Name: the use of gas chromatography to analyze compositional changes of fatty acids in rat liver tissue during pregnancy
Uploaded: 2014-03-13T13:45:00
Description: Watch this Scientific Journal Video about The Use of Gas Chromatography to Analyze Compositional Changes of Fatty Acids in Rat Liver Tissue during Pregnancy at JoVE.com

作者要感谢马萨纳罗梅乌 - 纳达尔在大鼠研究的贡献。

1。动物规程<ol><li>所有的动物工作应按照内政部动物（科学程序）法（1986）进行。 </li><li>交配10周龄以一夫一妻制繁殖旧Wistar大鼠，并通过阴道塞的外观确认怀孕。记录这是怀孕的第1天，并开始试验性的饮食。对于处女女，分别容纳各组大鼠，并开始试验性的饮食。 </li><li>喂养试验饲料20天安乐死后大鼠用CO 2窒息颈椎脱位。 </li><li>用解剖镊子和剪刀以暴露腹腔，并通过切割韧带，其?...

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Chem. 52 (12), 2265-2272 (2006).</li><li>Lauritzen, L., Carlson, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maternal+fatty+acid+status+during+pregnancy+andlactation+and+relation+to+newborn+and+infant+status.">Maternal fatty acid status during pregnancy andlactation and relation to newborn and infant status.</a> Matern. Child Health. 7 (2), 41-58 (2011).</li><li>Kelsall, C. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+dysfunction+induced+in+offspring+by+maternal+dietary+fat+involves+altered+arterial+polyunsaturated+fatty+acid+biosynthesis.">Vascular dysfunction induced in offspring by maternal dietary fat involves altered arterial polyunsaturated fatty acid biosynthesis.</a> PLoS One. 7 (4), (2012).</li><li>Karpe, F., Dickmann, J. R., Frayn, K. 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M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fatty+acid+content+of+marine+oil+capsules.">Fatty acid content of marine oil capsules.</a> Lipids. 25 (9), 523-528 (1990).</li><li>Folch, J., Lees, M., Sloane-Stanley, G. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+for+the+isolation+and+purification+of+total+lipids+from+animal+tissues.">A simple method for the isolation and purification of total lipids from animal tissues.</a> J. Biol. Chem. 226 (1), 497-509 (1957).</li><li>Burdge, G. C., Wright, P., Jones, E. A., Wootton, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+separation+of+phosphatidylcholine,+triacylglycerol,+non-esterified+fatty+acids+and+cholesterol+esters+from+plasma+by+solid-phase+extraction.">A method for separation of phosphatidylcholine, triacylglycerol, non-esterified fatty acids and cholesterol esters from plasma by solid-phase extraction.</a> Br. J. Nutr. 84 (5), 781-787 (2000).</li><li>Sepp&#228;nen-Laakso, T., Laakso, I., Hiltunen, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+fatty+acids+by+gas+chromatography,+and+its+relevance+to+research+on+health+and+nutrition.">Analysis of fatty acids by gas chromatography, and its relevance to research on health and nutrition.</a> Anal. Chim. Acta. 465 (1), 39-62 (2002).</li><li>Beesley, T. E., Buglio, B., Scott, R. P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Quantitative chromatographic analysis. , (2000).</li><li>Bartle, K. D., Myers, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=History+of+gas+chromatography.">History of gas chromatography.</a> Trends Anal. Chem. 21 (9), 9-10 (2002).</li><li>Childs, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The effect of gender, pregnancy and diet upon rat tissue fatty acid composition and immune function. , 378 (2008).</li><li>Harris, S. W., Pottala, J. V., Ramachandran, S. V., Larson, M. G., Robins, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+erythrocyte+membrane+Trans+and+marine+fatty+acids+between+1999+and+2006+in+older+Americans.">Changes in erythrocyte membrane Trans and marine fatty acids between 1999 and 2006 in older Americans.</a> J. Nutr. 142 (7), 1297-1303 (2012).</li><li>Roberts, L. D., McCombie, G., Titman, C. M., Griffin, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+matter+of+fat:+An+introduction+to+lipidomic+profiling+method.">A matter of fat: An introduction to lipidomic profiling method.</a> J. Chromatogr. B. 871 (2), 174-181 (2008).</li></ol>

气相色谱法是一种精确的技术来用于脂肪酸分析，以及其高重复性认为该技术适于临床分析。适当的气相色谱柱，必须使用以使得能够识别的感兴趣的脂肪酸，具有变化的固定相，柱的长度和内径的极性可用的列。在分析这种方法的使用石英毛细管柱提供了良好的热稳定性和高的，由于其高的表面惰性和良好的分辨率8保留时间的重现性。 本协议?...

