气相色谱法测定猪凝血酶干粉中磷酸三丁酯残留量.docVIP

气相色谱法测定猪凝血酶冻干粉中磷酸三丁酯残留量 　　摘 要 建立了气相色谱法测定猪凝血酶中磷酸三丁酯残留量，并进行了方法学验证。采用聚乙二醇毛细管柱Agilent HP-INNOWAX（30 m × 250 mm × 0.25 mm），溶剂为正己烷；气化室温度190 ℃，分流比1∶1，进样量1 ml；FID检测器温度210 ℃；载气为高纯氮气，流速为3 ml/min；柱温150 ℃，运行时间13 min。结果显示该方法专属性强；标准曲线的线性范围为2.406～19.25 mg/ml；低、中、高3个浓度水平的平均回收率分别为97.1%、101.1%和98.2%，总平均回收率为99.8%，总相对标准偏差为2.37%；中间精密度总标准偏差为5.01%；检测限为0.762 mg/ml，定量限为2.54 mg/ml；3批样品的磷酸三丁酯残留量均低于《中华人民共和国药典》2015年版三部的限量要求（中国论文网 /6/viewhtm　　关键词 气相色谱 猪凝血酶 磷酸三丁酯 ??留 　　中图分类号：O657.71； R926 文献标识码：A 文章编号：1006-1533（2017）21-0076-05 　　Determination of the residual tri-n-butyl phosphate in porcine thrombin by gas chromatography 　　LIU Lu*， ZHAO Yingying， ZHANG Jundong 　　（Shanghai Haohai Biological Technology Co.， Ltd， Shanghai 200052， China） 　　ABSTRACT A gas chromatography method was established to determine the residual Tri-n-butyl phosphate in porcine thrombin and was completely evaluated. An Agilent HP-INNOWAX column （30 m×250 mm×0.25 mm） was adopted and the operation procedures were as follows： vaporizing chamber temperature 190 ℃， split ratio 1：1， sample volume 1 ml； FID detector temperature 210 ℃； carrier gas： highly pure nitrogen at 3 ml/min； column temperature 150 ℃ and running for13 min. The results showed that the method had strong specificity and the standard curve was linear over the range of 2.406～19.246 mg/ml. The average recoveries at low， medium and high levels were 97.1%， 101.1% and 98.2%， respectively and the overall average recovery was 99.8%， and the overall RSD value was 2.37% and RSD value of the mid-precision was 5.01%. The limit of detection was 0.762 mg/ml and the limit of quantitation was 2.54 mg/ml. The residual TNBP values from three batches of porcine thrombin were apparently lower than the limit value set by Chinese Pharmacopoeia （ 　　磷酸三丁酯（GC，纯度99.9%，批号：BCBN5037V）购自上海凌峰化?W试剂有限公司；磷酸三丙酯（GC，纯度99%，批号：38196TJV）购自美国Sigma-Aldrich公司；正己烷（HPLC，批号购自上海迈瑞尔化学技术有限公司；高氯酸（AR，批号购自永华化学科技（江苏）有限公司；水为纯化水。 　　1.2 主要仪器 　　安捷伦7890B气相色谱仪含氢火焰离子化检测器（FID）（美国安捷伦科技有限公司）；ME104