Name: nmr spectroscopy as a robust tool for the rapid evaluation of the lipid profile of fish oil supplements
Uploaded: 2017-05-01T11:45:00
Description: Watch this Scientific Journal Video about NMR Spectroscopy as a Robust Tool for the Rapid Evaluation of the Lipid Profile of Fish Oil Supplements at JoVE.com

この作品は、オハイオ州立大学の健康ディスカバリーテーマのための食品とオハイオ州立大学食品科学省によってサポートされていました。著者は、ペンシルベニア州立大学でオハイオ州立大学のNMR施設とNMR施設に感謝したいと思います。

1 NMRサンプル調製注意：注意は、使用する前に、関連するすべての物質安全データシート（MSDS）を参照してください。試料調製に使用される重水素化クロロホルム（CDCl 3）は有毒です。ヒュームフードや個人用保護具の使用を含む試料の調製を行うときに、すべての適切な安全対策を使用してください（安全眼鏡、手袋、白衣、完全長ズボン、靴、つま先を閉じ?...

<ol>
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B., 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=Standardless+1H+NMR+determination+of+pharmacologically+active+substances+in+dietary+supplements+and+medicines+that+have+been+illegally+traded+over+the+internet.">Standardless 1H NMR determination of pharmacologically active substances in dietary supplements and medicines that have been illegally traded over the internet.</a> Drug Test. Anal. 5 (6), 400-411 (2013).</li><li>Berger, S., Braun, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> 200 and more NMR experiments: a practical course. , (2004).</li><li>Knothe, G., Kenar, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+the+fatty+acid+profile+by+1H-NMRspectroscopy.">Determination of the fatty acid profile by 1H-NMRspectroscopy.</a> Eur. J. Lipid Sci. Technol. 106, 88-96 (2004).</li><li>Sacchi, R., Medina, J. I., Aubourg, S. P., Paolillo, I. G. L., Addeo, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+High-Resolution+13C+NMR+Analysis+of+Lipids+Extracted+from+the+White+Muscle+of+Atlantic+Tuna+(Thunnus+alalunga).">Quantitative High-Resolution 13C NMR Analysis of Lipids Extracted from the White Muscle of Atlantic Tuna (Thunnus alalunga).</a> J. Agric. Food Chem. 41 (8), 1247-1253 (1993).</li><li>Dais, P., Misiak, M., Hatzakis, E. <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+marine+dietary+supplements+using+NMR+spectroscopy.">Analysis of marine dietary supplements using NMR spectroscopy.</a> Anal. Methods. 7 (12), 5226-5238 (2015).</li><li>Pickova, J., Dutta, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cholesterol+Oxidation+in+Some+Processed+Fish+Products.">Cholesterol Oxidation in Some Processed Fish Products.</a> J. Anal. Oil Chem. Soc. 80 (10), 993-996 (2003).</li><li>Siddiqui, N., Sim, J., Silwood, C. J. L., Toms, H., Iles, R. 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P., Gao, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Z-restored+spin-echo+13C+1D+spectrum+of+straight+baseline+free+of+hump,+dip+and+roll.">Z-restored spin-echo 13C 1D spectrum of straight baseline free of hump, dip and roll.</a> Magn. Reson. Chem. 46, 432-435 (2008).</li><li>Tengku-Rozaina, T. M., Birch, E. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Positional+distribution+of+fatty+acids+on+hoki+and+tuna+oil+triglycerides+by+pancreatic+lipase+and+13C+NMR+analysis.">Positional distribution of fatty acids on hoki and tuna oil triglycerides by pancreatic lipase and 13C NMR analysis.</a> Eur. J. Lipid Sci. Technol. 116 (3), 272-281 (2014).</li><li>Berry, S. E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Triacylglycerol+structure+and+interesterification+of+palmitic+and+stearic+acid-rich+fats:An+overview+and+implications+for+cardiovascular+disease.">Triacylglycerol structure and interesterification of palmitic and stearic acid-rich fats:An overview and implications for cardiovascular disease.</a> Nutr. Res. Rev. 22 (1), 3-17 (2009).</li><li>Hunter, J. 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トラブルシューティングのための変更と戦略 スペクトル品質 。 NMR信号の線幅及びNMRスペクトルの分解能は、従って、磁場の均一性を最適化するためのプロセスである、シミングに大きく依存します。ルーチン分析のために、1Dシミングが適切であると3Dシミングは、それが定期的にNMR員によって行われることを考えると、必要とされません。そうでない場合、...

