Multi-component plasma quantitation of anti-hyperglycemic pharmaceutical compounds using liquid chromatography-tandem mass spectrometry.

Type-2 diabetes is a disorder characterized by disrupted insulin production leading to high blood glucose levels. To control this disease, combination therapy is often used. Hypoglycemic agents such as metformin, glipizide, glyburide, repaglinide, rosiglitazone, nateglinide, and pioglitazone are widely prescribed to control blood sugar levels. These drugs provide the basis for the development of a quantitative multianalyte bioanalytical method. As an example, a highly sensitive and selective multi-drug method based on liquid chromatography tandem mass spectrometry (LC-MS/MS) was developed. This rapid, automated method consists of protein precipitation of 20 microL of plasma coupled with gradient HPLC elution of compounds using 10 mM ammonium formate buffer and 0.1% formic acid in acetonitrile as the mobile phases. MS/MS detection was performed using turbo ion spray in the positive ion multiple reaction monitoring (SRM) mode. A lower limit of quantitation (LLQ) in a range of 1.0-5.0 ng/mL was achieved for all analytes. The linearity of the method was observed over a 500-fold dynamic range. Drug recoveries ranged from 86.2 to 94.2% for all analytes of interest. Selectivity, sample dilution, intra-day and inter-day accuracy and precision, and stability assessment were evaluated for all compounds.

Determination of acepromazine, ketamine, medetomidine, and xylazine in serum: multi-residue screening by liquid chromatography-mass spectrometry.

A large variety of drugs are administered to large and small animals by veterinary clinicians for sedation, anesthesia, muscle relaxation, and analgesia. The present paper reports a simple and rapid multi-residue detection and quantitation method for four chemically different drugs: medetomidine, xylazine, ketamine, and acepromazine. Chromatographic separation was carried out on a liquid chromatography-mass spectrometry instrument with a C18-reversed-phase column. Fragmentation patterns were determined with atmospheric pressure chemical ionization mass spectrometry set to operate in a positive selective ion monitoring mode. The method was determined to be linear over the range of concentrations tested (2.0-100.0 ng/mL). Accuracy, precision, and specificity were evaluated and the method was determined to be applicable to detection of medetomidine, xylazine, ketamine, and acepromazine in serum samples of multiple animal species (canine, equine, and bovine). Matrix limits of quantitation were determined to be 5.0 ng/mL for all four analytes, and recoveries ranged between 82.0 and 118%, with a 3.0-18.3% relative standard deviation.