复方盐酸伊立替康和氟尿嘧啶脂质体注射液包封率测定方法研究

目的 测定复方脂质体注射液中盐酸伊立替康与氟尿嘧啶包封率。方法 采用凝胶柱层析法，以pH 6.5的磷酸盐缓冲溶液为洗脱剂，柱高400 mm，洗脱流速为1.0 mL·min^-1。在建立的HPLC-DAD色谱条件下同时测定。使用Kromasil C₁₈柱（250 mm×4.6 mm，5 μm），以甲醇-pH 6.6磷酸缓冲盐（5∶95）为流动相A，以甲醇-pH 6.6磷酸缓冲盐（60∶40）为流动相B，梯度洗脱。柱温35℃，流速1.0 mL·min^-1。检测波长分别为255，268 nm。结果 该方法能有效分离复方脂质体注射液中包封药物和游离药物。3种不同试验水平下，盐酸伊立替康柱回收率分别为98.9%，100.5%，100.4%；氟尿嘧啶柱回收率分别为98.2%，100.5%，100.2%。在液相色谱条件下，复方脂质体注射液中盐酸伊立替康与氟尿嘧啶能很好地分离检出，其线性浓度范围分别为10.7~107.0 μg·mL^-1（r=0.999 9，n=5），3.9~38.6 μg·mL^-1（r=1.000 0，n=5），呈良好的线性关系；盐酸伊立替康低、中、高浓度加样回收率分别为99.7%，99.0%，99.8%，RSD分别为0.20%，0.48%，1.38%；氟尿嘧啶低、中、高浓度加样回收率分别为100.3%，100.6%，99.8%，RSD分别为0.72%，0.09%，0.67%。结论 本方法简单，准确，重复性好，可用于测定复方脂质体中盐酸伊立替康、氟尿嘧啶2种药物的包封率。

Study on Determination Method of Encapsulation Rate of Irinotecan Hydrochloride and 5-Fluorouracil in Compound Liposome Injection

OBJECTIVE To determine encapsulation rate of irinotecan hydrochloride and 5-fluorouracil in compound liposome injection. METHODS The encapsulation rate was determined by sephadex column chromatography method with the pH 6.5 phosphate buffer solution as eluent. The sephadex column was 400 mm and the eluent flow rate was 1.0 mL·min^-1. Under the established HPLC-DAD chromatographic condition, it was used with Kromasil-C₁₈(250 mm×4.6 mm, 5 μm) by methanol-pH 6.6 phosphate buffer(5:95) as mobile phase A and methanol-pH 6.6 phosphate buffer(60:40) as mobile phase B. The column temperate was kept at 35℃. The flow rate was 1.0 mL·min^-1. The detector wavelength were set at 255, 268 nm, respectively. RESULTS The encapsulation drug and free drug could be separated effectively in compound liposome injection by this way. The recoveries of 3 concentrations of sephadex column for irinotecan hydrochloride were 98.9%, 100.5% and 100.4%, and for 5-fluorouracil were 98.2%, 100.5% and 100.2%. Under the applied chromatographic condition, irinotecan hydrochloride and 5-fluorouracil were separated from compound liposome injection. The calibration curves were linear in the range of 10.7-107.0 μg·mL^-1 for irinotecan hydrochloride(r=0.999 9, n=5) and 3.9-38.6 μg·mL^-1 for 5-fluorouracil(r=1.000 0, n=5). The average recoveries for irinotecan hydrochloride were 99.7%, 99.0% and 99.8%, and RSD were 0.20%, 0.48% and 1.38%. The average recoveries for 5-fluorouracil were 100.3%, 100.6% and 99.8%, and RSD were 0.72%, 0.09% and 0.67%. CONCLUSION The method is simple, accurate and reproducible. It can be used for the determination of encapsulation rate of irinotecan hydrochloride and 5-fluorouracil in compound liposome injection.