title: Multiplex quantification of lamprey specific bile acid derivatives in environmental water using UHPLC-MS/MS.
abstract: Larval and adult sea lampreys (Petromyzon marinus) release bile salts and acids into the surrounding aquatic environment. Some of these bile salts and acids, such as petromyzonol sulfate (PZS), 3-keto petromyzonol sulfate (3k PZS), petromyzonamine disulfate (PADS), petromyzosterol disulfate (PSDS), and 3-keto allocholic acid (3k ACA), may function as pheromones. To examine the release and distribution patterns of these metabolites, which this study has termed bile acid derivatives, we developed a novel UHPLC-MS/MS method that was characterized by simple sample preparation, baseline separation, and short analysis time for all studied compounds. These five analytes were separated in 7 min using a reversed-phase C18 column containing 1.7 μm particles and a gradient elution at pH 8.9. Once separated, the analytes were subjected to electrospray ionization-mass spectrometry (negative ion mode) and collision-induced dissociation tandem mass spectrometry (CID-MS/MS) using the multiple reaction monitoring (MRM) mode. Deuterated 3k PZS ([(2)H(5)]3k PZS) was added as the internal standard (IS) to the sample prior to solid phase extraction (SPE). Among the three types of SPE sorbent tested, mixed-mode cation-exchange and reversed-phase sorbent for bases (MAX) and acids (MCX), and reversed-phase C18 sorbent (Sep-pak), the best recoveries (84.1-99.7%) were obtained with MCX cartridges. The calibration curves of all five analytes were linear between 0.15 and 1200 ng/mL, with R(2)≥0.9997. This method had a precision of relative standard deviation (RSD) ≤9.9% and an accuracy of deviation (DEV) ≥92.5%. The developed method was successfully used to quantify bile acid derivatives found in streams where lampreys spawn (SD<1.4) and water conditioned with male sea lampreys (SD<4.8). Utilizing this method provides a routine analysis of lamprey bile acid derivatives and may prove useful for sea lamprey population estimates in future studies and applications.
authors: Department of Fisheries and Wildlife, Michigan State University, East Lansing, MI 48824, United States.
authors: Ke
authors: K
authors: Li
authors: Huiyong
authors: H
authors: Wang
authors: Cory O
authors: CO
authors: Brant
authors: SangChun
authors: S
authors: Ahn
authors: Weiming
authors: W
authors: Li
mesh_terms: D000818:Animals; D001647:Bile Acids and Salts / Q000032:analysis* / Q000378:metabolism; D002776:Cholestanes / Q000032:analysis; D002793:Cholic Acids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase; D016018:Least-Squares Analysis; D057230:Limit of Detection; D008297:Male; D048428:Petromyzon / Q000378:metabolism*; D011760:Pyrrolidinones / Q000032:analysis; D015203:Reproducibility of Results; D045483:Rivers / Q000737:chemistry; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001647:Bile Acids and Salts; D002776:Cholestanes; D002793:Cholic Acids; D011760:Pyrrolidinones; C507613:petromyzonamine disulfate; C500317:petromyzonol sulfate