HPLC-Q-TOF-MS-MS测定桑椹中多酚类物质

采用高效液相色谱与四极杆飞行时间串联质谱（high performance liquid chromatography of quadrupole time of flight-tandem mass spectrometry,HPLC-Q-TOF-MS-MS）联用技术定性检测桑椹中多酚类物质,新鲜桑椹样品经体积分数80%丙酮溶液超声辅助提取15 min后,采用C18固相萃取小柱分离纯化,纯化后的样品进行质谱鉴定。采用HPLC-Q-TOF-MS-MS对多酚进行分析：初步鉴定了桑椹中存在14 种多酚类物质,主要以酚酸、花色苷和黄酮的形式存在。其中6 种酚酸：3-O-咖啡酰奎宁酸、4-O-咖啡酰奎宁酸、二聚绿原酸、二聚4-O-咖啡酰奎宁酸、绿原酸顺式异构体、3,5-O-二咖啡酰基奎宁酸。4 种花色苷：飞燕草-3-半乳糖苷、飞燕草-3-葡萄糖苷、矢车菊-3-葡萄糖苷、矢车菊-3-芸香糖苷;3 种黄酮：芦丁、鞣花酸己糖苷和槲皮素3-O-（6’-O-丙二酰）葡萄糖苷,1 种白藜芦醇衍生物。HPLC-Q-TOF-MS-MS可以鉴定出桑椹的多酚类物质。     

Comparison between ultra-homogenisation and ultrasound for extraction of phenolic compounds from teff (Eragrostis tef (Zucc.))

This study investigated the effect of the solvent composition on the extraction of phenolic compounds from teff grains and compared the efficiency between the techniques homogeniser-assisted extraction (HAE) and ultrasound-assisted extraction (UAE). The solvents used were water, ethanol and methanol, and the response analysed was the total phenolic content (TPC). The quantitative profile of polyphenols extracted was also determined by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-TQD). According to the optimisation, the maximum response was estimated at solvent ratios of 51:40:9 and 49:26:25 (water/ethanol/methanol, v/v/v) for HAE and UAE, respectively. Under optimal conditions, the extraction technique did not significantly influence the TPC extracted, but the TFC (total flavonoid content) and the antioxidant capacity were significantly higher when UAE was used. The phenolic acids p-coumaric and protocatechuic and the flavonoids quercetin, rutin and myricetin were the main polyphenols extracted. Both techniques studied were efficient in extracting polyphenols from whole teff grains.     

沙棘果油提取工艺的正交试验优化及其脂肪酸组分测定

利用超声波辅助索氏提取技术对沙棘果肉提取工艺进行优化,并采用气相色谱-飞行时间质谱技术,考察果油脂肪酸组分。通过进行提取温度、回流时间、料液比、超声时间4 个因素的单因素试验和正交试验,结果表明：沙棘果油的最佳提取条件为料液比1∶70（g/mL）、提取温度55 ℃、超声辅助提取30 min、索氏提取回流时间6 h,沙棘果油提取率为28.14%,其中共检测出24 种脂肪酸,其中主要包括棕榈油酸和棕榈酸,相对含量分别为35.56%和59.37%。因此利用超声波辅助索氏法提取沙棘果油,可以为医药和工业生产提供更优质和丰富的原材料。     

Isolation of Anticomplementary Substances from Cucurbita Moschata Duch

Chloroform and ethyl acetate extracts of the edible part of Cucurbita moschata Duch, which is commonly used as an oriental medicine as well as a popular food source, showed significant anticomplementary activities on the classical pathway of the complement system. Bioassay‐guided chromatographic separation using silica gel, Sephadex LH‐20, octadecyl silica gel, thin layer chromatography, and high performance liquid chromatography was performed to isolate the anticomplementary substances. Among the isolated substances, apigenin, phytosterols, and mixture of fatty acids, which were identified by nuclear magnetic resonance and gas chromatography‐mass spectrometry, presented the strongest activities. Especially, phytosterols and mixture of saturated fatty acids isolated from the ethyl acetate extracts had 0.74 and 0.67 mg/ml as IC₅₀, respectively, indicating that the ethyl acetate extraction is the best way to isolate the anticomplementary substances from Cucurbita moschata Duch.     

