A Validated Enantioselective Assay for the Determination of Ibuprofen in Human Plasma Using Ultra Performance Liquid Chromatography with Tandem Mass Spectrometry (UPLC-MS/MS)

Abstract

A modified ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the quantitation of ibuprofen enantiomers in human plasma. Ibuprofen and flurbiprofen (internal standard) were extracted from human plasma at acidic pH, using a single-step liquid-liquid extraction with methyl-tert-butyl ether. The enantiomers of ibuprofen and flurbiprofen were derivatized to yield the corresponding diastereomers. Chromatographic separation was achieved using a phenyl column with a run time of 20 min. (R)-and (S)-ibuprofen were quantitated at the multiple reaction monitoring (MRM) transition of m/z 360.2  232.1, and (R)-and (S)-flurbiprofen were monitored at the MRM transition of m/z 398.3  270.1. The method was validated for accuracy, precision, linearity, range, limit of quantitation (LOQ), limit of detection (LOD), selectivity, absolute recovery, matrix effect, dilution integrity, and evaluation of carry-over. Accuracy for (R)-ibuprofen ranged between -11.8% and 11.2%, and for (S)-ibuprofen between -8.6% and -0.3%. Precision for (R)-ibuprofen was ≤ 11.2%, and for (S)-ibuprofen ≤ 7.0%. The calibration curves were weighted (1/X 2 , n = 7) and were linear with r 2 for (R)-ibuprofen ≥ 0.988 and for (S)-ibuprofen ≥ 0.990. The range of the method was 50 to 5000 ng/mL with the LOQ of 50 ng/mL, and LOD of 1 ng/mL, for (R)-and (S)-ibuprofen requiring 100 µL of sample. The method was applied successfully to a pharmacokinetic study with the administration of a single oral dose of ibuprofen capsules to human subjects.

Statistical approach for selection of regression model during validation of bioanalytical method

“…Therefore before defining the concentration range, it is essential to have information about the expected concentrations of the analyte in biological matrix. Considering that different dosage strength (200 mg, 400 mg, 800 mg etc) of IBP are available on the market and the information obtained from the literature data (Canaparo et al, 2000;Bonato et al 2003;Szeitz et al, 2010), a 500-fold concentration range was required (0.1 -50 mg/L).…”

Enantioselective analysis of ibuprofen enantiomers in mice plasma and tissues by high‐performance liquid chromatography with fluorescence detection: Application to a pharmacokinetic study

“…High-performance liquid chromatography (HPLC) coupled with UV detection is the most frequently analytical technique used. 11,13,[15][16][17][18][19][20][21][22][23][24] In addition, HPLC methods with fluorescence 1,25 or mass spectrometry [26][27][28][29] detection was applied. Methods using HPLC with UV detection are characterized by a relatively low cost, wide availability, and high precision and accuracy, but they are the least sensitive.…”

Bioanalytical method for the estimation of co‐administered esomeprazole, leflunomide and ibuprofen in human plasma and in pharmaceutical dosage forms using micellar liquid chromatography

“…The British pharmacopoeia introduces an acid base titration for the analysis of ibuprofen (British Pharmacopoeia Commission, ). Several different analytical methods has been reported for the determination of ibuprofen, including spectrophotometry (Małgorzata & Stefan, ; Snezana, Gordanaz, Alksandran, Biljanab, & Valentina, ; Vu, Dong, Nguyen, & Hue, ; Wahbi, Hassan, & Khamis, ; Yousry, Sayed, & Ibrahim, ), HPLC methods (Anwer, Syed, & Fotios, ; Peikova, Georgieva, & Tsvetkova, ), UPLC (Dantu, Shakil, & Mukkanti, ), UPLC/MS/MS (Szeitz, Edginton, Peng, Cheung, & Riggs, ) and GC–MS (Bilal & Ali, ). Esomeprazole is co‐administered to reduce stomach acid secretion that may result in peptic ulcer (Walker & Whittlesea, ).…”

“…Gordanaz, Alksandran, Biljanab, & Valentina, 2008;Vu, Dong, Nguyen, & Hue, 2014;Wahbi, Hassan, & Khamis, 2005;Yousry, Sayed, & Ibrahim, 2011), HPLC methods (Anwer, Syed, & Fotios, 2006;Peikova, Georgieva, & Tsvetkova, 2014), UPLC (Dantu, Shakil, & Mukkanti, 2011), UPLC/MS/MS (Szeitz, Edginton, Peng, Cheung, & Riggs, 2010) and GC-MS (Bilal & Ali, 2014). Esomeprazole is co-administered to reduce stomach acid secretion that may result in peptic ulcer (Walker & Whittlesea, 2008).…”