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目的:建立HPLC同时测定绿原酸、连翘酯苷A和黄芩苷的血药浓度,探析注射用双黄连粉针中3种成分在大鼠体内的药代动力学特征。方法:采用HPLC同时测定绿原酸、连翘酯苷A和黄芩苷含量,以葛根素为内标,流动相乙腈(A)-0.1%磷酸(B)梯度洗脱(0~10 min,10%~20%A;10~35 min,20%~35%A;35~40 min,35%A;40~40.5 min,35%~10%A;40.5~55 min,10%A),绿原酸、连翘酯苷A、黄芩苷和葛根素的检测波长分别为328,328,277,249 nm。结果:绿原酸和连翘酯苷A在0.2~20 mg·L-1、黄芩苷在2.0~300 mg·L-1线性关系良好(r>0.999 4);日内和日间精密度的RSD均<13.6%,低、中、高3个质量浓度样品的方法回收率在88.3%~109.3%,提取回收率均>80.8%。绿原酸、连翘酯苷A和黄芩苷的主要药动学参数AUC0-t分别为(8.406±1.175),(9.696±2.349),(145.35±22.02)mg·h·L-1;MRT0-t依次为(0.619±0.105),(0.634±0.115),(0.456±0.068)h;t1/2 Z分别为(0.532±0.064),(0.732±0.357),(0.542±0.175)h;Vz分别为(0.384±0.050),(0.673±0.422),(0.324±0.072)L·kg-1;CL Z分别为(0.504±0.067),(0.634±0.150),(0.426±0.066)L·h-1·kg-1。结论:该方法灵敏、简便、准确,适用于大鼠血浆中绿原酸、连翘酯苷A和黄芩苷的测定及注射用双黄连粉针的药代动力学研究。
OBJECTIVE: To establish HPLC for the simultaneous determination of plasma concentrations of chlorogenic acid, forsythoside A and baicalin, and to investigate the pharmacokinetics of three components in Shuanghuanglian injection for injection in rats. Methods: Simultaneous determination of chlorogenic acid, forsythoside A and baicalin by HPLC was carried out with puerarin as an internal standard. The mobile phase was eluted with a gradient of acetonitrile (A) -0.1% phosphoric acid (B) (0-10 min, 10 % To 20% A; 10 to 35 minutes, 20% to 35% A; 35 to 40 minutes 35% A; 40 to 40.5 minutes 35% to 10% A; 40.5 to 55 minutes 10% A) Protogenic acids, forsythoside A, baicalin and puerarin detection wavelength were 328,328,277,249 nm. Results: The linear relationship between chlorogenic acid and forsythoside A was 0.2 ~ 20 mg · L-1 and baicalin was 2.0-300 mg · L-1 (r> 0.999 4). The intra-and inter-day precision RSD The average recoveries of the three samples with low, medium and high concentration were 88.3% -109.3% and the recovery rates were both> 80.8%. The main pharmacokinetic parameters AUC0-t of chlorogenic acid, forsythoside A and baicalin were (8.406 ± 1.175), (9.696 ± 2.349) and (145.35 ± 22.02) mg · h · L -1, respectively. MRT0- t were (0.619 ± 0.105), (0.634 ± 0.115) and (0.456 ± 0.068) h respectively; t1 / 2 Z were (0.532 ± 0.064), (0.732 ± 0.357) and (0.384 ± 0.050), (0.673 ± 0.422) and (0.324 ± 0.072) L · kg-1, respectively; CL Z were 0.504 ± 0.067, 0.634 ± 0.150 and 0.426 ± 0.066 kg-1. Conclusion: The method is sensitive, simple and accurate and is suitable for the determination of chlorogenic acid, forsythoside A and baicalin in rat plasma and the pharmacokinetics of Shuanghuanglian injection.