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A simple and precise method to detect sterol esterification activity of lecithin/cholesterol acyltransferase by high-performance liquid chromatography

Analytical and bioanalytical chemistry, 2018-02, Vol.410 (6), p.1785-1792 [Peer Reviewed Journal]

Springer-Verlag GmbH Germany, part of Springer Nature 2018 ;COPYRIGHT 2018 Springer ;Analytical and Bioanalytical Chemistry is a copyright of Springer, (2018). All Rights Reserved. ;Springer-Verlag GmbH Germany, part of Springer Nature 2018. ;ISSN: 1618-2642 ;EISSN: 1618-2650 ;DOI: 10.1007/s00216-017-0834-4 ;PMID: 29307006

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Title:
A simple and precise method to detect sterol esterification activity of lecithin/cholesterol acyltransferase by high-performance liquid chromatography
Author: Wang, Yu ; Wang, Siming ; Zhang, Lijiao ; Zeng, Jie ; Yang, Ruiyue ; Li, Hongxia ; Tang, Yueming ; Chen, Wenxiang ; Dong, Jun
Subjects: Acyltransferase ; Adult ; Aged ; Analytical Chemistry ; Apolipoproteins ; Biochemistry ; Calibration ; Cardiovascular diseases ; Characterization and Evaluation of Materials ; Chemical properties ; Chemical vapor deposition ; Chemistry ; Chemistry and Materials Science ; Cholesterol ; Chromatography ; Chromatography, High Pressure Liquid - economics ; Chromatography, High Pressure Liquid - methods ; Coefficient of variation ; Cofactors ; Dehydrocholesterols - blood ; Dehydrocholesterols - isolation & purification ; Dehydrocholesterols - metabolism ; Enzyme Assays - economics ; Enzyme Assays - methods ; Esterification ; Ethanol ; Female ; Food Science ; Health risks ; Heart diseases ; Hexanes ; High density lipoprotein ; High performance liquid chromatography ; Humans ; Incubation ; Laboratory Medicine ; Lecithin ; Linear functions ; Lipid metabolism ; Lipids ; Liquid chromatography ; Low density lipoprotein ; Male ; Metabolism ; Methods ; Middle Aged ; Monitoring/Environmental Analysis ; Observations ; Phosphatidylcholine-Sterol O-Acyltransferase - blood ; Phosphatidylcholine-Sterol O-Acyltransferase - isolation & purification ; Phosphatidylcholine-Sterol O-Acyltransferase - metabolism ; Phosphocholine ; Reagents ; Reproducibility ; Reproducibility of Results ; Research Paper ; Risk assessment ; Sterols ; Sterols - blood ; Sterols - isolation & purification ; Sterols - metabolism ; Substrate Specificity ; Substrates ; Triglycerides ; Ultraviolet absorption ; Young Adult
Is Part Of: Analytical and bioanalytical chemistry, 2018-02, Vol.410 (6), p.1785-1792
Description: The measurement of lecithin: cholesterol acyltransferase (LCAT, EC 2.3.1.43) activity is important in high-density lipoprotein (HDL) metabolism study and cardiovascular disease (CVD) risk assessment. However, current methods suffer from complex design and preparation of exogenous substrate, low reproducibility, and interference of cofactors. In this study, we developed a simple and precise high performance liquid chromatography (HPLC) method for the measurement of LCAT activity. By using 7-dehydrocholesterol (7-DHC) and 1,2-didecanoyl- sn -glycero-3-phosphocholine(10:0PC) as substrates, and an LCAT activating peptide (P642) as activator and emulsifier, the substrate reagent was easily made by vortex. The substrate reagent was mixed with serum samples (50:1, v / v ) and incubated at 37 °C for 1 h. After incubation, the lipid was extracted with hexane and ethanol. With a conjugated double bond and ultraviolet absorption, 7-DHC and its esterification product could be separated and analyzed by a single HPLC run without calibration. LCAT activity was a linear function of the serum sample volume and the intra- and total assay coefficients of variation (CV) less than 2.5% were obtained under the standardized conditions. The substrate reagent was stable, and assay result accurately reflected LCAT activity. LCAT activities in 120 healthy subjects were positively correlated with triglyceride ( P < 0.05), fractional esterification rate of HDL cholesterol (FER HDL ) ( P < 0.0001), and negatively correlated with apolipoprotein AI (apoAI) ( P < 0.05) and HDL cholesterol (HDL-C) ( P < 0.001). These results suggest that this method is sensitive, reproducible, and not greatly influenced by serum components and added substances, and will be a useful tool in the lipid metabolism study and the risk assessment of CVD.
Publisher: Berlin/Heidelberg: Springer Berlin Heidelberg
Language: English
Identifier: ISSN: 1618-2642
EISSN: 1618-2650
DOI: 10.1007/s00216-017-0834-4
PMID: 29307006
Source: MEDLINE
ProQuest Central

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A simple and precise method to detect sterol esterification activity of lecithin/cholesterol acyltransferase by high-performance liquid chromatography

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