alone. VEGF decreases with therapeutic intervention. Conclusion: Elevation of VEGF levels is diagnostic for POEMS' syndrome, and should be followed to assess response to therapy. In addition, the presence of HHV-6 should be considered in each individual to ensure personalized therapeutic intervention

P152 Urinary 8-Hydroxydeoxyguanosine as a Biomarker of Microangiopathic Complications in Type 2 Diabetic Patients Shereen Amer Background :Reactive oxygen species (ROS) produced either endogenously or exogenously can attack lipid, protein and nucleic acid simultaneously in the living cells. Increased oxidative stress induced by hyperglycemia may contribute to the pathogenesis of diabetic complications. Urinary 8hydroxydeoxyguanosine (8-OHdG) has been reported to serve as a sensitive biomarker of oxidative DNA damage. Objective : To evaluate urinary 8-hydroxydeoxyguanosine (8-OHdG) as a marker for diabetic microangiophatic complications and to correlate its levels with the severity of diabetic nephropathy and retinopathy. Subjects and Methods: The study included 50 patients with type 2 diabetes mellitus and 30 non-diabetic age and sex matched control subjects. The patients group is further classified into 3 groups according to Albumin/creatinine ratio in urine. Patients were also classified into 3 groups according to retinopathy, using Davis classification. Random urine sample was used to measure urinary 8-hydorxydeoxyguanosine in pg/mL (8-OHdG), urine creatinine and urinary albumin excretion (UAE) rate in all patients and control subjects. Both 8-

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 OHdG and UAE rate were assayed by immunoassays. Assessment of glycemic control in patients was achieved by measurement of HbA1c. All of the patients underwent direct opthalmoscopy and photography with pupils dilated. Results: There was a highly significant difference between different groups of type 2 diabetic patients classified according to retinopathy, and controls as regards 8-OHdG (p<0.01). Using ROC curve, the diagnostic utility of urinary 8-OH-dG in discrimination of diabetic patients with retinopathy from those without retinopathy at a cutoff level of 34.4, ug/g creatinine had a diagnostic sensitivity of 92.9%, specificity of 86.4% and efficacy of 90%. Conclusion: Measuring Urinary 8hydroxydeoxyguanosine (8-OHdG) is a novel convenient method for evaluating oxidative DNA damage. Diabetic patients, especially those with advanced nephropathy and retinopathy had significantly higher that such changes may contribute to the development of microvascular complications of diabetes. P153 Rapid detection of extended spectrum beta-lactamase activity using electrospray LC-MS/MS Michael Jarvis, Sciex, Toronto, ON Alex Romaschin, St Michael's Hospital Objectives: Despite recent advances in accelerated bacterial identification by MALDI-TOF MS, characterization of bacterial antibiotic susceptibility remains a time intensive procedure often requiring 8-24 hours for completion. In patients with severe bacterial sepsis and septic shock the documented rate of mortality averages 7% per hour in the absence of effective antibiotic treatment. Timely identification of effective earliest generation antibiotics is important for therapeutic efficacy and antibiotic stewardship minimizing risk of resistance. In this work, we have developed a procedure (provisional patent filed) which rapidly screens for betalactamase activity. Design and Methods: Bacterial isolates are removed from agar culture plates and suspended in Mueller-Hinton broth at a concentration of 0.5 McFarlane units per mL containing a mixture of sentinel antibiotics (ampicillin, cefazolin, cefotaxime and imipenem). Following a 1-hour incubation at 37C, the mixture is deproteinized with methanol and the resulting supernatant subjected to a 4-minute LC-MS/MS analysis (SCIEX 3200 QTRAP), using a reverse phase C-18 column. Antibiotics and hydrolysis products are quantified using MRM transitions. Results: Fifty bacterial isolates with varying antibiotic susceptibilities were tested against a reference method (Vitek 2, gram negative panel). A 100% concordance with the reference method was achieved by quantitatively monitoring the disappearance of antibiotic or appearance of hydrolysis product. Differentiation between AmpC and Type A organisms was also achieved. Conclusions: The LC-MS/MS method presented here provides the ability to rapidly screen for antibiotic resistance, via the determination of beta-lactamase activity, in as little as 90 minutes. Keywords: Antibiotic resistance, beta-lactamase activity, LC-MS/MS P154 Establishment of a reference interval for hs-cTnI in umbilical cord blood. Stephen Hill, McMaster University Tapas Modal, Department of Paediatrics, McMaster University Kaaran Gupta, Department of Paediatrics, McMaster University George Radovanovic, Department of Paediatrics, McMaster

