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-2 alpha and 13, 14-dihydro-15-keto prostaglandin F-2 alpha levels in human plasma

作者：Komaba Junji^*; Matsuda Dai; Shibakawa Kimio; Nakade Susumu; Hashimoto Yoshitaka; Miyata Yasuyuki; Ogawa Mikio

来源：Biomedical Chromatography, 2009, 23(3): 315-323.

DOI：10.1002/bmc.1117

• prostaglandin
• LC/MS/MS
• multi-dimensional LC

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(2 alpha) and its metabolite 13,14-dihydro-15-keto PGF(2 alpha) (F-2 alpha-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(2 alpha) and F-2 alpha-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(2 alpha) and F-2 alpha-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(2 alpha), and F-2 alpha-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(2 alpha) and 9.48 pg/mL for F-2 alpha-M.

• 出版日期2009-3