以質譜法分析人類代謝體於刑事體液鑑別之研究

The Study of Human Metabolome Analysis by Mass Spectrometry on Forensic Biofluid Identification

10.6134/tjfm.201912_11(1_2).0003

張宇翔(Yu-Hsiang Chang)；謝幸媚(Hsing-Mei Hsieh)；王勝盟(Sheng-Meng Wang)；陳怡靜(Yi-Ching Chen)；蔡麗琴(Li-Chin Tsai)

液相層析 ； 液相層析四極柱飛行時間質譜術 ； 體液代謝體 ； 體液鑑定 ； Liquid Chromatography ； LC-Q-ToF Mass Spectrometry ； Biofluid Metabolome ； Biofluid Identification

台灣法醫學誌

11卷1&2期（2019 / 12 / 01）

12 - 18

繁體中文

本研究以液相層析四極柱飛行時間質譜儀進行體液中特異性代謝物分析，以應用於鑑別體液之種類。以6種體液（尿液、精液、周邊血、經血、唾液與汗液）各6名個體之檢體作為研究材料，由評估6種萃取溶劑於體液代謝體分析之適用性結果發現，甲醇／丙酮（v/v 3:7）於體液間可萃取出較大之代謝體範圍（經血除外），故以此萃取溶劑作為後續各體液於代謝體組成之探討。各體液代謝體中以精液能檢測到最多種類之代謝物（81.83±9.45種），唾液最少（42.50±3.39種）；而相同體液於6名個體間均可偵測到之共有代謝物比率分析中發現，汗液之共有比率最低（29.96±4.65%），最高者為唾液（52.03±3.98%）。本研究於體液特異性代謝物標記搜尋中，共篩選出3種具尿液特異性之代謝物、6種具精液特異性之代謝物、1種具血液特異性之代謝物以及1種可用以分辨周邊血與經血之代謝物。這些具有體液特異性潛力之代謝物，並未發現於其他體液間會出現偽陽性的結果，且於個體內及個體間的試驗中亦均具有再現性。本研究以液相層析四極柱飛行時間質譜儀所建立之人類體液鑑別系統，應具有應用於刑事體液鑑別之潛力。

The study aims to detect biofluid-specific metabolites by LC-Q-ToF mass spectrometer for forensic biofluid identification. Firstly, the applicability of 6 solvents for extraction of 6 different biofluids (urine, semen, venous blood, menstrual blood, saliva and sweat), each of them sourced from 6 individuals, were evaluated. The most abundant metabolites except for the menstrual blood were observed by using methanol/acetone (v/v 3:7) as extraction solvent, and it was thus taken as the extraction solvent in the following experiments. Among these 6 different biofluids, the most abundant metabolites were observed in semen (81.83 ± 9.45) while in saliva was the least (42.50 ± 3.39). Furthermore, metabolomic mutuality ratio for inter-individual similarity showed the lowest percentage for sweat (29.96 ± 4.65%) and the highest for saliva (52.03 ± 3.98%). For biofluid-specific metabolite markers searching, 3 urine-specific metabolites, 6 semen-specific metabolites and 1 blood-specific metabolic markers were observed. Additionally, 1 metabolite could be used to distinguish between venous and menstrual blood. All metabolites claimed above to be biofluid-specific markers showed reproducibility in both intra-individual and inter-individual tests, and no inter-biofluid false-positive results were observed. The human biofluid identification system established in this study by LC-Q-ToF mass spectrometry could be a potential technique for practical forensic biofluid identification.

1. Donfack, J,Wiley, A(2015).Mass spectrometry-based cDNA profiling as a potential tool for human body fluid identification.Forensic Sci Int Genet,16,112-120.
2. Luan, H,Liu, LF,Tang, Z(2015).Comprehensive urinary metabolomic profiling and identification of potential noninvasive marker for idiopathic Parkinson’s disease.Sci Rep,5,13888.
3. Scano, P,Locci, E,Noto, A(2013).1H NMR metabolite fingerprinting as a new tool for body fluid identification in forensic science.Magn Reson Chem,51,454-462.
4. Sekula, P,Goek, ON,Quaye, L(2016).A metabolome-wide association study of kidney function and disease in the general population.J Am Soc Nephrol,27,1175-1188.
5. Smith, CA,O’Maille, G,Want, EJ(2005).METLIN: A metabolite mass spectral database.Ther Drug Monit,27,747-751.
6. Van Steendam, K,De Ceuleneer, M,Dhaenens, M,Van Hoofstat, D,Deforce, D(2013).Mass spectrometry-based proteomics as a tool to identify biological matrices in forensic science.Int J Legal Med,127,287-298.
7. Vuckovic, D(2012).Current trends and challenges in sample preparation for global metabolomics using liquid chromatography-mass spectrometry.Anal Bioanal Chem,403,1523-1548.
8. Wang, Q,Gao, P,Wang, X,Duan, Y(2014).The early diagnosis and monitoring of squamous cell carcinoma via saliva metabolomics.Sci Rep,4,6802.
9. Wishart, DS,Jewison, T,Guo, AC(2013).HMDB 3.0—The Human Metabolome Database in 2013.Nucleic Acids Res,41,D801-D807.