Abstract
A simple, rapid, and sensitive liquid chromatography tandem mass spectro-metric (LC–MS/MS) assay method has been developed and fully validated for the simultaneous quantification of atorvastatin and aspirin in human plasma using a polarity switch. Proguanil and furosemide were used as the internal standards for the quantification of atorvastatin and aspirin, respectively. The analytes were extracted from human plasma by the liquid–liquid extraction technique using methyl tert-butyl ether. The reconstituted samples were chromatographed on a Zorbax XDB Phenyl column by using a mixture of 0.2% acetic acid buffer, methanol, and acetonitrile (20:16:64, v/v) as the mobile phase at a flow rate of 0.8 mL/min. Prior to detection, atorvastatin and aspirin were ionized using an ESI source in the multiple reaction monitoring (MRM) mode. The ions were monitored at the positive m/z 559.2→440.0 transition for atorvastatin and the negative m/z 179.0→136.6 transition for aspirin. The calibration curve obtained was linear (r2 ≥ 0.99) over the concentration range of 0.20–151 ng/mL for atorvastatin and 15.0–3000 ng/mL for aspirin. The method validation was performed as per FDA guidelines and the results met the acceptance criteria. A run time of 3.0 min for each sample made it possible to analyze more than 300 human plasma samples per day. The proposed method was found to be applicable to clinical studies.
Highlights
Hyperlipedemia is the elevation in the bloodstream of lipids including fats, fatty acids, cholesterol, cholesterol esters, phospholipids, and triglycerides
Good response was found in the positive ionization mode for atorvastatin and the negative ionization mode for aspirin
A negative–to–positive ionization switch mode was used to detect the two analytes in order to achieve the best sensitivity for aspirin and atorvastatin
Summary
Hyperlipedemia is the elevation in the bloodstream of lipids including fats, fatty acids, cholesterol, cholesterol esters, phospholipids, and triglycerides. Statins competitively inhibit the enzyme HMG–CoA reductase, which is involved in the rate–limiting step of cholesterol biosynthesis in the liver, resulting in the up–regulation of the low–density lipoprotein (LDL) receptor and the lowering of LDL cholesterol in the blood [3, 4]. 134523-00-5) is a potent and competitive inhibitor of the enzyme HMG–CoA reductase, the rate–limiting enzyme in cholesterol biosynthesis in the liver. Atorvastatin induces a significant reduction in total cholesterol (TC), LDL–C, and plasma triglycerides (TG) [5, 6]. At low doses (˂100 mg), aspirin is employed as an antithrombotic agent to selectively inhibit cyclooxygenase–dependent platelet aggregation [9]
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