University Janet Simons, Depot of Pathology & Molecular Medicine, McMaster University Rubennna Khan, Department of Paediatrics, McMaster University Objectives: to establish a reference interval for hs-cTnI in umbilical cord blood and to correlate cord hs-cTnI with the risk of developing pre-determined adverse outcomes before two weeks of life. Design and Methods: We recovered cord blood from 256 newborns after all clinical testing was completed and analysed hs-cTnI using the Abbott Architect system. We reviewed charts to collect data including gestational age, birth weight, delivery type (caesarean section, vaginal, vacuum assisted), APGAR score at 1 and 5 minutes NICU admission, establishment of feeding, age at discharge, presence of respiratory distress syndrome. Results: The mean and median hs-cTnI was 5.9 and th th th 13.0 ng/L respectively. The 90 , 95 and 99 percentile cut points were 27.5, 54.3 and 172.0 ng/L respectively. The th proportion of neonates above the 90 percentile admitted to the NICU was significantly greater than the general population (52 th vs. 21% p < 0.006). Neonates above the cTnI 90 percentile had significantly lower mean and median birthweights than th those below the 90 perceintile. The proportion of infants with low (4536g) birhtweight was, however, not significantly different between hs-cTnI groups. No significant difference was noted for APGAR score, age at establishment of feeding or discharge, birth type. th Conclusions: In our study, the 99 percentile cut point in cord blood is significantly greater than for adults populations (172 vs 30 ng/L). This may be an artefact due to th the small sample size. The 90 percentile of 27.4 ng/L may be more representative of a decision point. P155 A high-throughput broad-spectrum LC/MS assay for psychoactive substances in urine Danijela Konforte, LifeLabs Medical Laboratories, Toronto, ON Difei Sun, LifeLabs Medical Laboratories, Toronto, ON Yuyong Ke, Bioanalytical Laboratory Services, Toronto, ON Jan Palaty, LifeLabs Medical Laboratories, Toronto, ON Objectives a) a robust LC/MS assay on 3 dual-column triple-quadrupole instruments for 150 drugs/metabolites in urine, after enzymatic hydrolysis and solid phase extraction; b) middleware to optimize MS detection, automate reporting and reduce the need for manual review. Design Selection of optimal LC and MS conditions for target compounds ranging from morphine to THCA was followed by determination of optimal hydrolysis, extraction and LC separation conditions. Validation parameters included sensitivity, specificity, linearity (ion ratio), recovery/matrix effect, and precision. Results Up to 4 MRMs were determined for 180 targets by infusion. Chromatography was optimized on a partiallyporous biphenyl-type column with an inter-injection time of 7 min. Optimized sample preparation of 30 min hydrolysis with abalone enzyme gave essentially quantitative hydrolysis of benzodiazepines and opioids (except codeine: ~30%) without affecting 6-acetylmorphine. Automated extraction on Agilent Plexa-PCX mixed-mode plates gave an extraction efficiency of 65-80% for most compounds (60% for THCA) with an average matrix effect of about 18%. Cut-off values were in the range of 5-50 ng/mL. Linearity of ion ratios was typically 2-3 orders of magnitude. Morphine glucuronide-d3 and two fluorinated benzylpiperidine-type compounds were used to verify hydrolysis efficiency and crosstalk contamination for every well. Through criteria based on signal/noise, retention times,

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 ion ratio and the presence of specific metabolites, the middleware achieved high concordance with manual review. Conclusion The combination of rapid selective hydrolysis, automated solid phase extraction, data-dependent LC/MS and customized middleware permits ≥300 samples to be processed daily by 1-2 technologists. P156 LC-MS/MS quantification of 11-nor-9-carboxy- Δ9tetrahydrocannabinol and 11-nor-9-carboxy- Δ9tetrahydrocannabinol-glucuronide in urine S.M. Hossein Sadrzadeh, Calgary Laboratory Services Joshua Buse, Calgary Laboratory Services Jessica Boyd, Calgary Laboratory Services Objectives: Development of a LC-MS/MS method for the quantitative analysis of THCCOOH and THCCOOHglucuronide in patient urine, which includes investigating THCCOOH and THCCOOH-glucuronide interferences. Design and Methods: A low volume and sensitive LCMS/MS method has been developed for the quantification of THCCOOH and THCCOOH-glucuronide in 200 µL of urine on an Agilent 1290/6460 LC-MS/MS without the need for hydrolysis. 3.5 minute chromatographic separation utilized an acetonitrile/water gradient on a Restek Ultra Biphenyl II (2.1x100mm,5µm) column. Detection relied upon multiple reaction monitoring (MRM); THCCOOH(343.2à299.1/245.1), THCCOOH-D3(346.2à302.1/248.1) THCCOOH-glucuronide (519.2à343.2/299.2) THCCOOH-glucuronideD3(522.2à346.2/302.1). Quantification relied upon the ratio between the cps intensity of the MRM quantifier transitions of the analyte and internal standard, while analyte confirmation relied upon the ratio between the analyte’s MRM transitions. Results: Preliminary results show that both THCCOOH and THCCOOH-glucuronide were chromatographically separated with excellent peak resolution. The LLOQ was assessed to be 10 ng/mL for THCCOOH using both calibrators prepared in blank urine and patient samples (GC-MS confirmation). The LLOQ of THCCOOH-glucuronide was assessed as 10 ng/mL using calibrators prepared in blank urine. Linearity for both analytes was from 10 to 2000 ng/mL. Analysis of patient samples resulted in the observation of an unidentified compound that interfered with THCCOOH and THCCOOH-glucuronide; we are investigating this finding. Conclusion: We developed an accurate and cost effective LC-MS/MS method to quantify both THCCOOH and THCCOOH-glucuronide in patient urine with a linearity of 102000 ng/mL; work is in progress to fully evaluate this assay and will be presented at the meeting. P157 Usefulness of estimated average glucose (eAG) in glycemic control and cardiovascular risk reduction Yu Chen, Horizon Health Network Angela McGibbon, Dr. Everett Chalmers Regional Hospital, Horizon Health Network George Stoica, Research Services, Horizon Health Network Claire Jardine, Department of Health, Government of New Brunswick Objectives: Although debates have been constant over several years since the introduction of estimated average glucose (eAG), even among the medical and scientific society guidelines, there is lack of evidence to support or against the utility of eAG. One of the 8 regional health authority (RHA) zones in the province of New Brunswick implemented eAG in January 2010. The aim of this study was to evaluate the clinical outcomes of population glycemic control and cardiovascular risk levels before and after the implementation of eAG in this