气相色谱（GC）是一种用于识别和量化的脂肪酸掺入脂质池和细胞膜1,2期间补充或生理条件，例如肥胖（及其相关的疾病，如糖 ​​尿病），或者怀孕3行之有效的技术- 5。它也适合于分析中的食物的种类和脂肪的数量。当表征实验饲料，以及确保食品行业符合规定，这是很有用的。例如，GC可以用来确认商品如膳食补充剂中的身份和脂肪酸的量，以确保标签是正确和法规粘附到6,7。脂肪酸的分析可以提供有价值的见解在健康和疾病，饮食变化的影响，并改变生理状态8的影响脂质代谢。使用GC在怀孕期间，研究的样本提供了重要的变动脂肪酸和复合脂质稳态3的信息。 提前的色谱分离的，脂质通常从样品用脂质中的溶解度在氯仿和甲醇的混合溶剂萃取。加入氯化钠，以促进该混合物的成含有相9,10水溶液和有机脂质的分离。感兴趣的复合脂质类可以从总脂提取物固相萃取（SPE）分离。这种分离方法洗脱根据其极性或结合亲和力的脂质类。 Triacyglycerols（TAG）和胆甾醇酯（CE）是首先被洗脱的馏分合并，进一步类，磷脂酰胆碱（PC），磷脂酰乙醇胺（PE），和非酯化脂肪酸（NEFA）通过增加洗脱溶剂的极性洗脱。 TAG的从CE分离利用的TAG只能到一个新的SPE cartri绑定DGE，允许CE被洗脱。 TAG然后可以通过增加洗脱溶剂9,10的极性进行洗脱。这种方法允许多个样品有较高的产率比同时分离与薄层层析来实现，这意味着相对小尺寸的样品（ 如 &lt;100微升的血浆或血清，&lt;100 mg组织）可以分析11,12。 GC是一个行之有效的技术在20世纪50年代首次描述，有人提出，在当时的液 - 液系统中的流动相可以被替换为蒸气。它最初是用于石油分析，但迅速扩大到其他领域，如氨基酸分析和脂类生物化学，这仍然是主要的利益。前进中的GC设备和技术，例如从以前使用填充柱的毛细管柱的发展导致了我们目前的技术，其中的脂肪酸是能够被在导致GC被常规用于鉴定和定量脂肪酸在广泛调查13的较低的温度下更有效地分离。 GC需要的脂肪酸，使他们有可能成为充分挥发在合理的温度被洗脱不热分解进行衍生化。这通常涉及到含有氢以形成酯，硫酯或酰胺的用于分析的官能团的取代。甲酯通常被研究的衍生物，它是由甲基化制备。在该方法中复合脂质的酯键被水解以释放游离脂肪酸，它们transmethylated以形成脂肪酸甲基酯（FAME）。 FAME的所得到的轮廓，由GC测定，被称为的脂肪酸组成和可能不同的实验组9,10之间容易地进行比较。该技术允许individ两者的比例UAL脂肪酸和它们的浓度进行测量。 除了使用气相色谱，用于分析在营养学和食品工业中的脂肪酸，该技术可以在宽范围的分析领域中使用。例如，使用气相色谱环境分析包括通过杀虫剂和土壤测量水质污染分析测定氯苯的内容。在毒理学，GC也被用于识别非法物质在尿和个体的血液样品，这样的运动性能增强剂12和分离烃类的复杂混合物的能力，使得在石油工业中这种技术流行的用于石油化学分析12。 妊娠与显著改变母体组织中的脂肪酸组合物，特别是ω-3的含量（N-3）和ω-6（N-6）多不饱和脂肪酸ACI相关DS（PUFA）3。在目前的研究中，我们通过举例说明其使用从饲喂低，高脂肪的饮食不同油源和处女孕鼠取肝组织中脂肪酸组成的分析中脂肪酸的测定采用气相色谱。这里提供的试验日粮是一种低脂肪的大豆油为主的饮食，高脂肪的大豆油为主的饮食习惯（130.9克总脂肪/公斤总脂肪）或高脂肪的亚麻子油为主的饮食习惯（130.9克总脂肪/公斤饮食），提供了20天。这些饮食的全部营养和脂肪酸组合物先前已被14所述。豆油日粮中含有丰富的亚油酸（18:2的n-6）和含有一些α-亚麻酸（18点03的n-3），而亚麻油饮食中富含α-亚麻酸。这些高脂肪的饮食代表的亚油酸与α-亚麻酸（8:1和1:1分别口粮）不同的口粮。该方法对于个别脂质类和用GC分析的隔离是我们LL建立和验证，以及先前已10但没有在此找到详细的技术说明公布。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Methanol:</td>
			<td>Fisher Scientific</td>
			<td>M/4056/17</td>
			<td>&#39;CAUTION&#39; Fumes - HPLC Grade</td>
		</tr>
		<tr>
			<td>Chloroform:</td>
			<td>Fisher Scientific</td>
			<td>C/4966/17</td>
			<td>&#39;CAUTION&#39; Fumes - HPLC Grade</td>
		</tr>
		<tr>
			<td>BHT:</td>
			<td>Sigma- Aldrich</td>
			<td>W218405</td>
			<td>&#39;CAUTION&#39; Dust fumes - Anhydrous</td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma- Aldrich</td>
			<td>S9888</td>
			<td>Anhydrous</td>
		</tr>
		<tr>
			<td>Hexane</td>
			<td>Fisher Scientific</td>
			<td>H/0406/17</td>
			<td>&#39;CAUTION&#39; Fumes - HPLC Grade</td>
		</tr>
		<tr>
			<td>Glacial acetic acid</td>
			<td>Sigma- Aldrich</td>
			<td>695084</td>
			<td>&#39;CAUTION&#39; Burns - 99.85%</td>
		</tr>
		<tr>
			<td>Sulfuric acid</td>
			<td>Sigma- Aldrich</td>
			<td>339741</td>
			<td>&#39;CAUTION&#39; Burns - 99.999%</td>
		</tr>
		<tr>
			<td>Potassium carbonate</td>
			<td>Sigma- Aldrich</td>
			<td>209619</td>
			<td>99% ACS Reagent grade</td>
		</tr>
		<tr>
