Establishment of atomic absorption spectrophotometry for determination of calcium content in plasma-derived FⅧ products and evaluation of uncertainty
10.13200/j.cnki.cjb.004488
- VernacularTitle:血浆源性人凝血因子Ⅷ制品中钙含量原子吸收分光光度检测方法的建立及不确定度评价
- Author:
ZHANG Ting
- Publication Type:Journal Article
- Keywords:
Plasma-derived FⅧ(pdFⅧ);
Atomic absorption spectrophotometry;
Calcium content;
Uncertainty
- From:
Chinese Journal of Biologicals
2025;38(05):581-587
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an atomic absorption spectrophotometry for the determination of calcium content in humanplasma-derived FⅧ(pdFⅧ) products, and to evaluate the uncertainty of the method, so as to provide a reliable method forthe determination of calcium content in blood products. Methods After adding lanthanum chloride solution as ionizationinhibitor to pdFⅧ, the calcium content was detected by atomic absorption spectrophotometry. The gas flow rate(1. 2, 1. 4,1.6,1.8,2.0L/min),burnerheight(9,11,12,13mm),lampcurrent(100%,90%,85%,80%),andslitwidth(0.2,0.5,1.0nm)were optimized, and the method was verified for specificity, linearity range, limit of detection(LOD), limit of quantitation(LOQ),accuracy,precisionandstability.ThecalciumcontentinsixbatchesofpdFⅧsampleswasdeterminedbytheestablishedmethod.Theuncertaintyofthemethodwasevaluated,andtakingpdFⅧsampleofbatchnumber202404026asanexample,thecalcium content was determined, and the effects of various sources of uncertainty on the detection results were determined.Results The optimum gas flow rate was 1. 8 L/min, the combustion gas height was 12 mm, the lamp current was 85%,and the slit width was 0. 5 nm. Lanthanum chloride did not interfere with the determination of calcium content in pdFⅧ. Inthe concentration range of 1. 0-5. 0 μg/mL, the calcium working standard solution had a good linear relationship with A_(422.7),with the regression equation of: y = 0. 016 6 x-0. 000 8, r = 0. 998 7. The LOD was 0. 127 2 μg/mL, and the LOQ was0. 385 5 μg/mL. The recovery rates of low, medium and high concentration spiked samples were all in the range of 85%-110%. The RSDs of precision and stability verification were both less than 2%. The calcium content of six batches of pdFⅧsamples was not more than 50 μg/mL. The uncertainty was mainly derived from the preparation of standard solution. Thecalcium content of pdFⅧ sample with batch number 202404026 was(42. 11 ± 1. 736 1) μg/mL, which met the requirement(≤ 50 μg/mL), indicating that the sources of uncertainty showed no influence on the determination results of calcium concen-tration. Conclusion The established atomic absorption spectrophotometry has good accuracy, precision and stability, andthe reliability of detection results is ensured by uncertainty evaluation, which can be used for the accurate determination ofcalcium content in pdFⅧ.
