title: A highly sensitive method for the quantification of fludrocortisone in human plasma using ultra-high-performance liquid chromatography tandem mass spectrometry and its pharmacokinetic application.
abstract: A simple and high sensitive ultra-high-performance liquid chromatography tandem mass spectrometry method for the determination of fludrocortisone in human plasma was developed and validated as per guidelines. The analyte and internal standard (IS), fludrocortisone-d5 , were extracted from human plasma via liquid-liquid extraction using tert-butyl methyl ether. The chromatographic separation was achieved on a Chromolith RP18e column using a mixture of acetonitrile and 2 mm ammonium formate (70:30, v/v) as the mobile phase at a flow rate of 0.7 mL/min. Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique, operating in multiple reaction monitoring and positive ion mode. The precursors to product ion transitions monitored for fludrocortisone and IS were m/z 381.2 → 343.2 and 386.2 → 348.4, respectively. The assay was validated with linear range of 40-3000 pg/mL. The intra- and inter-day precisions (relative standard deviation) were within 0.49-7.13 and 0.83-5.87%, respectively. The proposed method was successfully applied to pharmacokinetic studies in humans.
authors.affiliation: Bioanalytical Department, QPS Bioserve India Pvt. Ltd, Balanagar, Hyderabad-, 500 037, India.
authors.forename: Jagadeesh
authors.identifier: http://orcid.org/0000-0002-2130-8771
authors.initials: J
authors.lastname: Banda
authors.affiliation: Bioanalytical Department, QPS Bioserve India Pvt. Ltd, Balanagar, Hyderabad-, 500 037, India.
authors.forename: Ramalingam
authors.initials: R
authors.lastname: Lakshmanan
authors.affiliation: Bioanalytical Department, QPS Bioserve India Pvt. Ltd, Balanagar, Hyderabad-, 500 037, India.
authors.forename: Shiva Prasad
authors.initials: SP
authors.lastname: Vvs
authors.affiliation: Bioanalytical Department, QPS Bioserve India Pvt. Ltd, Balanagar, Hyderabad-, 500 037, India.
authors.forename: Srinibas Patro
authors.initials: SP
authors.lastname: Gudla
authors.affiliation: Bioanalytical Department, QPS Bioserve India Pvt. Ltd, Balanagar, Hyderabad-, 500 037, India.
authors.forename: Rosaiah
authors.initials: R
authors.lastname: Prudhivi
mesh_terms: D000893:Anti-Inflammatory Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005438:Fludrocortisone / Q000097:blood*; D006801:Humans; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction / Q000379:methods; D008297:Male; D008738:Methyl Ethers / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000893:Anti-Inflammatory Agents; D008738:Methyl Ethers; C043243:methyl tert-butyl ether; D005438:Fludrocortisone
keywords: UHPLC-MS/MS; fludrocortisone; human plasma; method validation; pharmacokinetics