Simultaneous quantification of prostaglandins in human synovial cell-cultured medium using liquid chromatography/tandem mass spectrometry

Takabatake, Masaru; Hishinuma, Takanori; Suzuki, Naoto; Chiba, Shuya; Tsukamoto, Hiroki; Nakamura, Hironori; Saga, Toshihide; Tomioka, Y.; Kurose, Akira; Sawai, Takashi; Mizugaki, Michinao

doi:10.1054/plef.2002.0381

Cited by 53 publications

(44 citation statements)

References 12 publications

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“…Although a few LC/MS or LC/MS/MS (either with chromatographic separation or flow injection analysis) methods have been used for identification and quantification of either single eicosanoid [29], or various PGs [30], or PGs together with HETEs [31][32][33][34][35][36], or various DiHETrEs [37], or HETEs together with EETs [38,39] or HETEs, EETs, together with DiHETrEs [40] in various biological matrices in a single analysis, we could not identify a published mass spectrometric-based method for the simultaneous identification and quantification of PGs, DiHETrEs, HETEs, EETs, and parent compound AA in biological matrices, including brain tissue. In addition, the method validation data were not available in most of these methods.…”

Section: Currently Gc/ms Gc/ms/msmentioning

confidence: 99%

A liquid chromatography/mass spectrometric method for simultaneous analysis of arachidonic acid and its endogenous eicosanoid metabolites prostaglandins, dihydroxyeicosatrienoic acids, hydroxyeicosatetraenoic acids, and epoxyeicosatrienoic acids in rat brain tissue

Yue

Jansen

Strauss

et al. 2007

Journal of Pharmaceutical and Biomedical Analysis

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A sensitive, specific, and robust liquid chromatography/mass spectrometric (LC/ MS) method was developed and validated that allows simultaneous analysis of arachidonic acid (AA) and its cyclooxygenase, cytochrome P450, and lipoxygenase pathway metabolites prostaglandins (PGs), dihydroxyeicosatrienoic acids (DiHETrEs), hydroxyeicosatetraenoic acids (HETEs) and epoxyeicosatrienoic acids (EETs), including PGF 2α , PGE 2 , PGD 2 , PGJ 2 ,14,11,8,5,14,11,8,9-EET, and 5,6-EET in rat brain tissues. Deuterium labeled PGF 2α -d 4 , PGD 2 -d 4 , 15(S)-HETE-d 8 , 14,15-EET-d 8 , 11,12-EET-d 8 , 8,9-EET-d 8 , and AA-d 8 were used as internal standards. Solid phase extraction was used for sample preparation. A gradient LC/MS method using a C18 column and electrospray ionization source under negative ion mode was optimized for the best sensitivity and separation within 35 min. The method validation, including LC/MS instrument qualification, specificity, calibration model, accuracy, precision (without brain matrix and with brain matrix), and extraction efficiency were performed. The linear ranges of the calibration curves were 2-1000 pg for PGs, DiHETrEs, HETEs, and EETs, 10-2400 pg for PGE 2 and PGD 2 , and 20-2000 ng for AA, respectively.

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Section: Currently Gc/ms Gc/ms/msmentioning

confidence: 99%

A liquid chromatography/mass spectrometric method for simultaneous analysis of arachidonic acid and its endogenous eicosanoid metabolites prostaglandins, dihydroxyeicosatrienoic acids, hydroxyeicosatetraenoic acids, and epoxyeicosatrienoic acids in rat brain tissue

Yue

Jansen

Strauss

et al. 2007

Journal of Pharmaceutical and Biomedical Analysis

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“…In summary, the software processes raw data and tabulates an AA/AA-d 8 doublet "hit" list according to a set of criteria as follows. Minimum intensity values are set for (i) the (M-H) Ϫ peak, (ii) the max([M-H]ϩd 5,6,7,8 ) Ϫ peak, (iii) the ratio of max([M-H]ϩd 5,6,7,8 ) Ϫ peak to the corresponding non-deuterated control peak. In the example shown in Fig.…”

Section: Aa-dmentioning

confidence: 99%

Arachidonate-derived Dihomoprostaglandin Production Observed in Endotoxin-stimulated Macrophage-like Cells

