Preparation of biological fluids for simultaneous analysis of prostaglandin cyclo-oxygenase synthesized compounds by gas chromatography with electron capture detection

Charles Leffler, Dominic M. Desiderio, Claire E. Wakelyn

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

A method for isolating climax products of the arachidonic acid cascade from biological fluids is described which allows simultaneous measurement of PGs (PGE2, PGD2, 6-keto-PGF1α, TXB2, 6-keto-PGE1, 6, 15-diketo-13,14-dihydro-PGF1α) by electron capture detection of pentafluorobenzyloxime methyl ester trimethylsilyl (TMS) ether derivatives. PGs are adsorbed onto Amberlite XAD-2 from acidified solution and nonadhering substances removed by sequential administration of water, then petroleum ether. PGs are extracted into methanol. Following evaporation and reconstitution in water, the PGs are extracted into ethyl acetate at pH 3 and the ethyl acetate extracts are purified by lipidex column chromatography. Derivatization to pentafluorobenzyloxime methyl ester TMS ethers of PGs in the sample is followed by separation on either glass packed-columns or SCOT capillary columns, and detection by an electron capture detector. PGA2, added to the unpurified sample, is used as an internal standard for quantification. The methods have performed well on all biological fluids thus far examined. Examples of chromatographs from urine, Krebs-perfused lung effluents, and blood are shown.

Original languageEnglish (US)
Pages (from-to)227-241
Number of pages15
JournalProstaglandins
Volume21
Issue number2
DOIs
StatePublished - Jan 1 1981

Fingerprint

Prostaglandin-Endoperoxide Synthases
Gas chromatography
Gas Chromatography
Esters
Electrons
Prostaglandin D2
Column chromatography
Fluids
Ethers
Water
Cascades (fluid mechanics)
Dinoprostone
Arachidonic Acid
Ether
Glass
Methanol
Chromatography
Effluents
Evaporation
Blood

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Endocrinology

Cite this

Preparation of biological fluids for simultaneous analysis of prostaglandin cyclo-oxygenase synthesized compounds by gas chromatography with electron capture detection. / Leffler, Charles; Desiderio, Dominic M.; Wakelyn, Claire E.

In: Prostaglandins, Vol. 21, No. 2, 01.01.1981, p. 227-241.

Research output: Contribution to journalArticle

@article{afb195b6c08c4d1b8c42c12a208b5525,
title = "Preparation of biological fluids for simultaneous analysis of prostaglandin cyclo-oxygenase synthesized compounds by gas chromatography with electron capture detection",
abstract = "A method for isolating climax products of the arachidonic acid cascade from biological fluids is described which allows simultaneous measurement of PGs (PGE2, PGD2, 6-keto-PGF1α, TXB2, 6-keto-PGE1, 6, 15-diketo-13,14-dihydro-PGF1α) by electron capture detection of pentafluorobenzyloxime methyl ester trimethylsilyl (TMS) ether derivatives. PGs are adsorbed onto Amberlite XAD-2 from acidified solution and nonadhering substances removed by sequential administration of water, then petroleum ether. PGs are extracted into methanol. Following evaporation and reconstitution in water, the PGs are extracted into ethyl acetate at pH 3 and the ethyl acetate extracts are purified by lipidex column chromatography. Derivatization to pentafluorobenzyloxime methyl ester TMS ethers of PGs in the sample is followed by separation on either glass packed-columns or SCOT capillary columns, and detection by an electron capture detector. PGA2, added to the unpurified sample, is used as an internal standard for quantification. The methods have performed well on all biological fluids thus far examined. Examples of chromatographs from urine, Krebs-perfused lung effluents, and blood are shown.",
author = "Charles Leffler and Desiderio, {Dominic M.} and Wakelyn, {Claire E.}",
year = "1981",
month = "1",
day = "1",
doi = "10.1016/0090-6980(81)90140-4",
language = "English (US)",
volume = "21",
pages = "227--241",
journal = "Prostaglandins",
issn = "0090-6980",
publisher = "Elsevier BV",
number = "2",

}

TY - JOUR

T1 - Preparation of biological fluids for simultaneous analysis of prostaglandin cyclo-oxygenase synthesized compounds by gas chromatography with electron capture detection

AU - Leffler, Charles

AU - Desiderio, Dominic M.

AU - Wakelyn, Claire E.

PY - 1981/1/1

Y1 - 1981/1/1

N2 - A method for isolating climax products of the arachidonic acid cascade from biological fluids is described which allows simultaneous measurement of PGs (PGE2, PGD2, 6-keto-PGF1α, TXB2, 6-keto-PGE1, 6, 15-diketo-13,14-dihydro-PGF1α) by electron capture detection of pentafluorobenzyloxime methyl ester trimethylsilyl (TMS) ether derivatives. PGs are adsorbed onto Amberlite XAD-2 from acidified solution and nonadhering substances removed by sequential administration of water, then petroleum ether. PGs are extracted into methanol. Following evaporation and reconstitution in water, the PGs are extracted into ethyl acetate at pH 3 and the ethyl acetate extracts are purified by lipidex column chromatography. Derivatization to pentafluorobenzyloxime methyl ester TMS ethers of PGs in the sample is followed by separation on either glass packed-columns or SCOT capillary columns, and detection by an electron capture detector. PGA2, added to the unpurified sample, is used as an internal standard for quantification. The methods have performed well on all biological fluids thus far examined. Examples of chromatographs from urine, Krebs-perfused lung effluents, and blood are shown.

AB - A method for isolating climax products of the arachidonic acid cascade from biological fluids is described which allows simultaneous measurement of PGs (PGE2, PGD2, 6-keto-PGF1α, TXB2, 6-keto-PGE1, 6, 15-diketo-13,14-dihydro-PGF1α) by electron capture detection of pentafluorobenzyloxime methyl ester trimethylsilyl (TMS) ether derivatives. PGs are adsorbed onto Amberlite XAD-2 from acidified solution and nonadhering substances removed by sequential administration of water, then petroleum ether. PGs are extracted into methanol. Following evaporation and reconstitution in water, the PGs are extracted into ethyl acetate at pH 3 and the ethyl acetate extracts are purified by lipidex column chromatography. Derivatization to pentafluorobenzyloxime methyl ester TMS ethers of PGs in the sample is followed by separation on either glass packed-columns or SCOT capillary columns, and detection by an electron capture detector. PGA2, added to the unpurified sample, is used as an internal standard for quantification. The methods have performed well on all biological fluids thus far examined. Examples of chromatographs from urine, Krebs-perfused lung effluents, and blood are shown.

UR - http://www.scopus.com/inward/record.url?scp=0019434703&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0019434703&partnerID=8YFLogxK

U2 - 10.1016/0090-6980(81)90140-4

DO - 10.1016/0090-6980(81)90140-4

M3 - Article

VL - 21

SP - 227

EP - 241

JO - Prostaglandins

JF - Prostaglandins

SN - 0090-6980

IS - 2

ER -