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AuthorPetrucci, Giovanna
AuthorHatem, Duaa
AuthorLangley, Ruth
AuthorCleary, Siobhan
AuthorGentry-Maharaj, Aleksandra
AuthorPitocco, Dario
AuthorRizzi, Alessandro
AuthorRanalli, Paola
AuthorZaccardi, Francesco
AuthorHabib, Aida
AuthorRocca, Bianca
Available date2024-04-29T10:36:39Z
Publication Date2024-03-06
Publication NameScientific Reports
Identifierhttp://dx.doi.org/10.1038/s41598-024-55720-3
CitationPetrucci, G., Hatem, D., Langley, R., Cleary, S., Gentry-Maharaj, A., Pitocco, D., ... & Rocca, B. (2024). Effect of very long-term storage and multiple freeze and thaw cycles on 11-dehydro-thromboxane-B2 and 8-iso-prostaglandin F2α, levels in human urine samples by validated enzyme immunoassays. Scientific Reports, 14(1), 5546.
ISSN2045-2322
URIhttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85187189714&origin=inward
URIhttp://hdl.handle.net/10576/54451
AbstractBiological samples are often frozen and stored for years and/or thawed multiple times, thus assessing their stability on long-term storage and repeated freeze–thaw cycles is crucial. The study aims were to assess:—the long-term stability of two major enzymatic and non-enzymatic metabolites of arachidonic acid, i.e. urinary 11-dehydro-thromboxane-(Tx) B2, 8-iso-prostaglandin (PG)F2α, and creatinine in frozen urine samples;—the effect of multiple freeze–thaw cycles. Seven-hundred and three urine samples measured in previously-published studies, stored at −40 °C, and measured for a second time for 11-dehydro-TxB2 (n = 677) and/or 8-iso-PGF2α (n = 114) and/or creatinine (n = 610) were stable over 10 years and the 2 measurements were highly correlated (all rho = 0.99, P < 0.0001). Urine samples underwent 10 sequential freeze–thaw cycles, with and without the antioxidant 4-hydroxy-2,2,6,6-tetramethylpiperidin-1-oxyl (10 mM); urinary 11-dehydro-TxB2 and creatinine were stable across all cycles (11-dehydro-TxB2: 100.4 ± 21%; creatinine: 101 ± 7% of baseline at cycle ten; n = 17), while 8-iso-PGF2α significantly increased by cycle 6 (151 ± 22% of baseline at cycle ten, n = 17, P < 0.05) together with hydrogen peroxide only in the absence of antioxidant. Arachidonic acid metabolites and creatinine appear stable in human urines stored at −40 °C over 10 years. Multiple freeze–thaw cycles increase urinary 8-iso-PGF2α in urine samples without antioxidants. These data are relevant for studies using urine samples stored over long-term and/or undergoing multiple freezing–thawing.
SponsorThis work was supported by the AsCaP collaboration (C569/A24991#2) and in part by the institutional Linea D1 2021 and Linea D1 2022 Grants to GP and BR.
Languageen
PublisherSpringer Nature
Subjectenzyme immunoassay
antioxidant
TitleEffect of very long-term storage and multiple freeze and thaw cycles on 11-dehydro-thromboxane-B(2) and 8-iso-prostaglandin F(2α,) levels in human urine samples by validated enzyme immunoassays
TypeArticle
Issue Number1
Volume Number14
dc.accessType Open Access


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