Open AccessAn efficient method for determination of components in docosahexaenoic acid-phosphatidylcholine using pre-column derivatization HPLC
Author(s) -
Li LIU,
Tiantian Zhang,
Xiaoli Zhang,
Jiao Wang,
Wenfang Shu,
Xinyu Qi,
Binxia Zhao,
Binglin LI
Publication year - 2021
Publication title -
revue roumaine de chimie
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.135
H-Index - 21
ISSN - 0035-3930
DOI - 10.33224/rrch.2020.65.10.10
Subject(s) - derivatization , chromatography , chemistry , high performance liquid chromatography , docosahexaenoic acid , phosphatidylcholine , phospholipid , reagent , elution , hydrolysis , triethylamine , methanol , acetonitrile , fatty acid , organic chemistry , biochemistry , polyunsaturated fatty acid , membrane
The content of docosahexaenoic acid-phosphatidylcholine (DHA-PC) in the phospholipid product is closely related to its nutritional value and health function. An efficient high performance liquid chromatography (HPLC) was firstly proposed and successfully applied to the determination of DHA-PC. DHA-PC was pre-treated by alkaline hydrolysis, acidification and then the obtained components underwent derivatization using α-bromoacetophenone as a derivatization reagent, triethylamine as a catalyst, to make them have ultraviolet (UV) absorption. Major components of DHA-PC could reach baseline separation within a short time (50 min). A gradient elution method using acetonitrile and methanol/water (1:1, volume ratio) at 0.55 mL/min, UV detection at 244 nm and a column temperature of 50 oC was found to optimally separate the components of DHA-PC. The intra-assay precisions (n = 6) and stabilities (n = 6) relative standard deviations (RSDs) for the major components were 2.5%-3.2% and 1.9%-4.0%, respectively. The average recoveries were 97.8%-106%. The results indicated that this method was rapid, accurate and reliable to analyze the content of DHA-PC in the phospholipid product. This method could be used not only to detect the content of DHA-PC in the phospholipid product, but to analyze the content of various fatty acids in mixed PC.