Harkewicz

Fahy

Andreyev

et al. 2007

Journal of Biological Chemistry

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Eicosanoids, including the prostaglandins, leukotrienes, hydroxyeicosatetraenoic acids, epoxyeicosatetraenoic acids, and related compounds, are biosynthetic, bioactive mediators derived from arachidonic acid (AA), a 20:4(n-6) fatty acid. We have developed a comprehensive and sensitive mass spectral analysis to survey eicosanoid release from endotoxin-stimulated RAW 264.7 macrophage-like cells that is capable of detecting over 70 diverse eicosanoids and eicosanoid metabolites, should they be present. We now address the question: Are biologically significant eicosanoids being overlooked? Herein, we illustrate a general approach to diverse isotope metabolic profiling of labeled exogenous substrates using mass spectrometry (DIM-PLES/MS), demonstrated for one substrate (AA) and its resultant products (eicosanoids). RAW cells were incubated in medium supplemented with deuterium-labeled AA. When the cells are stimulated, two sets of eicosanoids are produced, one from endogenous AA and the other from the supplemented (exogenous) deuterium-labeled form. This produces a signature mass spectral "doublet" pattern, allowing for a comprehensive and diverse eicosanoid search requiring no previous knowledge or assumptions as to what these species may be, in contrast to traditional methods. We report herein observing unexpected AA metabolites generated by the cells, some of which may constitute novel bioactive eicosanoids or eicosanoid inactivation metabolites, as well as demonstrating differing metabolic pathways for the generation of isomeric prostaglandins and potential peroxisome proliferator-activated receptor activators. Unexpectedly, we report observing a series of 1a,1b-dihomologue prostaglandins, products of adrenic acid (22:4(n-6)), resulting from the two-carbon elongation of AA by the RAW cells.Starting in the early 1960s with the first structural characterization of the prostaglandins (1), mass spectrometry (MS) 2 has played a critical role in the biochemical study of eicosanoids. More recent advances in electrospray ionization-MS coupled to high-performance liquid chromatography have offered extremely sensitive and quantitative assays for most of the eicosanoids (2), without the need for chemical derivatization prior to analysis as was required by earlier gas chromatography electron ionization-MS methods (3). It is important to recognize, however, that while advances in high-performance LC-MS methods have offered extensive capabilities for surveying a number of different eicosanoid species in a single analysis, to date most efforts have focused on a specific eicosanoid or eicosanoid class, and additionally, with advanced knowledge and assumptions as to the identity of these species (4 -11). Recently, however, investigators have begun to explore the development of theoretical databases and algorithms based on virtual liquid chromatography-UV spectroscopy-tandem mass spectrometry (MS/MS) spectra and chromatograms for identifying potential lipid mediators without synthetic or authentic products as standards (12).System...

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“…To overcome the limitations of GC-MS and immunoassays for prostaglandin measurements, LC-MS [15][16][17] and LC-MS-MS [18][19][20][21][22][23][24][25][26][27] based methods have evolved as powerful tools for measuring prostaglandins in biological samples because of their high sensitivity, high selectivity and simplicity of sample preparation. Although five of these LC-MS or LC-MS-MS methods have measured PGD 2 [15][16][17][18][19], none have controlled for the inherent chemical instability of PGD 2 which is critical for determining accurate levels and for comparison with the values of other, more stable eicosanoids.…”

Section: Introductionmentioning

confidence: 99%

An improved LC–MS/MS method for the quantification of prostaglandins E2 and D2 production in biological fluids

Cao

Xiao

Park

et al. 2008

Analytical Biochemistry

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We report an improved LC-MS-MS assay that accurately measures prostaglandins D 2 (PGD 2 ) and E 2 (PGE 2 ) in cell culture supernatants and other biological fluids. The limit of detection for each prostaglandin was 20 pg/mL (0.20 pg; 0.55 fmol on-column), and the inter-day and intra-day coefficients of variation were less than 5%. Both d 4 -PGE 2 and d 4 -PGD 2 were used as surrogate standards to control for differential loss and degradation of the analytes. Stability studies indicated that sample preparation time should be less than 8 h to measure PGD 2 accurately, whereas preparation time did not affect PGE 2 measurement due to its greater stability in biological samples. As an application of the method, PGD 2 and PGE 2 were measured in culture supernatants from A549 cells and RAW 264.7 cells. The human lung alveolar cell line A549 was found to produce PGE 2 but no PGD 2 while the murine macrophage cell line RAW 264.7 produced PGD 2 and only trace amounts of PGE 2 . This direct comparison showed that COX-2 gene expression can lead to differential production of PGD 2 and PGE 2 by epithelial cells and macrophages. Since PGE 2 is anti-asthmatic and PGD 2 is pro-asthmatic, we speculate that the balance of production of these eicosanoids by epithelial cells and macropahges in the lung contributes to the pathogenesis of COPD, bronchiectasis, asthma, and lung cancer.

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Simultaneous quantification of prostaglandins in human synovial cell-cultured medium using liquid chromatography/tandem mass spectrometry

Cited by 53 publications

References 12 publications

A liquid chromatography/mass spectrometric method for simultaneous analysis of arachidonic acid and its endogenous eicosanoid metabolites prostaglandins, dihydroxyeicosatrienoic acids, hydroxyeicosatetraenoic acids, and epoxyeicosatrienoic acids in rat brain tissue

A liquid chromatography/mass spectrometric method for simultaneous analysis of arachidonic acid and its endogenous eicosanoid metabolites prostaglandins, dihydroxyeicosatrienoic acids, hydroxyeicosatetraenoic acids, and epoxyeicosatrienoic acids in rat brain tissue

Arachidonate-derived Dihomoprostaglandin Production Observed in Endotoxin-stimulated Macrophage-like Cells

An improved LC–MS/MS method for the quantification of prostaglandins E2 and D2 production in biological fluids

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