Quantitative Analysis of Bioactive Compounds from Aromatic Plants by Means of Dynamic Headspace Extraction and Multiple Headspace Extraction‐Gas Chromatography‐Mass Spectrometry

Seven monoterpenes in 4 aromatic plants (sage, cardamom, lavender, and rosemary) were quantified in liquid extracts and directly in solid samples by means of dynamic headspace‐gas chromatography‐mass spectrometry (DHS‐GC‐MS) and multiple headspace extraction‐gas chromatography‐mass spectrometry (MHSE), respectively. The monoterpenes were 1st extracted by means of supercritical fluid extraction (SFE) and analyzed by an optimized DHS‐GC‐MS. The optimization of the dynamic extraction step and the desorption/cryo‐focusing step were tackled independently by experimental design assays. The best working conditions were set at 30 °C for the incubation temperature, 5 min of incubation time, and 40 mL of purge volume for the dynamic extraction step of these bioactive molecules. The conditions of the desorption/cryo‐trapping step from the Tenax TA trap were set at follows: the temperature was increased from 30 to 300 °C at 150 °C/min, although the cryo‐trapping was maintained at ?70 °C. In order to estimate the efficiency of the SFE process, the analysis of monoterpenes in the 4 aromatic plants was directly carried out by means of MHSE because it did not require any sample preparation. Good linearity (r² > 0.99) and reproducibility (relative standard deviation % <12) was obtained for solid and liquid quantification approaches, in the ranges of 0.5 to 200 ng and 10 to 500 ng/mL, respectively. The developed methods were applied to analyze the concentration of 7 monoterpenes in aromatic plants obtaining concentrations in the range of 2 to 6000 ng/g and 0.25 to 110 μg/mg, respectively.     

Original article: Extraction of lipid from scallop (Patinopecten yessoensis) viscera by enzyme‐assisted solvent and supercritical carbon dioxide methods

In the present study, lipid was extracted from scallop (Patinopecten yessoensis) viscera by using the enzyme‐assisted solvent method and the supercritical carbon dioxide (SC‐CO₂) method. Soxhlet extraction with ethyl ether produced a yield of 23.7 ± 0.6 g of lipid 100 g^?1 of dry matter. Enzyme‐assisted solvent extraction allowed recovering 60.6 ± 1.5% of P. yessoensis viscera lipid from the samples treated with papain, whereas a lipid recovery rate of 78.3 ± 0.6% was achieved by SC‐CO₂ extraction. The lipid extracted was divided into the unsaponifiable fraction (sterol) and the saponifiable fraction (fatty acid) and analysed by gas chromatography mass spectrometry. Results indicated that the fatty acid composition and sterol composition for lipids extracted by different methods were slightly different. Eicosapentaenoic acid and docosahexaenoic acid were dominant polyunsaturated fatty acids accounting for 35–40% of the total fatty acid.     

Identification of free fatty acids and some other volatile flavour compounds from Swiss cheese using on-line supercritical fluid extraction-gas chromatography

On-line supercritical CO₂ extraction - gas chromatography was applied to the isolation and identification of free fatty acids and other volatile compounds of young and ripe Swiss cheese (Emmental) produced in Finland. Extractions were carried out using a micro-cartridge at 40 °C temperature and 10 MPa pressure and the volatile fractions were analysed by DB-WAX column (polyethylene glycol phase) with flame ionization detection and mass spectrometry. The total time of analysis was less than 2.5 h. Acetic acid and propionic acid predominated over the C12–C18 acids, the longer chain fatty acids increasing in concentration during the ripening of the cheese. Due to the high proportions of fatty acids, further fractionation is required for analysis of the less abundant aroma compounds such as alcohols, carbonyls and lactones.     

微波辅助提取棉籽油及其脂肪酸组成分析

以棉仁为原料,采用微波辅助提取法提取棉籽油,通过单因素试验和正交试验,研究了提取剂、微波功率、料液比、提取时间对棉籽得油率的影响,确定了棉籽油的最佳提取条件,并采用气相色谱-质谱法(GC-MS)分析了棉籽油的脂肪酸组成。结果表明,棉籽油提取的最佳条件为:提取剂为正己烷、微波功率500W、料液比1:7、提取时间5min,在该条件下棉籽油得率为32.50%。气相色谱-质谱分析表明,棉籽油含有10种脂肪酸,其中主要含有棕榈酸37.82%、亚油酸44.24%、油酸11.61%,还含有锦葵酸0.30%。     

