RP-HPLC法测定眼房水中三种不同剂型匹罗卡品含量  

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

作　　者：王向军[1] 陈滨[2] 姚彤炜[1] 张惠成[2] 曾苏[1] 

机构地区：[1]浙江大学药学院药物分析与药物代谢研究室,浙江杭州310031 [2]浙江省杭州市第一人民医院,浙江杭州310006

出　　处：《浙江大学学报（医学版）》2004年第1期33-36,共4页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省自然科学基金 (30 2 6 82 ) ;浙江省经贸委课题资助

摘　　要：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 KH2 PO4(10 mmol/ L)缓冲液 (p H 2 .5 ) -乙腈 (98/ 2 ,v/ v) ,流速 :1.0 ml/ min,检测波长 :2 15 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 5 0μl,分别测定给药后 5、10、30、4 0、6 0、90、12 0、180、2 4 0及 36 0 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～ 2 0μg/ ml范围内线性良好 (r=0 .9998) ,平均提取回收率为 6 8.1%± 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 5 0μl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。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 μl of eye drops containing 1% generic pilocarpine,group 2 1% mixture pilocarpine solution consisting of aqueous sample and liposome and group 3 1% 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 The aqueous humor was withdrawn at 5,10,30,40,60,90,120,180,240 and 360 min.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. Results: The linear calibration curve was obtained in the concentration range of 0.1~20 μg/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.

关 键 词：毛果芸香碱 投药和剂量 色谱法 高效液相 兔 

分 类 号：R927.2[医药卫生—药学]