RHA zone; and to compare the overall outcomes of this zone with 7 RHA zones that did not implement eAG. Methods: Data (845,762 HbA1c values and 614,376 LDLc values) was extracted from the all diabetic patients in provincial Diabetes Registry from 2008 to 2014. Medians and Inter Quartile Ranges were computed using the R software for each quarter of 2008-2014. Kruskal-Wallis statistic and Wilcoxon signed-rank test were conducted. Results: The overall HbA1c and LDL-c levels of the RHA zone implemented eAG showed no statistically significant difference before and after 2010; and the overall HbA1c and LDL-c levels showed no statistically significant difference compared with other RHA zones. There is no significant difference before and after the implementation of eAG in the zone on each of the stratified glycemic control level (e.g. < 8%, 8-8.9%, ≥ 9%) and cardiovascular risk level (e.g. <3.5 mmol/L, 3.5-4.9 mmol/L, ≥5 mmol/L). Conclusion: The utilization of eAG has demonstrated no significant impact on glycemic control and cardiovascular risk reduction. Key words: estimated average glucose, glycemic control, cardiovascular risk, Diabetes P158 TPO utilization: is Northern Alberta Choosing Wisely? Vilte E Barakauskas, Pathology and Laboratory Medicine, BC Children's Hospital Anita Ko, DynaLIFEdx, Edmonton, AB Mathew Estey, DynaLIFEdx and the University of Alberta Amanda VanSpronsen, Laboratory Medicine and Pathology, University of Alberta Don Zhang, DynaLIFEDx, Edmonton, AB Oksana Babenko , University of Alberta Roberta Martindale, University of Alberta Objectives: Canadian Choosing Wisely and Alberta TOP guidelines recommend anti-thyroperoxidase antibodies (TPO) not be ordered routinely or repetitively. This study characterizes TPO ordering and result patterns in Northern Alberta to determine alignment with recommendations. Design and Methods: TPO results were extracted retrospectively from DynaLIFE laboratories LIS for a 3-year period. The University of Alberta REB approved this study. Frequency of repeat orders and significant changes in TPO results were determined and compared by gender and patient location (urban/rural postal code) using Microsoft Excel and Graphpad Prism. Results: A total of 87106 TPO results from 61757 patients were evaluated. Average patient age was 44 years, 71% of patients were female, 10% lived in rural Alberta. Repeat TPO testing occurred in 24% of patients. For one- third, repeat results were identical. For the remainder, average difference between results was 28.5 kU/L. Only 14.5% of repeats differed based on RCV criteria (35 kU/L) while clinical interpretation (antibody positivity) changed in only 3.7%. There was little difference in frequency of repeat testing between gender (24.9% of males vs 23.6% of females) and patient location (25.6% rural vs 23.8% urban). Significant differences between repeat TPO values were associated with 2 gender (X =23.08,p<0.001) but not patient location (p>0.05). Conclusions: Despite recommendations, a significant amount of TPO and repeat testing occurs in Northern Alberta. Our data support current recommendations, given that >95% of repeat results did not change clinical interpretation. Further analysis to identify which physicians order repeat TPOs and for which clinical indications, is needed. Keywords: utilization, TPO, thyroperoxidase, thyroid testing P159

