- Email: [email protected]
Quantitative LC analysis of cyclosporine A in ocular samples
S136
Abstracts
Poster – [A-10-245-1] Extraction of crocin from saffron (Crocus sativus) using molecularly imprinted polymer solid-phase extraction Ahmad Mohajeri Seyed, Hossein Hosseinzadeh, Fariborz Keyhanfar, Javad Aghamohammadian Mashhad University of medical science.Mashhad, Iran E-mail addresses: [email protected] (A. Mohajeri Seyed), [email protected] (J. Aghamohammadian) A new molecularly imprinted polymer for extraction of crocin from saffron stigmas was prepared using gentiobiose (a glycoside moiety in crocin structure) as a template. Crocin binding to gentiobiose imprinted polymer (Gent-MIP) was studied in comparison with a blank nonimprinted polymer in aqueous media. Affinity of the Gent-MIP for the crocin was more than the nonimprinted polymer at all concentrations. In Scatchard analysis, the number of binding sites in each gram of polymer (maximum binding sites) and dissociation constant of crocin to binding sites were 18.4 mmol/g polymer and 11.2 mM, respectively. The Gent-MIP was then used as the sorbent in an SPE method for isolation and purification of crocin from methanolic extract of saffron stigmas. The recovery of crocin, safranal and picrocrocin was determined in washing and elution steps. The Gent-MIP had significantly higher affinity for crocin than other compounds and enabled selective extraction of crocin with a high recovery (84%) from a complex mixture. The results demonstrated the possibility of using a part of a big molecule in preparing a molecularly imprinted polymer with a good selectivity for the main structure. Keywords: Saffron, Crocin, Molecularly imprinted polymer, Gentibiosis, HPLC
doi:10.1016/j.clinbiochem.2011.08.326
Poster – [A-10-245-2] Quantitative LC analysis of cyclosporine A in ocular samples Javad Aghamohammadian, Toka Banaee, Navid Mosallaei, Ahmad Mohajeri Seyed Mashhad, Iran E-mail addresses: [email protected] (J. Aghamohammadian), [email protected] (A. Mohajeri Seyed) Introduction: An isocratic reversed-phase HPLC method with ultraviolet detection at 205 nm has been developed for analysis of cyclosporine A (CyA) in rabbit ocular samples. Neither internal standard nor extraction was needed for sample preparation. Acetonitrile (ACN; 1 mL) was added to 250 μL aqueous and vitreous samples to precipitate proteins. The supernatant was dried and the residue was reconstituted in mobile phase and injected for HPLC analysis. Chromatography was performed on an octadecyl silane-A (ODSA) C18 (4.6 × 250 mm, 5 μm) column. The column temperature was fixed at 70 °C and the mobile phase was ACN 65%, methanol 20% and water 15% at a flow rate of 1.5 mL min− 1. The calibration curve for CyA in rabbit ocular samples was linear over the concentration range 0.2 and 10 μg mL− 1 with a correlation coefficient of 0.9992. Intra-day and inter-day precision were 4.61–7.83% and 5.27–10.70%, respectively. Intra-day and inter-day accuracy were 89.2–108% and 83.4–111%, respectively. The limits of detection (LOD) and quantification (LOQ) were 5.7 and 38 ng mL− 1, respectively. Conclusion: The method was successfully used for analysis of CyA in real aqueous and vitreous humor samples from New Zealand albino rabbits. The method is therefore suitable for analysis of CyA in ocular samples.
