Simultaneous extraction of acetylsalicylic acid and salicylic acid from human plasma and simultaneous estimation by liquid chromatography and atmospheric pressure chemical ionization/tandem mass spectrometry detection

Abstract

A simple analytical method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in atmospheric chemical ionization mode (APCI) for the simultaneous estimation of acetylsalicylic acid (ASA, CAS 50-78-2) and its active metabolite salicylic acid (SA, CAS 69-72-7) in human plasma has been developed and validated. ASA and SA were analyzed simultaneously despite differences in plasma concentration ranges of ASA and SA after oral administration of ASA. In spite of having different chemical, ionization and chromatographic properties, ASA and SA were extracted simultaneously from the plasma sample using acetonitrile protein precipitation followed by liquid-liquid extraction. The analytes were separated on a reversed phase column with rapid gradient program using mobile phase consisting of ammonium acetate buffer and methanol. The structural analogue diclofenac was used as an internal standard. The multiple reaction monitoring (MRM) transitions m/z 179 --> 137 for ASA, m/z 137 --> 65 for SA and m/z 294 --> 250 for IS were used. The assay exhibited a linear dynamic range of 0.02-10 microg/mL for ASA and 0.1-50 microg/mL for SA. The between-batch precision (%CV) ranged from 2.1 to 7.9% for ASA and from 0.2 to 5.2% for SA. The between-batch accuracy ranged from 95.4 to 96.7% for ASA and from 94.6 to 111.3% for SA. The validated method was successfully applied for the evaluation of pharmacokinetics of ASA after single oral administration of 650 mg test formulation versus two 325 mg reference formulations of ASA in human subjects.

Robust and sensitive LC/MS-MS method for simultaneous detection of acetylsalicylic acid and salicylic acid in human plasma

“…As this method used duplicate sample preparation for the analysis of the analytes, it was time consuming for both the sample preparation and the analysis. Nirogi et al [23] developed a two-step extraction procedure for the simultaneous extraction of ASP and SAL. They used a protein precipitation method followed by a solvent extraction step for sample preparation.…”

“…The number of the published LC-MS/MS methods that are suitable for the simultaneous determination of ASP and SAL from plasma is still limited. Bae et al [22] applied sample preparation methods separately for the parent compound and its deacetylated metabolite, whereas others reported multi-step sample preparation (protein precipitation followed by a liquid-liquid extraction) [23], one-step solidphase extraction [24], or even direct injection of the supernatant of precipitated plasma into the LC-MS/MS system [25]. To the best of our knowledge, no published LC-MS/MS method with a one-step liquid-liquid extraction is available for the simultaneous extraction and quantification of ASP and SAL in human plasma.…”

Low dose aspirin estimation: an application to a human pharmacokinetic study

“…The reported bioanalytical methods for the simultaneous quantification of ASP and SAL in biological matrices were based on HPLC (Peng et al ., ; Buskin et al ., ; Bakar and Niazi, ; Mays et al ., ; O'Kruk et al ., ; Shen et al ., ; Kees et al ., ; McMahon and Kelly, ; Pirola et al ., ), GC (Walter et al ., ; Rance et al ., ), capillary electrophoresis (Hansen et al ., ) column switching followed by online SPE (McMahon and Kelly, ) and LC‐MS/MS (Bae et al ., ; Xu et al ., ; Nirogi et al ., ). All the earlier reported methods used sodium fluoride/potassium fluoride as enzyme inhibitors to prevent the enzymatic hydrolysis and the collection procedure required a set temperature of 4°C for the sample processing.…”

Development and Validation of a Method for the Simultaneous Quantification of Aspirin, Salicylic Acid, Rosuvastatin, Rosuvastatin Lactone and N-Desmethyl Rosuvastatin in Human Plasma Using UPLC-MS/MS-API-5500

“…Literature survey shows that RP-HPLC stability indicating methods for rosuvastatin alone or with combination [2], LC-MS/MS methods for plasma for rosuvastatin alone or with metabolites/combination of other drugs [3][4][5][6][7][8][9][10][11][12][13][14][15][16], LC-MS/MS methods for plasma for aspirin alone or with metabolites/combination of other drugs [17][18][19][20][21][22][23][24][25] are reported. It is noted that method for simultaneous estimation of RVT with metabolites and ASP with metabolite is not available.…”