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Demonstration of product equivalence between BD and Greiner Bio-One blood collection tubes Dana Bailey, Dynacare, Brampton, ON Michelle Seaton, Dynacare, Brampton, ON Adam S. Ptolemy, Dynacare, Brampton, ON Hui Li, Dynacare, Brampton, ON Gayle Waite, Dynacare, Brampton, ON Joel Goodman, Dynacare, Brampton, ON Objectives: Perform end-user verification of Greiner BioOne (GBO) versus BD blood collection tubes. Design and Methods: Matched samples for over 100 chemistry and hematology tests from ~1000 patient volunteers were collected in GBO and BD tubes. Each analyte was screened using 10 specimens analyzed in duplicate and the difference assessed by paired t-test. If statistically significant (p<0.05), the observed average bias was compared to the allowable bias. Assays were deemed equivalent if either the test results were not statistically different or the average results were statistically different but less than the allowable bias. If an unacceptable bias was observed, an additional 10 specimens were collected and the sample set was evaluated using linear regression. In addition, the mean percent difference of serum indices and percent packed cell volume were evaluated across various centrifugation conditions. Finally, qualitative assessments regarding colour, clarity, and quality of gel barrier separation were made. Results: For the majority of analytes, equivalent performance was observed. Exceptions included: 1) dihydrotestosterone, where the tube type was changed to a red top serum tube; 2) navy top serum tubes, where the GBO product failed to produce an adequate serum specimen; and 3) magnesium, where despite a significant difference between tubes, no impact to the reference interval was observed. Centrifugation speeds of 1500-1800 xg (serum) or 1500-2000 xg (plasma) produced equivalent quantitative and qualitative results. Conclusions: This investigation demonstrates that, for most analytes, GBO blood collection tubes are equivalent to BD comparative tubes as per our acceptance criteria. P160 Erroneous HbA1c results in a patient with elevated HbC and HbF Joy Adekanmbi, University of Alberta Trefor Higgins, DynaLIFEDx, Edmonton, AB Karina Rodriguez-Capote, DynaLIFEDx and the University of Alberta Dylan Thomas, DynaLIFEDx, Edmonton, AB Tara Dixon, University of Alberta Jessica Gifford, Calgary Laboratory Services Richard Krause, Calgary Laboratory Services Mathew Estey, DynaLIFEdx and the University of Alberta Background HbA1c is used in the diagnosis and monitoring of diabetes mellitus (DM). Interference from hemoglobin variants is a well-described phenomenon, particularly with HPLC-based methods. While immunoassays may give more reliable HbA1c results in the presence of some variants, these methods are susceptible to negative interference from high concentrations of HbF. We report a case where an accurate HbA1c result could not be obtained by any available method due to the presence of an unusual hemoglobinopathy. Methods HbA1c was measured by HPLC (BioRad Variant II and Tosoh), immunoassay (Siemens DCA 2000+ and Roche Integra) and capillary electrophoresis (Sebia). Hemoglobin fractionation was performed by HPLC (BioRad Variant II) and electrophoresis at alkaline and acid pH (Sebia

Hydrasis). Serum fructosamine was assayed on the Siemens Advia 1800. Results HbA1c analysis by HPLC and capillary electrophoresis gave no result due to the absence of HbA. High concentrations of HbC (58%) and HbF (34%) were noted and confirmed by hemoglobin fractionation using HPLC and electrophoresis. Analysis by immunoassay yielded HbA1c results of 5.9% (Siemens DCA 2000+) and 5.1% (Roche Integra), which were inconsistent with the patient’s urine dipstick (3+ glucose) and fructosamine (538 umol/L) results. Conclusion This case illustrates that reliable HbA1c results may be unobtainable in the presence of some hemoglobinopathies. HPLC and capillary electrophoresis did not produce results, but did alert the laboratory to the presence of an unusual hemoglobinopathy. Immunoassays generated falsely low results due to elevated HbF, which could lead to missed diagnoses and under treatment of patients with DM. P161 Reproducibility of 3 autonomously operated cartridgebased Radiometer ABL 90s derived from the analysis of 4 months of external quality control Goudreau Bobbi-Lynn, University of Alberta Hospital Linda Sheen. Janet Thomson, Anna Füzéry, Alberta Health Services Introduction. Previously, the reproducibility of cartridgebased blood gas analyzers has been established by calculating the standard deviation of the internally-assayed quality control (QC). The imprecision of external QC of these analyzers greatly exceeded the internal QC. The reproducibility of external QC materials is a superior surrogate of cartridge and analytic stability and we assayed 3 levels of QC material over 4 months on three ABL90 systems. To simulate the serial reporting of blood gas data from multiple ABL90s, we merged and reanalyzed the QC data. Methods. Over 4 months, we ran 3 levels of Radiometer AutoCheck QC on ABL90s housed in the Adult ICU, Neonatal ICU and Emergency Department at Grey Nuns Hospital. The standard deviations of the controls were calculated for the instruments separately, then for the instruments two and three at a time. Sigma was obtained by dividing biologic variation by the reproducibility (both expressed as CV). Results. The Table summarizes the performance of the non-oximetry tests. Most assays are 3 sigma or higher. Only for hemoglobin was there a significant decrease in sigma with reporting by two or three different ABL90s. Conclusions. The reproducibility of the ABL90 is very acceptable, both separately or used in tandem.

