Development and validation of an on-line two-dimensional reversed-phase liquid chromatography-tandem mass spectrometry method for the simultaneous determination of prostaglandins E2and F2αand 13,14-dihydro-15-keto prostaglandin F2αlevels in human plasma
| Autor: | Junji Komaba, Dai Matsuda, Yoshitaka Hashimoto, Yasuyuki Miyata, Kimio Shibakawa, Mikio Ogawa, Susumu Nakade |
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| Rok vydání: | 2009 |
| Předmět: |
Pharmacology
Analyte Chromatography Calibration curve Metabolite medicine.medical_treatment Electrospray ionization Clinical Biochemistry General Medicine Reversed-phase chromatography Tandem mass spectrometry Biochemistry Analytical Chemistry chemistry.chemical_compound chemistry Drug Discovery medicine Solid phase extraction Molecular Biology Prostaglandin E |
| Zdroj: | Biomedical Chromatography. 23:315-323 |
| ISSN: | 1099-0801 0269-3879 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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