HPLC法测定米铂白蛋白结合型纳米制剂中米铂的含量

Determination of human serum albumin-bound miriplatin by HPLC

目的建立米铂白蛋白结合型纳米制剂中米铂含量测定的高效液相色谱(HPLC)法。方法采用HPLC测定米铂白蛋白结合型纳米制剂中米铂的含量。色谱条件:辛烷基硅烷键合硅胶色谱柱(250 mm×4.6 mm,5μm),甲醇-乙腈-水(91:1:8)洗脱,流速1.0 ml/min,检测波长210 nm,柱温30℃,进样量20μl。结果建立了简便可行的米铂白蛋白结合型纳米制剂样品前处理方法;建立并验证了HPLC方法,线性范围3.63~130.80μg/ml(r=0.9995);低、中、高3个浓度的平均回收率(n=3)分别为(99.46±0.24)%、(99.20±1.38)%、(98.30±0.26)%;日内、间精密度RSD分别为1.62%(n=6)和1.32%(n=6);流速在0.99~1.01 ml/min范围内变化时米铂峰面积的RSD为1.01%(n=9),柱温在28~32℃之间变化时米铂峰面积的RSD为1.14%(n=9)。结论本文建立的含量测定方法简便易行,专属性强,灵敏度高,耐用性好;可定量检测米铂白蛋白结合型纳米制剂中米铂的含量。

Objective To establish a high performance liquid chromatography(HPLC)for the quantification of miriplatin in human serum albumin-bound miriplatin nanoparticle(HSA-miriplatin).Methods The analysis was performed on an octylsilane bonded silica column(250 mm×4.6 mm,5μm)at the flow rate of 1.0 ml/min with SPD detector at 210 nm.The mobile phase consisted of methanol,acetonitrile and water(91:1:8).Results Free miriplatin was extracted from HSA-miriplatin by methanol,and the linear range of miriplatin was 3.63-130.80μg/ml with a correlation coefficient of r=0.9995.The average recovery rates were(99.46±0.24)%,(99.20±1.38)%,(98.30±0.26)%(n=3)at low,medium,high concentrations,respectively.The relative standard deviation(RSD)values reflecting inter-day and intra-day precision were 1.62%(n=6)and 1.32%(n=6),respectively.The method was robust with slight change of flow rate(0.99-1.01 ml/min)and column temperature(28-32℃)with RSD of 1.01%(n=9)and 1.14%(n=9),respectively.Conclusion A simple,selective,sensitive and robust HPLC method is established for determination of miriplatin loaded in HSA-miriplatin.