Accurate and sensitive detection of Catechol-O-methyltransferase activity by liquid chromatography with fluorescence detection. 2020

Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
School of Life Science and Medicine, Dalian University of Technology, Panjin 124221, China; Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.

Catechol-O-methyltransferase (COMT) is a major drug metabolizing enzyme in humans. COMT expression is directedly associated with various mental diseases and cancers due to its essential role in catalyzing metabolic inactivation of endogenous catecholamines and catechol estrogens. However, a practical method to precisely measure COMT activities in biological samples is lacking. In the current study, we established a liquid chromatography-fluorescence detection (LC-FD) method based on fluorometric detection of the methylated product of 3-BTD, a fluorescent probe for COMT, to sensitively quantify COMT activities in human erythrocytes and cell homogenates. Assay validation of the established LC-FD based method was conducted for selectivity and sensitivity, range of linearity, precision and accuracy, recovery, biological matrices effect and stability. The limit of quantification for 3-BTMD (the methylated product of 3-BTD by COMT) of this method was 0.0083 nM, which is nearly 10-fold lower than that for previously published methods. The method was precise with intra- and inter-day relative standard deviation (RSD) lower than 5%. In addition, this method showed an excellent anti-interference ability with no effects of the endogenous substances on the fluorometric detection of 3-BTMD. The practical use of this method was established by its successful application for the measurement of COMT activities in individual human erythrocytes (n = 13), and in cell homogenates generated from four different human cell lines. Our results suggest that this method will be of great value in accurately determining the native activity of COMT in biological samples, which is beneficial for a complete understand of the role of COMT both in physiological and pathological conditions.

UI MeSH Term Description Entries
D002394 Catechol O-Methyltransferase Enzyme that catalyzes the movement of a methyl group from S-adenosylmethionone to a catechol or a catecholamine. Catechol Methyltransferase,Catechol-O-Methyltransferase,Catechol O Methyltransferase,Methyltransferase, Catechol,O-Methyltransferase, Catechol
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D002853 Chromatography, Liquid Chromatographic techniques in which the mobile phase is a liquid. Liquid Chromatography
D003374 Coumarins Synthetic or naturally occurring substances related to coumarin, the delta-lactone of coumarinic acid. 1,2-Benzopyrone Derivatives,1,2-Benzopyrones,Coumarin Derivative,Coumarine,1,2-Benzo-Pyrones,Benzopyran-2-ones,Coumarin Derivatives,Coumarines,1,2 Benzo Pyrones,1,2 Benzopyrone Derivatives,1,2 Benzopyrones,Benzopyran 2 ones,Derivative, Coumarin,Derivatives, 1,2-Benzopyrone,Derivatives, Coumarin
D004912 Erythrocytes Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN. Blood Cells, Red,Blood Corpuscles, Red,Red Blood Cells,Red Blood Corpuscles,Blood Cell, Red,Blood Corpuscle, Red,Erythrocyte,Red Blood Cell,Red Blood Corpuscle
D005456 Fluorescent Dyes Chemicals that emit light after excitation by light. The wave length of the emitted light is usually longer than that of the incident light. Fluorochromes are substances that cause fluorescence in other substances, i.e., dyes used to mark or label other compounds with fluorescent tags. Flourescent Agent,Fluorescent Dye,Fluorescent Probe,Fluorescent Probes,Fluorochrome,Fluorochromes,Fluorogenic Substrates,Fluorescence Agents,Fluorescent Agents,Fluorogenic Substrate,Agents, Fluorescence,Agents, Fluorescent,Dyes, Fluorescent,Probes, Fluorescent,Substrates, Fluorogenic
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D013050 Spectrometry, Fluorescence Measurement of the intensity and quality of fluorescence. Fluorescence Spectrophotometry,Fluorescence Spectroscopy,Spectrofluorometry,Fluorescence Spectrometry,Spectrophotometry, Fluorescence,Spectroscopy, Fluorescence
D015203 Reproducibility of Results The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results. Reliability and Validity,Reliability of Result,Reproducibility Of Result,Reproducibility of Finding,Validity of Result,Validity of Results,Face Validity,Reliability (Epidemiology),Reliability of Results,Reproducibility of Findings,Test-Retest Reliability,Validity (Epidemiology),Finding Reproducibilities,Finding Reproducibility,Of Result, Reproducibility,Of Results, Reproducibility,Reliabilities, Test-Retest,Reliability, Test-Retest,Result Reliabilities,Result Reliability,Result Validities,Result Validity,Result, Reproducibility Of,Results, Reproducibility Of,Test Retest Reliability,Validity and Reliability,Validity, Face
D016014 Linear Models Statistical models in which the value of a parameter for a given value of a factor is assumed to be equal to a + bx, where a and b are constants. The models predict a linear regression. Linear Regression,Log-Linear Models,Models, Linear,Linear Model,Linear Regressions,Log Linear Models,Log-Linear Model,Model, Linear,Model, Log-Linear,Models, Log-Linear,Regression, Linear,Regressions, Linear

Related Publications

Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
May 2005, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
December 2002, Biomedical chromatography : BMC,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
February 1984, Analytical biochemistry,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
December 1981, Journal of chromatography,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
March 1989, Biomedical chromatography : BMC,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
March 1980, Analytical chemistry,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
January 1985, Analytical biochemistry,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
January 1995, Journal of chromatography. B, Biomedical applications,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
May 2001, Se pu = Chinese journal of chromatography,
Yang-Liu Xia, and Hui-Lin Pang, and Shi-Yang Li, and Yong Liu, and Ping Wang, and Guang-Bo Ge
August 2003, Analytical biochemistry,
Copied contents to your clipboard!