梅花鹿鹿油的提取及成分检测

采用超临界CO2提取法从梅花鹿(Cervus nippon)鹿脂肪中提取鹿油,探讨提取工艺对鹿油提取率的影响。以鹿脂肪为原料,通过单因素试验和正交试验分别研究提取温度、提取压力、提取时间以及CO2流量对鹿油提取率的影响。研究结果表明,最佳的提取工艺为:提取温度45℃、提取压力40 MPa、提取时间150 min、CO2流量14 L/h。此时鹿油的提取率为44.81%。通过气相色谱-质谱法(gas chromatography mass spectrometry,GC-MS)联用对鹿油的成分进行检测,确定其含有16种脂肪酸,其中饱和脂肪酸和不饱和脂肪酸相对含量分别为69.840 9%和30.159 1%,主要脂肪酸有十六烷酸、十八烷酸及十八碳烯酸。     

Comparison of Gender Differences in Nutritional Value and Key Odor Profile of Hepatopancreas of Chinese Mitten Crab (Eriocheir Sinensis)

The nutritional value and key odor profile of hepatopancreas of Chinese mitten crab (Eriocheir sinensis) was evaluated, and gender differences in terms of edible yield, proximate composition, different lipid fractions, fatty acid composition, and key odor compounds were compared. Total lipids were separated into different lipid fractions using silicic acid columns. And odorants were detected by monolithic material sorptive extraction coupled with gas chromatography‐mass spectrometry‐olfactometry (GC‐MS‐O). Furthermore, detected frequency and odor intensity method of GC‐O were applied to select key odor compounds in steamed E. sinensis. The results showed male crabs (34.06%) had a higher edible yield than female crabs (32.42%). Hepatopancreas was the most abundant portion in both genders, which contained high lipid content. Higher contents of triacylglycerols, sterols, free fatty acids, and phosphatidylethanolamine in female crabs resulted in a significant higher total lipid content in females (40.05%) than in males (37.94%). Meanwhile, total monounsaturated fatty acids and polyunsaturated fatty acids of female crabs had significant higher content than male crabs, and the ratio of docosahexaenoic acid to eicosapentaenoic acid of female crabs (1.26) showed higher value than male crabs (1.18), which indicated that female crabs had higher nutritional value than male crabs. For further analyses, unique odor compounds detected in female crabs showed a plant/vegetable‐like odor, while sweety, corn‐like odor in male crabs. These suggested that hepatopancreases of female crabs are healthier than male crabs for human consumption, however, they may be favored by different consumers individually due to their unique aroma profiles.     

Comparison of aromatic and phenolic compounds in cherry wines with different cherry cultivars by HS‐SPME‐GC‐MS and HPLC

To compare the influence of cultivars on the production of aroma compounds and polyphenols, eight cherry varieties (six sweet and two sour) were fermented into cherry wines, and their compositions were analysed. Results showed that by headspace solid microextraction coupled to gas chromatography–mass spectrometry (HS‐SPME‐GC‐MS), a total of twenty‐first aromatic compounds, mostly consisting of esters, acids, terpenoids and C₁₃‐norisoprenoids, were identified, and eleven polyphenols including five anthocyanins and six phenolic acids were quantified using HPLC. May Duck and Early Richmond wines contained significantly higher levels of phenolic acids (>42 mg L^?1) that were associated with high contents of chlorogenic and neochlorogenic acids and also relatively high amounts of octanoic acid and β‐damascenone (odour activity values ≥1000). Hongdeng, Zhifuhong and Bing wines were characterised by higher concentrations of anthocyanins (>41 mg L^?1) and also relatively higher levels of isopentyl acetate. Lapins and Governer Wcod wines were distinguished by lower concentrations of polyphenols and relatively higher contents of ethyl 3‐methylbutanoate.     

两种海藻物提取物的制备及脂肪酸GC-MS分析

制备三列凹顶藻和松节藻醇提物，采用气相色谱。质谱联用仪（GC／MS）对这两种海藻提取物脂肪酸进行了分析，各分离出17个和18个峰，鉴定了12种脂肪酸。用峰面积归一化法得出各类脂肪酸的相对百分含量。凹顶藻提取物中含饱和脂肪酸4种，总体百分含量为18．095％：单不饱和脂肪酸2种，总百分含量为3．910％；多不饱和脂肪酸种类较多，共有6种，总百分含量为7．437％。松节藻醇提物中4种饱和脂肪酸的含量为51．842％，2种单不饱和脂肪酸含量为13．124％，6种多不饱和脂肪酸含量为27．375％。     

