Studies On Determination Of Sex Hormones In Antler Velvet And Antler Velvet Healthy Products

Sex hormones can be used to regulate body metabolism and physiologicalfunction and have high biological activity in very low concentrations. The abuse ofsex hormone is harmful to human and animal health, social and ecologicalenvironment. Therefore the detection of sex hormone levels in velvet antler and itshealthy products, especially the simultaneous detection of a variety of sex hormones issignificantly important.In this thesis18kinds of sex hormones, includingestriol,corticosterone,17β-estradiol,estrone,17α-ethinylestradiol,17α-estradiol,17α-hydroxyprogesterone,medroxyprogesterone,19-nortestosterone,testosteron,megestrol-17-acetate, norethisterone acetate, chloromadinon17-acetate,medroxyprogesterone-17-acetate,progesterone,androsterone,testosteron-17-propionateand17β-Estradiol-benzoate were determined.15kinds of velvet antler and its healthyproducts from home and abroad were analyzed.The LC-MS/MS was developed to detect sex hormones in velvet antler.Theultrasonic extraction was coupled with solid phase extraction (SPE) to treat to thesamples.The extraction conditions, including extraction solvent type of solid phaseextraction column were examined and optimized. The recoveries are64.17~126.67%and RSDs are lower than20%, the detection limits are0.500~2.00μg/kg. InLC-MS/MS multiple reaction monitoring (MRM) mode was applied.7kinds of velvetantler samples were analyzed, the content of17α-hydroxyprogesterone in the sampleis4.40μg/kg.The content of progesterone in three samples are1.30μg/kg、8.00μg/kgand10.50μg/kg respectively, and the content of testosterone propionate is1.20μg/kgin sample. The analytical results obtained in different laboratories illustrate that themethod has good repeatability.The LC-MS/MS was developed to detect sex hormones in velvet antler healthyproducts. The ultrasonic extraction was coupled with C18solid phase extractionpurification. The recoveries are71.24~107.40%, RSDs are0.88~16.20%, LODs are0.20~1.50μg/kg.8kinds of velvet antler healthy products were analyzed.17α- hydroxyprogesterone content was10.60μg/kg in one sample and sex hormones inother samples are not detectable.The GC-MS/MS was developed to detect sex hormones in velvet antler. Theultrasonic extraction was coupled with SPE to treat the samples. The derivativereagent was seven fluorine acetic anhydride and derivative conditions were examinedand optimized. The MRM mode was applied. A precursor/product ion and twoprecursor/product ions were applied for qualitative and quantitative detection,respectively. The recoveries are62.25~95.58%, RSDs are lower than18%, LODs are1.00~30.00μg/kg.7kinds of velvet antler were analyzed, estriol are1.10μg/kg and1.02μg/kg in two samples, respectively.17α-hydroxyprogesterone is4.62μg/kg inone sample. Progesterone are2.51μg/kg、8.64μg/kg and10.40μg/kg in threesamples respectively. LC-MS/MS are not significantly different.The GC-MS/MS was developed to detect sex hormones in antler velvet healthyproducts. The recoveries are78.17~109.17%, RSDs are1.93~16.47%, LODs are0.50~10.00μg/kg.8kinds of velvet antler healthy products are analyzed.17α-hydroxyprogesterone content was9.90μg/kg in one sample.When LC-MS/MS was applied the expanded uncertainty is calculated. The effect ofa variety of uncertainties, including sex hormone standard purity, the preparation ofstandard solution, the amount of the sample, sample extraction, purification of SPEand LC-MS/MS determination, on the testing results were examined such as theevaluation, extension and synthesis.Based on sources of measurement uncertainties,the expanded uncertainty was obtained and is0.171.