P162 Evaluation of ABL90® whole blood bilirubin as a screening tool for neonatal hyperbilirubinemia Leslie Trowski, BC Children's Hospital Li Wang, BC Children's Hospital Objectives: Our previous evaluations on whole blood bilirubin in healthy newborns identified both bilirubin calibrator and hemoglobin accuracy can affect blood gas analyzer performances. Two questions from that study were raised. One is to compare whole blood bilirubin to BuBc from VITROS 5600® (VITROS) analyzer (not total bilirubin), the other is preanalytical factors such as clots in samples could affect the accuracy of the hemoglobin (Hb) and subsequently the plasma equivalent bilirubin results. In order to answer these questions,

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 we evaluated another vendor’s blood gas instrument with the same research study protocol. Methods: 157 consecutive samples obtained from health newborn babies less than 14 days postnatal age who had bilirubin testing as per usual care, were included. Bilirubin was measured on specimens in the central laboratory, first on the whole blood using the ABL90® (ABL), and then on the VITROS using plasma obtained from the remaining blood, and the leftover plasma were run by ABL again. Statistics were performed with EXCEL. Data and results: The correlation between plasma bilirubin by VITROS and by ABL (Y=0.9907x+0.2812, 2 R =0.9734) is slightly better than the correlation between plasma bilirubin by VITROS and whole blood bilirubin by ABL 2 (Y=1.0479x-9.787, R =0.9559). The Bland-Altman plots found an overall negative bias with a mean difference of -1.19 mmol/L (95% CI:-22.06 to 19.67 mmol/L) and -2.25mmol/L (95% CI: 30.24 to 25.74 mmol/L) respectively. Conclusions: An acceptable correlation was observed between whole blood bilirubin measured on the ABL90® and plasma BuBc on the VITROS 5600®. Keywords: newborns, hyperbilirubinemia, blood gas instrument, whole blood bilirubin P163 Reference change value (RCV) for monoclonal immunoglobulins measurement by agarose gel-based serum protein electrophoresis Ronald A Booth, University of Ottawa, The Ottawa Hospital & EORLA Christopher McCudden, The Ottawa Hospital, EORLA, University of Ottawa Background: Monoclonal immunoglobulin (MIg) quantitation is central to diagnosis, prognosis and follow-up of multiple myeloma and MGUS. Partial response is 50% decrease with 90% decrease considered very-good-partial response. Relapse is defined as 25% (or 5g/L absolute) increase in MIg. However, these recommendations are based on expert opinion. Only a single study has investigated biological-variation and percent change. Here, we provide additional evidence for MIg RCV. Methods: Following CLSI-EP05 recommendations we determined precision of high- and low-concentration MIg on the Sebia Hydrasys 2-Scan Gel Electrophoresis System running Phoresis-8.6.3 software. Imprecision of MIg quantitation was estimated between 4 technological staff over 5 days. MIg was manually quantitated while other fractions were quantitated automatically by the software. RCVs were calculated using previously established biological variability. Results: Precision of fractions other than MIg were calculated using 1yr of QC results. Analytical CV (CVa) and RCVs are presented in the table. Conclusions: We used QC results and real-world conditions to assess imprecision of serum protein fractions including low and high MIg fractions. The calculated RCV of 27% for MIg is similar to that shown previously and slightly greater than the established criteria for relapse. Criteria could be tightened to increase sensitivity for detection of response.

P164 Improving specimen stability for ammonia testing Jessica Gifford, Calgary Laboratory Services Dennis Orton, Calgary Laboratory Services

Lawrence de Koning, Calgary Laboratory Services Isolde Seiden-Long, Calgary Laboratory Services Objectives: Samples for ammonia measurement are collected on ice and run within 1 hour to minimize falsely high results. We developed a method to preserve specimens for ammonia analysis in the absence of these conditions. Methods: Platelet removal and chemical enzyme inhibition were used in specimens with normal or high [>3x upper limit of normal (>3xULN)] levels of alanine aminotransferase (ALT) and/or gamma glutamyl-transferase (GGT). Following plasma separation, enzyme inhibition and/or platelet removal (1500 x g for 15min), samples were incubated at room temperature (RT) or 4°C. Ammonia was measured using a Randox GLDH ammonia assay. Average rate of change was compared between groups using the Student’s T test. Results: At RT, samples with normal ALT/GGT had a mean increase of 2.6 µmol/L/h versus 6.8 µmol/L/h for >3xULN ALT/GGT samples (p3xULN ALT/GGT (5.5 µmol/L/h, p=0.09) or normal samples (0.8 µmol/L/h, p=0.06) vs controls. Inhibitors were more effective for >3xULN ALT/GGT samples (3.1 µmol/L/h, pp=0.16) vs controls. A combination of inhibitors, platelet removal, and refrigeration effectively eliminated the rate of increase for 3XULN ALT/GGT (0.3 µmol/L/h, pp=0.03) samples vs controls. Conclusions: Using chemical inhibitors, platelet removal, and refrigeration, in vitro formation of plasma ammonia was nearly eliminated in normal or 3XULN ALT/GGT samples. P165 Can transcutaneous bilirubin measurement safely replace plasma bilirubin in well newborns greater than 35 weeks gestation? Stephen Hill, McMaster University Laurie Yamamoto, Maternal/Fetal Health, Hamilton Health Sciences Janet Simons, Depot of Pathology & Molecular Medicine, McMaster University Meghan Jameson, School of Nursing, McMaster University Jordan Field, School of Nursing, McMaster University Eunice Antwi-Boasiako, School of Nursing, McMaster University Objectives: 1) Evaluate the ability of transcutaneous bilirubin (TcB) measurement to safely replace plasma bilirubin (TSB) measurement. 2) Estimate the number of TSB measurements that can be replaced by TcB. 3) Estimate the number of heel pricks that can be avoided in well newborns. Design and Methods: The Provincial Council for Maternal Child Health (Ontario) recommends universal bilirubin screening at 24-72 hours of life. TcB is acceptable if the measurement is more than 50 umol/L below the age and risk adjusted phototherapy cutpoint. In a prospective observational trial of 304 well newborns greater than 35 weeks gestation, we used a Dräger JM-105 transcutaneous bilimeter to measure forehead and sternum TcB at the time of each blood collection for TSB. Blood was collected within 30 minutes of TcB. We used linear regression and Altman-Bland plots to compare correlation and agreement. Results: 274 babies and 362 data sets were included. Table 1 presents agreement and correlation. TcB (sternum) and TcB (forehead) could replace 85.9% and 82.3% of TSB respectively. TcB sternum and forehead concordance is 93.6%. Conclusions: TcB, either forehead or sternum, can replace most TSB measurements in well newborns 35 or greater weeks gestation.

