Determination of Fluvoxamine in Human Plasma by FPLC-MS/MS / 中国药师

Objective:To establish an FPLC-MS/MS method for the determination of fluvoxamine in human plasma. Methods:The separation was performed on an Inertsil? ODS-SP column(2. 1 × 100 mm, 3 μm). The mobile phase was acetonitrile-2 mmol· L-1 ammonium acetate (45: 55, v/v) containing 0. 1% formic acid at a flow rate of 0. 3 ml·min-1 . Lansoprazole was used as the internal standard( IS) . Electrospray ionization ( ESI) source was applied and operated in a positive ion model. Multiple reaction moni-toring (MRM) model with the transitions of fluvoxamine m/z 319. 1→m/z 69. 8 and lansoprazole m/z 370. 2→m/z 252. 1 was used to quantify fluvoxamine and IS, respectively. Results:In human plasma, the standard curve was linear within the range of 1-100 μg· L-1 . The lower limit of quantification of fluvoxamine( LLOQ) was 1μg·L-1 . The intra-day RSD was less than 5%, the inter-day RSD was less than 10%, and the method recovery was 85%-95%. Conclusion:The method is simple, sensitive, accurate and reproduci-ble. It is applicable in the pharmacokinetic study of fluvoxamine for clinical pharmacokinetics and bioequivalence studies.

Determination of Fluvoxamine in Human Plasma by FPLC-MS/MS / 中国药师

Objective:To establish an FPLC-MS/MS method for the determination of fluvoxamine in human plasma. Methods:The separation was performed on an Inertsil? ODS-SP column(2. 1 × 100 mm, 3 μm). The mobile phase was acetonitrile-2 mmol· L-1 ammonium acetate (45: 55, v/v) containing 0. 1% formic acid at a flow rate of 0. 3 ml·min-1 . Lansoprazole was used as the internal standard( IS) . Electrospray ionization ( ESI) source was applied and operated in a positive ion model. Multiple reaction moni-toring (MRM) model with the transitions of fluvoxamine m/z 319. 1→m/z 69. 8 and lansoprazole m/z 370. 2→m/z 252. 1 was used to quantify fluvoxamine and IS, respectively. Results:In human plasma, the standard curve was linear within the range of 1-100 μg· L-1 . The lower limit of quantification of fluvoxamine( LLOQ) was 1μg·L-1 . The intra-day RSD was less than 5%, the inter-day RSD was less than 10%, and the method recovery was 85%-95%. Conclusion:The method is simple, sensitive, accurate and reproduci-ble. It is applicable in the pharmacokinetic study of fluvoxamine for clinical pharmacokinetics and bioequivalence studies.

Comparison of fast protein liquid chromatography (FPLC) with HPLC, electrophoresis & microcolumn chromatography techniques for the diagnosis of β-thalassaemia.

Background & objectives: β-thalassaemia is a genetic disorder and an important health problem around the world. Quantitative haemoglobin A2 (HbA2) levels are used for the diagnosis of β-thalassaemia. The conventional methods are high performance liquid chromatography (HPLC), electrophoresis, and microcolumn chromatography techniques. We established a fast protein liquid chromatography (FPLC) method, to measure quantitatively of HbA2 levels, and compared its efficacy with conventional methods. Methods: The FPLC method, using a DEAE Sepharose, Hi Trap anion-exchange column chromatography technique was set up for HbA2 measurement. In this study, 220 blood samples were screened for haemoglobin type by FPLC technique and also using HPLC, microcolumn chromatography and electrophoresis. Results: The FPLC results were highly correlated (r = 0.985, P<0.001) with those of HPLC for quantification of HbA2 as well as cellulose acetate electrophoresis (r = 0.977) and microcolumn chromatography (r = 0.980). The FPLC method showed 100 per cent sensitivity and specificity, positive and negative predictive value for β-thalassaemia diagnosis. In addition, the FPLC method was simple, rapid, low cost and reproducible. The HbA2/E range of FPLC for β-thalassaemia was 6-10 per cent, HbE trait was 10-40 per cent, β-thalassaemia/HbE was 40-60 per cent and homozygous HbE was more than 60 per cent. Interpretation & conclusions: Our findings suggested that FPLC method could be used as a cost-effective method for routine β-thalassaemia diagnosis.

Purification, identification and characterization of the major allergen from Linnaeus / 中国免疫学杂志

Objective:To purify and characterize the major antigenic components of sand swimming crab,and provide theoretical evidences for the research of standardization of allergenic vaccine.Methods:The crude extract of Linnaeus was prepared using classical method, and electrophoresis of SDS-PAGE was used to separate the proteins of each group. The allergen proteins of 26 crab were identified using Western blot. To distinguish between the primary allergen proteins and secondary allergen proteins, FPLC(Gel Chromatography and ion exchange chromatography) was used to filtrate and identify the allergen protein.Results:SDS-PAGE analysis revealed that sand swimming crab proteins were composed of at lease 9 discrete protein bands, which molecular weight ranging from 13 000 to 90 000. Major bands were of 20 900, 24 200, 27 100, 29 200, 33 700, 38 900, 48 700, 74 700, 89 100 respectively. Western blot assay indicated that the crude extract reacted with sera obtained from 26 crab allergenic subjects and contained 5 allergen bands altogether, and the bands of 74 400 and 48 700 were the major allergenic components. The positive rates of the two major allergen proteins were both 100%, indicating the allergenic components maintained the immunocompetence after yielding from chromatography.Conclusion:The 74 400 and 48 700 bands are the major allergens of sand swimming crab. The major allergen proteins can be purified by chromatography.