HPLC法测定他克莫司软膏剂含量及含量均匀度

Content determination and content uniformity of tacrolimus ointment by HPLC

目的建立他克莫司软膏剂含量测定方法。方法用四氢呋喃∶水∶无水乙醇(5∶2∶1)先、后在60℃水浴和冰浴中提取软膏,应用反相高效液相色谱法测定了两种规格共6批(30g,每支0.03%;30g,每支0.1%)他克莫司软膏剂的含量及含量均匀度。色谱条件:TSKgelODS-80TM柱(150mm×4.6mm,5μm);柱温(55±1)℃,流动相为水∶异丙醇∶四氢呋喃(5∶2∶2),流速0.6ml/min;紫外检测器,检测波长220nm。结果他克莫司的色谱峰与其它杂质峰能够较好的分离,柱理论塔板数按他克莫司峰计为5000,规格为30g,每支0.03%软膏的80%加样平均回收率为100.6%(RSD=0.5%,n=3),100%加样平均回收率为100.2%(RSD=1.6%,n=3),120%平均回收率为98.7%(RSD=2.3%,n=3);规格为30g,每支0.1%软膏的80%加样平均回收率为99.3%(RSD=0.5%,n=3),100%加标平均回收率为100.8%(RSD=1.6%,n=3),120%加标平均回收率为99.3%(RSD=0.7%,n=3);0.03%规格软膏的线性方程为y=6989786x+105.4(r=0.9999),浓度范围0.0075～0.0225mg/ml;0.1%规格的线性方程为y=6995488x-64.2(r=0.9999),浓度范围0.025～0.075mg/ml。他克莫司的检测限为0.002mg/ml,定量限为0.006mg/ml。结论方法简便,快速,回收率高,重现性好,可用于他克莫司软膏剂(0.03%,0.1%)含量测定,能够对药品质量进行控制。

Objective The content determination of tacrolimus ointment and its content uniformity were studied by using reversed-phase high performance liquid chromatography. Methods In this paper, for quantitative analyses of tacrolimus ointment （two strengths, 30g： 0.1%/tube and 30g： 0.03%/tube）, a mixture of THF, water and absolute ethanol （5 ： 2 ： 1） in water bath （60℃） and followed by ice bath was used to extract the ointment to prepare the sample solution. The separation was carried out on a TSK gel ODS-80^TM （150mm×4.6mm, 5μm） with PDA detector at 220nm, the temperature of column was 55℃. The mobile phase consisted of water ： isopropanol ： THF （5 ： 2 ： 2）, at a flow rate of 0.6ml/min. Result Good resolution was achieved between the peak of tacrolimus and the relative impurities. The 80%, 100% and 120% sample added recoveries for one strength （30g： 0.03%/tube） were 100. 6%, 100.2% and 98.7% respectively, and for another strength （30g： 0.1%/tube） were 99.3%, 100.8% and 99.3% respectively. Conclusion This HPLC method for content determination of tacrolimus ointmentis was simple, rapid, reproducible, and with high recovery. It was also feasible to assay and determinate the content uniformity of tacrolimus ointment （0.03% and 0.1%）.