Free amino acids,biogenic amines,and ammonium profiling in tobacco from different geographical origins using microwave‐assisted extraction followed by ultra high performance liquid chromatography

This work describes a rapid, stable, and accurate method for determining the free amino acids, biogenic amines, and ammonium in tobacco. The target analytes were extracted with microwave‐assisted extraction and then derivatized with diethyl ethoxymethylenemalonate, followed by ultra high performance liquid chromatography analysis. The experimental design used to optimize the microwave‐assisted extraction conditions showed that the optimal extraction time was 10 min with a temperature of 60°C. The stability of aminoenone derivatives was improved by keeping the pH near 9.0, and there was no obvious degradation during the 80°C heating and room temperature storage. Under optimal conditions, this method showed good linearity (R ² > 0.999) and sensitivity (limits of detection 0.010–0.081 μg/mL). The extraction recoveries were between 88.4 and 106.5%, while the repeatability and reproducibility ranged from 0.48 to 5.12% and from 1.56 to 6.52%, respectively. The newly developed method was employed to analyze the tobacco from different geographical origins. Principal component analysis showed that four geographical origins of tobacco could be clearly distinguished and that each had their characteristic components. The proposed method also showed great potential for further investigations on nitrogen metabolism in plants.     

Ultrasound‐assisted extraction and preliminary purification of proanthocyanidins and chlorogenic acid from almond (Prunus dulcis) skin

An aqueous solution of polyethylene glycol (PEG) as a green solvent was employed for the first time to develop the ultrasound‐assisted extraction of proanthocyanidins (PA) and chlorogenic acid (CA) from almond skin. The optimized extraction parameters were determined based on response surface methodology, and corresponded to an ultrasound power of 120 W, a liquid‐to‐solid ratio of 20:1 (mL/g), and a PEG concentration of 50% (v/v). Under these optimized conditions, the extraction yields of PAs and CA from almond skin were 32.68 ± 0.22 and 16.01 ± 0.19 mg/g, respectively. Compared with organic solvent extraction, PEG solution extraction produced higher yields. Different macroporous resins were compared for their performance in purifying PAs and CA from almond skin extract. Static adsorption/desorption experimental results demonstrated that AB‐8 resin exhibits excellent purification performance at pH 4. Under the optimized dynamic adsorption/desorption conditions on the AB‐8 column, the total recovery of purification for PAs and CA was 80.67%. The total content of PAs and CA in the preliminarily purified extract was 89.17% (with respective contents of 60.90 and 28.27%).     

Separation of triacylglycerols and free fatty acids in microalgal lipids by solid-phase extraction for separate fatty acid profiling analysis by gas chromatography

Microalgal lipids were separated into two fractions, triacylglycerols (TAGs) and free fatty acids (FFAs), by solid-phase extraction employing sodium carbonate as the sorbent and dichloromethane (20% by volume) in n-hexane as the extracting solvent. The TAG fraction was then saponified, followed by acidification, extraction and tert-butyldimethylsilyl esterification. The FFA fraction was directly acidified, extracted and derivatized. From the lipid extracts of eight microalgal species examined, a total of 13 fatty acids were detected in the TAG fractions and nine were found in the FFA fractions, with at much higher total TAG content in all microalgae. Oleic acid was the most prominent fatty acid in three species, α-linolenic acid was more abundant in two others, and palmitic acid was present in highest concentration in the remaining three species.     

Territorial origin of olive oil: representing georeferenced maps of olive oils by NMR profiling

ABSTRACT Proton NMR profiling is nowadays a consolidated technique for the identification of geographical origin of food samples. The common approach consists in correlating NMR spectra of food samples to their territorial origin by multivariate classification statistical algorithms. In the present work, we illustrate an alternative perspective to exploit territorial information, contained in the NMR spectra, which is based on the implementation of a geographic information system (GIS). Nuclear magnetic resonance spectra are used to build a GIS map permitting the identification of territorial regions having strong similarities in the chemical content of the produced food (terroir units). These terroir units can, in turn, be used as input for labeling samples to be analyzed by traditional classification methods. In this work, we describe the methods and the algorithms that permit to produce GIS maps from NMR profiles and apply the described method to the analysis of the geographical distribution of olive oils in an Italian region. In particular, we analyzed by ¹H NMR up to 98 georeferenced olive oil samples produced in the Abruzzo Italian region. By using the first principal component of the NMR variables selected according to the Moran test, we produced a GIS map, in which we identified two regions incidentally corresponding to the provinces of Teramo and Pescara. We then labeled the samples according to the province of provenience and built an LDA model that provides a classification ability up to 99% . A comparison between the variables selected in the geostatistics and classification steps is finally performed. Copyright © 2016 John Wiley & Sons, Ltd.     