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016

P166 Long-term stability of analytical performance for a single lot of glucose test strips over an 18-month period in a large academic healthcare organization Nicole White-Al Habeeb, Department of Laboratory Medicine and Pathobiology, University of Toronto Sousan Bagherpoor, University Health Network, Toronto Christine Cursio, University Health Network, Toronto Paul Yip Department of Laboratory Medicine and Pathobiology, University of Toronto Objectives: Point-of-care testing (POCT) glucose meters are widely used in healthcare facilities for patient glycemic management. Often a single lot of test strips with long shelf life is sequestered. We reviewed the long-term, analytical performance of the Nova StatStrip® glucose meter. Design and Methods: Glucose measurements were performed using a single lot of strips and quality (QC) material including all devices (>200) and trained operators (>5000) between Sept 2014 to Feb 2016 at UHN in Toronto. QC and linearity materials were from Nova Biomedical. Whole blood patient samples were compared to the Siemens RAPIDPoint® 500 blood gas system. Results: Monthly QC data showed steady mean but increased coefficient of variation (CV) over the study period. After the first month, Level 1 QC (mean 3.3 mmol/L), rose from 6.2% CV (n= 210) to 7.7% CV (n= 1,701) at time of expiration. Similarly Level 3 QC (mean 15.3 mmol/L,) rose from 4.4% CV (n= 203) to 5.9% CV (n= 1,628), indicating an increase in imprecision over time. Linearity verification revealed a negative bias of 5.5% compared to target values in January 2016. Finally, while a negligible mean bias (n= 20) was originally observed in comparison to the laboratory analyzer, we found an overall negative bias of -8.1% (range -3.1% to -12.3%, n= 30) in the final month of use. Conclusions: We identified notable deterioration in glucose test strip performance that increased imprecision and bias when approaching the expiry. This highlights the need for greater vigilance in monitoring the quality of glucose measurements when the lot is nearing their stated lifespan. Keywords: Point-of-care testing, glucose meter, quality assurance P167 A computer program to extract and chart clinical laboratory results Nicole White-Al Habeeb, Department of Laboratory Medicine and Pathobiology Angela W.S. Fung, Department of Laboratory Medicine and Pathobiology, University of Toronto Dorothy Truong, Department of Laboratory Medicine and Pathobiology, University of Toronto Tracy Teodoro-Morrison Joshua Raizman, Newborn Metabolic Screening and Biochemical Genetics Laboratory, University of Alberta Hospital Irvin Bromberg, Department of Pathology and Laboratory Medicine, Mount Sinai Hospital Objectives: A patient’s laboratory history is helpful when interpreting results. We developed a computer program that

