000076056 001__ 76056
000076056 005__ 20240118091946.0
000076056 0247_ $$2doi$$a10.1016/j.jpba.2017.12.033
000076056 0248_ $$2sideral$$a104148
000076056 037__ $$aART-2018-104148
000076056 041__ $$aeng
000076056 100__ $$0(orcid)0000-0001-8830-6554$$aMendiara, Isabel$$uUniversidad de Zaragoza
000076056 245__ $$aDetermination of total plasma oxysterols by enzymatic hydrolysis, solid phase extraction and liquid chromatography coupled to mass-spectrometry
000076056 260__ $$c2018
000076056 5060_ $$aAccess copy available to the general public$$fUnrestricted
000076056 5203_ $$aThe potential use of cholesterol esterases was tested to avoid alkaline hydrolysis for cleavage of plasma esterified oxysterols. The enzymatic hydrolysis was optimized by testing two sources of enzyme—Pseudomonas and bovine pancreas, presence of surfactants, incubation time and amount of enzyme. Free forms of 4ß-, 7-, 24-, 25- and 27-hydroxycholesterol (HC) as well 7-ketocholesterol (7-KC) were analyzed by liquid chromatography and mass-spectrometry using the deuterated internal standard, 25-HC(d6). Enzymatic hydrolysis was more effective using the Pseudomonas enzyme and in presence of surfactants. Compared to alkaline hydrolysis, it generated a cleaner chromatographic baseline and better recovery of the internal standard. Oxysterols were assayed with detection limits between 7 and 31 pg/mL. Interassay coefficients of variation were lower than 10% and extraction recovery efficiencies, higher than 90%. The procedure was used to characterize plasma levels of Cyp7b1-deficient rat, where it showed increased plasma levels of 7, 24 and 25-HC. Due to the low volume of sample required, it may be used in other animal models, particularly rodents, as well as in pediatric samples where sample amount is always a problem. Thus, the proposed new method offers mild enzymatic processing that greatly facilitates oxysterol determinations to delineate their role in physiopathology.
000076056 536__ $$9info:eu-repo/grantAgreement/ES/DGA/T10$$9info:eu-repo/grantAgreement/ES/ISCIII/CB06-03-1012$$9info:eu-repo/grantAgreement/ES/MINECO/SAF2013-41651-R$$9info:eu-repo/grantAgreement/ES/MINECO/SAF2016-75441-R
000076056 540__ $$9info:eu-repo/semantics/openAccess$$aby-nc-nd$$uhttp://creativecommons.org/licenses/by-nc-nd/3.0/es/
000076056 590__ $$a2.983$$b2018
000076056 591__ $$aPHARMACOLOGY & PHARMACY$$b105 / 266 = 0.395$$c2018$$dQ2$$eT2
000076056 591__ $$aCHEMISTRY, ANALYTICAL$$b24 / 84 = 0.286$$c2018$$dQ2$$eT1
000076056 592__ $$a0.786$$b2018
000076056 593__ $$aAnalytical Chemistry$$c2018$$dQ1
000076056 593__ $$aClinical Biochemistry$$c2018$$dQ1
000076056 593__ $$aSpectroscopy$$c2018$$dQ1
000076056 593__ $$aPharmaceutical Science$$c2018$$dQ1
000076056 593__ $$aDrug Discovery$$c2018$$dQ1
000076056 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/acceptedVersion
000076056 700__ $$0(orcid)0000-0001-8893-0963$$aDomeño, Celia$$uUniversidad de Zaragoza
000076056 700__ $$0(orcid)0000-0003-2685-5739$$aNerín, Cristina$$uUniversidad de Zaragoza
000076056 700__ $$aGeurts, Aron M.
000076056 700__ $$0(orcid)0000-0002-8251-8457$$aOsada, Jesús$$uUniversidad de Zaragoza
000076056 700__ $$0(orcid)0000-0002-8100-5596$$aMartínez-Beamonte, Roberto$$uUniversidad de Zaragoza
000076056 7102_ $$12009$$2750$$aUniversidad de Zaragoza$$bDpto. Química Analítica$$cÁrea Química Analítica
000076056 7102_ $$12008$$2640$$aUniversidad de Zaragoza$$bDpto. Produc.Animal Cienc.Ali.$$cÁrea Nutrición Bromatología
000076056 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000076056 7102_ $$12008$$2780$$aUniversidad de Zaragoza$$bDpto. Produc.Animal Cienc.Ali.$$cÁrea Tecnología de Alimentos
000076056 773__ $$g150 (2018), 396-405$$pJ. pharm. biomed. anal.$$tJournal of Pharmaceutical and Biomedical Analysis$$x0731-7085
000076056 8564_ $$s2208947$$uhttps://zaguan.unizar.es/record/76056/files/texto_completo.pdf$$yPostprint
000076056 8564_ $$s33133$$uhttps://zaguan.unizar.es/record/76056/files/texto_completo.jpg?subformat=icon$$xicon$$yPostprint
000076056 909CO $$ooai:zaguan.unizar.es:76056$$particulos$$pdriver
000076056 951__ $$a2024-01-18-09:10:30
000076056 980__ $$aARTICLE