A Rapid, Sensitive and Validated Ultra Performance Liquid Chromatography and Tandem Mass Spectrometry Method for Determination of Rifampicin in Rat Plasma: Application to Pharmacokinetic Study

Abstract

Objective: To develop and validate simple, sensitive and selective ultra-performance liquid chromatography and tandem mass spectrometry (UPLC-MS/MS) method for quantification of rifampicin (RIF) in rat plasma and its application to pharmacokinetics study. Methods:Precipitation method was used for the extraction of plasma samples, an aliquot of 25 µl plasma samples was extracted using acetonitrile precipitation technique. Chromatographic separation was performed usingWaters Acquity TM Results:The method had a short chromatography run time of 1.8 min with improved sensitivity over existing methods. Calibration curves been linear over the wide range of 1.97-5047.00 ng/ml. The between and within-batch precision and accuracy of the method was determined by using 4 quality control samples; the highest %CV observed was10.11. The mean recovery values are 74.26, 82.77 and 101.73 at low, medium and highquality control levels; respectively. UPLC columns, BEH C18 (50 mm× 2.1 mm, 1.7 µm) by a gradient mixture of acetonitrile and water (both containing 0.1 % formic acid) as a mobile phase at the flow rate of 0.7 ml/min.The analyte was protonated in the positive ESI (electrospray ionization interface) and detected in MRM (multiple reactions monitoring) modes using the transition m/z 308.60-455.30. Conclusion:It was concluded that the developed and validated UPLC-MS/MS method was sensitive,specific, precise, linear, and rapid. Therefore, the method can be used for quantification of RIFin rat plasma with various advantages over the reported methods. RIF is widely recommended by US-FDAguidance for industry on drug interaction studies and the developed method can be used to explore drug interaction studies in drug discovery and development.

Advancements in Preprocessing and Analysis of Nitrite and Nitrate since 2010 in Biological Samples: A Review

“…HPLC systems are available in many laboratories and are typically used for nitrate or nitrite detection in biological specimens. Usually, the Griess reaction in combination with different extraction techniques are adopted [10,18,24,25,31,37,38,[58][59][60][61] (Table 4). He et al [24] developed a method for the reaction between p-nitroaniline and nitrite ion, with acid media containing diphenylamine, which was used in combination with HPLC-UV to detect nitrite levels in saliva specimens (the reaction principle is shown in Figure 9).…”

Validated high‐performance thin layer chromatographic method for simultaneous quantification of rifampicin and quercetin in liquisolid formulation using fractional factorial design in robustness study

“…Literature reported uncovers different HPTLC, HPLC, UPLC, LC-MS/MS, and UPLC-MS/MS strategies that are accessible for the estimation of Rifampicin exclusively and in combination with other drugs in various dosage forms [8][9][10][11][12][13][14][15][16][17]. A few techniques have been accounted for, like HPLC, HPTLC, LC-MS/MS, and UPLC-MS/MS methods for the estimation of Quercetin exclusively and in combination with other drugs in various dosage forms [18][19][20][21][22][23][24][25][26][27].…”

Advanced Amplitude Centering as an Invigorating Manipulation for Unified Wavelength Spectral Resolution of Ternary Mixtures

“…Analysis of complex mixtures containing three or more components represented a challenge for analysts. For resolving such mixtures, Analysts preferred using hyphenated instrumentations such as LC-MS, GC-MS, LC-NMR [1,2] , which consume lots of time and money. Unfortunately, the spectrophotometric methods, which provided a solution for complex ternary mixtures, were tedious, require several manipulating steps and exhibited several limitations [3][4][5].…”