Determination of free fatty acids by capillary zone electrophoresis

Capillary zone electrophoresis was investigated for the separation of free fatty acids as an alternative to well established techniques such as GC or HPLC. The analysis was performed with indirect UV detection in a counterelectroosmotic flow mode using a diethylbarbiturate carrier electrolyte at a pH between 10 and 11 in a mixed aqueous-organic solvent. Separation of saturated and unsaturated fatty acids could be achieved after bromination of the double bonds. Problems with wall adsorption of fatty acids could be overcome by increasing the temperature and using a high concentration of a zwitterionic reagent to inactivate the silica surface. Increased sensitivity could be achieved after preconcentration on alumina. The method was applied to the determination of free fatty acids in dairy products. The advantages compared to traditional methods include short analysis times and simple preparation steps.Dedicated to Univ. Prof. Dr. Karl Winsauer on the occasion of his 70th birthday     

Marine macroalgae analyzed by mass spectrometry are rich sources of polyunsaturated fatty acids

Algae from cold water (Canada) and warm water (China) were analyzed for their total lipid content, and for their fatty acid (FA) composition and content. The major findings are that FA from Canadian algae are generally richer in polyunsaturated FA (PUFA), with a higher n-3/n-6 FA ratio, and a higher degree of total unsaturation. The 18 C, 4 double bonds FA (18 : 4 stearidonic acid, morotic acid as synonym) was detected in greater amounts in cold water samples. The high levels of total PUFA, and especially of n-3 FA in Canadian algae, suggests their possible utilizations for nutritional purposes.     

Densitometric determination of unsaturated fatty acids with p-hydroxybenzaldehyde by HPTLC

A rapid method for the detection and quantltatlve determlnation of unsaturated fatty acids by TLC is presented. After separation on HPTLC-RP₁₈-plates, reaction wlth p-hydroxybenzaldehyde is carried out by a dipping procedure. Nanogram amounts of unsaturated fatty acids can be determined with good reproducibility.     

Separation of dietary omega‐3 and omega‐6 fatty acids in food by capillary electrophoresis

A lower dietary omega‐6/omega‐3 (n‐6/n‐3) fatty acid ratio (<4) has been shown to be beneficial in preventing a number of chronic illnesses. Interest exists in developing more rapid and sensitive analytical methods for profiling fatty acid levels in foods. An aqueous CE method was developed for the simultaneous determination of 15 n‐3 and n‐6 relevant fatty acids. The effect of pH and concentration of buffer, type and concentration of organic modifier, and additive on the separation was investigated in order to determine the best conditions for the analysis. Baseline separations of the 15 fatty acids were achieved using 40 mM borate buffer at pH 9.50 containing 50 mM SDS, 10 mM β‐cyclodextrin, and 10% acetonitrile. The developed CE method has LODs of <5 mg/L and good linearity (R² > 0.980) for all fatty acids studied. The proposed method was successfully applied to the determination of n‐3 and n‐6 fatty acids in flax seed, Udo® oils and a selection of grass‐fed and grain‐fed beef muscle samples.     

气相色谱法直接测定工业油酸中脂肪酸

建立了气相色谱直接测定工业油酸中未衍生化的6种常见脂肪酸含量的检测方法。样品用四氢呋喃(THF)溶解,通过氢火焰离子化检测器(FID)对目标化合物进行分析,外标法定量。在优化的气相色谱条件下,6种脂肪酸实现了有效分离,6种脂肪酸在5~5000 mg/L范围内定量曲线的相关系数(R2)均大于0.99991,表明线性关系良好,方法检出限(S/N≥3)为0.36~0.58 mg/L,相对标准偏差为3.4%~8.1%,加标回收率在84.7%~110.2%之间。选取4种工业油酸样品进行分析,实验结果表明,样品未经衍生化处理直接进样,本方法在10 min内能够对6种脂肪酸实现基线分离,为工业油酸的品质鉴定提供了一种快速有效的检测方法。     

The structure and properties of hybrid fluorous-hydrocarbon fatty acids

A series of long chain fatty acids is prepared with alternating blocks of fluorocarbon and hydrocarbon. This hybrid construction provides three domains (fluorous, hydrocarbon, hydrophilic) on the same molecule. The ability of the monocarboxylic acids to form monolayers on water is explored and, in selected cases, X-ray structure analysis of the dicarboxylic acids provides information on the conformation and solid state organisation of these molecules. The hybrid molecules display surface behaviour and show packing distances which are intermediate between alkyl and perfluoroalkyl chains.     

