Determination of tromethamine residues in human coagulation factor Ⅷ by high performance liquid chromatography method
Journal Title: Chinese Journal of Blood Transfusion - Year 2022, Vol 35, Issue 10
Abstract
Objective To develop and verify high performance liquid chromatography (HPLC) detection for tromethamine (Tris) residues in human coagulation factor Ⅷ. Methods Alanine was used as internal standard, and AQC for pre-column derivation. Inertsil® ODS-SP was adopted, and acetic acid-sodium acetate buffer and acetonitrile were used for gradient elution at the flow rate of 1 mL/min and column temperature of 37℃. The 25 μL sample was loaded and determined by UV-detector with detection wavelength at 248 nm. This method was then verified. Results Glycine, sodium citrate and calcium chloride showed no interference with the detection of tromethamine (Tris) residues. The recovery rate of spike samples was within 90.0%~100.8%. The RSD in repeatability test were 2.5%, 0.7% and 1.2%, respectively, and in intermediate precision test 0.8%. The tromethamine (Tris) at concentrates of (0.5~4.0)μg/mL showed good linear relationship to the peak area to internal standard, of which the regression equation was Y=0.012 9X-0.000 782, R=0.999 97.The quantitative detection limit was 0.04μg/mL. Conclusion The HPLC for determination of tromethamine (Tris) residues in human coagulation factor Ⅷ was successfully developed, which showed good linearity as well as high specificity, precision and accuracy.
Authors and Affiliations
Xiuliang CHEN, Erhua LUO, Xiaoming LIANG, Jiafei CHEN
Keywords
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- EP ID EP738372
- DOI 10.13303/j.cjbt.issn.1004-549x.2022.10.006
- Views 33
- Downloads 0
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