目的: 建立注射用重组人凝血因子Ⅷ中钙含量的原子吸收分光光度检测方法,并评定其不确定度。方法: 原子吸收分光光度法采用钙元素空心阴极灯光源;空气-乙炔火焰原子化器;检测波长422.7 nm;氧化镧溶液作为电离抑制剂。开展专属性、准确度、精密度、定量限、线性和范围、稳定性等考察;同时分析测定方法的不确定度。结果: 钙元素质量浓度在0.0~5.0 μg · mL-1范围内与吸收度线性关系良好,r=0.999 8;检测限为0.071 6 μg · mL-1;定量限为0.216 9 μg · mL-1。加标试样的回收率为100.8%~103.2%。重复性RSD为0.64%,溶液在8 h内稳定。6批样品含量测定结果均符合规定。不确定度评定结果为（117.12±4.82）μg · mL-1,k=2。结论: 建立原子吸收分光光度测定注射用重组人凝血因子Ⅷ中钙含量的方法,具有灵敏度高,分析速度快,重复性好,准确度高,节约分析成本等优点。同时对该方法进行了不确定度评定,可为该品种的质量控制提供量化评价指标。
Objective: To establish a atomic absorption spectroscopy (AAS) method for detecting calcium content in recombinant human coagulation factor Ⅷ for injection, and evaluate its uncertainty. Methods: A calcium hollow cathode lamp light source was used. Air acetylene flame atomizer. Detection wavelength 422.7 nm. 8.9% lanthanum oxide solution was used as an ionization inhibitor. Conduct inspections on specificity, accuracy, precision, quantification limit, linearity and range, stability, etc. in accordance with the validation guidelines for drug quality standard analysis methods in Part 9101 of the 2020 edition of the Chinese Pharmacopoeia. The established method was applyed to detect the content of 6 batches of samples. Results: There was a good linear relationship between calcium element and absorbance in the concentration range of 0.0-5.0 μg · mL-1, with r=0.999 8. The detection line was 0.071 6 μg · mL-1; The quantification limit was 0.216 9 μg · mL-1. The recovery rate of the spiked sample was 100.8% to 103. 2%. The repeatability RSD was 0.64%, and the solution was stable within 8 hours. The content determination results of 6 batches of samples all complied with the regulations. The uncertainty assessment result was (117.12±4.82) μg · mL-1, k=2. Conclusion: AAS method is established for the detection of calcium content in recombinant human coagulation factor Ⅷ for injection.This method has the advantages of high sensitivity, fast analysis speed, good repeatability, high accuracy, and cost savings. At the same time, the uncertainty of this method is evaluated, which can provide quantitative evaluation indicators for the quality control of this variety.
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