Orthogonal Comparison of GC-MS and (1)H NMR Spectroscopy for Short Chain Fatty Acid Quantitation.
Orthogonal Comparison of GC-MS and (1)H NMR Spectroscopy for Short Chain Fatty Acid Quantitation.
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DOI：
10.1021/acs.analchem.7b00848
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发表时间：
2017-08-01
影响因子：
7.4
通讯作者：
Patterson AD
中科院分区：
文献类型：
--
作者：
Cai J;Zhang J;Tian Y;Zhang L;Hatzakis E;Krausz KW;Smith PB;Gonzalez FJ;Patterson AD
短链脂肪酸（SCFAs）是宿主生理和代谢的重要调节因子，可能与肥胖和相关代谢疾病有关。对scfa的兴趣增加，部分原因是由于认识到微生物群如何产生scfa可能向宿主发出信号的重要性。因此，需要可靠的、可重复的和负担得起的SCFA分析方法来进行准确的鉴定和定量。在本研究中，采用气相色谱-质谱联用（GC - MS）和1H核磁共振（NMR）谱两个独立的平台，比较了四种不同的SCFA（乙酸、丙酸和丁酸）提取和定量方法。对所有方法的灵敏度、回收率、可重复性、基质效应和小鼠粪便样品的有效性进行了测定。GC - MS丙基酯化法测定乙酸和丁酸具有较高的灵敏度（LOD < 0.01 μ mL−1,LOQ < 0.1 μ mL−1）和回收率（100 μ mL−1 SCFA混合标准峰in回收率为99.4% ~ 108.3%，250 μ mL−1 SCFA混合标准峰in回收率为97.8% ~ 101.8%）。与GC - MS方法相比，核磁共振方法通过相对于内部标准的定量或使用校准曲线的定量产生更好的重复性和最小的基质效应。所有方法均产生了良好的校准曲线线性（R2 > 0.99）和可比较的粪便SCFA浓度测量。最后，这些方法被用于定量从常规饲养（convr）和无菌（GF）小鼠中获得的粪便SCFAs。利用1H NMR和弹量热法对粪便进行代谢组学分析，进一步验证了这些方法。
Short chain fatty acids (SCFAs) are important regulators of host physiology and metabolism and may contribute to obesity and associated metabolic diseases. Interest in SCFAs has increased in part due to the recognized importance of how production of SCFAs by the microbiota may signal to the host. Therefore, reliable, reproducible, and affordable methods for SCFA profiling are required for accurate identification and quantitation. In the current study, four different methods for SCFA (acetic acid, propionic acid, and butyric acid) extraction and quantitation were compared using two independent platforms including gas chromatography coupled with mass spectrometry (GC−MS) and 1H nuclear magnetic resonance (NMR) spectroscopy. Sensitivity, recovery, repeatability, matrix effect, and validation using mouse fecal samples were determined across all methods. The GC−MS propyl esterification method exhibited superior sensitivity for acetic acid and butyric acid measurement (LOD < 0.01 μg mL−1, LOQ < 0.1 μg mL−1) and recovery accuracy (99.4%−108.3% recovery rate for 100 μg mL−1 SCFA mixed standard spike in and 97.8%−101.8% recovery rate for 250 μg mL−1 SCFAs mixed standard spike in). NMR methods by either quantitation relative to an internal standard or quantitation using a calibration curve yielded better repeatability and minimal matrix effects compared to GC−MS methods. All methods generated good calibration curve linearity (R2 > 0.99) and comparable measurement of fecal SCFA concentration. Lastly, these methods were used to quantitate fecal SCFAs obtained from conventionally raised (CONV-R) and germ free (GF) mice. Results from global metabolomic analysis of feces generated by 1H NMR and bomb calorimetry were used to further validate these approaches.
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影响因子：
4.1
作者：
Arellano, M;ElKaddouri, S;Puig, P
通讯作者：
Puig, P
影响因子：
2.2
作者：
FRANKEL, W;LEW, J;ROMBEAU, J
通讯作者：
ROMBEAU, J
DOI：
10.1038/ijo.2015.84
发表时间：
2015-09
期刊：
International journal of obesity (2005)
影响因子：
--
作者：
Byrne CS;Chambers ES;Morrison DJ;Frost G
通讯作者：
Frost G
抗氧化药物tempol促进肠道菌群中的功能代谢变化。
影响因子：
4.4
作者：
Cai J;Zhang L;Jones RA;Correll JB;Hatzakis E;Smith PB;Gonzalez FJ;Patterson AD
通讯作者：
Patterson AD
影响因子：
17.1
作者：
Blumberg R;Powrie F
通讯作者：
Powrie F