			<td>Potassium bicarbonate</td>
			<td>Sigma- Aldrich</td>
			<td>237205</td>
			<td>99.7% ACS Reasgent grade</td>
		</tr>
		<tr>
			<td>Ethyl acetate</td>
			<td>Fisher Scientific</td>
			<td>10204340</td>
			<td>&#39;CAUTION&#39; Fumes - 99+% GLC SpeciFied</td>
		</tr>
		<tr>
			<td>Toluene</td>
			<td>Fisher Scientific</td>
			<td>T/2300/15</td>
			<td>&#39;CAUTION&#39; Fumes</td>
		</tr>
		<tr>
			<td>Diethyl ether</td>
			<td>Sigma- Aldrich</td>
			<td>309966</td>
			<td>&#39;CAUTION&#39; Fumes</td>
		</tr>
		<tr>
			<td>Nitrogen (oxygen free) cylinder</td>
			<td>BOC</td>
			<td>44-w</td>
			<td>&#39;CAUTION&#39; Compressed gas - explosion risk</td>
		</tr>
		<tr>
			<td>Aminopropyl silica SPE cartridges</td>
			<td>Agilent</td>
			<td>12102014</td>
			<td>Cartridge - Bead mass 100mg</td>
		</tr>
		<tr>
			<td>Silica gel SPE cartidges</td>
			<td>Agilent</td>
			<td>14102010</td>
			<td>Cartridge - Bead mass 100mg</td>
		</tr>
		<tr>
			<td>Molecular seives</td>
			<td>Sigma- Aldrich</td>
			<td>334324</td>
			<td>Pellets, AW-300, 1.6mm</td>
		</tr>
		<tr>
			<td>Glass pasteur pipettes</td>
			<td>Fisher Scientific</td>
			<td>FB50251</td>
			<td></td>
		</tr>
		<tr>
			<td>Borosilicate glass test tube</td>
			<td>Fisher Scientific</td>
			<td>FB59527</td>
			<td>Non-screw cap</td>
		</tr>
		<tr>
			<td>Screw thread glass test tubes</td>
			<td>Fisher Scientific</td>
			<td>FB59555</td>
			<td>13mm</td>
		</tr>
		<tr>
			<td>Caps for screw thread test tubes</td>
			<td>Fisher Scientific</td>
			<td>FB51353</td>
			<td>To fit 13mm tube</td>
		</tr>
		<tr>
			<td>Solida phase extraction (SPE) tank</td>
			<td>Agilent</td>
			<td></td>
			<td>VacElut 20 Manifold</td>
		</tr>
		<tr>
			<td>Luer stopcocks for SPE tank</td>
			<td>Agilent</td>
			<td>12131005</td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum pump</td>
			<td>Sigma- Aldrich</td>
			<td>2656-194GB-1EA</td>
			<td></td>
		</tr>
		<tr>
			<td>GC vials</td>
			<td>Kinesis</td>
			<td>STV 12-03TS</td>
			<td>Short thread 9mm, TPX 0.2ml fused glass insert</td>
		</tr>
		<tr>
			<td>GC vial lids</td>
			<td>Kinesis</td>
			<td>SCC09-0.2B</td>
			<td>Short thread 9mm blue</td>
		</tr>
		<tr>
			<td>GC inlet liners</td>
			<td>SGE Analytical science</td>
			<td>092002</td>
			<td>Split/splitless Focus liner ID 4mm, OD 6.3mm length 78.5mm</td>
		</tr>
		<tr>
			<td>GC septa</td>
			<td>SGE Analytical science</td>
			<td>041856</td>
			<td>11mm, MN material</td>
		</tr>
		<tr>
			<td>GC column</td>
			<td>SGE Analytical science</td>
			<td>054612</td>
			<td>Length 30m, ID 0.22mm, Film thickness 0.25&#181;m</td>
		</tr>
		<tr>
			<td>Gas chromatograph</td>
			<td></td>
			<td></td>
			<td>HP 6890 series</td>
		</tr>
	</tbody>
</table>