食餌中のn-3脂肪酸の摂取は、心疾患1、2、3、4、炎症性疾患および糖尿病5のように、いくつかの状態に対して有益であることが証明されています。西洋食は、n -3脂肪酸の貧しい考えられているので、魚油サプリメントの消費量は、nを改善することが推奨され-6 / N -3消費者の栄養1のバランス。魚油サプリメントの消費量の最近の増加にもかかわらず、質問は、これらの製品のいくつかの安全性、信頼性、および品質について残ります。魚油サプリメントの迅速かつ正確な組成分析は、適切にこれらの商用製品の品質を評価し、消費者の安全を確保するために不可欠です。 魚油サプリメントを評価するための最も一般的な方法論Sは、ガスクロマトグラフィー（GC）及び赤外分光法（IR）です。これらは非常に敏感な方法ですが、彼らはいくつかの欠点がある6。 GC分析は、個々の化合物の分離および誘導体が7必要であり、脂質酸化を分析8,9の間に発生する可能性があるため、時間がかかり（4~8時間）です。 IR分光法を定量することができるがIRバンドは、単一化合物10に起因することが可能な例外はあるが、予測モデルは、部分最小二乗回帰（PLSR）を使用して構築する必要があります。 PLSR解析11の時間を増大させる多数のサンプルの分析を必要とします。このため、魚油多数のサンプルの正確かつ迅速な分析を可能にする新しい分析手法の開発に関心が高まっています。このようオフィとしての組織国立衛生研究所（NIH）と食品医薬品局（FDA）でのサプリメントのCE（ODS）は、これらの新しいメソッド12、13を開発するための公式分析化学者協会（AOAC）と協力してきました。 スクリーニングおよびそのような栄養補助食品などの多成分マトリックスを評価するための最も有望な分析方法の一つは、核磁気共鳴（NMR）分光法14、15です。 NMR分光法は、いくつかの利点を有する：それは非破壊的かつ定量的な技術は、それはサンプル調製に最小限必要であり、優れた精度と再現性を特徴とします。それは、溶媒を少量しか使用しているためまた、NMR分光法は、環境に配慮した方法論です。 NMR分光法の主な欠点は、他のanalytiと比較して相対的に低い感度でありますCALの方法は、しかし、そのような強い磁場、種々の直径の極低温プローブ、高度なデータ処理、及び多目的パルスシーケンス及び技術として計測における最近の技術進歩は、nM範囲まで感度が増加しています。 NMR計測は、高コストではあるが、NMR分光計の長寿命およびNMRの多くのアプリケーションは、長期的には、分析のコストを下げます。この詳細なビデオプロトコルは、フィールドに新しい実践者が1 Hおよび魚油サプリメントの13の C NMR分光分析に関連した問題を回避するためのものです。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Avance III 850 NMR instrument</td>
			<td>Bruker</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Avance III 500 NMR instrument</td>
			<td>Bruker</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>TCI 5mm probe</td>
			<td>Bruker</td>
			<td></td>
			<td>Helium cooled inverse (proton deetected) NMR probe featuring three independent channels (1H, 13C, 15N)</td>
		</tr>
		<tr>
			<td>BBO prodigy 5mm probe</td>
			<td>Bruker</td>
			<td></td>
			<td>Nitrogen cooled observe (X-nuclei detected) probe, featuring two channels; one for 1H and 19F detectionand one for X-nuclei (covering from 15N to 31P)</td>
		</tr>
		<tr>
			<td>Spinner turbin</td>
			<td>Bruker</td>
			<td></td>
			<td>NMR spinners are made by polymer materials and they have a rubber o-ring to hold the NMR tube securely in place</td>
		</tr>
		<tr>
			<td>Topspin 3.5</td>
			<td>Bruker</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>deuterated chloroform</td>
			<td>Sigma-Aldrich</td>
			<td>&#160;865-49-6</td>
			<td>99.8 atom % D, contains 0.03 TMS</td>
		</tr>
		<tr>
			<td>2,6-Di-tert-butyl-4-methylphenol,</td>
			<td>Sigma-Aldrich</td>
			<td>&#160;128-37-0</td>
			<td>purity &#62;99%</td>
		</tr>
		<tr>
			<td>Fish oil samples</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>NMR tubes</td>
			<td>New Era</td>
			<td>NE-RG5-7</td>
			<td>5mm OD Routine &#8220;R&#8221; Series NMR Sample Tube</td>
		</tr>
		<tr>
			<td>BSMS</td>
			<td>Bruker</td>
			<td></td>
			<td>Bruker Systems Management System; control system device</td>
		</tr>
	</tbody>
</table>

