An ultra-performance liquid chromatography-tandem mass spectrometry method for the determination of obeticholic acid in rat plasma and its application in preclinical pharmacokinetic studies

Obeticholic Acid: An Insight Into a Quantitative Determination and Methodological Validation Through Nuclear Magnetic Resonance

“…As standard expository methodology for more up to date medications or formulations may not be available in pharmacopeias, hence it is fundamental need to create novel analytical procedures which should be precise and accurate. The proposed method is relatively fast, simple, precise, selective and sensitive sufficient for all observed related compounds of obeticholic acid compared with previously published studies mainly had used HPLC-MS [12][13][14] to define the concentration of obeticholic acid in plasma, and its methods were not explained whatsoever because they cannot perform broad method validation, detailed method optimization as according to ICH parameters. The main advantage of this proposed method was that no reference standard of drug was required for quantification and method validation, this high simplicity, reliability, simultaneously measurement enable it to apply for quantitation of obeticholic acid in future as well as existing commercial products.…”

“…NMR experiments were performed on Bruker Avance-III HD 400 MHz and Avance-II 400 MHz ultra-shield spectrometer (JCL/ANAL/NMR/02 and 03) equipped with a 5 mm multinuclear broad-band-observe (BBO) probe-head and 5 mm 1 H- 13 C dual probehead respectively. Other instruments were also used for drug characterization, sample preparations and comparison of assay result that are FT-IR instrument: Perkin Spectrum Two FT-IR 114570 spectrometer (GCD FIR 02 2020), LC-MS: ultra-highperformance liquid chromatography-mass spectrometry (ID: UHPLC-MS-01) equipped with a dual agilent jet stream electrospray ionization (AJS-ESI) and evaporative light scattering detector (ELS1 A), Melting point: Buchi Melting Point (QCD/MPT-01), weighing machine: mettler Toledo XS 205 dual range (JCL/ANAL/BALANCE/03) and Sonicator: Ultrasonicator (JCL/ANAL/US/01).…”

“…Structure of the analyte drug and the internal standard shown in fig. 2, a various experiments such as 13 C NMR, APT, 1 H-1 H COSY and 1 H- 13 C HMQC were performed in DMSO-d6 for the structural characterization of obeticholic acid drug. From these experiments it was noted that there are three hydroxyl (OH) protons are present in molecule which was further ensured by performing D2O exchange/shake analysis in which all exchangeable protons disappeared.…”

“…Although, the proposed method is relatively fast, simple, precise, selective and sensitive sufficient for all observed related compounds of obeticholic acid compared with previously published studies mainly had used HPLC-MS [12][13][14] to define the concentration of obeticholic acid in plasma, and its methods were not explained whatsoever because they didn't perform comprehensive method validation, detailed method optimization as per international council for harmonisation (ICH) parameters like Malz et al, El-Sheikh et al and Harahap et al performed [16][17][18]. Thus, the research has been undertaken and this paper reflect the work of developing advantageous and competitive selective NMR method for the determination of the drug in formulation as well as in active pharmaceutical ingredient (API) samples that complies well with the validation requirements in the pharmaceutical industry as per standard guidelines by ICH guidelines Q2 (R1) [19].…”

“…gov, number NCT01265498 and NCT02177136) [10,11]. A numerous analytical method has been reported over time for the determination of obeticholic acid in biological fluids or individually in which mainly includes chromatographic-based HPLC/lC-MS [12][13][14] and other analysis like volumetric with visual end-point detection [15].…”