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Titolo:
Hydrolysis of conjugated steroids by the combined use of beta-glucuronidase preparations from Helix pomatia and Ampullaria: Determination of urinary cortisol and its metabolites
Autore:
Shibasaki, H; Tanabe, C; Furuta, T; Kasuya, Y;
Indirizzi:
Tokyo Univ Pharm & Life Sci, Sch Pharm, Dept Med Chem & Clin Pharm, Hachioji, Tokyo 1920392, Japan Tokyo Univ Pharm & Life Sci Hachioji Tokyo Japan 1920392 o 1920392, Japan
Titolo Testata:
STEROIDS
fascicolo: 11, volume: 66, anno: 2001,
pagine: 795 - 801
SICI:
0039-128X(200111)66:11<795:HOCSBT>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
APPARENT MINERALOCORTICOID EXCESS; DILUTION MASS-SPECTROMETRY; 11-BETA-HYDROXYSTEROID DEHYDROGENASE; HUMAN PLASMA; IN-VIVO; TETRAHYDROCORTISONE;
Keywords:
cortisol; cortisol metabolites; conjugated steroids; hydrolysis; beta-glucuronidase;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
23
Recensione:
Indirizzi per estratti:
Indirizzo: Kasuya, Y Tokyo Univ Pharm & Life Sci, Sch Pharm, Dept Med Chem & Clin Pharm, 1432-1Horinouchi, Hachioji, Tokyo 1920392, Japan Tokyo Univ Pharm & Life Sci 1432-1 Horinouchi Hachioji Tokyo Japan 1920392
Citazione:
H. Shibasaki et al., "Hydrolysis of conjugated steroids by the combined use of beta-glucuronidase preparations from Helix pomatia and Ampullaria: Determination of urinary cortisol and its metabolites", STEROIDS, 66(11), 2001, pp. 795-801

Abstract

This study describes the enzymatic hydrolysis of urinary conjugates of cortisol, cortisone, tetrahydrocortisol, allotetrahydrocortisol, and tetrahydrocortisone with beta -glucuronidase preparations from Helix pomatia and Ampullaria. The objective of the present studies was to find optimal hydrolysis conditions for these conjugated steroids. Assay of the isolated steroids was carried out by GC-MS using deuterium-labeled compounds as internal standards. The allotetrahydrocortisol conjugate was clearly the hardest to hydrolyze with enzyme from Helix pomatia and required increased enzyme concentration and prolonged incubation. Hydrolysis of a urine sample for 2.0 h withthe simultaneous use of 3400 units/ml Ampullaria and 5400 units/ml Helix pomatia enzymes in 0.5 M acetate buffer at 55 degreesC achieved more complete cleavage of the urinary conjugates of the five steroids examined. It is thus advantageous to use the Ampullaria and Helix pomatia enzymes in combination to obtain the highest yield in the urinary corticosteroid assay. (C) 2001 Elsevier Science Inc. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/03/20 alle ore 20:06:25