AUTOMATED REPORTING ON THE QUALITY OF HOPS AND HOP PRODUCTS

The quality of a hop variety or a hop product can readily be assessed by a fully automated sequence of selective extraction, fractionation and quantitative analysis. To illustrate the elegance of the method, nine hop varieties and three hop extracts were compared with respect to the content of important marker compounds in the hop oils and of the hop acids. Supercritical fluid extraction at different densities of carbon dioxide was applied to extract selectively, the hop oils and the hop acids, respectively. The hop oils were further fractionated into an apolar and a polar fraction by solid phase extraction and consecutive elution with n-hexane and ethyl acetate. Separation and identification were achieved by capillary gas chromatography coupled to mass spectrometry. Myrcene, β;-caryophyllene, α;-humulene and β;-farnesene in the apolar fraction, linalool, undecan-2-one, tridecan-2-one and humuladienone in the polar fraction were selected for quantitative evaluation of the respective hop oils. Sulphur-containing compounds were revealed by capillary gas chromatography using sulphur-selective atomic emission detection. Complete separation and quantification of all hop α;-acids and β;-acids was effected by microemulsion electrokinetic chromatography coupled to diode array detection .     

Bioactive compounds present in the Mediterranean sofrito

Sofrito is a key component of the Mediterranean diet, a diet that is strongly associated with a reduced risk of cardiovascular events. In this study, different Mediterranean sofritos were analysed for their content of polyphenols and carotenoids after a suitable work-up extraction procedure using liquid chromatography/electrospray ionisation-linear ion trap quadrupole-Orbitrap-mass spectrometry (LC/ESI-LTQ-Orbitrap-MS) and liquid chromatography/electrospray ionisation tandem triple quadrupole mass spectrometry (LC/ESI-MS-MS). In this way, 40 polyphenols (simple phenolic and hydroxycinnamoylquinic acids, and flavone, flavonol and dihydrochalcone derivatives) were identified with very good mass accuracy (<2 mDa), and confirmed by accurate mass measurements in MS and MS² modes. The high-resolution MS analyses revealed the presence of polyphenols never previously reported in Mediterranean sofrito. The quantification levels of phenolic and carotenoid compounds led to the distinction of features among different Mediterranean sofritos according to the type of vegetables (garlic and onions) or olive oil added for their production.     

Exclusive quantification of methyl-branched fatty acids and minor 18:1-isomers in foodstuff by GC/MS in the SIM mode using 10,11-dichloroundecanoic acid and fatty acid ethyl esters as internal standards

Minor fatty acids (iso- and anteiso-fatty acids, vaccenic acid, elaidic acid) in foodstuff (seafood, milk, and dairy products) were analyzed by gas chromatography with electron ionisation mass spectrometry in the selected ion monitoring mode (GC/EI-MS-SIM). For this purpose, lipids were obtained by accelerated solvent extraction (ASE) and the fatty acid constituents were converted into methyl esters. Instead of the determination of the relative contribution of the minor fatty acids to the sum of all fatty acids detected (the so-called 100% method), we exclusively quantified the minor fatty acids, which was possible by using two types of internal standards (IS-1 and IS-2). For recovery checks during the extraction and/or the transesterification step we added the novel 10,11-dichloroundecanoic acid (DC-11:0) as IS-1. DC-11:0, which has never been detected in foodstuff, was synthesized by electrophilic addition of chlorine to 10-undecenoic acid (11:1n-10). The novel IS eluted in the range of 23:0 from the polar GC column used and showed the same properties as fatty acids in foodstuff during sample preparation. Recovery rate of DC-11:0 was generally >96% in the various samples analyzed. Ethyl esters (FAEE) of a12:0, a14:0, a15:0, a16:0, a17:0, and a18:0 (IS-2) were added to both the external standard (a quantitative mixture of methyl esters of methyl-branched fatty acids and 18:1n-9trans) in order to determine their response factors relative to FAEE and to the food samples. With this technique, (only) methyl-branched fatty acids (MBFAs) as well as vaccenic acid (18:1n-7) and elaidic acid (18:1n-9trans) were quantified in a range of dairy products (including twelve cheeses) and seafood. All samples were analyzed in triplicates, and good standard deviations (concentrations 0.002–5 g/100 g; standard deviations 0.00–0.03) were obtained in all cases. MBFAs were detected in all samples analyzed. The highest content of MBFAs (3.0 g/100 g) was determined in red-smear of romadur cheese. In all except two cheeses, i17:0 was the most abundant MBFA. The highest amount of 18:1n-9trans was found in feta (2.84 g/100 g) whereas 0.03 g/100 g in big eye snapper (Pricanthus tayenus) marked the lowest record of this minor fatty acid. Seal oil contained the highest amount of 18:1n-7 with 5.00 g/100 g, whereas emmental cheese was the sample with the lowest content of this monoenoic fatty acid. The combination of suitable IS and a sensitive GC/EI-MS-SIM method proved to be well suited for the quantification of minor fatty acids in foodstuff. When only a set of fatty acids is going to be analyzed, this method is less time consuming compared to “100% methods” and less prone to false results due to the higher selectivity of GC/MS compared to GC in combination with flame ionisation detection (GC/FID).     

