Title:Determination of three Unsaturated Fatty Acids in Pressure Ulcer Rats Using A UPLC-MS/MS Method
Volume: 17
Issue: 8
Author(s): Fuman Cai, Yuwei Dong, Shaosheng Lou, Zeping Ma, Ting Wu, Liuqing Yang, Congcong Wen*Chenxiang Wang*
Affiliation:
- Laboratory Animal Center, Wenzhou Medical University, Wenzhou,China
- Department of Pharmacy, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou,China
Keywords:
UPLC-MS/MS, pressure ulcer, docosahexaenoic acid, eicosapentaenoic acid, arachidonic acid, rat.
Abstract:
Background: The serum levels of Docosahexaenoic Acid (DHA), Eicosapentaenoic Acid
(EPA) and Arachidonic Acid (AA) under the state of Pressure Ulcers (PUs) are still unclear.
Introduction: In order to investigate serum levels of DHA, EPA, and AA in PUs rats, an ultraperformance
liquid chromatography-tandem mass spectrometry (UPLC-MS/ MS) method was developed
and validated.
Methods: Chromatographic separation of DHA, EPA, AA was carried out on a BEH C18 column and
gradient elute consisted of 5 mM ammonium acetate-0.1% formic acid and acetonitrile. Subsequently,
fifty rats were divided into five groups (n=10), four PU groups (A-D) underwent various pressure and
release time protocols, with group E as the control. The concentrations of DHA, EPA, AA from five
groups were determined by using a validated method.
Results: The results showed there was good linearity for DHA (327.3/283.4), EPA (301.2/257.0), and
AA (303.1/258.9) within 0.05-6.4 μg/mL. In control group, the levels of DHA, AA and EPA were
1.16±0.68, 0.59±0.19 and 0.78±0.21 μg/mL. At the end of modeling, concentrations of DHA, EPA and
AA were increased after long and persistent pressure (>8 h). Especially, the level of DHA was significantly
higher (P<0.01) than that of control group.
Conclusion: A stable, reliable and accurate UPLC-MS/MS for determination of DHA, EPA, AA in
blood was developed. Serum concentrations of DHA, EPA and AA were altered differently after long
and persistent pressure (>8 h), and DHA is a remarkable one.