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Screening and determination of potential xanthine oxidase inhibitors from perilla frutescens leaves using ultrafiltration liquid chromatography-mass spectrometry
Drug Metabolism and Pharmacokinetics 32 (2017) S108eS111
Contents lists available at ScienceDirect
Drug Metabolism and Pharmacokinetics journal homepage: http://www.journals.elsevier.com/drug-metabolism-andpharmacokinetics
Late-breaking abstracts LB1 DETERMINATION OF KRI-60522 AS A POTENT DIACYLGLYCEROL ACYLTRANSFERASE 1 INHIBITOR BY LIQUID CHROMATOGRAPHYe TANDEM MASS SPECTROMETRY AND ITS APPLICATION TO RATS PHARMACOKINETIC STUDY So Hee Im, Seong Soon Kim, Jeong Im Ryu, Jin Sook Song, Sunjoo Ahn, Myung Ae Bae. Korea Research Institute of Chemical Technology, Daejeon, South Korea A simple, rapid and sensitive method for assaying a KRI-60522 as a potent diacylglycerol acyltransferase 1 (DGAT1) inhibitor was to develop and validate in rat plasma using liquid chromatography-electronic spay ionization tandem mass spectrometry (LC-ESI-MS/MS). Plasma samples from rats were prepared by simple protein precipitation and injected onto the LCeMS/MS system for quantification. KRI-60522 and disopyramide as an internal standard (IS) were separated by a reversed phase C18 column (2.1 mm 50 mm, 3 mm). A mobile phase was composed of 10 mM ammonium formate in water and acetonitrile (ACN) (v/v) by a linear gradient system, increasing the percentage of ACN from 2% at 0.5 min to 98% at 2.5 min with 5 min total run time. The ion transitions monitored were m/z 539 > 299.3 for KRI-60522 and m/z 340 > 239 for IS in the multiple reaction monitoring (MRM) mode. The detector response was specific and linear for KRI-60522 at concentrations within the range 0.005-5 mg/mL and the correlation coefficient (R2) was greater than 0.999 and the signal-to-noise ratios for the samples were 10. The intra- and inter-day precision and accuracy of the method ranged from 0.40 to 11.2% were determined to be within the acceptance criteria for assay validation guidelines. The matrix effects were approximately 107 and 103% from rat plasma for KRI-60522 and disopyramide, respectively. Furthermore, KRI-60522 was stable under various handling and processing conditions. KRI-60522 concentrations were readily measured in rat plasma samples after intravenous administration. This analytical method was successfully applied to a pharmacokinetic study of KRI-60522 in rats; intravenous administration of 5 mg/kg of RI-60522 yielded an elimination half-life (t1/2) of 2.83 ± 1.07 h and a steady-state volume of distribution (Vss) of 11.1 ± 4.69 L/ kg, with a clearance of 5.03 ± 0.27 L/h/kg. LB2 DETERMINATION OF THE ANTIMALARIAL DRUG PIPERAQUINE AND ITS METABOLITES IN HUMAN PLASMA BY LC-MS/MS Mohd Yusmaidie Aziz, Michael Ashton, Kurt-Jurgen Hoffmann. Unit for Pharmacokinetics and Drug Metabolism, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden Introduction: The antimalarial piperaquine (PQ) and some of its metabolites are suggested to be CYP3A inhibitors. To understand their potential for drug-drug interactions (DDI), pharmacokinetic profiles are required. Thus, determination of PQ and its metabolites in human plasma is essential. Method: A Sciex API 4000 triple quadropole mass spectrometer with an electrospray ionization source operated in the positive ion mode was used as detector. Detection was by multiple reaction monitoring (MRM) transition (PQ: 535/228; carboxylic PQ; 320/205; N-oxide PQ: 551/258; PQ-d6: 541/294) Separation was on a C18 column with gradient elution. Sample preparation was done with 1% formic acid acidification.