nmr spectroscopy as a robust tool for the rapid evaluation of the lipid profile of fish oil supplements

西洋の食事は、したがって、魚油サプリメントの消費はこれらの必須栄養素の摂取量を増やすことが推奨され、nは -3脂肪酸に乏しいです。この研究の目的は、二つの異なるNMR器具を利用した高分解能1 H及び13 C NMR分光法を用いてカプセル化された魚油サプリメントの定性・定量分析を実証することです。 500 MHzおよび850 MHz装置。両方のプロトン（1 H）および炭素（13 C）NMRスペクトルは、魚油サプリメントの主要な構成成分の定量的決意のために使用することができます。魚油サプリメント中の脂質の定量は、当該1Dスペクトルに適切なNMRシグナルの積分によって達成されます。 1 H及び13 C NMRにより得られた結果は、2つの核と2つの機器間の分解能と感度の差にもかかわらず、互いによく一致しています。 1 H NMRプランスペクトルは1時間に10分から続く13の C NMR分析とは対照的に、1分未満に記録することができるようにSAより迅速な分析は、13 C NMRに比べ。 13 C NMRスペクトルは、しかし、はるかに有益です。個々の脂肪酸のより大きな数の定量的データを提供することができ、グリセロール骨格上の脂肪酸の位置分布を決定するために使用することができます。両方の核を精製または分離工程を必要とせずにただ1つの実験において定量的情報を提供することができます。磁界の強度はほとんどが13 C NMRに対するその低い解像度に1 H NMRスペクトルに影響を与える、しかし、低コストのNMR機器を効率的に食品業界および品質管理研究所による標準的な方法として適用することができます。

1 H及び13の C NMRスペクトルは、二つのNMR機器を用いて、市販の魚油サプリメントのために収集しました。 850 MHzおよび500 MHzの分光計。これらのスペクトルは、ドコサヘキサエン酸（DHA）とエイコサペンタエン酸（EPA）、例えばNなどのn -1アシル鎖及び栄養的に重要な指標として、他の化合物などの魚油の成分の定量的決意のために使用す?...

Watch this Scientific Journal Video about NMR Spectroscopy as a Robust Tool for the Rapid Evaluation of the Lipid Profile of Fish Oil Supplements at JoVE.com

NMR Spectroscopy as a Robust Tool for the Rapid Evaluation of the Lipid Profile of Fish Oil Supplements

ここで、高分解能1 H及び13 C核磁気共鳴（NMR）分光法は、カプセル化された魚油サプリメントの定量的および定性的分析のための迅速かつ信頼性の高いツールとして使用しました。

魚油サプリメントの脂質プロファイルの迅速な評価のための強力なツールとしてNMR分光法

The western diet is poor in n-3 fatty acids, therefore the consumption of fish oil supplements is recommended to increase the intake of these essential ...

