Deeper insights into the stability of oxylipins in human plasma across multiple freeze-thaw cycles and storage conditions

IF 3.1 3区 医学 Q2 CHEMISTRY, ANALYTICAL Journal of pharmaceutical and biomedical analysis Pub Date : 2024-11-29 DOI:10.1016/j.jpba.2024.116587
Maria Moran-Garrido , Sandra M. Camunas-Alberca , Jorge Sáiz , Ana Gradillas , Ameer Y. Taha , Coral Barbas
{"title":"Deeper insights into the stability of oxylipins in human plasma across multiple freeze-thaw cycles and storage conditions","authors":"Maria Moran-Garrido ,&nbsp;Sandra M. Camunas-Alberca ,&nbsp;Jorge Sáiz ,&nbsp;Ana Gradillas ,&nbsp;Ameer Y. Taha ,&nbsp;Coral Barbas","doi":"10.1016/j.jpba.2024.116587","DOIUrl":null,"url":null,"abstract":"<div><div>Oxylipins are signaling lipids derived from the oxidation of polyunsaturated fatty acids (PUFAs). In lipidomic studies, human plasma may be subjected to various storage conditions and freeze-thaw cycles, which may impact the analysis of these compounds. In this study, we used liquid chromatography coupled with mass spectrometry (LC-MS) to examine the influence of up to five freeze-thaw cycles (FTCs) on free and total (mostly esterified) oxylipins in human plasma and the influence of temperature and storage duration (4 °C for up to 120 h and –20 °C and –80 °C for 1–98 days) in the presence or absence of butylated hydroxytoluene (BHT) on extracted oxylipins stored in LC-MS amber vials. In fresh plasma subjected to several FTCs, approximately 48 % of the detected free oxylipins were significantly altered by the third cycle, with increases in cytochrome P450 (CYP450) and lipoxygenase (LOX)-derived compounds and reductions in trihydroxylated oxylipins. In contrast, multiple FTCs did not significantly alter esterified oxylipins. At 4 °C, the extracted oxylipins did not change significantly for up to 120 h (5 days). Oxylipin levels remained stable for 98 days at –80 °C but decreased by 98 days at –20 °C. The antioxidant activity of butylated hydroxytoluene (BHT) did not influence oxylipin stability at 4 °C for 120 h or at –80 °C for 98 days, but it reduced oxylipin degradation at –20 °C at 98 days. Conversely, prostaglandin F<sub>2α</sub> (PGF<sub>2α</sub>) exhibited substantial increases at –20 °C and –80 °C, independent of BHT. This study demonstrates that (i) unlike free oxylipins, the esterified oxylipin pool remains stable following repeated FTCs, (ii) extracted oxylipins are stable at 4 °C for up to 120 h and at –80 °C for up to 98 days, but not at –20 °C for 98 days, and (iii) BHT may minimize oxylipin degradation of sample extracts stored at –20 °C. This study provides a framework for measuring oxylipins under various freeze-thaw and storage conditions.</div></div>","PeriodicalId":16685,"journal":{"name":"Journal of pharmaceutical and biomedical analysis","volume":"255 ","pages":"Article 116587"},"PeriodicalIF":3.1000,"publicationDate":"2024-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of pharmaceutical and biomedical analysis","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0731708524006290","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0

