UPLC-MS/MS同时测定巴戟天中4个环烯醚萜苷的含量
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摘要
目的:建立UPLC-MS/MS法同时测定巴戟天中4个环烯醚萜苷(水晶兰苷、去乙酰车叶草苷酸、车叶草苷酸和车叶草苷)含量。方法:巴戟天以甲醇回流提取;采用Xbridge BEH C18色谱柱(150 mm×4.6 mm,5μm),乙腈-0.3%甲酸水为流动相,梯度洗脱;电喷雾离子源,负离子多反应监测模式监测,内标法测定。结果:水晶兰苷、去乙酰车叶草苷酸,车叶草苷酸和车叶草苷质量浓度在.015~79.65、0.030~96.00、0.042~21.00、0.018~37.50μg·m L~(-1)范围内与峰面积呈现良好的线性关系(R20≥0.994 9);平均加样回收率分别为99.4%、98.5%、98.1%、97.7%,RSD分别为2.3%、2.5%、3.5%、4.1%;不同产地巴戟天中车叶草甘酸和车叶草甘的含量差异较大;结论:该方法可靠,简便,可用于巴戟天中环烯醚萜苷的快速检测。
Objective:To establish a UPLC-MS/MS method for quantitative analysis of monotropein,deacetyl asperulosidic acid,asperulosidic acid and asperuloside in Morindae Officinalis Radix. Methods:Morindae Officinalis Radix was extracted with methanol by reflux extraction. The assay was performed on an Xbridge BEH C18(150 mm×4.6 mm,5 μm) column and the sample was eluted with a gradient mobile phase consisting of acetonitrile and 0.3% formic acid. The MS system was operated by using an electrospray ionization(ESI) in the negative ion mode,and the scan mode was in multiple reaction ion monitoring(MRM) mode. Internal standard method was used in the experiment. Results:The linear ranges of monotropein,deacetyl asperulosidic acid,asperulosidic acid and asperulo-side were 0.015-79.65 μg·m L~(-1),0.030-96.00 μg·m L~(-1),0.042-21.00 μg·m L~(-1) and 0.018-37.50 μg·m L1,respectively(R2≥ 0.994 9). The average recoveries were 99.4%,98.5%,98.1% and 97.7% with RSD of 2.3%,2.5%,3.5% and 4.1%,respectively. The contents of asperulosidic acid and asperuloside in Morindae Officinalis Radix collected from different areas in China exhibited great variation. Conclusion:The proposed method is reliable and simple,which can be used in rapid detection of iridoid glycosides in Morindae Officinalis Radix.
Objective:To establish a UPLC-MS/MS method for quantitative analysis of monotropein,deacetyl asperulosidic acid,asperulosidic acid and asperuloside in Morindae Officinalis Radix. Methods:Morindae Officinalis Radix was extracted with methanol by reflux extraction. The assay was performed on an Xbridge BEH C18(150 mm×4.6 mm,5 μm) column and the sample was eluted with a gradient mobile phase consisting of acetonitrile and 0.3% formic acid. The MS system was operated by using an electrospray ionization(ESI) in the negative ion mode,and the scan mode was in multiple reaction ion monitoring(MRM) mode. Internal standard method was used in the experiment. Results:The linear ranges of monotropein,deacetyl asperulosidic acid,asperulosidic acid and asperulo-side were 0.015-79.65 μg·m L~(-1),0.030-96.00 μg·m L~(-1),0.042-21.00 μg·m L~(-1) and 0.018-37.50 μg·m L1,respectively(R2≥ 0.994 9). The average recoveries were 99.4%,98.5%,98.1% and 97.7% with RSD of 2.3%,2.5%,3.5% and 4.1%,respectively. The contents of asperulosidic acid and asperuloside in Morindae Officinalis Radix collected from different areas in China exhibited great variation. Conclusion:The proposed method is reliable and simple,which can be used in rapid detection of iridoid glycosides in Morindae Officinalis Radix.
