Determination Of Drug-time Curve And Study On Intestinal Bacterial Metabolism In Rats Of Anemarrhena Extract

Anemarrhenae is the dried rhizome of Anemarrhena asphodeloides Bunge belonged to Liliaceae, possesses lots of functions such as clearing heat, moisturizing dryness-syndrome quenching thirst, eliminating annoyance and so on. Quite a lot of clinical researches showed that saponins of Anemarrhena and sapogenin are the main active components, therefore, the researches on pharmacectic compounds of Anemarrhenae had been taken more and more attention to in and abroad. Our team prepared the timosaponin of Anemarrhena into XingShu tablets, which had been certificated and already started industrial scale researches. We had established stable production progress, quality standards of extracts from Anemarrhenae and its preparation, studies on pharmacodynamics and acute toxicity. This assay aimed at learning the biotransformation, absorption, metabolism, distribution and discharge of Anemarrhenae in vivo, which could help improving the bioavailability, designing the schedule of using the drug and new preparation. Generally speaking, we did research on the pharmacokinetics of Anemarrhenae.First of all, the HPLC-ELSD method was utilized to determine the content of the timosaponin B-II, there was satisfying linearization between the logarithm of the content of timosaponin B-II and the logarithm of the peak area, the regression equation was y=1.6427x+1.9001(r=0.9999), the mean recovery rate was 98.26% and the RSD=3.56%(n=6). The contents of the several batches were over 55-76%.Through the in vivo and in vitro experiments of intestinal bacterial metabolism in rats, we found that the extract of Anemarrhenae could be metabolized to three sapogenins, and were identified as Sarsasapogenin and its isomers by TLC and LC-MS. While the chemical hydrolyses of the extract only obtained two sapogenins. Then the content of the Sarsasapogenin was determined, which was one of the metabolite, according to the results, we learned that the yield of Sarsasapogenin increased abundantly with the extension of the metabolism time.Following, the LC-MS method was established to analyzed the concentration of the timosaponin B-II absorbed into rat serum after oral administration of Anemarrhenae, the results showed there was satisfying linearization between the concentration and the peak area in 4.95-1980ng/ml, the recovery rate was over 70%. Finally, the pharmacokinetics of timosaponin B-II was carried out, the main parameters were as follows: AUC0-24=3233.65±594.61ug.L-1.h-1, t1/2=5.01±1.81h, Tmax=1.15±0.49h, Cmax=953.15±186.75 ng.mL-1.