Verapamil drug metabolism studies by automated in-tube solid phase microextraction.
Verapamil is a common calcium antagonist described with antianginal, antihypertensive and antiarrythmic properties. The metabolites of verapamil have also shown pharmacological properties and therefore sample preparation and analysis techniques capable of metabolic screening for verapamil are important. In-tube SPME is a relatively new method integrating sample extraction, concentration and introduction into one single step without the use of organic solvents. The capability of in-tube SPME in bioanalysis has been reviewed but there has been no application described in the field of drug metabolism. Since automation and interfacing of in-tube SPME coupled to liquid chromatography-mass spectrometry (LC-MS) is possible, we confirm in this study that it is a powerful method to monitor the main metabolites of verapamil in various biological matrices like plasma, urine and cell culture media. Further, we show that it could also be used in routine pharmacokinetics measurements. An in-tube SPME LC-MS method was developed to extract and analyze the metabolic profile of verapamil from biological matrices. The detection limit for verapamil, gallopamil, norverapamil and PR22 were 52, 53, 65 and 83 ng/ml (UV detection) and 5, 6, 6 and 8 ng/ml (MS detection), respectively. The precision of the method was calculated in various biological matrices and the average % R.S.D. (N=5) for verapamil, gallopamil, norverapamil and PR22 was 3.9, 3.7, 3.8 and 4.3% (MS detection), respectively. The linear dynamic range was determined to be 100-800 ng/ml (UV detection) with a total sample preparation and analysis time of 34 min.
Fraunhofer Institute of Toxicology and Aerosol Research, Drug Research and Clinical Inhalation, Hanover D-30625, Germany.
M
M
Walles
W M
WM
Mullett
K
K
Levsen
J
J
Borlak
G
G
Wünsch
J
J
Pawliszyn
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000706:statistics & numerical data; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods / Q000706:statistics & numerical data; D051381:Rats; D017207:Rats, Sprague-Dawley; D014700:Verapamil / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*
D014700:Verapamil