Objective To investigate the preparation process of the benchmark sample of Yinchenhao Decoction (YD, 茵陈蒿汤). Methods An HPLC method was established to simultaneous determine the content of 11 components in YD, including chlorogenic acid, 1-caffeoylquinic acid (1-CQA), geniposide, shanzhiside, genipin gentiobioside (GG), crocin I, crocin II, jasminoside B, aloe emodin-8-O-glucopyranoside (AE8G), rhein-8-O-glucoside (R8G) and gallic acid. Using the extraction amount or transfer rate of these 11 components as indicators, single factor investigations were conducted on the preparation of benchmark sample of YD, including the decocting methods, quantity of water addition, decoction time, decoction times, and concentration temperature. Taking the quantity of water addition, decoction time, and concentration temperature as the evaluation factors, the two levels set for the single factor investigations mentioned above were optimized and combined by the orthogonal experimental method, and the optimized process was verified through amplification experiments. Results The single-factor investigation indicated that the extraction amounts of six, nine, and 10 components were higher under the conditions of co-decoction, decocting with 18-fold and 15-fold water for 30 min and 20 min, respectively, compared to the conditions of pre-decocting Yinchen (Artemisiae Scopariae Herba) followed by decocting with 12-fold and 10-fold water for 60 min and 40 min (P < 0.05, 0.01). Compared with the total amount extracted three times, the extraction amount of each component in the first two decoctions were all close to or exceeded 70%. The transfer rates of two components were significantly higher under reduced-pressure concentration at 50  ℃ than at 80  ℃ (P < 0.01). The optimal preparation process using orthogonal experiment for the benchmark sample of YD was as follows: decocting the three herbs together twice, with 18-fold and 15-fold volume water, for 30 and 20 min, respectively, then concentrating at 50 ℃ under reduced pressure. The results of the validation experiment magnified by 10 was consistent with lab-scale results. Conclusion The optimized process is stable, simple, and repeatable, which can be used for the preparation of substance benchmark of classical prescription YD.

classical prescription  /  Yinchenhao Decoction  /  Artemisiae Scopariae Herba  /  Gardeniae Fructus  /  Rhei Radix et Rhizoma  /  benchmark sample  /  preparation process  /  1-caffeoylquinic acid  /  geniposide  /  shanzhiside  /  genipin gentiobioside  /  crocin I  /  crocin II  /  aloe emodin-8-O-glucopyranoside  /  rhein-8-O-glucoside  /  chlorogenic acid  /  jasminoside B  /  gallic acid