Method for establishing fingerprint of leech and standard fingerprint of leech

Abstract

The invention provides a method for establishing a fingerprint of leech. The method comprises the following specific steps: preparing the leech into a test solution; and performing separation detection through a high performance liquid chromatograph under certain conditions to obtain the fingerprint of the leech. The invention also provides a standard fingerprint of the leech, which is obtained through the method. The fingerprint can provide a reliable basis for leech identification and internal quality control. The method is simple and convenient to operate, high in stability, high in repeatability and large in quantity of characteristic peaks, and is capable of comprehensively and accurately evaluating the quality of leech medicine.

Description

technical field [0001] The invention relates to a method for establishing a leech fingerprint and a leech standard fingerprint obtained by the method, belonging to the field of traditional Chinese medicine analysis. Background technique [0002] Leeches are the dried whole body of leech Whitmania pigra Whitman, Hirudo nipponica Whitman or willow-leaf leech Whitmania acronulata Whitman of the leech family. They are caught in summer and autumn, scalded to death with boiling water, dried in the sun or dried at low temperature. Flat, returns liver meridian, slightly poisonous. Traditional Chinese medicine believes that leech is a traditional blood-breaking medicine, which has the effect of removing blood stasis, dredging meridians, and benefiting water channels. Leeches are widely used clinically. The clinical diseases treated with leeches mainly include: chronic nephritis, varicose veins, prostatitis, infertility, diabetes, cancer, heart disease, liver cirrhosis, hypertension,...

AI Technical Summary

Problems solved by technology

For using fingerprints to control the quality of leech medicinal materials, there are existing patents and literature reports in China. For example, Chinese patent CN1844911A discloses the establishment method of leech fingerprints and discloses characteristic fingerprints. The components in the leech, the test period is long, the pteridine components in the leech cannot be effectively extracted by normal saline or water, and there are many irrelevant impurities, which will affect the comprehensiveness and accuracy of the fingerprints; only hypoxanthine is used as a reference, and for the Other important efficacy and quality marker components in leech, such as uracil, xanthine, uridine, leechamine C, leechamine B, and leechamine A, etc. are not involved, and cannot accurately and comprehensively reflect the quality of leech medicinal materials; During the elution process, during the time period when the mobile phase does not contain an organic phase, ordinary C 18 C in column packing 18 The random coiling of the chain will lead to changes in the retention value of the components, resulting in instability of the chromatographic system

Yuan Xiaohuan et al. used HPLC fingerprints to control the quality of leech injections and compound leech injections, and established the fingerprints of leech injections. However, the limitations of the methods for establishing leech fingerprints in the two documents are the same as those of Chinese patent CN1844911A. Or the compound injection only uses hypoxanthine as a reference substance, while other important medicinal ingredients and quality marker ingredients in leeches, such as uracil, xanthine, uridine, leechamine C, leechamine B, leechamine A, etc. None of them are involved, and cannot accurately and comprehensively reflect the quality of leech injection or compound leech injection; and during the gradient elution process, when the mobile phase does not contain an organic phase, ordinary C 18 C in filler 18 The random coiling of the chain will lead to changes in the retention value of the components, resulting in instability of the chromatographic system (Yang Xiaohuan, Yang Xudong, Cui Rongjun, et al. HPLC fingerprint of leech injection [J]. China Pharmaceutical Industry Journal, 2008, 39 (7): 530~532.), (Yang Xiaohuan, Yang Xudong, Wang Chuntao, etc. Study on HPLC Fingerprint of Compound Leech Injection[J]. Chinese Journal of Traditional Chinese Medicine, 2008, 33(15): 1825~1827.)

Xu Changgen and others studied the establishment method of the fingerprint of leech freeze-dried powder injection, and obtained its fingerprint, but it only uses hypoxanthine solution as a reference, which cannot comprehensively and accurately reflect other important leeches in the leech freeze-dried powder injection. The quality of the active ingredients; during the gradient elution process, when the mobile phase contains a very small proportion (less than 0.15%) of the organic phase, ordinary C 18 C in filler 18 The random coiling of the chain will lead to changes in the retention value of the components, resulting in an unstable chromatographic system; and there are too few characteristic peaks on its fingerprints, and the separation is not good (Xu Changgen, Luo Shishu, Xie Yanhua. Evaluation of fingerprints of leech freeze-dried powder injection [J ]. Journal of Fourth Military Medical University, 2009, 30 (22): 2667~2669.)

Images