Development and validation of an LC-MS method for quantitation of ergot alkaloids in lateral saphenous vein tissue.
A liquid chromatography-mass spectrometry (LC-MS) method for simultaneous quantitation of seven ergot alkaloids (lysergic acid, ergonovine, ergovaline, ergocornine, ergotamine, ergocryptine, and ergocristine) in vascular tissue was developed and validated. Reverse-phase chromatography, coupled to an electrospray ionization source, was used to separate and ionize alkaloids. Singly protonated molecular ions for each alkaloid and methysergide (internal standard), were detected by single-ion monitoring (SIM). Calibration curves were obtained over a linear range of 0.1 to 40 pmol on column with correlation coefficients better than 0.994. Method recoveries were 68.4% to 111.0%. Intra-assay precision was 3.4% to 16.1%. Matrix effects were observed and overcome by introducing matrix components into calibrant solutions to create matrix-diluted standards. Limits of detection and quantitation were 0.05 pmol and 0.1 pmol, respectively. Method ruggedness tests resulted in recoveries of 86.1 to 122% with an interassay precision of 7.9% to 22.8%. These results indicate that this method is suitable for quantitation of alkaloids extracted from in vitro-exposed vascular tissue.
Smith
Darrin
D
Department of Chemistry, Eastern Kentucky University, Richmond, Kentucky 40475, USA. darrin.smith@eku.edu
Smith
Lori
L
Shafer
Wilson
W
Klotz
James
J
Strickland
James
J
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004876:Ergot Alkaloids / Q000032:analysis*; D005260:Female; D057230:Limit of Detection; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D012501:Saphenous Vein / Q000737:chemistry*
D004876:Ergot Alkaloids