Inhibitive activities detection of monoamine oxidases (MAO) A and B inhibitors in human liver MAO incubations by UPLC-ESI-MS/MS.
A sensitive UPLC-ESI-MS/MS method was developed for determining and screening of inhibitors of monoamine oxidase (MAO) by using mix MAO enzymes prepared from human liver. 5-Hydroxytryptamine (5-HT) and 2-phenethylamine (2-PEA) were used as substrates for MAO-A and MAO-B in incubations, and 5-hydroxyindole-3-acetic acid (5-HIAA) and phenylacetic acid (PAA) resulting from 5-HT and 2-PEA were used as markers to evaluate inhibitive activities of test compounds on MAO-A and MAO-B. Proper separation was achieved for positive multiple reaction monitoring of 5-HIAA (m/z 192.1→146.1) and negative multiple reaction monitoring PAA (m/z 135.0→91.0) via isocratic elution (0.1% fromic acid:acetonitrile=60:40) on a HSS T3 column following a simple precipitation of proteins for sample treatment. The relative standard deviations of intra- and inter-day precisions were ranged from 1.74% to 6.76% and 0.77% to 9.35%. The mean accuracies for the quality control samples were 101.37±6.60% and 101.39±2.85%, respectively. This method exhibited characteristics of small total reaction volume (100μl), short analysis time (3.5min), highly sensitivity, low cost and without matrix effect (103.56±2.33% to 112.63±8.57% for 5-HIAA and 105.68±8.75% to 112.76±4.67% for PAA). The developed method was successfully applied for detection of the MAO-A and MAO-B inhibitive activities by model drugs, including pargyline, clorgyline, as well as β-carboline alkaloids from Peganum harmala.
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China; Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
Bo
B
Jiang
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China.
Shuping
S
Li
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China.
Wei
W
Liu
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China.
Yadi
Y
Yang
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China.
Wenxia
W
Chen
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China.
Dandan
D
He
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China.
Xuemei
X
Cheng
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China.
Zhengtao
Z
Wang
Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China. Electronic address: chenwansheng@smmu.edu.cn.
Wansheng
W
Chen
Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, The MOE Key Laboratory for Standardization of Chinese Medicines and The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Shanghai R&D Centre for Standardization of Chinese Medicines, Shanghai 201203, China. Electronic address: wchcxm@hotmail.com.
Changhong
C
Wang
D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D008099:Liver / Q000201:enzymology*; D008995:Monoamine Oxidase / Q000378:metabolism*; D008996:Monoamine Oxidase Inhibitors / Q000032:analysis* / Q000494:pharmacology; D011786:Quality Control; D012015:Reference Standards; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013379:Substrate Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
D008996:Monoamine Oxidase Inhibitors; D008995:Monoamine Oxidase; C579458:monoamine oxidase A, human
5-HIAA; Inhibitor; MAO-A; MAO-B; PAA; UPLC–ESI MS/MS