Determination of corticosterone from rat hair samples by HPLC-MS method

Ferencz Elek, Boda Ferenc, Gáll Zsolt, Kolcsár Melinda, Donáth-Nagy Gabriella, Vancea Szende
{"title":"Determination of corticosterone from rat hair samples by HPLC-MS method","authors":"Ferencz Elek, Boda Ferenc, Gáll Zsolt, Kolcsár Melinda, Donáth-Nagy Gabriella, Vancea Szende","doi":"10.2478/orvtudert-2019-0008","DOIUrl":null,"url":null,"abstract":"Abstract Corticosterone is an adrenocortical steroid hormone with glucocorticoid and mineralocorticoid effects. Based on previous studies, the plasma level of corticosterone correlates with the stress exposure of rodents. Because the half-life of corticosterone in blood is short, its plasma concentration can be used as an acute stress marker. But hair is accumulating the systemic and locally produced corticosterone, therefore it can be used to study chronic stress. However, the accurate quantification of corticosterone is an analytical challenge owing to the very low amount of hormone found in a complicated biological matrix. The high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) can provide the required selectivity and sensitivity for this purpose. Currently published methods for corticosterone quantification involve complicated sample preparation and long run time. Accordingly, the aims of the study were to simplify the extraction method of the corticosterone from rat hair samples and to develop an optimized HPLC-MS method for the accurate quantification. The rat hair samples were washed with methanol, dried and cut, then extracted with methanol at room temperature for 24 hours. The lipids were precipitated with formic acid aqueous solution and eliminated by centrifugation. The corticosterone was separated from other compounds with reverse phase chromatography using acetonitrile and 0,1% aqueous solution of formic acid as mobile phase. The detection was performed in positive SIM mode measuring the 347 m/z molecular ion. A six point calibration was performed in the range of 0,5-20 ng/ml, the accuracy was tested with quality control samples at two different concentration level. The total run time is only 4,2 minutes and the lower limit of quantification (LLOQ) is 0,5 ng/ml, with 10 pg absolute sensitivity. By determining the quantity of the hormone for a well-defined hair region, based on the speed of hair growth, we can characterize the retrospective stress exposure of the animals in different conditions.","PeriodicalId":9334,"journal":{"name":"Bulletin of Medical Sciences","volume":"19 1","pages":"27 - 34"},"PeriodicalIF":0.0000,"publicationDate":"2019-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bulletin of Medical Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.2478/orvtudert-2019-0008","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 1

Abstract

Abstract Corticosterone is an adrenocortical steroid hormone with glucocorticoid and mineralocorticoid effects. Based on previous studies, the plasma level of corticosterone correlates with the stress exposure of rodents. Because the half-life of corticosterone in blood is short, its plasma concentration can be used as an acute stress marker. But hair is accumulating the systemic and locally produced corticosterone, therefore it can be used to study chronic stress. However, the accurate quantification of corticosterone is an analytical challenge owing to the very low amount of hormone found in a complicated biological matrix. The high performance liquid chromatography coupled with mass spectrometry (HPLC-MS) can provide the required selectivity and sensitivity for this purpose. Currently published methods for corticosterone quantification involve complicated sample preparation and long run time. Accordingly, the aims of the study were to simplify the extraction method of the corticosterone from rat hair samples and to develop an optimized HPLC-MS method for the accurate quantification. The rat hair samples were washed with methanol, dried and cut, then extracted with methanol at room temperature for 24 hours. The lipids were precipitated with formic acid aqueous solution and eliminated by centrifugation. The corticosterone was separated from other compounds with reverse phase chromatography using acetonitrile and 0,1% aqueous solution of formic acid as mobile phase. The detection was performed in positive SIM mode measuring the 347 m/z molecular ion. A six point calibration was performed in the range of 0,5-20 ng/ml, the accuracy was tested with quality control samples at two different concentration level. The total run time is only 4,2 minutes and the lower limit of quantification (LLOQ) is 0,5 ng/ml, with 10 pg absolute sensitivity. By determining the quantity of the hormone for a well-defined hair region, based on the speed of hair growth, we can characterize the retrospective stress exposure of the animals in different 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
HPLC-MS法测定大鼠毛发样品中皮质酮的含量
皮质酮是一种具有糖皮质激素和矿皮质激素作用的肾上腺皮质类固醇激素。根据以往的研究,血浆皮质酮水平与啮齿动物的应激暴露有关。由于皮质酮在血液中的半衰期短,其血浆浓度可作为急性应激标志物。但是头发会积累全身和局部产生的皮质酮,因此它可以用来研究慢性压力。然而,皮质酮的准确定量是一项分析挑战,因为在复杂的生物基质中发现的激素量非常低。高效液相色谱-质谱联用技术(HPLC-MS)可提供所需的选择性和灵敏度。目前公布的皮质酮定量方法涉及复杂的样品制备和较长的运行时间。因此,本研究的目的是简化大鼠毛发样品中皮质酮的提取方法,并建立一种优化的高效液相色谱-质谱法以准确定量。将大鼠毛发样品用甲醇洗涤、干燥、切割,然后在室温下用甲醇提取24小时。脂质用甲酸水溶液沉淀,离心除去。以乙腈和0.1%甲酸水溶液为流动相,采用反相色谱法分离皮质酮和其他化合物。在正SIM模式下进行检测,测量347 m/z分子离子。在0.5 ~ 20 ng/ml范围内进行了6点校准,并以两种不同浓度水平的质控样品进行了准确度测试。总运行时间仅为4.2 min,定量下限(LLOQ)为0.5 ng/ml,绝对灵敏度为10 pg。根据毛发生长的速度,在一个明确的毛发区域确定激素的数量,我们可以描述动物在不同条件下的回顾性应激暴露。
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
自引率
0.00%
发文量
0
期刊最新文献
Quality by design-based method development for the determination of related substances of ezetimibe by high performance liquid chromatography Determination of the sun protection factor of sunscreens Alzheimer’s disease beyond the amyloid accumulation Radioactive background radiation measurement in Mureş county Study of pulmonary complications in patients referred to the intensive care unit
×
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