rapidly fetches test results from the laboratory information system (LIS) and displays them in tabular and graphical formats. Design and Methods: We wrote a Visual Basic for Applications (VBA) macro in Microsoft Excel 2010 that can retrieve laboratory data via network connection (ODBC) from the LIS, collating the data by clinically oriented test panels displayed and optionally charted together. Results: Using minimal input, such as an order#, chart#, case#, or insurance#, the program quickly retrieves clinical laboratory data for tests in the selected panels. Clinically oriented panels, such as Immunology, Anemia, Bone, Endocrine, etc., allow listing multiple tests per table in horizontal or vertical layout, optionally consolidating synonym tests. Generating charts from a tabulated panel takes about a second. For the Immunology panel the program plots IgG, IgA, IgM, IgD and IgE concentration changes in parallel to the M band estimates as well as the Hemoglobin, Protein, and Albumin concentrations. Kappa and Lambda free light chain concentrations are charted as trends along with the K:L ratio, and separately as Kappa vs. Lambda similar to Binding Site educational materials but enhanced by arrows indicating change directions. The program also has the essential ability to display age and sex-adjusted reference ranges for each analyte. Conclusions: The program quickly retrieves and displays clinical laboratory data. The desired test panels are easily selected according to the user’s preference, facilitating interpretation of result trends. Keywords: patient trends, bioinformatics, laboratory data P168 Inappropriate Utilization of Specific Biochemical Tests Continues in Ontario Canada Dana Bailey, Dynacare, Brampton, ON Mike Godwin, Dynacare, Brampton, ON Gayle Waite, Dynacare, Brampton, ON Hui Li, Dynacare, Brampton, ON Adam S. Ptolemy, Dynacare, Brampton, ON Background: The Ontario Health Technology Advisory Committee (OHTAC) has made the following laboratory testing recommendations: severely restrict access to Ontario Health Insurance Plan (OHIP) insured vitamin D testing (December 2010); remove ferritin (January 2012), vitamin B12 (January 2012) and chloride (January 2013) from the Provincial laboratory requisition (check boxes removed so tests must be hand-written); and restrict insured folate and aspartate aminotransferase (AST) orders to a small set of clinical conditions and/or specialist physicians (January 2013). Objective: Evaluate the impact of each OHTAC recommendation on physician ordering through a retrospective review of vitamin D, ferritin, vitamin B12, chloride, folate and AST test volumes from our Ontario-based community reference laboratory. Design and Method: Volumes of the above-named tests from January, 2005 to July, 2015 were reviewed. Quantitative trends for each test’s monthly volume, pre and post-OHTAC recommendations were analyzed. Results: Relative to November, 2010 (N=19117), OHIPinsured vitamin D test volume decreased 89% in December, 2010 (N=2143) but then edged up an average of 3.8% per month until July, 2015. Average monthly volumes of ferritin and vitamin B12 increased 2.7% and 9.6% per month respectively, after their removal from the OHIP requisition, while chloride volume decreased an average of 0.4% per month from January 2013 (N=89443) to July 2015 (N=63652). Folate (RBC) and AST testing volume decreased an average of 0.9% and 0.3% per month following the OHTAC recommendation restrictions.

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 Conclusion: Despite evidence-based recommendations and efforts to influence ordering, inappropriate test utilization continues to impact the Ontario health-care system. P169 Serum25-Hydroxy-Vitamin D Testing is Unjustifiably Increasing in Ontario Canada Dana Bailey, Dynacare, Brampton, ON Mike Godwin, Dynacare, Brampton, ON Gayle Waite, Dynacare, Brampton, ON Hui Li, Dynacare, Brampton, ON Adam S. Ptolemy, Dynacare, Brampton, ON Background: The Ontario Health Insurance Plan (OHIP) laboratory requisition requires clinicians to classify patients as insured or uninsured for 25-hydroxy-vitamin D (vitamin D) testing. From December 2010 to July 2015 the volume of OHIP insured vitamin D tests performed by our community reference laboratory increased an average of 3.8% per month, while uninsured testing decreased an average of 0.2% per month. Objective: Determine if the observed trends in test ordering are clinically justified by reviewing patient vitamin D statuses from all OHIP insured and uninsured testing performed between December 2010 and July 2015. Design/Method: Patient vitamin D statuses were respectively partitioned by gender and age (e.g., ≤19, 20-34, 35-49, 50-64, 65-79 and ≥80y) and defined as: deficiency, 250 nmol/L. Results: N=372796 and N=134752 unique OHIP insured and uninsured vitamin D tests were reviewed. Relative to OHIP insured male and female patients; the respective prevalence of vitamin D insufficiency was higher in uninsured patients (50 and 43% vs. 55 and 52%). These patients also had a relatively lower prevalence of vitamin D sufficiency (47 and 55% vs. 42 and 45%). Regardless of OHIP insurance status; the prevalence of vitamin D insufficiency and toxicity trended downward with age, while sufficiency levels consistently trended upward. Conclusion: The observed patient vitamin D statuses do not justify routine vitamin D testing of healthy adults and children. Medically unnecessary vitamin D testing is increasingly being performed and covered by OHIP. P170 Development and validation of a computerized alert for identification of acute kidney injury. Michael Knauer, Department of Laboratory Medicine and Pathobiology, University of Toronto Daniel Beriault, Department of Laboratory Medicine and Pathobiology, University of Toronto Stephen Cochlin, University Health Network, Toronto Abhijat Kitchlu, University of Toronto Emilie Chan, University of Toronto Jennifer Rodrigues, University of Toronto Sushil Ratnaparkhe, University of Toronto Rory McQuillan, UHN and University of Toronto Paul Yip, UHN and Univesrity of Toronto Objectives: Acute kidney injury (AKI) is an often unrecognized condition that contributes to poor patient outcomes. Electronic alerts have been used to help clinicians recognize AKI based on serial changes in serum creatinine. The objective of this study was to develop and validate a computer algorithm based on the Acute Kidney Injury Network criteria for identifying patients with AKI. Design and Methods: Currently AKI needs to be recognized by clinicians and referred to Nephrology for treatment and follow-up. We developed a computerized algorithm to calculate AKIN stage and highlight AKI cases