the use of gas chromatography to analyze compositional changes of fatty acids in rat liver tissue during pregnancy

气相色谱（GC）是用于从组织，细胞，血浆/血清识别和量化脂质的脂肪酸含量，得到具有高精度和高再现性的结果高度敏感的方法。代谢和营养研究的GC允许更改下列措施中的脂肪酸浓度或在改变生理状态，如妊娠的评估。固相萃取（SPE），使用氨丙基硅胶管允许各大类脂质包括甘油三酯，磷脂不同，和胆固醇酯（CE）的分离。气相色谱结合固相萃取是用来分析的变化，行政长官在部分已被送入各种高和低脂肪饮食的处女和孕鼠的肝脏中脂肪酸的组成。有肝经行政长官的ω-3和ω-6脂肪酸含量显著饮食/妊娠互动效应，这表明怀孕的女性有不同的反应，饮食manipulat离子比被认为是处女的女性之一。

这种方法的成功依赖于精确的按照协议和有关使用清洁溶剂和试剂，以减少“噪声”，并且可以出现在色谱污染。污染的样品是更具挑战性的分析，从而降低了计算曲线下的面积的精度。如果该协议被成功地遵循的色谱图清晰对称的，明确定义的峰值和最小的背景噪声， 如图3应获得。如果已发生污染的色谱将显示额外的峰，并表现出如图4所示的非对称的（倾斜）的峰。...