The overall goal of this method is to determine the lipid profile of fish oil supplements using Nuclear Magnetic Resonance spectroscopy. This method can help answer key questions for lipid analysis such as the lipid contents of fish oil supplements and the positional distribution of various fatty acids on the glycerol skeleton. The main advantage of this technique is that it makes the rapid and robust analysis of several different molecules feasible in one snapshot without using separation and purification steps.To prepare the samples for proton and carbon NMR analysis, first extract 120 microliters, which is around 110 milligrams of fish oil, from a dietary capsule using a syringe and place it in a 4-mL glass vial. Record the weight of the fish oil. Use 500 microliters of a BHT stock solution to dissolve 100 milligrams plus or minus 10 milligrams of fish oil.After dissolving the oil, transfer all of the solution directly into a high quality, five millimeter NMR tube and attach a cap. Analyze the samples within 24 hours of preparing the samples. Insert the NMR tube into a spinner turbine.Place the spinner and the tube on the top of a graded depth gauge and gently push the top of the tube until its bottom part touches the bottom of the gauge. Then place the NMR tube in an open spot of the sample case. Note the slot number the sample is placed in.To load the sample in the NMR instrument, return to the control computer and type SX-number, where number is the slot in the sample case holding the sample. Wait for the deuterium signal of deuterated chloroform to appear on the screen. As soon as the deuterium signal is visible, type lock"on the command line and select deuterated chloroform from the solvents list to lock the sample using its deuterium resonance.Type bsmsdisp"in the command line to ensure spinning is not active. If the spin button is green, click it to deactivate spinning. After typing the command new"to create a new data set, enter a name for the data set in the name tab and the experiment number in the experiment number tab titled EXPNO.Use number one in the process number tab titled PROCNO. Write the title of the experiment in the TITLE tab. In the Experiment tab, hit Select and choose the C13IG parameter file.Then hit Set selected item in editor"and click OK.Type getprosol"in the command line to obtain the standard parameters for the current NMR probe and solvent. Then type the command atma"to perform automatic tuning and to matching of the probe for both carbon and proton nuclei. Next, perform one-dimensional gradient shimming to achieve a highly homogeneous magnetic field and thus optimum line shape for the NMR signals.To do so, use the standard automatic procedure for 1D shimming simply by sequently executing the commands qu topshim 1dfast ss;qu topshim tuneb ss;and qu topshim report"in the command line. After parameter optimization as described in the text protocol, acquire one dimensional carbon NMR spectra. To do so, go to the carbon data set and use an inverse gated decoupled pulse sequence by typing pulprog zgig"in the command line.Type commands on the command line to set up the spectral with nppm, the center of the RF transmitter, the number of scans, the number of dummy scans, the number of data points, and the pulse duration for a ninety degree pulse angle. Type digmod baseopt"in the command line to acquire a spectrum with an improved base line. Then set a relaxation delay of 60 seconds for the 850 megahertz instrument by typing di 60s"in the command line.Set the receiver gain to an appropriate value using the command rga"for automatic calculation of receiver gain. Finally, start the acquisition by typing the pulse acquire command zg"in the command line. To process the data, first type si 64K"in the command line to apply zero filling and set the size of the rail spectrum to 64K.Then set the line broadening parameter to 1.0 hertz by typing lb 1.0"in the command line to apply a waiting function with a line broadening factor of 1.0 hertz prior to 4ea transform. Execute 4ea transformation by typing efp"in the command line. Next perform automatic phase correction by typing the command apk"in the command line.If additional phase adjustments are required to further improve the spectrum, click on the Process tab. Then click on the Adjust Phase icon and the Phase Correction icons for zero order and first order phase correction. While clicking on the zero order and first order phase correction icons, drag the mouse until all of the signals are in positive absorption mode.Apply and save the phase correction values by clicking the Return and Save button to exit the Phase Correction mode. Apply a polynomial forth order function for baseline correction upon integration by typing the command abs n"in the command line. To report chemical shifts in ppm from TMS, zoom in on the TMS signal, then click on the Calibration icon and place the cursor with the red line on top of the NMR signal to be referenced, TMS in this case.To adjust the threshold intensity, use the middle mouse button if needed. Left click and type in 0. When using BHT as an internal standard, integrate the peak at delta 151.45 extending five hertz from each side of the peak.Then set the integral equal to the micromoles of BHT per 0.5 milliliters of the stock solution. Integrate the peaks of interest extending fiver hertz from each side of the peak. If there is a need to focus on a region, click on the highlight icon to deactivate it.Then left click and drag to zoom in on the region. Click on the highlight icon again to make the integration function active and move to the next peak. Left click and drag the cursor through the integral.Click the Return Save Regions button. Shown here is a representative spectrum of the Carbon NMR analysis in the carbonyl carbon region. The NMR signals of EPA and DHA on the sn-1, 3 and sn-2 positions are shown.These signals can be used for the quantitative determination of EPA and DHA. Shown here is a representative spectrum of the proton NMR analysis. The NMR signals of EPA and DHA that can be used for their determination are shown.The proton NMR spectrum is characterized by a narrower spectral width as compared to the carbon NMR spectrum and thus a lower spectral resolution. Once mastered, this technique can be done in less than 30 minutes if it is performed properly. After its development, this technique paved the way for researchers in the field of lipid analysis to rapidly analyze various lipid components that appear in multi-component matrices, such as dietary supplements, vegetable oil, and biofuels.After watching this video, you should have a good understanding of how to apply proton and carbon, NMR spectroscopy for the analysis of the lipid profile of fish oil supplements. Don't forget that working with strong magnetic fields produced by NMR spectrometers can be extremely hazardous to pacemakers and surgical protheses as well as electronic items such as credit cards and watches.

