超高效液相色谱-质谱联用技术检测阿胶中马皮源成分
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摘要
目的:建立阿胶中马皮源成分的检测方法,为阿胶药材质量控制提供技术支持。方法:利用高分辨质谱分析方法获得马皮中专属特征肽段,采用胰蛋白酶对阿胶进行酶解,经超高效液相色谱-三重四极杆质谱对马源寡肽段进行检测。电喷雾离子化(ESI),正离子模式下,进行多反应监测,选择马皮特征分子离子峰m/z 386.4(双电荷)→377.3,322.3作为检测离子对。结果:方法学试验考察表明,所建立方法专属性、稳定性、重复性及耐用性均良好,对照物质马源寡肽A线性范围为0.010 86～1.086μg·mL~(-1),相关系数为0.999 9。以所建立的方法对专项抽验的56批阿胶进行检验,结果 28批样品检出马皮源成分。结论:所建立的方法准确可靠,专属性强,可用于阿胶中的马皮源成分的检测。
Objective:To establish an analysis method for specific identification of equine-hide gelatin in Ejiao.Method:The UPLC-MS was used to find the marker peptides for identification of equine-hide gelatin in Ejiao.The sample was analyzed on UPLC coupled with triple quadrupole mass spectrometry after digesting by trypsin.Choosing the characteristic molecular peaks of m/z 386.4 → 377.3,322.3 as target ion pair,collagen marker peptides were detected by mass spectrometry with positive electrospray ionization and multiple reaction monitoring(MRM).Results:The methodological evaluation showed that this method had good specificity,stability,repeatability and durability.The linear ranges of peptide A equine fell within the ranges of 0.010 86-1.086μg·mL~(-1) with the correlation coefficient of 0.999 9.The characteristic molecular peaks of equine-hide gelatin were detected in 28 batches of Ejiao.Conclusion:The present method is specific,precise,reliable,and suitable for identification of equine-hide gelatin in Ejiao.
Objective:To establish an analysis method for specific identification of equine-hide gelatin in Ejiao.Method:The UPLC-MS was used to find the marker peptides for identification of equine-hide gelatin in Ejiao.The sample was analyzed on UPLC coupled with triple quadrupole mass spectrometry after digesting by trypsin.Choosing the characteristic molecular peaks of m/z 386.4 → 377.3,322.3 as target ion pair,collagen marker peptides were detected by mass spectrometry with positive electrospray ionization and multiple reaction monitoring(MRM).Results:The methodological evaluation showed that this method had good specificity,stability,repeatability and durability.The linear ranges of peptide A equine fell within the ranges of 0.010 86-1.086μg·mL~(-1) with the correlation coefficient of 0.999 9.The characteristic molecular peaks of equine-hide gelatin were detected in 28 batches of Ejiao.Conclusion:The present method is specific,precise,reliable,and suitable for identification of equine-hide gelatin in Ejiao.
引文
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[3]程显隆,李文杰,张小龙,等.UPLC-QTOF-MS结合主成分分析法用于龟甲胶、鹿角胶中添加牛皮源成分的检测研究[J].药物分析杂志,2012,32(6):931CHENG XL,LI WJ,ZHANG XL,et al.Identification of bovinehide gelatin from glue of tortoise shell and deer-horn gelatin by UPLC-QTOF-MS and principal component analysis[J].Chin JPharm Anal,2012,32(6):931
[4]张贵锋,刘涛,王前,等.中药阿胶的质量控制方法研究[J].药物生物技术,2009,16(3):250ZHANG GF,LIU T,WANG Q,et al.Investigation on quality control of E-jiao by mass spectrometry[J].Chin J Pharm Biotechnol,2009,16(3):250
[5]周芳妍,刘力.胶类中药质量控制方法研究进展[J].中国药业,2013,22(1):1ZHOU FY,LIU L.Progress on quality control of gelatin drugs in TCM[J].China Pharm,2013,22(1):1
[6]程显隆,李文杰,魏锋,等.动物胶类药材的鉴别方法研究进展[J].亚太传统医药,2011,7(3):167CHENG XL,LI WJ,WEI F,et al.Advances in studies on identificated method of gelatins[J].Asia-Pacific Tradit Med,2011,7(3):167
[7]李辉虎,任刚,陈利民,等.胶类药材真伪鉴别与质量评价技术研究进展[J].中国中药杂志,2018,43(1):15LI HH,REN G,CHEN LM,et al.Research progress on identification and quality evaluation of glues medicines[J].China JChin Mater Med,2018,43(1):15
[8]2011012,阿胶中铬(Cr)含量的补充检验方法[S].20112011012,Determination Method for Chromium in Donkey-hide Glue[S].2011
[9]2012001,阿胶中牛皮源含量的补充检验方法[S].20122012001,Identification of Bovine-hide Gelatin in Donkey-hide Glue[S].2012
[10]尤金花,周祥山,秦玉峰,等.一种胶类中药及其制品中马种和骡种源性成分的检测方法:中国,CN104280468A[P].2015-01-14YOU JH,ZHOU XS,QIN YF,et al.Method for Detecting Horsebreed and Mule-breed Derived Ingredients in Gelatin-type Traditional Chinese Medicine and Products:China,CN104280468A[P].2015-01-14
[11]石峰,巩丽萍,李雪,等.一种马和骡共有特征性多肽及其应用:中国,CN106198790A[P].2016-12-07SHI F,GONG LP,LI X,et al.Common Characteristic Polypeptide of Horse and Mule and Application Thereof:China,CN106198790A[P].2016-12-07
[12]张全芳,步讯,刘艳艳.一种快速鉴定驴皮、马皮和骡子皮的检测试剂盒:中国,CN104046700A[P].2014-09-17ZHANG QF,BU X,LIU YY.Detection Kit for Quickly Identifying Donkey Skin,Horse Skin and Mule Skin:China,CN104046700A[P].2014-09-17