Danshensu And Protocatechuic Aldehyde In Vivo Pharmacokinetic Study In Rabbits

AIM Salivia Miltiorrhiza Injection (SMI) is a chinese herbal preparation used widely in medical practice, Because of its action of promoting blood circulation of coronary arteries, improving the ischemia of myocardiurn and others. Danshensu (Dhpi) and Protocatechuic Aldehyde (Pal) are two active components in salivia miltiorrhiza injection. To study their pharmacokinetics rules and to provide pharrnacokinetics parameters for clinical application of salivia miltiorrhiza injection, This work aim at developing an reversed-phase high performance liquid chromatogrophic (RP-HIPLC) method for the separation and determination of two active component in SMI in rabbit plasma simultaneously, then, applying it to the pharmacokinetics study of Danshensu and protoeateehuic aldehyde, at acquiring the pharmacokinetics process and parameters of the two active components in preparation. METHODS 1. The separation and analytical methods of the plasma drug concerntrations: Shimadzuts HPLC systerrn was used, plasma drug concemtrations were determined by external standard method . The analytical column is kromasil C18and C18 precolumn; the mobile phase is composed of methanol-water-acetic acid (19:80:1 V/V, pH 3.0), the flow rate is lml/min; the LIV detector is set at 280nm. 2. The pharmacokinetics study: Blood samples were collected before and at various time after injecting vein a single dose of salivia rniltiorrhiza injection in rabbits. Then, prepared the sample on demand and determined them by RP-HPLC. The concerntration-time curves were analyzed with 3P87 program to determine the compartment models and pharmacokinetic parameters. RESULTS 1. The retention time of danshensu is 7.877min,The linear range is 0.326 it g/rnl -~ 72.74 i-i glint, Recovery coefficient of its three concerntration(high,middle,low) is respectively 1O0.3%~ 100.1%~ 99.71%, The coefficient of variation (RSD stand for it) are 3.39%~ 2.37%~ 2.3600 for intra-day and 4.57%~. 3.21%~ 3.63% for interday; The retention time of Protocatechuic aldehyde is 9.047mm, The linear range is 0.0329 Ii glint 18.18 ii g/inl, Recovery coefficient of its three concerntration(high,rniddle,low) is respectively 97~50 o.~ 96.530 o~. 96.11%, The coefficient of variation (RSD) are 1 .45%~ 5.01%~ 4.85% for intra-day and 1 .20%~ 4.95%~. 4.5 6% for interday. The test conclusion demonstrate: they are both good for the demand of biology sample analysis that the effect of separation and accuracy of the RP-1-IIPLC method. 2. The test proved that concentration-time curves of Danshensu and protocatechuic aldehyde in plasma ai~e both fitted to two-compartment model. Their pharmacokinetics parameters are showed as following: Danshensu kj~0.038?.002min?, t112~19.18?1.809mm, W0.324 ?0.049L,CLs~0.012 ?0.OO1Lmuii? AUC~1604 ?30.l9mg.min/L; Protocatechuic aldehyde k~~0.055?.004 miii? t,12~49.22?.953 mm, V~ 4.497 ?0.262L, CLs~0.011 ?0.001 Lrniii? AUC~433.4 ?8.332mg.mirt/L. CONCLUSION 1. The RP-HIPLC method is simple, sensitive and accurate that is developed in this work and used to determined plasma drug concerntration by external standard method. It will provide a scientific basis for the Pharmacokinetics studies of salivia miltiorrhiza injection (SMI). 2. Bharmacokinetics parameters of active components provide references for rational clinical application of salivia miltiorrhiza injection.