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dc.contributor.authorHäkkinen, Merja R
dc.contributor.authorMurtola, Teemu
dc.contributor.authorVoutilainen, Raimo
dc.contributor.authorPoutanen, Matti
dc.contributor.authorLinnanen, Tero
dc.contributor.authorKoskivuori, Johanna
dc.contributor.authorLakka, Timo
dc.contributor.authorJääskeläinen, Jarmo
dc.contributor.authorAuriola, Seppo
dc.date.accessioned2019-01-18T13:41:36Z
dc.date.available2019-01-18T13:41:36Z
dc.date.issued2019
dc.identifier.urihttps://erepo.uef.fi/handle/123456789/7355
dc.description.abstractThis study describes a validated LC–MS/MS method for assaying 23 steroids within a single run from 150 μl of human plasma, serum or prostatic tissue homogenate. Isotope-labeled steroids were used as internal standards. Samples were extracted with toluene, and ketosteroids were derivatized with hydroxylamine prior to LC–MS/MS analysis. The steroids were separated on a C18 column and methanol was used as an organic solvent with the addition of 0.2 mM ammonium fluoride to improve underivatized estradiol (E2) ionization. Certified reference serums as well as plasma samples, and homogenates of prostate tissue were utilized in the method validation. The specificity of the method was inspected with a total of 27 steroids. The validation proved that the method was suitable for the quantitative analysis of a wide panel of androgens (testosterone, T (3.3 pM-13 nM); androstenedione, A4 (3.3 pM-13 nM); 5α-androstanedione, DHA4 (13 pM-13 nM); dehydroepiandrosterone, DHEA (67 pM-133 nM); dihydrotestosterone, DHT (33 pM-33 nM); 11-ketodihydrotestosterone, 11KDHT (13 pM-13nM); 11-ketotestosterone, 11KT (33 pM-6.7 nM); 11β-hydroxyandrostenedione, 11bOHA4 (33 pM-13 nM); 11β-hydroxytestosterone, 11OHT (13 pM-33 nM)), as well as estrogens (estrone, E1 (3.3 pM-13 nM)), progestagens (17α-hydroxypregnenolone, 17OHP5 (32 pM-127 nM); 17α-hydroxyprogesterone, 17OHP4 (67 pM-133 nM); progesterone, P4 (3.3 pM-13 nM); pregnenolone, P5 (6.6 pM-13 nM)), and glucocorticoids (cortisol, F (33 pM-134 nM); cortisone E (66 pM-131 nM); corticosterone, B (33 pM-67 nM); 11-deoxycortisol, S (33 pM-66 nM); 21-hydroxyprogesterone, 21OHP4 (32 pM-13 nM)). Furthermore, E2 (335 pM-134 nM) and 11α-hydroxyandrostenedione, 11aOHA4 (33 pM-33 nM) could be analyzed if the concentration in the sample was high enough. In addition, aldosterone, A (128 pM-64 nM) and 11-ketoandrostenedione, 11KA4 (33 pM-13 nM) could be analyzed semiquantitatively. The limits of quantification for all compounds ranged from 0.9 to 91 pg/ml, and from 0.009 to 0.9 pg/mg tissue. Compared to our previous method, this new method also permits the analysis of the more challenging steroids, like DHT, DHEA and P5, and a panel of 11-ketosteroids.
dc.language.isoenglanti
dc.publisherElsevier BV
dc.relation.ispartofseriesJOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
dc.relation.urihttp://dx.doi.org/10.1016/j.jpba.2018.11.035
dc.rightsCC BY-NC-ND https://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectsteroids
dc.subjectandrogens
dc.subjecttissue
dc.subjectprostate
dc.subjectserum
dc.subjectLC–MS
dc.titleSimultaneous analysis by LC-MS/MS of 22 ketosteroids with hydroxylamine derivatization and underivatized estradiol from human plasma, serum and prostate tissue
dc.description.versionfinal draft
dc.contributor.departmentSchool of Pharmacy, Activities
dc.contributor.departmentSchool of Medicine / Biomedicine,School of Medicine / Clinical Medicine
uef.solecris.id58713079en
dc.type.publicationTieteelliset aikakauslehtiartikkelit
dc.rights.accessrights© Elsevier B.V.
dc.relation.doi10.1016/j.jpba.2018.11.035
dc.description.reviewstatuspeerReviewed
dc.format.pagerange642-652
dc.publisher.countryAlankomaat
dc.relation.issn0731-7085
dc.relation.issue164
dc.relation.volume164
dc.rights.accesslevelopenAccess
dc.type.okmA1
uef.solecris.openaccessEi


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