Results: 100 mL human plasma was required for the quantitation. The method has a linear calibration range (weighted 1/x) of 1 nM e 625 nM with a runtime of 7 min per sample. LOQ was 0.5 nM. No degradation was observed at room temperature within 24 hours up to 3 months plasma storage at -80 C. Intra- and interday precision and accuracy were below 15%. Recovery was 80% with no significant matrix effects. Conclusion: A sensitive and rapid LC-MS/MS method was developed for the first time for piperaquine metabolites. The method was applied to plasma samples from healthy volunteers given piperaquine. LB3 SCREENING AND DETERMINATION OF POTENTIAL XANTHINE OXIDASE INHIBITORS FROM PERILLA FRUTESCENS LEAVES USING ULTRAFILTRATION LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY Shin Hwa Kwon 1, Zhiqiang Wang 2, Set Byeol Kim 2, Min Joo Ahn 3, Yanymee N. Guillen Quispe 2, Soo Kyeong Lee 2, 4, Soon Sung Lim 1, 2, 4. 1 Institute of Natural Medicine, Hallym University, Chuncheon, South Korea; 2 Department of Food Science and Nutrition, Hallym University, Chuncheon, South Korea; 3 Department of Life Science, Hallym University, Chuncheon, South Korea; 4 Institue of Korean Nutrition, Hallym University, Chuncheon, South Korea
Perilla frutescens leaves are a typical Chinese herb with a large number of polyphenols existing in all parts of it. The most common methods for screening and isolating polyphenols are mostly labor intensive and time consuming. In this study, a new assay based upon ultrafiltration liquid chromatography was developed for the rapid screening of ligands for xanthine oxidase. The P. frutescens leaves extract was found to contain polyphenols with xanthine oxidase binding activities. Four polyphenols were isolated from the P. frutescens using by Sephadex LH-20 open column chromatography. Four compounds were identified as caffeic acid (Compound 1, base peak 180 m/z; 180 [M]+; 163 [M-OH]+; 134 [M-COOH]+), oleanolic acid (Compound 2, base peak 248 m/z; 203 [M-COOH] + ), luteolin (Compound 3, base peak 286 m/z; 153 [M-133]+), apigenin (Compound 4, base peak 270 m/z; 242 [M-28]+; 153 [M-89]+) using electron ionization mass spectrometry (EIMS), 1H NMR. Among them, compound 3 inhibited xanthine oxidase with IC50 value of 10.70 mM more than positive control (allopurinol with IC50 value of 17.64 mM). Therefore, ultrafiltration liquid chromatography is not only a powerful tool for screening and isolating xanthine oxidase inhibitors in complex samples but is also a useful platform for discovering bioactive compounds for the prevention and treatment of gout. LB4 STRUCTURE-BASED VIRTUAL SCREENING AND BIOLOGICAL EVALUATION OF THE FLAVONOIDS THAT TARGET CYP4A AND ANGIOGENESIS Chenlong Wang, Xuewei Chen, Tianjian Yu, Chong Shu, Chengpeng Fan, Jing Yang. School of Basic Medical Sciences, Wuhan University, Wuhan, China Cytochrome P450 (CYP) 4A enzymes play important roles in angiogenesis, and flavonoids inhibit tumor angiogenesis. Here, virtual screening combined with model organism approach was applied to identify the flavonoids that target CYP4A and angiogenesis. The 16839 flavonoid compounds from
Contents lists available at ScienceDirect
Drug Metabolism and Pharmacokinetics journal homepage: http://www.journals.elsevier.com/drug-metabolism-andpharmacokinetics
Late-breaking abstracts LB1 DETERMINATION OF KRI-60522 AS A POTENT DIACYLGLYCEROL ACYLTRANSFERASE 1 INHIBITOR BY LIQUID CHROMATOGRAPHYe TANDEM MASS SPECTROMETRY AND ITS APPLICATION TO RATS PHARMACOKINETIC STUDY So Hee Im, Seong Soon Kim, Jeong Im Ryu, Jin Sook Song, Sunjoo Ahn, Myung Ae Bae. Korea Research Institute of Chemical Technology, Daejeon, South Korea A simple, rapid and sensitive method for assaying a KRI-60522 as a potent diacylglycerol acyltransferase 1 (DGAT1) inhibitor was to develop and validate in rat plasma using liquid chromatography-electronic spay ionization tandem mass spectrometry (LC-ESI-MS/MS). Plasma samples from rats were prepared by simple protein precipitation and injected onto the LCeMS/MS system for quantification. KRI-60522 and disopyramide as an internal standard (IS) were separated by a reversed phase C18 column (2.1 mm 50 mm, 3 mm). A mobile phase was composed of 10 mM ammonium formate in water and acetonitrile (ACN) (v/v) by a linear gradient system, increasing the percentage of ACN from 2% at 0.5 min to 98% at 2.5 min with 5 min total run time. The ion transitions monitored were m/z 539 > 299.3 for KRI-60522 and m/z 340 > 239 for IS in the multiple reaction monitoring (MRM) mode. The detector response was specific and linear for KRI-60522 at concentrations within the range 0.005-5 mg/mL and the correlation coefficient (R2) was greater than 0.999 and the signal-to-noise ratios for the samples were 10. The intra- and inter-day precision and accuracy of the method ranged from 0.40 to 11.2% were determined to be within the acceptance criteria for assay validation guidelines. The matrix effects were approximately 107 and 103% from rat plasma for KRI-60522 and disopyramide, respectively. Furthermore, KRI-60522 was stable under various handling and processing conditions. KRI-60522 concentrations were readily measured in rat plasma samples after intravenous administration. This analytical method was successfully applied to a pharmacokinetic study of KRI-60522 in rats; intravenous administration of 5 mg/kg of RI-60522 yielded an elimination half-life (t1/2) of 2.83 ± 1.07 h and a steady-state volume of distribution (Vss) of 11.1 ± 4.69 L/ kg, with a clearance of 5.03 ± 0.27 L/h/kg. LB2 DETERMINATION OF THE ANTIMALARIAL DRUG PIPERAQUINE AND ITS METABOLITES IN HUMAN PLASMA BY LC-MS/MS Mohd Yusmaidie Aziz, Michael Ashton, Kurt-Jurgen Hoffmann. Unit for Pharmacokinetics and Drug Metabolism, Sahlgrenska Academy at University of Gothenburg, Gothenburg, Sweden Introduction: The antimalarial piperaquine (PQ) and some of its metabolites are suggested to be CYP3A inhibitors. To understand their potential for drug-drug interactions (DDI), pharmacokinetic profiles are required. Thus, determination of PQ and its metabolites in human plasma is essential. Method: A Sciex API 4000 triple quadropole mass spectrometer with an electrospray ionization source operated in the positive ion mode was used as detector. Detection was by multiple reaction monitoring (MRM) transition (PQ: 535/228; carboxylic PQ; 320/205; N-oxide PQ: 551/258; PQ-d6: 541/294) Separation was on a C18 column with gradient elution. Sample preparation was done with 1% formic acid acidification.
