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通过超高效液相色谱与电喷雾电离四极杆飞行时间质谱检测器联用对两种吸血水蛭的肽进行分析。

Peptides from two sanguinovorous leeches analyzed by ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometric detector.

作者信息

Xiao Ling, Nie Jing, Li Danping, Chen Keli

机构信息

College of Pharmacy, Hubei University of Chinese Medicine, Wuhan, China ; Department of TCM, Hubei Provincial Institute for Food and Drug Control, Wuhan, China.

Department of TCM, Hubei Provincial Institute for Food and Drug Control, Wuhan, China.

出版信息

Pharmacogn Mag. 2015 Jan-Mar;11(41):32-7. doi: 10.4103/0973-1296.149699.

Abstract

BACKGROUND

Hirudo nipponica Whitman and Poecilobdella manillensis Lesson fall into the family of Hirudinidae Whitman, both of them are sanguinovorous leeches and used a anticoagulant medicines in China. Their medicinal parts are the dried bodies. However, the peptides in the dried body of the two leeches have not been very clear up to now.

OBJECTIVE

To analyze the peptides from two sanguinovorous leeches, H. nipponica and P. manillensis.

MATERIALS AND METHODS

In this article it is reported that the peptides were obtained from anticoagulant active extracted parts of dried bodies of the two leeches and their molecular weights were analyzed by ultra-performance liquid chromatography with electrospray ionization quadrupole time-of-flight mass spectrometry mass spectrometric detector online.

RESULTS

Three peptide components were identified from H. nipponica with their molecular weight separately 14998, 15988, and 15956, six peptide components were identified from P. manillensis with molecular weight 9590, 13642, 14998, 17631, 15988, and 16567. Two of peptides from P. manillensis have the same molecular weight 14998 and 15988 as that in H. nipponica.

CONCLUSION

And the two peptides are the main peaks in the base peak ion chromatogram because they occupied a large ratio of total base peak area. Hence the composition of the extracted active part of the two leeches are very close, difference is in that the extract of P. manillensis has more small peptide peaks, but the extract of H. nipponica has not. Furthermore, the tryptic digestion hydrolysates of the extracted active part of each sample were analyzed and the results showed that there were four peaks which only exist in P. manillensis, but not in Hirudo nipponia. They may be the identified peak between the two leeches. This work support the viewpoint that P. manillensis can be used as a medicinal leech as H. nipponia and these peptide components of dried bodies of the two species leeches are a basis for their chemical identification and further investigations in active action.

摘要

背景

日本医蛭和菲牛蛭隶属于医蛭科,均为吸血水蛭,在中国作为抗凝血药物使用。其药用部位为干燥体。然而,至今这两种水蛭干燥体中的肽类成分尚不清楚。

目的

分析两种吸血水蛭(日本医蛭和菲牛蛭)中的肽类成分。

材料与方法

本文报道从两种水蛭干燥体的抗凝血活性提取物部位获得肽类成分,并采用超高效液相色谱-电喷雾电离四极杆飞行时间质谱联用仪在线分析其分子量。

结果

从日本医蛭中鉴定出3种肽类成分,分子量分别为14998、15988和15956;从菲牛蛭中鉴定出6种肽类成分,分子量分别为9590、13642、14998、17631、15988和16567。菲牛蛭中有2种肽类成分与日本医蛭中的2种肽类成分分子量相同,分别为14998和15988。

结论

这两种肽类成分在总离子流图中所占比例较大,是主要峰。因此,两种水蛭提取物活性部位的组成非常接近,不同之处在于菲牛蛭提取物中小肽峰较多,而日本医蛭提取物中没有。此外,对各样本提取物活性部位的胰蛋白酶消化水解产物进行分析,结果显示有4个峰仅存在于菲牛蛭中,而日本医蛭中没有。它们可能是两种水蛭的鉴别峰。本研究支持菲牛蛭可作为日本医蛭入药的观点,两种水蛭干燥体的这些肽类成分是其化学鉴别及进一步研究活性作用的基础。

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