title: Development and validation of an on-line two-dimensional reversed-phase liquid chromatography-tandem mass spectrometry method for the simultaneous determination of prostaglandins E(2) and F(2alpha) and 13,14-dihydro-15-keto prostaglandin F(2alpha) levels in human plasma.
abstract: We developed and validated an on-line reverse-phase two-dimensional LC/MS/MS (2D-LC/MS/MS) system for simultaneous determination of the levels of prostaglandin (PG) E(2) as well as PGF(2alpha) and its metabolite 13,14-dihydro-15-keto PGF(2alpha) (F(2alpha)-M) in human plasma. Analytes were extracted by a three-step solid-phase extraction. Samples were then analyzed by on-line 2D-LC/MS/MS with electrospray ionization in negative mode. The 2D-LC system is composed of two reverse-phase analytical columns with a trapping column linking the two analytical columns. While an acidic buffer was used for both separation dimensions, differing organic solvents were employed for each dimension: methanol for the first and acetonitrile for the second to increase resolving power. The 2D-LC/MS/MS method was highly selective and sensitive with a significantly lower limit of quantitation (0.5 pg/mL for PGE(2) and 2.5 pg/mL for PGF(2alpha) and F(2alpha)-M, respectively). Linearity of the 2D-LC/MS/MS system was demonstrated for the calibration ranges of 0.5-50 pg/mL for PGE(2) and 2.5-500 pg/mL for PGF(2alpha) and F(2alpha)-M, respectively. Acceptable precision and accuracy were obtained throughout the calibration curve ranges. This highly selective and sensitive method was successfully utilized to determine the endogenous levels of PGE(2), PGF(2alpha), and F(2alpha)-M in plasma samples from six (four male and two female) normal volunteers. The mean concentrations for each analyte were 0.755 pg/mL for PGE(2), 5.70 pg/mL for PGF(2alpha) and 9.48 pg/mL for F(2alpha)-M.
authors.affiliation: Pharmacokinetic Research Laboratories, Ono Pharmaceutical Co. Ltd, Ibaraki, Japan. junji.komaba@ono-usa.com
authors.forename: Junji
authors.initials: J
authors.lastname: Komaba
authors.forename: Dai
authors.initials: D
authors.lastname: Matsuda
authors.forename: Kimio
authors.initials: K
authors.lastname: Shibakawa
authors.forename: Susumu
authors.initials: S
authors.lastname: Nakade
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Hashimoto
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Miyata
authors.forename: Mikio
authors.initials: M
authors.lastname: Ogawa
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000097:blood*; D015232:Dinoprostone / Q000097:blood*; D004867:Equipment Design; D005260:Female; D006801:Humans; D016014:Linear Models; D008297:Male; D008956:Models, Chemical; D055550:Protein Stability; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C008228:15-keto-13,14-dihydroprostaglandin F2alpha; D015237:Dinoprost; D015232:Dinoprostone