Study On UPLC-MS/MS Methods For The Multicomponent Simultaneous Analysis Of Traditional Chinese Medicine

Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) results in improving resolution and providing additional selectivity. Compared with many other separation and identification techniques in complex samples analysis, the strength of this technique lies in the selectivity, high sensitivity and fast screening capabilities. It is a powerful tool for analysis of complex samples, such as traditional Chinese medicines. In this thesis, the development of UPLC and MS/MS and the applications of UPLC-MS/MS in analysis of traditional Chinese medicines were reviewed, and then new methods for simultaneous determination of effective components in traditional Chinese medicines by UPLC-MS/MS were established. The main contents are as follows:1. A method for simultaneous determination of paeoniflorin, tetrahydropalmatine, jatrorrhizine, berberine, palmatine, evodiamine, saikosaponin C, saikosaponin A and saikosaponin D in Jiaweizuojin pills was established by UPLC-MS/MS. The UPLC separation was using0.2%(V/V) formic acid aqueous solution and methanol as mobile phase with the gradient elution. The mass spectrometer was operated under the positive ion mode with the ESI source and in the multiple reaction monitoring (MRM) mode. Under the optimum conditions, the detection limits were5.0,0.20,0.45,0.54,0.45,1.0,10,1.0,1.5μg·L-1for paeoniflorin, tetrahydropalmatine, jatrorrhizine, berberine, palmatine, evodiamine, saikosaponin C, saikosaponin A and saikosaponin D, respectively. The average recoveries of the nine effective components were between99.3%and107%with all relative standard deviations not more than2.6%. The developed method is simple, rapid and accurate, and suitable for the quality control of the nine effective components in Jiaweizuojin pills.2. A method for simultaneous determination of chlorogenic acid, caffeic acid, scutellarin, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, schisandrin, ophiopogonin D, schisantherin, deoxyschizandrin, y-schisandrin in Dengzhanshengmai capsules was established by UPLC-MS/MS. The negative and positive ion modes converted to detect the eleven compounds in the MRM mode. Eleven effective components in Dengzhanshengmai capsules were baseline separated in30minutes. Under the optimized conditions, the calibration curves were good linear (r>0.9990) within the scope of the mass concentration and peak area. The recoveries of the eleven effective components ranged from96.5%to105%. The developed method is simple, rapid and accurate, and suitable for the quality control of the eleven effective components in Dengzhanshengmai capsules. 3. A method for simultaneous determination of catechins, epicatechin, liquiritin, tetrahydropalmatine, peimine, peiminine, notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1, glycyrrhizic acid, glycyrrhetinic acid in Kuiyangling capsules was established by UPLC-MS/MS. Eleven effective components in Kuiyangling capsules were baseline separated in30minutes. Under the optimum conditions, the detection limits were1.0,1.0,5.0,0.050,0.050,0.10,0.50,2.0,0.80,5.0,1.0μg·L-1for catechins, epicatechin, liquiritin, tetrahydropalmatine, peimine, peiminine, notoginsenoside R1, ginsenoside Rg1, ginsenoside Rb1, glycyrrhizic acid, glycyrrhetinic acid, respectively. The average recoveries of the eleven effective components were between98.8%and105%. The developed method is simple, rapid and accurate, and suitable for the quality control of the eleven effective components in Kuiyangling capsules.4. A method was established for the rapid separation and simultaneous determination of twelve effective components in Tongtian Oral liquid by UPLC-MS/MS. The mass spectrometer was operated under the positive ion mode with the ESI source. Under the optimum conditions, the detection limits were0.80,1.0,5.0,0.20,4.0,0.20,0.40,5.0,5.0,0.30,0.30,1.5μg·L-1for catechins, epicatechin, paeoniflorin, prim-o-glucosylcimifugin, liquiritin, cimifugin,5-O-methyvisammioside, glycyrrhizic acid, aristolochic acid A, imperatorin, isoimperatorin, glycyrrhetinic acid, respectively. The average recoveries of the twelve effective components were between98.0%and106%with all relative standard deviations not more than3.0%. The developed method is simple, rapid and accurate, and suitable for the quality control of the twelve effective components in Tongtian Oral liquid.5. A method for simultaneous determination of paeoniflorin, quercitin, paeonol, astragaloside IV, astragaloside II, astragaloside I, cryptotanshinone, tanshinone ⅡA in Yiganning pills was established by UPLC-MS/MS. The UPLC separation was using0.1%(V/V) formic acid aqueous solution and acetonitrile as mobile phase with the gradient elution. Under the optimum conditions, eight effective components in Yiganning pills were baseline separated in21minutes. Under the optimum conditions, the detection limits were4.0,2.0,15,2.0,3.5,0.50,3.5,0.10,0.10μg·L-1for paeoniflorin, quercitin, paeonol, astragaloside IV, astragaloside II, astragaloside I, cryptotanshinone, tanshinone IIA, respectively. The average recoveries of the eight effective components were between95.5%and106%. The developed method is simple, rapid and accurate, and suitable for the quality control of the eight effective components in Yiganning pills.