Abstract

Oxylipins are signaling lipids derived from the oxidation of polyunsaturated fatty acids (PUFAs). In lipidomic studies, human plasma may be subjected to various storage conditions and freeze-thaw cycles, which may impact the analysis of these compounds. In this study, we used liquid chromatography coupled with mass spectrometry (LC-MS) to examine the influence of up to five freeze-thaw cycles (FTCs) on free and total (mostly esterified) oxylipins in human plasma and the influence of temperature and storage duration (4 °C for up to 120 h and –20 °C and –80 °C for 1–98 days) in the presence or absence of butylated hydroxytoluene (BHT) on extracted oxylipins stored in LC-MS amber vials. In fresh plasma subjected to several FTCs, approximately 48 % of the detected free oxylipins were significantly altered by the third cycle, with increases in cytochrome P450 (CYP450) and lipoxygenase (LOX)-derived compounds and reductions in trihydroxylated oxylipins. In contrast, multiple FTCs did not significantly alter esterified oxylipins. At 4 °C, the extracted oxylipins did not change significantly for up to 120 h (5 days). Oxylipin levels remained stable for 98 days at –80 °C but decreased by 98 days at –20 °C. The antioxidant activity of butylated hydroxytoluene (BHT) did not influence oxylipin stability at 4 °C for 120 h or at –80 °C for 98 days, but it reduced oxylipin degradation at –20 °C at 98 days. Conversely, prostaglandin F (PGF) exhibited substantial increases at –20 °C and –80 °C, independent of BHT. This study demonstrates that (i) unlike free oxylipins, the esterified oxylipin pool remains stable following repeated FTCs, (ii) extracted oxylipins are stable at 4 °C for up to 120 h and at –80 °C for up to 98 days, but not at –20 °C for 98 days, and (iii) BHT may minimize oxylipin degradation of sample extracts stored at –20 °C. This study provides a framework for measuring oxylipins under various freeze-thaw and storage conditions.
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
深入了解人体血浆中多种冻融循环和储存条件下氧磷脂的稳定性。
氧脂素是由多不饱和脂肪酸(PUFAs)氧化产生的信号脂质。在脂质组学研究中,人血浆可能受到不同的储存条件和冻融循环,这可能会影响这些化合物的分析。在这项研究中,我们使用液相色谱-质谱联用(LC-MS)检测了多达五次冻融循环(FTCs)对人血浆中游离和总(主要是酯化)氧脂的影响,以及温度和储存时间(4°C保存120 h, -20°C和-80°C保存1-98 天)在存在或不存在丁基羟基甲苯(BHT)的情况下对提取的氧脂储存在LC-MS琥珀瓶中的影响。在新鲜血浆中,大约48% %检测到的游离氧脂素在第三个周期中显著改变,细胞色素P450 (CYP450)和脂氧合酶(LOX)衍生化合物增加,三羟基化氧脂素减少。相反,多个FTCs没有显著改变酯化的氧化脂。在4°C下,提取的氧磷脂在120 h(5天)内没有显著变化。在-80°C下,氧脂素水平保持稳定98天,而在-20°C下,氧脂素水平下降98天。丁基羟基甲苯(BHT)的抗氧化活性在4℃下持续120 h或在-80℃下持续98 d不影响氧化脂质的稳定性,但在-20℃下持续98 d降低了氧化脂质的降解。相反,前列腺素F2α (PGF2α)在-20°C和-80°C时显著升高,与BHT无关。本研究表明:(i)与游离的氧脂不同,酯化的氧脂池在重复FTCs后保持稳定;(ii)提取的氧脂在4°C下可稳定至120 h,在-80°C下可稳定至98天,但在-20°C下不能稳定至98天;(iii) BHT可以最大限度地减少在-20°C下储存的样品提取物的氧脂降解。本研究为测定不同冻融和储存条件下的氧化脂质提供了一个框架。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
CiteScore
6.70
自引率
5.90%
发文量
588
审稿时长
37 days
期刊介绍: This journal is an international medium directed towards the needs of academic, clinical, government and industrial analysis by publishing original research reports and critical reviews on pharmaceutical and biomedical analysis. It covers the interdisciplinary aspects of analysis in the pharmaceutical, biomedical and clinical sciences, including developments in analytical methodology, instrumentation, computation and interpretation. Submissions on novel applications focusing on drug purity and stability studies, pharmacokinetics, therapeutic monitoring, metabolic profiling; drug-related aspects of analytical biochemistry and forensic toxicology; quality assurance in the pharmaceutical industry are also welcome. Studies from areas of well established and poorly selective methods, such as UV-VIS spectrophotometry (including derivative and multi-wavelength measurements), basic electroanalytical (potentiometric, polarographic and voltammetric) methods, fluorimetry, flow-injection analysis, etc. are accepted for publication in exceptional cases only, if a unique and substantial advantage over presently known systems is demonstrated. The same applies to the assay of simple drug formulations by any kind of methods and the determination of drugs in biological samples based merely on spiked samples. Drug purity/stability studies should contain information on the structure elucidation of the impurities/degradants.
期刊最新文献
Editorial Board The cutting edge of surveillance: Exploring high-resolution mass spectrometry in wastewater-based epidemiology for monitoring forensic samples Impact of ultralow oxygen preservation on the quality of Atractylodis Macrocephalae Rhizoma evaluated by multi-chromatographic determination of small molecules and polysaccharides GC and HPLC-RID method development and validation for the determination of twelve related substances, including several novel compounds in tetrabenazine Utilizing copper-doped graphene quantum dots as a fluorescent sensor for determination of carbamazepine in exhaled breath condensate
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1