GC-MS分析大叶金花草中挥发性成分和脂肪酸

应用 GC-MS 联用技术对大叶金花草(Stenoloma chusana(L.)Ching)的挥发性成分和脂肪酸进行了检测,共检测出7种挥发性成分和32种脂肪酸,脂肪酸检出量为87.65%,其中饱和脂肪酸占脂肪酸总量的53.13%,不饱和脂肪酸占脂肪酸总量的46.87%.该研究为大叶金花草的进一步开发利用提供了科学依据.     

多级溶剂萃取-气相色谱法测定啤酒中的游离脂肪酸

啤酒中含量极低、极不稳定的游离脂肪酸是影响啤酒风味重要因素之一。本文采用溶剂萃取和气相色谱联用技术,对啤酒不良风味物质游离脂肪酸进行了检测。啤酒样品经3次萃取(有机溶剂萃取、碱液反萃取、有机溶剂萃取)后,通过甲醇-硫酸甲酯化处理,最后用正己烷萃取浓缩进行气相色谱(GC)分析。该方法中以辛酸、癸酸和月桂酸为代表的平均回收率分别为86.26%、92.19%和83.57%。辛酸、癸酸、月桂酸、棕榈酸在2.326~2326 mg/L,0.362~362 mg/L,0.250~250 mg/L,0.330~330 mg/L范围内线性良好,相关系数在0.9996~0.9999之间。     

Interpretation of surface-tension isotherms of n-alkanoic (fatty) acids by means of the van der Waals model

Here we apply the two-dimensional van der Waals model to interpret surface-tension isotherms of aqueous solutions of n-alkanoic (fatty) acids. We processed available experimental data for a homologous series of eight acids, from pentanoic to dodecanoic (lauric). Only three adjustable parameters have been varied to fit simultaneously all experimental curves. Excellent agreement between the theoretical model and the experiment has been obtained. The determined parameter values comply well with the molecular properties and allow one to calculate the surfactant adsorption, surface elasticity, and the surface pressure vs area isotherms. For the dodecanoic acid, the van der Waals model indicates the existence of a surface phase transition.     

Production of glycerides from glycerol and fatty acids by native lipase ofNigella sativa seed

The possible application of native lipase ofNigella sativa seed in the esterification of fatty acids to glycerol was investigated, and the effect of process parameters and the enzyme selectivity on the reaction were determined. For this aim, the esterification of oleic acid, sunflower oil fatty acids, and coco oil fatty acids with glycerol were studied.     

A high yield method for the direct amidation of long-chain fatty acids

The amidation of long-chain fatty acids is the key step for preparing surfactants with excellent interfacial activity. Gas chromatography–mass spectrometry was employed to detect the reactants and products in the direct amidation reactions. The conversion and the concentration of the amides in the reaction process were also investigated to determine the best catalyst, the reaction rate constants, and activation energy. It was identified that the amidation reaction of the long-chain phenyl fatty acid was a zero-order reaction and 3,4,5-trifluorophenylboronic acid was the most effective catalyst by which the activation energy reduced to 55.79 kJ/mol from 95.44 kJ/mol. The method can be applied to other long-chain fatty acids, saturated or unsatureated. The turning-over-temperature was 156°C, over which high yields can be achieved without any catalyst. These provide a reference for the preparation of long-chain fatty acid amides.     

