RP-HPLC法测定眼房水中三种不同剂型匹罗卡品含量

目的:建立反相高效液相色谱法(RP-HPLC)测定眼房水中匹罗卡品含量的方法.方法:RP-HPLC法采用ODS-C18柱,流动相:0.5%三乙胺的KH2PO4(10 mmol/L)缓冲液(pH 2.5)-乙腈(98/2,v/v),流速:1.0 ml/min,检测波长:215 nm.90只兔均分为3组,分别于每只兔眼滴入不同剂型滴眼剂50 μl,分别测定给药后5、10、30、40、60、90、120、180、240及360 min时眼房水中匹罗卡品滞留量,用二氯甲烷提取房水中药物.结果:RP-HPLC法适用于眼房水中匹罗卡品的含量测定,匹罗卡品质量浓度在0.1～20 μg/ml范围内线性良好(r=0.9998),平均提取回收率为68.1%±2.7%(n=9),日内精密度为2.87%,日间精密度为4.33%.3种不同剂型匹罗卡品50 μl滴眼后,药-时曲线下面积以1%脂质体为最高,且维持时间最长.结论:本方法易行,重现性好,线性范围广.将匹罗卡品滴眼液的剂型改为脂质体可以明显提高匹罗卡品的生物利用度,具有较好的应用前景.

Determination of three formulations of pilocarpine in rabbit ocular aqueous by RP-HPLC

OBJECTIVE: To develop an RP-HPLC method for assay of pilocarpine in rabbit ocular aqueous humor. METHODS: The RP-HPLC method was performed on a column of ODS-C(18) with the mobile phase consisting of 0.5% of triethylamine (TEA) of phosphate solutions (10 mmol/L, pH 2.5) and acetonitrile (98/2,v/v). The detection wavelength was 215 nm and flow rate was 1.0 ml/min. Ninety albino rabbits were divided into 3 groups (30 in each):group 1 received 50 microl of eye drops containing 1% generic pilocarpine, group 21% mixture pilocarpine solution consisting of aqueous sample and liposome and group 31% liposome pilocarpine, respectively. The aqueous humor was withdrawn at 5, 10, 30, 40, 60, 90, 120, 180, 240 and 360 min. Pilocarpine was extracted from aqueous humor with dichloromethane. RESULT: The linear calibration curve was obtained in the concentration range of 0.1 - 20 microg/ml. The average recovery was (68.1+/-2.7)% (n=9). Inter-day and intra-day RSD were 4.33% and 2.87%, respectively. In three formations 1% liposome pilocarpine was the best for the areas under curve and measurable amounts. CONCLUSION: The RP-HPLC method is simple and reliable for pilocarpine measurement in ocular aqueous. Liposome formulation can significantly increase the bioavailability of pilocarpine in ocular aqueous.