Watch this Scientific Journal Video about The Use of Gas Chromatography to Analyze Compositional Changes of Fatty Acids in Rat Liver Tissue during Pregnancy at JoVE.com

The Use of Gas Chromatography to Analyze Compositional Changes of Fatty Acids in Rat Liver Tissue during Pregnancy

使用气相色谱怀孕期间分析脂肪酸组成的变化在大鼠肝组织

妊娠导致显著改变母体组织中的脂肪酸组合物。血脂可通过气相色谱法来获得允许在怀孕期间大鼠中识别和个别类脂质的脂肪酸定量饲喂各种高，低脂肪饮食。

Gas chromatography (GC) is a highly sensitive method used to identify and quantify the fatty acid content of lipids from tissues, cells, and plasma/serum, ...

Research

JoVE Journal

Chemistry

Determination of the Gas-phase Acidities of Oligopeptides

寡肽测定气相酸性

Amino acid residues located at different positions in folded proteins often exhibit different degrees of acidities. For example, a cysteine residue ...

科研

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Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography

通过超性能液相色谱对细菌细胞壁进行隔离和制备，进行组成分析

The bacterial cell wall is critical for the determination of cell shape during growth and division, and maintains the mechanical integrity of cells in the ...

Quantitative Detection of Trace Explosive Vapors by Programmed Temperature Desorption Gas Chromatography-Electron Capture Detector

定量检测痕量爆炸物蒸气通过程序升温脱附 - 气相色谱 - 电子捕获检测器

The direct liquid deposition of solution standards onto sorbent-filled thermal desorption tubes is used for the quantitative analysis of trace explosive ...

Use of Stopped-Flow Fluorescence and Labeled Nucleotides to Analyze the ATP Turnover Cycle of Kinesins

利用停流荧光和标记的核苷酸来分析驱动蛋白的ATP周转周期

The kinesin superfamily of microtubule associated motor proteins share a characteristic motor domain which both hydrolyses ATP and binds microtubules. ...

Radiolabeling and Quantification of Cellular Levels of Phosphoinositides by High Performance Liquid Chromatography-coupled Flow Scintillation

放射性标记和磷酸肌醇的细胞水平的定量高效液相色谱耦合流动闪烁

Phosphoinositides (PtdInsPs) are essential signaling lipids responsible for recruiting specific effectors and conferring organelles with molecular ...

Facile Preparation of 4-Substituted Quinazoline Derivatives

的4-取代喹唑啉衍生物简便制备

Reported in this paper is a very simple method for direct preparation of 4-substituted quinazoline derivatives from a reaction between substituted ...

Using Capillary Electrophoresis to Quantify Organic Acids from Plant Tissue: A Test Case Examining Coffea arabica Seeds

Using Capillary Electrophoresis to Quantify Organic Acids from Plant Tissue: A Test Case Examining Coffea arabica Seeds

利用毛细管电泳从植物组织量化有机酸：一个测试用例检查<em&gt;小粒种咖啡</em&gt;种子

Carboxylic acids are organic acids containing one or more terminal carboxyl (COOH) functional groups. Short chain carboxylic acids (SCCAs; carboxylic ...

Raman and IR Spectroelectrochemical Methods as Tools to Analyze Conjugated Organic Compounds

拉曼光谱和红外光谱电化学法作为分析共轭有机化合物的工具

In the presented work, two spectroelectrochemical techniques are discussed as tools for the analysis of the structural changes occurring in the molecule ...

Using Cyclic Voltammetry, UV-Vis-NIR, and EPR Spectroelectrochemistry to Analyze Organic Compounds

利用循环伏安法、紫外-近红外光谱和 EPR 光谱电化学对有机化合物进行分析

Cyclic voltammetry (CV) is a technique used in the analysis of organic compounds. When this technique is combined with electron paramagnetic resonance ...

A Two-Step Pyrolysis-Gas Chromatography Method with Mass Spectrometric Detection for Identification of Tattoo Ink Ingredients and Counterfeit Products

一种用于检测纹身油墨成分和假冒产品的两步热解-气相色谱法

Tattoo inks are complex mixtures of ingredients. Each of them possesses different chemical properties which have to be addressed upon chemical analysis. ...