Objective: To establish the HPLC fingerprint method and the separation method of tetrahydropalmatine enantiomers of Yuanhu Zhitong tablets, so as to evaluate the overall quality condition and improve the quality controllability of Yuanhu Zhitong tablets. Methods: HPLC fingerprint was performed on a Shiseido MG C18 chromatographic column (4.6 mm×250 mm, 5 μm).The mobile phase consisted of acetonitrile-tetrahydrofuran(A) and 0.6% glacial acetic acid solution (using triethylamine to adjust the pH value to 6.0, B) with gradient elution. The flow rate was 1 mL·min-1, the detection wavelength was 220 nm and the column temperature was 30 ℃. A chiral separation method of tetrahydropalmatine enantiomers in Yuanhu Zhitong tablets was established based on the heart-cutting two-dimensional liquid chromatography technology.The peak area ratio of L-tetrahydropalmatine(L-THP) to D-tetrahydropalmatine(D-THP) was used as an indicator to compare the difference between samples. Results: The fingerprint results showed that 156 batches of samples from 39 manufacturing enterprises could be divided into classes of A and B. Among them, 25 batches of samples from 7 manufacturing enterprises were classified as class B samples with similarities to the reference fingerprint below 0.8. And their fingerprint differences were as follows:except the fingerprint peak corresponding to DL-THP(DL-tetrahydropalmatine), other characteristic peaks from Corydalis yanhusuo W.T.Wang were small or even missing. The results of heart-cutting two-dimensional liquid chromatography showed that DL-THP was transferred to the second-dimensional chromatography through the heart-cutting valve and was separated as D-THP and L-THP. The ratio of the peak area of D-THP to L-THP were less than 1.0 in class B samples, while the opposite was found in class A samples, which was the most important difference between samples. Conclusion: There are some problems in the raw materials from some enterprises, and they may add chemicals(rotundine) illegally to reduce costs and comply with pharmacopoeia.The HPLC fingerprint method is simple, accurate, highly reproducible and sensitive, providing scientific basis for the overall quality control of Yuanhu Zhitong tablets.Taking the ratio of tetrahydropalmatine enantiomers as the evaluation index, the quality problems of Yuanhu Zhitong tablets are analyzed and clarified, which can be used as an important mean for its quality control.At the same time, it providesa new idea for quality control of other prescription preparations containing tetrahydropalmatine, or other Chinese medicinal materials and Chinese patent medicines containing enantiomer compounds.
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