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논문 기본 정보

자료유형
학위논문
저자정보

엄진희 (고려대학교, 고려대학교 대학원)

지도교수
천철홍
발행연도
2016
저작권
고려대학교 논문은 저작권에 의해 보호받습니다.

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이 논문의 연구 히스토리 (2)

초록· 키워드

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5-Androsten-3β-,17β-diol-7-one diacetate is an anabolic steroid derived from dehydroepiandrosterone. Anabolic steroids, intraglandular precursors of hormones, usually exhibit minimal hormonal effects by themselves. They are known to increase muscle growth and strength, and bodybuilders often use them. However, they can cause gynecomastia, virilism, and skin diseases as side effects. Even these side effects steroids are being developed. In this study, new metabolites and metabolic processes are investigated for determining safety of drugs. 5-Androsten-3β-,17β-diol-7-one diacetate as unknown metabolites was orally administrated to six healthy volunteers. And then samples were prepared from urine by way of PAD-1 adsorption, enzymatic hydrolysis with β-glucuronidase, and liquid/liquid extraction. As a result of optimizing the pretreatment, it was extracted with diethyl ether at pH 6.8 and derivatized with MSTFA/NH4I/DTE (500 : 2 : 1 (w/w/w)) at 60 ℃ for 15 min. The extracted 5-androsten-3β-,17β-diol-7-one diacetate and its metabolites were compared to blank urine, spiked urine, and dosed urine samples by LC/MS and GC/MS. 5-Androsten-3β-,17β-diol-7-one diacetate was not detected in dosed urine, but M1-5-androsten-3β, 7α, 17β?triol, M2-5-androsten-3β, 17β?diol-7-one, and M3-5-androsten-3β, 16α, 17β?triol-7-one were identified as metabolites. As a result, M1 and M2 were endogenous metabolites, and M3 was a new metabolite of 5-androsten-3β-,17β-diol-7-one diacetate. The results of this study on the metabolic pathway and metabolites of the drug (unknown steroid) by LC/MS and GC/MS can be used to determine whether steroid is taken.

목차

CONTENTS
Chapter 1
1. Introduction
1.1 Introduction 3
1.2 Metabolism of drugs 4
1.3 Analysis of metabolites 7
1.4 The purpose of this study 10
Chapter 2
2. Experiment
2.1 Chemicals and reagents 11
2.2 Instruments 12
2.3 Experiment methods 13
2.3.1 Sample preparation 13
2.3.1.1 Urine sampling 13
2.3.1.2 Purification 13
2.3.1.3 Hydrolysis 14
2.3.1.4 Extraction 15
2.3.1.5 Derivatization 16
2.3.2 Validation of Experimental methods 22
2.3.3 Analysis of 5-androsten-3β-,17β-diol-7-one diacetate and its metabolites using LC/TOF-MS 23
2.3.4 Analysis of 5-androsten-3β-,17β-diol-7-one diacetate and its metabolites using GC/MSD 23
Chapter 3
3. Results
3.1 Separation and extraction of 5-androsten-3β-,17β-diol-7-one diacetate in urine sample 25
3.1.1 The optimal extraction conditions for metabolites of 5-androsten-3β-,17β-diol-7-one in urine sample 25
3.1.2 The optimal derivatization conditions of 5-androsten-3β-,17β-diol-7-one metabolites from urine sample 27
3.1.3 Validation of analysis procedures 30
3.2 Analysis of 5-androsten-3β-,17β-diol-7-one diacetate and its endogenous metabolites by LC/MS 32
3.2.1 Identification of M1-5-androsten-3β, 7α, 17β -triol in urine sample 37
3.2.2 Identification of M2-5-androsten-3β, 17β?diol-7-one in urine sample 37
3.2.3 Identification of M3-5-androsten-3β, 16α, 17β?triol-7-one in urine sample 38
3.3 Analysis of 5-androsten-3β-,17β-diol-7-one diacetate and its endogenous metabolites by GC/MSD 38
3.3.1 Identification of M1-5-androsten-3β, 7α, 17β-triol in urine sample 39
3.3.2 Identification of M2-5-androsten-3β, 17β?diol-7-one in urine sample 40
3.3.3 Identification of M3-5-androsten-3β, 16α, 17β?triol-7-one in urine sample 41
3.4 Investigation of excretion of chemical species using endogenous metabolites of 5-androsten-3,17β-diol-7-one-diacetate 48
3.5 Investigation of excretion tendencies of metabolites of 5-androsten-3β, 17β?diol-7-one diacetate 50
Chapter 4
4. Conclusion 52
References 53

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