HPLC法测定血小板贮存袋增塑剂BTHC在混合浓缩血小板保存过程中的溶出量

目的：建立增塑剂丁酰柠檬酸三正己酯（butyryl trihexyl citrate，BTHC）的测定方法，并测定血小板贮存袋所含增塑剂BTHC在混合浓缩血小板保存过程中的溶出量，为该产品的安全性评价提供依据。方法：采用乙腈沉淀血小板悬液中的蛋白等杂质，离心所得上清液用高效液相色谱法（HPLC）测定BTHC的含量，并对测定方法进行方法学研究。结果：HPLC方法测定BTHC浓度在10~502 μg&#183;mL^-1范围内线性关系良好（r=0.999 9），灵敏度为0.04 μg，仪器精密度RSD<5%，回收率为98.6%&#177;8.4%，用该方法测定血小板贮存袋贮存一个治疗剂量的混合浓缩血小板时BTHC的溶出量，贮存第5天溶出量为28.10&#177;2.38 mg；贮存第7天溶出量36.63&#177;3.89 mg。结论：本法经方法学验证，可以用于测定BTHC在血小板悬液中的溶出量。

Objective: To develop a method for the determination of plasticizer Butyryl trihexyl citrate(BTHC) released from platelet storage containers into pooled platelet concentrates and to evaluate the safety of this product.Method: BTHC was obtained by centrifugation of pooled platelet concentrates using acetonitrile as protein precipitation agent, and the quantification of BTHC was achieved by HPLC.The method was validated by methodology study.Results: The methodology study of HPLC was presented that the concentration of BTHC 10-502 μg&#183;mL^-1 had a good linear relationship(r=0.999 9), the method sensitivity was 0.04 μg, instrumental precision showed RSD<5%, and recovery rate was 98.6%&#177;8.4%.By this method, the amount of BTHC releasing into platelet concentrates with one treatment dose from platelet storage bags was 28.10&#177;2.38 mg at the 5^th day of storage and 36.63&#177;3.89 mg at the 7^th day of storage.Conclusion: This method showed high accuracy, good reliability and stability for the determination of BTHC leaching into pooled platelet concentrates.