HPLC法测定阿司达莫缓释胶囊在人体内的血药浓度及其生物等效性研究

Determination of Dipyridamole and Salicylic Acid in Compound Dipyridamole Sustained-release Capsules by HPLC and Study on Its Bioequivalence

目的：建立人血浆中双嘧达莫和水杨酸的高效液相色谱（HPLC）测定法，研究2种阿司达莫缓释胶囊的生物等效性。方法：采用自身对照随机交叉给药方案，志愿者分别口服2种阿司达莫缓释胶囊后在不同时间点取血，血浆样品以新建立的HPLC法测定，应用不同的流动相经Diamonsic C18柱，分别测定水杨酸和双嘧达莫，检测波长分别为238nm和290nm。结果：水杨酸和双嘧达莫与血浆中内源性杂质分离度好，水杨酸、双嘧达莫线性范围分别为20～2000（r=0．9997）、25～4000（r：0．9999）μg·L^-1；方法回收率分别在94．7％～104．4％和87．7％～100．0％之间，日内RSD分别〈5．2％、2．2％，日间RSD分别〈9．0％、8．8％。单剂量后参比制剂和受试制剂中水杨酸的Cmax分别为（734．5±106．7）、（785．4±148．5）μg·L^-1，tmax分别为（0．9±0．2）、（0．8±0．3）h，AUC0-12分别为（2579．1±606．2）、（2571．8±700．0）μg·h·L^-1；双嘧达莫的Cmax分别为（1080．7±740．9）、（936．4±686．1）μg·L^-1，tmax分别为（2．4±0．7）、（2．8±0．7）h，AUC0-24分别为（4699．9±3166．8）、（4841．0±3201．9）μg·h·L^-1。受试制剂相对于参比制剂的生物利用度为（99．7±13．3）％。结论：本方法简便快速，灵敏度高，可用于双嘧达莫和阿司匹林在人体内药动学过程的研究。2种制剂生物等效。

OBJECTIVE： To develop an HPLC method for the determination of dipyridamole and salicylic acid in plasma and evaluate the bioequivalence of compound dipyridamole sustained - release capsules. METHODS： By self - control and randomized crossover way, the volunteers were assigned to receive two kinds of compound dipyridamole sustained release capsules. Then serum samples were taken at different time for the determination of salicylic acid and dipyridamole by HPLC on Diamonsic C18 column with different mobile phase. The detection wavelength was 238 nm and 290 nm, respectively. RESULTS： Salicylic acid and dipyridamole were separated well from endogenous foreign substances. The calibration curve of salicylic acid and dipyridamole were liner over the range of 20-2 000μg·L^-1 （r = 0.999 7）and 25-4 000 μg·L^-1（r = 0.999 9）, respectively. The method recovery rates were within 94.7%- 104.4% and 87.7%- 100.0%, respectively; the inter- day RSD were less than 5.2% and 2.2%, respectively and the intra day RSD were less than 9.0% and 8.8%, respectively. After single dose of the two formulations （test formulation vs. the reference formulation）, the pharmacokinetic parameters of salicylic acid were as follows： Cmax were （734.5±106.7）μg·L^-1and （785.4± 148.5）μg·L^-1,tmax were （0.9±0.2） h and （0.8±0.3） h; AUC0-12 were （2 579.1±606.2） μg·h·L^-1 and （2 571.8± 700.0）μg·h·L^-1; the pharmacokinetic parameters of dipyridamole were as follows： C were （1 080.7±740.9）μg·L^-1 and （936.4±686.1）μg·L^-1; tmax were （2.4±0.7） h and （2.8±0.7） h; AUC0-24 were （4 699.9± 3 166.8）μg·h·L^-1 and （4 841.0± 3 201.9） μg·h·L^-1, respectively. The relative bioavailability of test formulation versus the reference one was （99.7 ± 13.3） % . CONCLUSION： The method is simple, sensitive and rapid for the pharmacokinetic study of dipyridamole and salicylic acid in human plasma. The two formulations of dipyridamole sustained- release capsules are bioequivalent.