metabolomics

代谢组学（ 代谢组学（Metabonomics）技术： 技术： Metabolomics
— Detection of endogenous compounds (内源性物质) — Xenobiotics are interferences — Multi-sample statistical approach — Exact mass needed — MS/MS not necessary — High sensitivity — High resolution LC — Dedicated Application Manager: MarkerLynx?XS
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

代谢组学研究思路
分析 方法
数据 处理
寻找 标志 物
数据 库查 询
生物 功能 解析
分析方法是代谢组学研究的核心技术
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

A Manual Approach Think about doing a rigorous comparison of just these 4 spectra (~2,000 masses or metabolites in each).
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

MarkerLynx 能干什么？ 能干什么？
UPLC-QTF Raw data
MarkerLynx
Result file
Exact Mass – Retention Time Pairs (EMRT’S)
Extended statistics analysis
Multivariant statistical analysis PCA OPLS PLS
Element composition
S-Plot
Search database
Markers
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

常用的数据 常用的数据库
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

Human Metabolome Project (HMP)
The Human Metabolome Project 是在2005年1月投资的 一个 $7.5 million的加拿大基因组项目 2007年1月23号，ＨＭＰ项目由加大拿艾伯特大学的 Dr. David Wishart领导，完成了第一个人类代谢组数据库，数据库 包含了大约２５００种代谢物，１２００种药类和 ３５００种 食品 化合物 HMP是一个主要给科学机构提供化学组分数据信息的机构。它 并没有基金和资助来源去使用它们的原始数据来帮助疾病确认 和描述
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

KEGG
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BIOCYC
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ChemSpider
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探索银杏叶片的质量控制方法
样品由湖南农业大学提供 提取处理后样品分三组
银杏叶片采集于浏阳郊区,采集时间是2010年5月到9月份,批次间隔15 天 ? 新鲜叶片水提后样品 A1, A2, …… A10 ? 新鲜叶片醇提后样品 C1, C2, ….. C10
? 干燥后叶片水提后样品 B1, B2, …… B10
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

难以形象化样品间差异
B3 BPI A3 BPI
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整体样品组主成分分析(PCA)分数图(Scores plot)
Fresh Leaves Dry Leaves Fresh Alcohol
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

2010 年新鲜银杏叶片样品 (A)主成分分析(PCA) 分数图(Scores plot)
Spring
Fall
Summer
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

2010 新鲜银杏叶片(A) PCA Loadings Plot
High in Spring (A1, A2)
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Markers 强度随采集时间(春→秋)减弱
Tr (min) m/z 1.86 1.85 5.53 4.69 3.19 4.37 6.46 5.53 2.89 4.69 2.33 3.02
163.11 193.12 284.03 300.02 609.14 623.16 739.18 739.18 739.20 755.18 755.20 769.22
10A01 10A02 10A03 10A04 10A05 10A06 10A07 10A08 10A09 10A10
322.37 435.79 515.86 409.01 210.33 130.74 66.93 114.97 130.20 134.88
55.14 64.04 73.71 54.80 30.30 19.33 9.84 17.15 20.14 20.68
131.91 101.12 78.19 68.40 70.33 58.41 54.05 49.02 22.59 22.94
77.85 53.67 47.21 38.02 31.28 26.12 25.25 20.44 10.72 11.18
246.49 328.51 115.39 562.21 309.36 410.45 140.97 153.50 192.33 258.07 202.96 265.83 180.73 216.73 185.82 227.97 169.13 205.77 140.55 190.27 140.77 173.19 72.84 76.92 118.30 118.33 83.54 30.05 57.18 42.56 30.15 35.04 35.82 14.27 14.75 414.55 245.01 276.74 77.90 111.08
May 1st May 15th June 1st June 15th July 1st July 15th August 1st August 15rt
313.70 257.70 260.52 112.92 138.37 279.04 181.23 201.42 285.10 222.37 195.78 235.10 155.34 170.91 213.85 94.60 153.74 78.94 62.91 34.49 21.22 43.62 15.17 15.84 81.23 95.31 61.98 33.06 56.64
196.16 141.56 120.18 114.27 111.55 68.40 73.19 60.65 63.93
23.89 September 1st 24.53
September 15st
ChangeFold
2.39
2.67
5.75
6.96
3.20
2.78
7.82
5.04
4.23
6.42
8.90
6.26
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

OH OH OH O O H O H O OH O OH O OH OH O OH CH3 O OH OH
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MSE Data
Low CE
High CE
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

使用MassFragment软件进行结构确证
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Phytochemistry 2001, 58, P1201P1201 -1205
OH OH OH O O H O H O OH O OH O OH OH O OH CH3 O OH OH
Antioxidant Flavonol Glycoside
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011

Trending Plot 显示该Marker的变化趋势
OH OH OH O O H O H O OH O OH O OH OH O OH CH3 O OH OH
C36H36O18, MW 756.1901 Antioxidant Flavonol Glycoside
?2009 Waters Corporation | COMPANY CONFIDENTIAL ?2011