干旱区生态环境知识资源中心(Arid): 9种WADA禁用生长激素释放肽和生长激素促分泌素的HPLC-MS/MS检测及尿中稳定性研究

Arid

	9种WADA禁用生长激素释放肽和生长激素促分泌素的HPLC-MS/MS检测及尿中稳定性研究
其他题名	Detection of Nine WADA Prohibited GHRPs and GHS Using the HPLC- MS/MS Method and Their Stability in Human Urine
	申利 1; 杨辛兰 2; 张力思 1; 河春姬 1; 周鑫淼 1; 徐友宣 1; 闫宽 1
来源期刊	中国运动医学杂志
ISSN	1000-6710
出版年	2018
卷号	37 期号:5 页码:420-426
中文摘要	目的:选取世界反兴奋剂机构(WADA)禁用的7种生长激素释放肽(GHRP-1, GHRP-2, GHRP-4, GHRP-5, GHRP-6, Hexarelin和Alexamorelin)和2种生长激素促分泌素(Anamorelin和Ipamorelin)为研究对象,建立人尿中的HPLC-MS/MS检测方法,进行稳定性实验。方法:使用Oasis WCX固相提取小柱(1cc, 30mg)对尿样进行化学前处理,尿样加入内标后离心,取1mL加入小柱,依次用5% NH_4OH和20% CH_3CN清洗后,用含2%甲酸的水/乙腈(1/3)洗脱液洗脱,35℃氮气流下吹干,复溶后进样于LC-MS/MS。使用1290/6470液相色谱质谱仪和Zorbax 300SB-C18色谱柱进行定性分析,流动相采用含0.2%甲酸的水/乙腈体系。结果:检测限根据不同物质分布在0.01~0.5 ng/mL(S/N>3)。回收率分布在40%~76%之间,日内精密度和日间精密度均小于15%,尿中基质无明显干扰。室温,冷藏条件储存和反复冻融对Anamorelin、GHRP-2、GHRP-4和GHRP-5有明显影响。结论:建立了尿中9种生长激素释放肽和生长激素促分泌素的HPLC-MS/MS检测方法,方法简单,特异性、灵敏度均可满足WADA实验室国际标准和技术文件要求。现已应用于我实验室的常规检测。尿样在传送、检测及实验室长期保存过程中应尽量避免反复冻融。
英文摘要	Objective To introduce a practical high performance liquid chromatography-tandem mass spectrometry(HPLC- MS/MS)method for the detection of seven growth hormone- releasing peptides (GHRPs)including GHRP-1, GHRP-2, GHRP-4, GHRP-5, GHRP-6, Hexarelin and Alexamorelin and two growth hormone secretagogues(GHS)including anamorelin and ipamorelin, and study the stability of these nine substances in the human urine. Method The urine samples were purified and extracted by a solid phase extraction procedure using Oasis WCX column. The urine was first centrifuged and tak- en out 1 mL into a small column, cleaned by 5% NH_4OH and 20% CH_3CN respectively, eluated using the mixture of water and acetomitrile(1/3)with 2% formic acid, blow-dried in the nitrogen at 35℃ and finally redissolved to be injected into the LC-MS/MS. Result The limits of detection were between 0.01~0.5 ng/mL accordingly. The spiked recoveries at the low concentration(1 ng/mL), medium concentration(2 ng/mL)and high concentration(10 ng/mL)ranged between 40% and 76%. The intra- and interday precisions of the target substances at these three concentrations were all less than 15%. The indoor temperature, refrigeration condition and multigelation were observed to have significant impact on the anamorelin, GHRP-2,GHRP-4 and GHRP-5. Conclusion The method established in this study is sim- ple, and its specificity and sensitivity meets the international standard and technical documents for labo- ratories set up by the Wworld Anti-Doping Agency. It has been applied in our routine work. Multigelation should be avoided in the transport, detection and long-term laboratory storage of urine samples.
中文关键词	禁用物质 ; 生长激素释放肽 ; 生长激素促分泌素 ; 固相提取 ; 反复冻融
英文关键词	prohibited substances growth-hormone release peptides(GHRPs) growth hormone secre- tagogues(GHS) solid-phase extraction multigelation
语种	中文
国家	中国
收录类别	CSCD
WOS类目	MEDICINE GENERAL INTERNAL
WOS研究方向	General & Internal Medicine
CSCD记录号	CSCD:6261774
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/238871
作者单位	1.国家体育总局反兴奋剂中心, 北京 100029, 中国; 2.北京体育大学
推荐引用方式 GB/T 7714	申利,杨辛兰,张力思,等. 9种WADA禁用生长激素释放肽和生长激素促分泌素的HPLC-MS/MS检测及尿中稳定性研究[J],2018,37(5):420-426.
APA	申利.,杨辛兰.,张力思.,河春姬.,周鑫淼.,...&闫宽.(2018).9种WADA禁用生长激素释放肽和生长激素促分泌素的HPLC-MS/MS检测及尿中稳定性研究.中国运动医学杂志,37(5),420-426.
MLA	申利,et al."9种WADA禁用生长激素释放肽和生长激素促分泌素的HPLC-MS/MS检测及尿中稳定性研究".中国运动医学杂志 37.5(2018):420-426.