Research

JoVE Journal

Chemistry

Isolation and Chemical Characterization of Lipid A from Gram-negative Bacteria

グラム陰性菌からの単離とリピドAの化学特性評価

Lipopolysaccharide (LPS) is the major cell surface molecule of gram-negative bacteria, deposited on the outer leaflet of the outer membrane bilayer. LPS ...

化学

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Copper (I) binding by metallochaperone transport proteins prevents copper oxidation and release of the toxic ions that may participate in harmful redox ...

Methods to Identify the NMR Resonances of the 13C-Dimethyl N-terminal Amine on Reductively Methylated Proteins

Methods to Identify the NMR Resonances of the 13C-Dimethyl N-terminal Amine on Reductively Methylated Proteins

Nuclear magnetic resonance (NMR) spectroscopy is a proven technique for protein structure and dynamic studies. To study proteins with NMR, stable magnetic ...

HKUST-1 as a Heterogeneous Catalyst for the Synthesis of Vanillin

HKUST-1バニリンの合成のための不均一触媒として、

Vanillin (4-hydoxy-3-methoxybenzaldehyde) is the main component of the extract of vanilla bean. The natural vanilla scent is a mixture of approximately ...

Spin Saturation Transfer Difference NMR (SSTD NMR): A New Tool to Obtain Kinetic Parameters of Chemical Exchange Processes

Spin Saturation Transfer Difference NMR SSTD NMR: A New Tool to Obtain Kinetic Parameters of Chemical Exchange Processes

化学Exchangeプロセスの動力学的パラメーターを取得するために新しいツール：飽和移動差NMR（SSTD NMR）をスピン

This detailed protocol describes the new Spin Saturation Transfer Difference Nuclear Magnetic Resonance protocol (SSTD NMR), recently developed in our ...

Insights into the Interactions of Amino Acids and Peptides with Inorganic Materials Using Single-Molecule Force Spectroscopy

単一分子力分光法を用いて無機材料とアミノ酸とペプチドの相互作用への洞察

The interactions between proteins or peptides and inorganic materials lead to several interesting processes. For example, combining proteins with minerals ...

Coulomb Explosion Imaging as a Tool to Distinguish Between Stereoisomers

立体異性体を区別するためのツールとしてクーロン爆発イメージング

This article shows how the COLTRIMS (Cold Target Recoil Ion Momentum Spectroscopy) or the "reaction microscope" technique can be used to distinguish ...

Electrochemical Impedance Spectroscopy as a Tool for Electrochemical Rate Constant Estimation

電気化学インピー ダンス分光電気化学定数推定法のためのツールとして

Electrochemical impedance spectroscopy (EIS) was used for advanced characterization of organic electroactive compounds along with cyclic voltammetry (CV). ...

Color Spot Test As a Presumptive Tool for the Rapid Detection of Synthetic Cathinones

合成 Cathinones の急速な検出のための推定ツールとして色スポット テスト

Synthetic cathinones are a large class of new psychoactive substances (NPS) that are increasingly prevalent in drug seizures made by law enforcement and ...

A New Straightforward Method for Lipophilicity (logP) Measurement using 19F NMR Spectroscopy

A New Straightforward Method for Lipophilicity logP Measurement using 19F NMR Spectroscopy

新しい簡単な19F NMR 分光学を使用して親油性 (logP) 測定法

Fluorination has become an effective tool to optimize physicochemical properties of bioactive compounds. One of the applications of fluorine introduction ...