The Primary Study On Kinetic Process Of Harp Seal Oil In Rats With Liquid Chromatography And Mass Spectrometry

Harp seal oil is extracted from the thick layer of blubber beneath the pelt. EPA, DHA and DPA are the three main polyunsaturated fatty acids of harp seal oil. As an extremely rich source of Omega-3 long chain polyunsaturated fatty acids, study results showed that harp seal oil had the actions of anti-inflammation, anti-coagulation, anti-thrombus, improving the micro-circulation, decreasing blood pressure, regulating serum lipid level, anti-prostate hyperplasia , anti-tumor and so on. Until now, the study on harp seal oil has focused on analysis technics, pharmacology and the clinical trials. However, there are no republic reports on its kinetic process. In this paper, the methods for the extraction and determination of EPA, DHA and DPA in plasma were established, and the primary study on kinetic process of harp seal oil in rats was investigated.In our study, it is the first time to establish the method of extracting EPA,DHA and DPA from the plasma. After protein precipitation, liquid-liquid extraction and derivation, the analytes are determined by the form of methyl ester.A highly sensitive, specific and rapid LC/MS method was established for the determination of three main components including EPA, DHA and DPA in plasma. Methyl nonadecadienoate, a structurally related analog of analyte, was used as the internal standard. The extract was chromatographed on a Waters RP18 column. The mobile phase consisted of TFA-acetonitrile-water, at a flow rate of 0.3ml/min. A Shimadzu 2010A mass spectrometer equipped with atmospheric pressure chemical ionization source was used as detector and was operated in the positive ion mode. Selected ion monitoring mode was used to qualify EPAM,DHAM and DPAM. The assay was reproducible and linear for EPAM,DHAM and DPAM in the range of 50-10000ng/mL.The lower limit of qualification was 50ng/mL. Each sample was chromatographed only 15 minutes. The precision was within 7% .The extraction recovery was greater than 80%.The preclinical kinetic process of harp seal oil in rats was studied by the validated LC/MS method. After multiple oral administration of harp seal oil to rats in 9.0,4.5 and 2.7g/kg, plasma concentrations of the rats were determined by the method. The...