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顶空固相微萃取(SPME)和气相色谱-质谱联用(GC-MS)法测定亚洲雌象体内的5α-雄甾-2-烯-17-酮和-17β-醇:在生殖监测和分娩预测中的应用

Headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for the determination of 5alpha-androst-2-en-17-one and -17beta-ol in the female Asian elephant: application for reproductive monitoring and prediction of parturition.

作者信息

Dehnhard M, Hatt J-M, Eulenberger K, Ochs A, Strauss G

机构信息

Institute for Zoo Biology and Wildlife Research, PF 601103, D-10252 Berlin, Germany.

出版信息

J Steroid Biochem Mol Biol. 2003 Feb;84(2-3):383-91. doi: 10.1016/s0960-0760(03)00056-6.

Abstract

Asian elephants are not self-sustaining in captivity. The main reasons for this phenomenon are a low birth rate, an aging population, and poor calf-rearing. Therefore, it is essential that reproductive rates had to be improved and there is need for rapid quantitative measures to monitor reproductive functions focussing on estrous detection and the prediction of the period of parturition. The objective of this study was to develop a method which combines headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for analyses of 5alpha-androst-2-en-17beta-ol and -17-one to prognose estrous and to predict the period of parturition. SPME was carried out with a CTC Combi Pal system. The course of the luteal phase-specific substance 5alpha-androst-2-en-17beta-ol and -17-one followed a cyclic pattern in which the follicular and luteal phases could be clearly distinguished (mean estrous cycle length, 15+/-1.4 weeks). Based on daily urine samples, estrous prognosis might be possibly based on the initial 5alpha-androst-2-en-17beta-o1 increase at the end of the follicular phase. Parturition prognosis was performed in three elephant cows based on the 5alpha-androst-2-en-17beta-o1 drop to baseline levels 5-4 days prior parturition. Experiments revealed that 5alpha-androst-3alpha-ol-17-one and probably 5alpha-androst-3alpha-ol-17beta-ol are generated from sulfate conjugates by a thermal process.

摘要

亚洲象在圈养环境中无法自我维持种群数量。出现这种现象的主要原因是出生率低、种群老龄化以及幼崽饲养情况不佳。因此,提高繁殖率至关重要,并且需要快速的定量措施来监测生殖功能,重点是发情检测和分娩期预测。本研究的目的是开发一种结合顶空固相微萃取(SPME)和气相色谱 - 质谱联用(GC - MS)的方法,用于分析5α - 雄甾 - 2 - 烯 - 17β - 醇和 - 17 - 酮,以预测发情期和分娩期。SPME使用CTC Combi Pal系统进行。黄体期特异性物质5α - 雄甾 - 2 - 烯 - 17β - 醇和 - 17 - 酮的变化过程呈周期性模式,其中卵泡期和黄体期可明显区分(平均发情周期长度为15±1.4周)。基于每日尿液样本,发情预测可能基于卵泡期末期5α - 雄甾 - 2 - 烯 - 17β - 醇的初始增加。根据分娩前5 - 4天5α - 雄甾 - 2 - 烯 - 17β - 醇降至基线水平,对三头母象进行了分娩预测。实验表明,5α - 雄甾 - 3α - 醇 - 17 - 酮以及可能的5α - 雄甾 - 3α - 醇 - 17β - 醇是由硫酸盐结合物通过热过程产生的。

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