Keywords: Ocular sampels, Liquid chromatography, Cyclosporin A doi:10.1016/j.clinbiochem.2011.08.327
Poster – [A-10-250-1] Effect of granulocyte colony-stimulating factor on the activity of liver enzymes after using acute dosage of DMN or CCL4 in the liver tissue of rats Roya Abbasia, Durdi Qujegb, Hadi Parssianb, Sohrab Halalkhorb, Farideh Pheyzeeb, Hassan Taheryb, Mohssen Tatarb a Babol University of Medical Science, Babol, Iran b Babol Medical University, Iran E-mail addresses: [email protected] (R. Abbasi), [email protected] (D. Qujeg) Introduction: There are some reports in the literature showing that liver regeneration is important for the recovery process after liver damage. We studied the effect of G-CSF on the model of liver fibrosis caused by DMN, CCL4 in rats by measuring the activity of the index enzymes of the liver performance. Materials and methods: Acute liver damage was induced by injecting a dose (100–400 μg/kg) of CCL4 or DMN (100–400 μg/kg) in rats intraperitoneally. Liver damage and its regeneration were evaluated by measuring the activity of liver enzymes and histologically studies. Results: The serum ALT and AST activity level from CCl4 injection in rats receiving G-CSF at day 7 was expressed as Means ± S.E: (691.74 ± 54 U/L, 1211.97 ± 70 U/L) in G-CSF group and (837 ± 29 U/L,522.587 ± 12 U/L) in control group and the serum ALT and AST activity level from DMN injection in rats receiving G-CSF at day 7 expressed as Means ± S.E: (981.71 ± 19 U/L, 794.96 ± 55 U/L) in G-CSF group and (152.95 ± 55 U/L, 606.49 ± 23 U/L) in control group. Conclusion: This study supports the conclusion that G-CSF exerts a protective effect against toxic induced non lethal acute liver injury. Therefore, in future studies, it will be important to examine the effect of G-CSF in liver injury and regeneration. Keywords: G-CSF, DMN, CCL4, ALT, AST doi:10.1016/j.clinbiochem.2011.08.328
Poster – [A-10-256-1] Bovine serum albumin aggregation: An optimizing approach Amir Arasteh, Mehran Habibi Rezaei, Azadeh Ebrahim Habibi, Akbar Moosavi Movahedi Ali IBO, Tehran, Iran E-mail addresses: [email protected] (A. Arasteh), [email protected] (M. Habibi Rezaei), [email protected] (A. Ebrahim Habibi), [email protected] (A. Moosavi Movahedi Ali) Introduction: Many polypeptides and proteins with unrelated sequences and tertiary structures have been shown to be involved in amyloidoses. In fact, the consequential link between original protein structure at the native state and resultant conformation of amyloid protein, is supposed to be very important for realizing the aggregation process. Bovine serum albumin (BSA) is a ubiquitous protein involved in serum in vivo. BSA forms amyloid under destabilizing conditions in vitro and optimizing the aggregation conditions for producing the best one for the process could be very important to introduce a new biomaterial. Materials and methods: In this study, the optimization of process was assessed by various techniques such as circular dichroism (CD) spectroscopy, transmission electron microscopy and fluorescence
Abstracts
Poster – [A-10-245-1] Extraction of crocin from saffron (Crocus sativus) using molecularly imprinted polymer solid-phase extraction Ahmad Mohajeri Seyed, Hossein Hosseinzadeh, Fariborz Keyhanfar, Javad Aghamohammadian Mashhad University of medical science.Mashhad, Iran E-mail addresses: [email protected] (A. Mohajeri Seyed), [email protected] (J. Aghamohammadian) A new molecularly imprinted polymer for extraction of crocin from saffron stigmas was prepared using gentiobiose (a glycoside moiety in crocin structure) as a template. Crocin binding to gentiobiose imprinted polymer (Gent-MIP) was studied in comparison with a blank nonimprinted polymer in aqueous media. Affinity of the Gent-MIP for the crocin was more than the nonimprinted polymer at all concentrations. In Scatchard analysis, the number of binding sites in each gram of polymer (maximum binding sites) and dissociation constant of crocin to binding sites were 18.4 mmol/g polymer and 11.2 mM, respectively. The Gent-MIP was then used as the sorbent in an SPE method for isolation and purification of crocin from methanolic extract of saffron stigmas. The recovery of crocin, safranal and picrocrocin was determined in washing and elution steps. The Gent-MIP had significantly higher affinity for crocin than other compounds and enabled selective extraction of crocin with a high recovery (84%) from a complex mixture. The results demonstrated the possibility of using a part of a big molecule in preparing a molecularly imprinted polymer with a good selectivity for the main structure. Keywords: Saffron, Crocin, Molecularly imprinted polymer, Gentibiosis, HPLC
doi:10.1016/j.clinbiochem.2011.08.326
Poster – [A-10-245-2] Quantitative LC analysis of cyclosporine A in ocular samples Javad Aghamohammadian, Toka Banaee, Navid Mosallaei, Ahmad Mohajeri Seyed Mashhad, Iran E-mail addresses: [email protected] (J. Aghamohammadian), [email protected] (A. Mohajeri Seyed) Introduction: An isocratic reversed-phase HPLC method with ultraviolet detection at 205 nm has been developed for analysis of cyclosporine A (CyA) in rabbit ocular samples. Neither internal standard nor extraction was needed for sample preparation. Acetonitrile (ACN; 1 mL) was added to 250 μL aqueous and vitreous samples to precipitate proteins. The supernatant was dried and the residue was reconstituted in mobile phase and injected for HPLC analysis. Chromatography was performed on an octadecyl silane-A (ODSA) C18 (4.6 × 250 mm, 5 μm) column. The column temperature was fixed at 70 °C and the mobile phase was ACN 65%, methanol 20% and water 15% at a flow rate of 1.5 mL min− 1. The calibration curve for CyA in rabbit ocular samples was linear over the concentration range 0.2 and 10 μg mL− 1 with a correlation coefficient of 0.9992. Intra-day and inter-day precision were 4.61–7.83% and 5.27–10.70%, respectively. Intra-day and inter-day accuracy were 89.2–108% and 83.4–111%, respectively. The limits of detection (LOD) and quantification (LOQ) were 5.7 and 38 ng mL− 1, respectively. Conclusion: The method was successfully used for analysis of CyA in real aqueous and vitreous humor samples from New Zealand albino rabbits. The method is therefore suitable for analysis of CyA in ocular samples.
