Determination of clenbuterol, salbutamol, and cimaterol in bovine retina by electrospray ionization-liquid chromatography-tandem mass spectrometry.
An electrospray ionization-liquid chromatography-tandem mass spectrometry (ESI/LC/MS/MS) method was developed for the simultaneous determination of the beta-agonists clenbuterol, salbutamol, and cimaterol in bovine retina. The tissue was homogenized in alkaline buffer and spiked to give 10, 15, and 20 ng/g each of the 3 analytes together with the internal standards d6-salbutamol and d6-clenbuterol. The mixture was incubated with protease enzyme to release any protein-bound analytes and then made alkaline before extraction with isobutanol. The extract was dissolved in water and transferred to a clenbuterol immunoaffinity column. After washing, the analytes were eluted and analyzed by ESI/LC/MS/MS using a C18 column with acetic acid-methanol as mobile phase. No interferences were observed from the spiked retina extract at the various single-reaction monitoring modes. Average recoveries for clenbuterol, salbutamol, and cimaterol were 94, 85, and 87% with coefficients of variation (CVs) of 9.4, 9.9, and 8.6%, respectively. A correlation coefficient of r2 = 0.9999 was obtained for all analytes. The limits of detection for clenbuterol, salbutamol, and cimaterol, determined from 3 times the standard deviation of 7 replicates of the lowest spike, were 2.5, 3.5, and 2.0 ng/g with CVs of 8.9, 11.6, and 7.2%, respectively.
Lau
Joseph H W
JH
University of Western Sydney, Center for Complementary Medicine Research, Locked Bag 1797, Penrith South DC, NSW 1797, Australia.
Khoo
Cheang S
CS
Murby
John E
JE
D000318:Adrenergic beta-Agonists / Q000032:analysis*; D000420:Albuterol / Q000032:analysis*; D000818:Animals; D002021:Buffers; D002417:Cattle; D002976:Clenbuterol / Q000032:analysis*; D004983:Ethanolamines / Q000032:analysis*; D007120:Immunochemistry; D007202:Indicators and Reagents; D012015:Reference Standards; D012160:Retina / Q000737:chemistry*; D012996:Solutions; D021241:Spectrometry, Mass, Electrospray Ionization
D000318:Adrenergic beta-Agonists; D002021:Buffers; D004983:Ethanolamines; D007202:Indicators and Reagents; D012996:Solutions; D000420:Albuterol; D002976:Clenbuterol; C047411:cimaterol