| Tongshukoushuang Capsule consisited of eight commonly used Chinese herbs:Rheum tanguticum, Citrus aurantium, Artemisia capillaris, Paeonia suffruticosa, Gentianamcrophylla, Equisetum hiemale, Angelica sinensis, Prunella vudgaris, is one of traditional Chinese medicine recipe for removing damp-heat, turning turbid catharsis. It has been used clinically to the treatment of constipation, halitosis, painful gum cansed by colorectal damp-heat. In cunrrent quality standard, there is only using TLC to determine the contents of emodin and chtysophanol. In this thesis, HPLC method was employed in quality control of index components in Tongshukoushuang capsule, and also in studying pharmacokinetics of emodin and chtysophanol in rat.HPLC methods were established to determine emodin, rhein, chrysophanol, physcion, naringin, hesperidin, neohesperidin, chorogenic acid, gentiopicrin, ferulic acid, quercetin, luteolin, kaempferol and paeonol in Tongshukoushuang Capsule. The calibration curve was linear within the range of 0.70 to 7.0μg·mL-1 for emodin (r=0.9999),0.384 to 3.84μg·mL-1 for rhein (r=0.9996),1.803 to 18.03μg·mL-1 for chrysophanol (r=0.9996),0.504 to 5.04μg·mL-1 for physcion (r=0.9996),8.88 to 88.8μg·mL-1 for naringin (r=0.9997),7.60 to 76.0μg·mL-1 for hesperidin (r=0.9996),10.56 to 105.6μg·mL-1 for neohesperidin (r=0.9999),1.672 to 16.72μg·mL-1 for chorogenic acid (r= 0.9999),12.80 to 128.0μg·mL-1 for gentiopicrin (r= 0.9995),1.188 to 11.88μg·mL-1 for ferulic acid (r= 0.9996),1.60 to 16.0μug·mL-1 for quercetin 0.63 (r=0.9999),0.80 to 8.0μg·mL-1 for luteolin (r=0.9993),1.62 to 16.2μg·mL-1 for kaempferol (r= 0.9998),2.04 to 20.4μg·mL-1 for paeonol (r= 0.9999). The average recoveries were 99.3%(RSD= 1.5%) for emodin,100.7%(RSD= 0.8%) for rhein,100.9%(RSD= 0.9 %) for chrysophanol,99.1%(RSD=1.1%) for physcion,100.5%(RSD= 1.0%) for naringin, 100.5%(RSD= 1.0%) for hesperidin,100.3%(RSD=0.9%) for neohesperidin,100.1%(RSD = 1.3%) for chorogenic acid,99.6%(RSD= 0.9%) for gentiopicrin,99.6%(RSD= 0.9%) for ferulic acid,99.5%(RSD= 1.1%) for quercetin,100.4%(RSD= 1.4%) for luteolin,100.2% (RSD= 1.4%) for kaempferol and 98.9%(RSD= 0.38%) for paeonol. The methods are simple, reproducible and with good reproducibility, which provide the quantitative basis for the quality assessment for Tongshukoushuang capsule.The method of determine Emodin,chrysophanol in rat plasma after i.g. administration of Tongshukoushuang capsule was developed to study the pharmacokinetics by using HPLC-UV. The HPLC separation was achieved on a Diamonsil-C18 (200 mm×4.6 mm,5μm) column at 30 ℃. The mobile phase consisted of methol-0.4%phosphonic acid (83:17, v/v) at a flow rate of 1.1 mL·min-1. The UV detection wavelength was set at 254 nm. Linear calibration plot was obtained in the concentration range from 29.00 to 2900 ng·mL-1 (r=0.9985) of emodin,21.00 to 2100 ng·mL-1 (r 0.9977) of chrysophanol intra-day RSD and inter-day RSD of emodin were both less than 9.0%. Intra-day RSD and inter-day RSD of chrysophanol were both less than 6.0 %. The extraction recoveries of emodin were 75.7±4.2%,78.1±4.2%,76.1±2.9%.The extraction recoveries of chrysophano were 74.4±3.7%,72.0±4.7%,75.9±2.9%. The pharmacokinetic parameters of emodin was achieved as follows:Cmax was 1436±365.0 ng·mL-1, Tmaxwas 0.88±0.14 h, t1/2αwas 1.79±2.40, t1/2βwas 6.99±1.57 h, AUC0â†'t was 7578±727.9 ng·h·mL-1, AUC0â†'∞was 8548±1236 ng·h·mL-1. The pharmacokinetic parameters of chrysophanol was achieved as follows:Cmax was 1546±319.8 ng·mL-1, Tmax was 0.79±0.10 h, t1/2αwas 1.72±1.83, t1/2βwas 9.37±3.10 h, AUC0â†'t was 7696±693.9 ng·h·mL-1, AUC0â†'∞was 8563±758.4 ng·h·mL-1. Emodin, chrysophanol could be absorbed rapidly and its pharmacokinetics was consistent with two-compartment model. |
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