Keywords: Ocular sampels, Liquid chromatography, Cyclosporin A doi:10.1016/j.clinbiochem.2011.08.327
Poster – [A-10-250-1] Effect of granulocyte colony-stimulating factor on the activity of liver enzymes after using acute dosage of DMN or CCL4 in the liver tissue of rats Roya Abbasia, Durdi Qujegb, Hadi Parssianb, Sohrab Halalkhorb, Farideh Pheyzeeb, Hassan Taheryb, Mohssen Tatarb a Babol University of Medical Science, Babol, Iran b Babol Medical University, Iran E-mail addresses: [email protected] (R. Abbasi), [email protected] (D. Qujeg) Introduction: There are some reports in the literature showing that liver regeneration is important for the recovery process after liver damage. We studied the effect of G-CSF on the model of liver fibrosis caused by DMN, CCL4 in rats by measuring the activity of the index enzymes of the liver performance. Materials and methods: Acute liver damage was induced by injecting a dose (100–400 μg/kg) of CCL4 or DMN (100–400 μg/kg) in rats intraperitoneally. Liver damage and its regeneration were evaluated by measuring the activity of liver enzymes and histologically studies. Results: The serum ALT and AST activity level from CCl4 injection in rats receiving G-CSF at day 7 was expressed as Means ± S.E: (691.74 ± 54 U/L, 1211.97 ± 70 U/L) in G-CSF group and (837 ± 29 U/L,522.587 ± 12 U/L) in control group and the serum ALT and AST activity level from DMN injection in rats receiving G-CSF at day 7 expressed as Means ± S.E: (981.71 ± 19 U/L, 794.96 ± 55 U/L) in G-CSF group and (152.95 ± 55 U/L, 606.49 ± 23 U/L) in control group. Conclusion: This study supports the conclusion that G-CSF exerts a protective effect against toxic induced non lethal acute liver injury. Therefore, in future studies, it will be important to examine the effect of G-CSF in liver injury and regeneration. Keywords: G-CSF, DMN, CCL4, ALT, AST doi:10.1016/j.clinbiochem.2011.08.328
Poster – [A-10-256-1] Bovine serum albumin aggregation: An optimizing approach Amir Arasteh, Mehran Habibi Rezaei, Azadeh Ebrahim Habibi, Akbar Moosavi Movahedi Ali IBO, Tehran, Iran E-mail addresses: [email protected] (A. Arasteh), [email protected] (M. Habibi Rezaei), [email protected] (A. Ebrahim Habibi), [email protected] (A. Moosavi Movahedi Ali) Introduction: Many polypeptides and proteins with unrelated sequences and tertiary structures have been shown to be involved in amyloidoses. In fact, the consequential link between original protein structure at the native state and resultant conformation of amyloid protein, is supposed to be very important for realizing the aggregation process. Bovine serum albumin (BSA) is a ubiquitous protein involved in serum in vivo. BSA forms amyloid under destabilizing conditions in vitro and optimizing the aggregation conditions for producing the best one for the process could be very important to introduce a new biomaterial. Materials and methods: In this study, the optimization of process was assessed by various techniques such as circular dichroism (CD) spectroscopy, transmission electron microscopy and fluorescence