액체 크로마토그래피 텐덤 질량 분석기를 이용 한 감마 하이드록시낙산 경구 투여 마우스 모델 소변에서의 스테로이드 호르몬 프로파일 분석
Metabolomic profile analysis of urinary steroid hormones following intraperitoneal injection with gamma-hydroxybutyric acid in rats by liquid choromatography-tandem mass spectrometry
- 주제(키워드) Endogenous steroid hormone , Profiling analysis , Trimethylsilylation , Gas chromatography–tandem mass spectrometry , Ovarian cancer tissue
- 발행기관 아주대학교
- 지도교수 이광
- 발행년도 2018
- 학위수여년월 2018. 2
- 학위명 석사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000027480
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Endogenous steroid hormones have been reported as an important role to metabolism control, inflammation regulation and development of reproductive organ. These endogenous steroid hormones are related with human disease such as breast, ovary cancer. In this study profiling method of endogenous steroid hormones as trimethylsilyl (TMS) derivative in ovarian cancer tissue was developed by high resolution gas chromatography–tandem mass spectrometry (GC−MS/MS). Characteristic ions in MRM mode with electron ionization (EI) permitted the accurate and selective analysis of endogenous steroid hormones. In optimal conditions, this method showed good linearity (r ≥ 0.9989) with limit of detection (LOD) of 2.5−48.5 pg and limit of quantification (LOQ) of 8.5−161.6 pg. The repeatabilities of intra− and inter−days as a percentage of relative standard deviation (% RSD) varied from 0.2−10.0 (% RSD), respectively accuracy as a percentage relative error (% RE) was from -8.2 to 8.2 (% RE). The recovery ranged from 87.7 ± 6.4 % to 99.7 ± 9.7 %. When applied to tissues of 11 ovarian cancer patients, total eight endogenous steroid hormones of dehydroepiandrosterone, estradiol, testosterone, pregnenolone, estriol, progesterone, cortisone and cortisol were identified and determined by GC−MS/MS. Therefore, the present endogenous steroid hormone profiling method in ovarian tissue by GC−MS/MS will be useful for the monitoring and screening of ovarian cancer in clinical fields.
more목차
Ⅰ. INTRODUCTION 1
Ⅱ. EXPERIMENTAL 4
1. Chemicals and reagents 4
2. Preparation of standard solutions 4
3. Sample preparation of ovary cancer tissue 4
4. Gas chromatographytandem mass spectrometry 7
5. Method validation for analysis of steroid hormone standards 9
6. Sample preparation of steroid hormones profiling analysis from ovarian cancer tissue 11
Ⅲ. Results 13
1. Method development for profiling analysis of steroid hormones 13
(1) Optimization of TMS derivative 13
(2) Mass spectra of 10 steroid hormones with TMS derivative 13
(3) Optimization of matrix effects and extract solvents 21
(4) Optimization of multiple reactions monitoring (MRM) analysis condition 21
2. Method validation of profiling analysis of steroid hormones 28
3. Steroid hormones profiling analysis in ovarian cancer tissue 32
Ⅳ. Conclusion 38
REFERENCES 39