Results: 100 mL human plasma was required for the quantitation. The method has a linear calibration range (weighted 1/x) of 1 nM e 625 nM with a runtime of 7 min per sample. LOQ was 0.5 nM. No degradation was observed at room temperature within 24 hours up to 3 months plasma storage at -80 C. Intra- and interday precision and accuracy were below 15%. Recovery was 80% with no significant matrix effects. Conclusion: A sensitive and rapid LC-MS/MS method was developed for the first time for piperaquine metabolites. The method was applied to plasma samples from healthy volunteers given piperaquine. LB3 SCREENING AND DETERMINATION OF POTENTIAL XANTHINE OXIDASE INHIBITORS FROM PERILLA FRUTESCENS LEAVES USING ULTRAFILTRATION LIQUID CHROMATOGRAPHY-MASS SPECTROMETRY Shin Hwa Kwon 1, Zhiqiang Wang 2, Set Byeol Kim 2, Min Joo Ahn 3, Yanymee N. Guillen Quispe 2, Soo Kyeong Lee 2, 4, Soon Sung Lim 1, 2, 4. 1 Institute of Natural Medicine, Hallym University, Chuncheon, South Korea; 2 Department of Food Science and Nutrition, Hallym University, Chuncheon, South Korea; 3 Department of Life Science, Hallym University, Chuncheon, South Korea; 4 Institue of Korean Nutrition, Hallym University, Chuncheon, South Korea
Perilla frutescens leaves are a typical Chinese herb with a large number of polyphenols existing in all parts of it. The most common methods for screening and isolating polyphenols are mostly labor intensive and time consuming. In this study, a new assay based upon ultrafiltration liquid chromatography was developed for the rapid screening of ligands for xanthine oxidase. The P. frutescens leaves extract was found to contain polyphenols with xanthine oxidase binding activities. Four polyphenols were isolated from the P. frutescens using by Sephadex LH-20 open column chromatography. Four compounds were identified as caffeic acid (Compound 1, base peak 180 m/z; 180 [M]+; 163 [M-OH]+; 134 [M-COOH]+), oleanolic acid (Compound 2, base peak 248 m/z; 203 [M-COOH] + ), luteolin (Compound 3, base peak 286 m/z; 153 [M-133]+), apigenin (Compound 4, base peak 270 m/z; 242 [M-28]+; 153 [M-89]+) using electron ionization mass spectrometry (EIMS), 1H NMR. Among them, compound 3 inhibited xanthine oxidase with IC50 value of 10.70 mM more than positive control (allopurinol with IC50 value of 17.64 mM). Therefore, ultrafiltration liquid chromatography is not only a powerful tool for screening and isolating xanthine oxidase inhibitors in complex samples but is also a useful platform for discovering bioactive compounds for the prevention and treatment of gout. LB4 STRUCTURE-BASED VIRTUAL SCREENING AND BIOLOGICAL EVALUATION OF THE FLAVONOIDS THAT TARGET CYP4A AND ANGIOGENESIS Chenlong Wang, Xuewei Chen, Tianjian Yu, Chong Shu, Chengpeng Fan, Jing Yang. School of Basic Medical Sciences, Wuhan University, Wuhan, China Cytochrome P450 (CYP) 4A enzymes play important roles in angiogenesis, and flavonoids inhibit tumor angiogenesis. Here, virtual screening combined with model organism approach was applied to identify the flavonoids that target CYP4A and angiogenesis. The 16839 flavonoid compounds from