based upon patient creatinine results. The AKI cases identified by the computer algorithm were compared to the AKI case referrals to the AKI clinic at UHN between October 2014 and January 2015. Results: Over the four-month period 169 cases of AKI were referred to the AKI clinic, 161 of which were also identified by the computer AKIN staging algorithm. The computer algorithm and Clinicians assigned the same AKIN stage in 60% of the cases, while the AKIN stage assigned by the algorithm was higher, lower, or not assigned in 28%, 7% or 5% of cases respectively. The timing of the algorithm alert and clinical referral were also compared. In 62% of the cases the computer algorithm identified the AKI case at least 1 day before the recorded Clinical referral. In 24% of the cases the clinician and algorithm identified the case on the same day and in 14% of cases Clinicians identified the AKI case before the computer algorithm. Conclusions: AKI is a common occurrence in hospitalized patients and the implementation of a computer algorithm for recognizing AKI and alerting clinicians may improve the speed at detecting cases and improve outcomes. P171 A risk score for abnormal fecal fat excretion based on fecal weight and lipid droplets measured by photomicroscopy and automated image analysis Michael Korostensky, Calgary Laboratory Services Christopher Naugler, Calgary Laboratory Services Steven R. Martin, Alberta Children's Hospital Lawrence de Koning, Calgary Laboratory Services Background: The 72-hour quantitative fecal fat test is difficult for laboratories and patients, but is sometimes necessary to perform. We derived a risk score predicting abnormal fecal fat excretion based on simple laboratory results, including lipid droplets imaged by photomicroscopy and automatically analyzed by computer software. Methods: 72-hour quantitative fecal fat (mmol/day) was measured in 100 stool specimens (abnormal fat excretion: ≥ 21 mmol/day). Specimen wet mounts were prepared after staining with oil-red-o. Slides were image-captured at 55X using a Cellavision microscope camera, and analyzed for oil-red-o droplet number and surface area using an ImageJ macro. Logistic regression with backwards elimination was used to select significant predictors of abnormal fat excretion among sex and quintiles of age, droplet number, surface area, and fecal weight. A risk score (%) was created by multiplying significant coefficients by patient quintile values, and adding the products together. The c-statistic was used to assess predictive power of the risk score. Results: After backward elimination, droplet number quintile, surface area quintile and fecal weight quintile were significantly (p<0.05) associated with abnormal fat excretion. A risk score (min=4, max=91) based on these variables had a cstatistic of 0.77. The top vs the bottom score quintile was associated with a 24X greater odds of abnormal fecal fat output. A cut-point of 46 yielded a sensitivity of 60% and specificity of 84%. Conclusions: A risk score based on simple laboratory tests had acceptable predictive power for abnormal fat excretion. This score may be useful in reducing utilization of the 72-hour quantitative fecal fat test.

Abstracts of CSCC Conference 2016 Edmonton AB • June 19-22, 2016 P172 Increasing the specificity of steroid measurements by LCMS/MS using differential ion mobility spectrometry Evelyn McClure, Sciex, Toronto, ON Michael Jarvis, Sciex, Toronto, ON Objectives: Steroid researchers investigating inborn errors of metabolism are increasingly turning to LC-MS/MS as a tool due to the high selectivity and sensitivity of this technique. Despite the inherent selectivity of LC-MS/MS measurements, there nevertheless exists the possibility of interferences from related steroid compounds. We have explored the use of differential ion mobility spectrometry (DMS) to filter out interferences prior to the detection of 17hydroxyprogesterone (17-OHP), androstenedione (AND), and cortisol (COR) by MS/MS, thereby enabling the use of rapid LC gradients. Design and Methods: LC separation was performed using the SCIEX ExionLC™ 20AD system, with a reversed phase C18 (50x2.1mm, 1.8um) column. MS/MS detection was + accomplished using the SCIEX Triple Quad™ 6500 system, + equipped with SelexION® ion mobility technology. Sample preparation consisted of a liquid-liquid extraction using MTBE. Results: Using conventional LC-MS/MS, it was necessary to chromatographically separate isobaric steroids. When the analysis was performed by LC-DMS-MS/MS, it was possible to remove interferences from the MRM channels of the target analytes, enabling short LC run-times. For example, 11deoxycorticosterone is a known interference to 17-OHP, and chromatographic separation of the two is required using conventional LC-MS/MS; using DMS, the compounds could be differentiated without chromatographic separation. By LC-DMSMS/MS, accuracy for all 3 target compounds was > 90% and precision values for 17-OHP, AND, and COR were < 8%, 10% and 7%, respectively. These values were equivalent, or better, than those obtained using LC-MS/MS. Conclusions: Ion mobility separation removed interferences caused by isobaric steroid compounds, thereby enabling significantly shorter run-times for the analysis of several steroids.