引文
[1]中华人民共和国药典2015年版.一部[S].2015:81Ch P 2015.VolⅠ[S].2015:81
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[9]王玉磊,崔翰明,黄世敬,等.HPLC测定不同产地和批次巴戟天中主要环烯醚萜苷含量[J].中药材,2011,34(8):1187WANG YL,CUI HM,HUANG SJ,et al.Determination of main iridoid glucosides from Morindae Officinalis Radix of different producting areas by HPLC[J].J Chin Med Mater,2011,34(8):1187
[10]LI CM,DONG J,TIAN JC,et al.LC/MS/MS determination and pharmacokinetic study of iridoid glycosides monotropein and deacetylasperulosidic acid isomers in rat plasma after oral administration of Morinda officinalis extract[J].Biomed Chromatogr,2016,30:163
[11]徐吉银,梁英娇,丁平.炮制方法对巴戟天有效成分水晶兰苷含量的影响[J].中药材,2007,30(1):20XU JY,LIANG YJ,DING P.Determination of monotropein in Radix Morinda officinalis from different processed products by HPLC[J].J Chin Med Mater,2007,30(1):20
[12]陈黎,刘春霞,何秀丽,等.LC-MS/MS法同时测定中药白及中militarine、原儿茶酸和咖啡酸的含量[J].中国药师,2015,18(2):230CHEN L,LIU CX,HE XL,et al.Simultaneous determination of militarine,protocatechuic acid and caffeic acid in Bletilla striata by LC-MS/MS[J].China Pharm,2015,18(2):230
[2]刘瑾,徐吉银,罗进辉,等.不同产地巴戟天中水晶兰苷的含量测定[J].中成药,2010,32(3):517LIU J,XU JY,LUO JH,et al.Determination of monotropein in Morinda officinalis from different areas[J].Chin Tradit Pat Med,2010,32(3):517
[3]ZHANG HL,ZHANG QW,ZHANG XQ,et al.Chemical constituents from the roots of Morinda officinalis[J].Chin J Nat Med,2015,8(3):192
[4]景海漪,史辑,崔妮,等.巴戟天炮制前后寡糖类成分HPLC-CAD指纹图谱研究[J].中草药,2014,45(10):1412JING HY,SHI J,CUI N,et al.Fingerprints of oligosaccharides in Morinda officinalis before and after processing by HPLC-CAD[J].Chin Tradit Herb Drugs,2014,45(10):1412
[5]CHOI J,LEE K,CHOI M,et al.Antinociceptive anti-inflammatory effect of monotropein isolated from the root of Morinda officinalis[J].Biol Pharm Bull,2005,28:1915
[6]ZHANG ZG,ZHANG QY,YANG H,et al.Monotropein isolated from the roots of Morinda officinalis increases osteoblastic bone formation and prevents bone loss in ovariectomized mice[J].Fitoterapia,2016,110:166
[7]黄珍珍.巴戟天的化学成分及其生物活性研究[D].广州:广州中医药大学,2013HUANG ZZ.Studies on Chemical Constituents and Their Bioactivities of Morinda officinalis[D].Guangzhou:Guangzhou University of Chinese Medicine,2013
[8]仰铁锤,徐吉银,梁英娇,等.HPLC测定巴戟天中的水晶兰苷[J].华西药学杂志,2008,23(6):695YANG TC,XU JY,LIANG YJ,et al.Determination of monotropein from Radix Morinda officinalis by HPLC[J].West China J Pharm Sci,2008,23(6):695
[9]王玉磊,崔翰明,黄世敬,等.HPLC测定不同产地和批次巴戟天中主要环烯醚萜苷含量[J].中药材,2011,34(8):1187WANG YL,CUI HM,HUANG SJ,et al.Determination of main iridoid glucosides from Morindae Officinalis Radix of different producting areas by HPLC[J].J Chin Med Mater,2011,34(8):1187
[10]LI CM,DONG J,TIAN JC,et al.LC/MS/MS determination and pharmacokinetic study of iridoid glycosides monotropein and deacetylasperulosidic acid isomers in rat plasma after oral administration of Morinda officinalis extract[J].Biomed Chromatogr,2016,30:163
[11]徐吉银,梁英娇,丁平.炮制方法对巴戟天有效成分水晶兰苷含量的影响[J].中药材,2007,30(1):20XU JY,LIANG YJ,DING P.Determination of monotropein in Radix Morinda officinalis from different processed products by HPLC[J].J Chin Med Mater,2007,30(1):20
[12]陈黎,刘春霞,何秀丽,等.LC-MS/MS法同时测定中药白及中militarine、原儿茶酸和咖啡酸的含量[J].中国药师,2015,18(2):230CHEN L,LIU CX,HE XL,et al.Simultaneous determination of militarine,protocatechuic acid and caffeic acid in Bletilla striata by LC-MS/MS[J].China Pharm,2015,18(2):230