气相色谱法测定中华绒螯蟹中脂肪酸组成与含量

中华绒螯蟹中脂肪酸组成与含量的测定对评估其营养价值与品质具有重要意义,但面对种类繁多的脂肪酸提取试剂和甲酯化试剂,测定结果参差不齐,很难对中华绒螯蟹中丰富的脂肪酸准确定量。研究通过比较4种常见的脂肪提取试剂、2种脂肪酸甲酯化试剂,确定以氯仿-甲醇(1∶1, v/v)为提取试剂,含2%硫酸的甲醇溶液为甲酯化试剂,建立了测定中华绒螯蟹肌肉中脂肪酸组成与含量的气相色谱分析方法。实验按照程序升温的条件,采用DM-2560毛细管色谱柱(100 m×0.25 mm×0.20 μm)分离37种脂肪酸,氢火焰离子化检测器(FID)检测,外标法定量。37种脂肪酸在0.5~100.0 μg/mL范围内线性关系良好,其相关系数(R²)为0.9981~0.9999,检出限(LOD)与定量限(LOQ)分别为0.01~0.02 mg/100 g和0.04~0.06 mg/100 g;以棕榈酸和硬脂酸进行加标回收验证,在1、2、10 mg/100 g 3个加标水平下的加标回收率为76.0%~97.5%,相对标准偏差(RSD, n=5)为3.31%~7.90%。该方法应用于中华绒螯蟹肌肉中脂肪酸组成与含量的测定,肌肉中共测得31种脂肪酸,碳链长度为12~24,脂肪酸总含量为281.03 mg/100 g,其中油酸、二十二碳六烯酸、二十碳五烯酸等为中华绒螯蟹肌肉中主要脂肪酸。该方法操作简便,试剂、样品用量少,且定性可靠,定量准确,能检测较多的脂肪酸种类,适用于中华绒螯蟹肌肉中脂肪酸组成与含量的快速检测。     

Separation of regioisomers and enantiomers of triacylglycerols containing branched fatty acids (iso and/or anteiso)

A combination of two chromatographic and one enzymatic methods was used for identification of the molecular species of triacylglycerols (TAGs) from Streptomyces avermitilis. Streptomyces avermitliswas cultured on various carbon sources and the ratio of iso- (i-FAs), anteiso- (ai-FAs), and straight-chain- (n-FAs) fatty acids was modified by precursor-directed biosynthesis. Saturated TAGs were separated from other lipids (including TAGs containing unsaturated FAs) using Ag⁺ ion cartridges. Analysis of TAGs wereperformed by RP-HPLC/ESI⁺ tandem mass spectrometry. Both the synthetically prepared sn-TAGs and the natural mixture of TAGmolecular species of wereseparated and identified by tandem MS. The structures of synthetic TAGs werefurther confirmed by pancreatic lipase, which cleaves sn-TAGs into sn-2-monoacylglycerols. The retention times (tR) of the individual regioisomers and enantiomers were found to be depend on the structure of the TAGs. If one branched acyl (iso or anteiso) is present in the TAG molecule, then the elution order is enantiomer (n/n/br), opposite enantiomer (br/n/n), regioisomer (n/br/n). In the case where two branched acyls are in the TAG molecule, the order of the elution is different, that is, br/n/br, n/br/br, br/br/n. In all cases, it was further demonstrated that tandem MS of either synthetically prepared TAGs or TAGs obtained from natural material, that is, n-16:0/ai-15:0/n-16:0 and i-16:0/n-15:0/i-16:0 are identical. Unfortunately, it is not possible to distinguish by ESI⁺ tandem MS such TAGs, which differ only in the branching of the acyls. The results of our analyses of TAGs are in good agreement with previously published data in other streptomycetes.     

Untargeted metabolomics liquid chromatography-high resolution mass spectrometry approach for the geographical origin assessment of Italian dehydrated apples

Geographical provenience is nowadays a relevant aspect of the authenticity and the quality of many food commodities. Dehydrated apple cubes/slices represent an ingredient commonly used by food companies for bakery products. However, this apple-based matrix is not so known and studied from an analytical point of view. In the present work, seven compounds were identified as key molecules to distinguish between Italian and non-Italian samples, through an untargeted ultrahigh-pressure liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS) approach. This methodology was merged with multivariate statistical analysis, and the principal features were studied and identified considering several identification steps. Samples from 2020 and 2021 harvesting campaigns, with partial and total dehydration rates, with or without peel, and from different apple varieties were considered for the study, for a total of 91 samples. Afterward, the same analysis protocol was applied to an external set (n = 12 samples), included in the statistical models, searching for the key compounds identified in the training set. Interesting and significant results underlined the potentiality of the UHPLC-HRMS technology as a confirmatory strategy for the geographical origin assessment of dehydrated apple commodities.