TY - JOUR
T1 - [67] Quantitation of 15-Keto-13,14-dihydro-PGE2 in Plasma by GC-MS
AU - Hubbard, Walter C.
N1 - Funding Information:
This work was supported by Grant GM 15431 from the National Institutes of Health. B. Samuelsson, E. Granstr0m, K. Green, M. Hamberg, and S. Hammarstr0m, Annu. Rev. Biochem. 44, 695 (1975). 2 B. Samuelsson, M. Goldyne, E. GranstrOm, M. Hamberg, S. HammarstrOm, and C. Malmsten, Annu. Rev. Biochem. 47, 997 (1978). a E. Granstr0m and B. Samuelsson, Adv. Prostaglandin Thromboxane Res. 5, 1 (1978). 4 M. D. Mitchell, H. Sors, and A. P. F. Flint, Lancet 2, 558, (1977). 5 E. GranstrOm and H. Kindahl, Adv. Prostaglandin Thromboxane Res. 6, 181 (1980). 6 E. Granstr0m, M. Hamberg, G. Hanson, and H. Kindahl, Prostaglandins 19, 933 (1980). 7 F. A. Fitzpatrick, R. Aguire, J. E. Pike, and F. H. Lincoln, Prostaglandins 19, 917 (1980). s B. Samuelsson and K. Green, Biochem. Med. 11, 298 (1974). a W. C. Hubbard and J. T. Watson, Prostaglandins 12, 21 (1976). ~0 L. Levine, Prostaglandins 14, 1125 (1977). ~t D. Gordon, L. Myatt, A. Gordon-Wright, J. Hanson, and M. G. Elder, Prostaglandins 13, 399 (1977). ~2 A. H. Tashjian, Jr., E. F. Voelkel, and L. Levine, Biochem. Biophys. Res. Commun. 74, 199 (1977). ~ A. H. Tashjian, Jr., E. F. Voelkel, and L. Levine, Prostaglandins 14, 309 (1977). 14 H. W. Seyberth, W. C. Hubbard, O. Oelz, B. J. Sweetman, J. T. Watson, and J. A. Oates, Prostaglandins 14, 319 (1977). ~5 L. J. Roberts, II, W. C. Hubbard, Z. T. Bloomgarden, X. Y. Bertagna, T. J. McKenna, D. Rabinowitz, and J. A. Oates, Trans. Assoc. Am. Physicians 92, 286 (1979). 16 W. C. Hubbard, A. J. Hough, R. M. Johnson, and J. A. Oates, Prostaglandins 19, 881
PY - 1982/1/1
Y1 - 1982/1/1
N2 - Measurement of the 15-keto-13,14-dihydro metabolite of prostaglandin E2 (PGE2) in plasma is a more reliable index of the entry of PGE2 into the circulation than direct quantitation of PGE2. This chapter presents the procedures for preparation of 3,3,4,4-tetradeutero- 15K-H2-PGE2 for use as an internal standard for gas chromatography–mass spectrometry (GC-MS) assay of the PGE2 metabolite, for minimization of loss of endogenous 15K-H2-PGE2 in blood samples prior to addition of the tetradeuterated internal standard, and for stabilization of 15K-H2-PGE2 via derivatization for prevention of additional loss during purification. 3,3,4,4-Tetradeutero-15K-H2-PGE2 and 5,6,8,11,12,14-hexatritiated- 15K-H2-PGE2 ([3H]15K-H2-PGE2) are simultaneously prepared from 3,3, 4,4-tetradeutero-PGE2 and 5,6,8,1 l, 12,14,15-heptatritiated-PGE2 ([3H]PGE2), respectively, via incubation with the 100,000 g subcellular fraction of either swine kidney or guinea pig liver. In partial derivatization, the plasma extract containing 15K-H2-PGE2 is oximized and esterified. Quantitation of 15K-H2-PGE2 in plasma employing GC-MS is performed by simultaneously monitoring characteristic fragment ions of the bismethyloxime-methyl ester-trimethylsilyl ether derivative of 15K-H2- PGE2 and of 3,3,4,4-tetradeutero-15K-H2-PGE2.
AB - Measurement of the 15-keto-13,14-dihydro metabolite of prostaglandin E2 (PGE2) in plasma is a more reliable index of the entry of PGE2 into the circulation than direct quantitation of PGE2. This chapter presents the procedures for preparation of 3,3,4,4-tetradeutero- 15K-H2-PGE2 for use as an internal standard for gas chromatography–mass spectrometry (GC-MS) assay of the PGE2 metabolite, for minimization of loss of endogenous 15K-H2-PGE2 in blood samples prior to addition of the tetradeuterated internal standard, and for stabilization of 15K-H2-PGE2 via derivatization for prevention of additional loss during purification. 3,3,4,4-Tetradeutero-15K-H2-PGE2 and 5,6,8,11,12,14-hexatritiated- 15K-H2-PGE2 ([3H]15K-H2-PGE2) are simultaneously prepared from 3,3, 4,4-tetradeutero-PGE2 and 5,6,8,1 l, 12,14,15-heptatritiated-PGE2 ([3H]PGE2), respectively, via incubation with the 100,000 g subcellular fraction of either swine kidney or guinea pig liver. In partial derivatization, the plasma extract containing 15K-H2-PGE2 is oximized and esterified. Quantitation of 15K-H2-PGE2 in plasma employing GC-MS is performed by simultaneously monitoring characteristic fragment ions of the bismethyloxime-methyl ester-trimethylsilyl ether derivative of 15K-H2- PGE2 and of 3,3,4,4-tetradeutero-15K-H2-PGE2.
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U2 - 10.1016/0076-6879(82)86227-7
DO - 10.1016/0076-6879(82)86227-7
M3 - Article
C2 - 7132770
AN - SCOPUS:0020021866
SN - 0076-6879
VL - 86
SP - 571
EP - 578
JO - Methods in enzymology
JF - Methods in enzymology
IS - C
ER -