Central composite design for the optimization of supercritical carbon dioxide fluid extraction of fatty acids from Borago officinalis L. flower

Abstract: In the present study, fatty acids and essential oils of the flower of borage (Borago officinalis L.) were obtained by supercritical carbon dioxide fluid extraction under different conditions. The extracts obtained were compared to oils of borage flower oil isolated by hydrodistillation. The obtained oils were analyzed by gas chromatography mass spectrometry. The compounds were identified according to their retention indices and mass spectra. The experimental parameters of supercritical fluid extraction (SFE) were optimized using a central composite design after a full factorial experimental design. Extraction yields based on SFE varied in the range of 0.02% to 1.96% (w/w), and the oil yield based on the hydrodistillation was 0.05% (v/w). The optimum conditions of SFE were obtained at a pressure of 350 atm, a temperature of 65 °C, a methanol modifier volume of 100 μL, and static and dynamic extraction time of 10 min. Main components of the extracts under optimum SFE conditions were palmitic acid, linoleic acid, γ‐linolenic acid, and oleic acid. The results indicated that by using the suitable extraction conditions, SFE is more effective than the conventional hydrodistillation method in the extraction of fatty acids and the preservation of its quality. Practical Application: SFE is a good technique for the extraction of oils from plants. The extraction yields by SFE are more than the conventional method. SFE is used on a large scale for production of essential oils and pharmaceutical products from plants.     

多酚类化合物检测分析方法研究进展

摘要：多酚类化合物是广泛存在于植物中的一类酚羟基化合物, 具有抗氧化、清除自由基、抑制癌症、降低血糖和血脂、延缓衰老、增强免疫力等功能, 其应用和开发价值很高。由于多酚类物质组成复杂, 结构不稳定, 易氧化, 对其检测技术提出了更高的要求。近年来, 光谱、色谱和质谱法技术不断发展和进步, 使其检测方法更先进、更灵敏、更稳定、更可靠。本研究在检索大量文献基础上, 综述了近年来国内外多酚类化合物检测分析方法, 对目前主要的检测分析技术光谱法(分光光度法、原子吸收光谱法、近红外光谱法)、色谱法(高效液相色谱法、气相色谱法)、质谱法(高效液相色谱-质谱法、气相色谱-质谱法、超高效液相飞行时间质谱法)进行对比, 旨在比较各种检测技术的优劣, 为多酚类化合物检测技术开发提供参考。     

响应面试验优化超临界CO_2萃取胭脂萝卜籽油工艺及其脂肪酸组成分析

以胭脂萝卜籽为原材料,采用响应面试验优化超临界CO2萃取胭脂萝卜籽油工艺,并对胭脂萝卜籽油脂肪酸组成进行分析。首先Plackett-Burman设计对影响超临界萃取的7个因素进行筛选,获得影响提取率的3个主要因素:萃取压力、萃取温度、萃取时间;然后用最陡爬坡路径逼近最大响应区域;最后通过Box-Behnken设计进行三因素三水平试验及响应面分析,确定最佳萃取工艺条件为萃取压力34 MPa、萃取温度44℃、萃取时间91 min,提取率预测值为93.11%,验证值为(93.09±0.80)%。采用气相色谱-质谱联用分析脂肪酸组成,结果表明胭脂萝卜籽油各脂肪酸组成与菜籽油相似,不饱和脂肪酸和单不饱和脂肪酸含量分别为89.01%和67.50%。     

基于UPLC-Q-TOF-MS技术分析黄刺玫果的化学成分

采用超高效液相色谱-串联四级杆-飞行时间质谱（UPLC-Q-TOF-MS）技术对黄刺玫果乙醇提取物的化学成分进行定性分析。在负离子模式下扫描,根据色谱峰分子离子峰的精确分子量及元素组成、主要碎片信息、保留时间,并结合刺玫果相关文献和在线数据库检索,从黄刺玫果中共鉴定出53个化合物,包括29个多酚、15个黄酮、2个有机酸和7个脂肪酸化合物。分析结果显示,黄刺玫果乙醇提取物中没食子多酚、鞣花鞣质多酚和黄酮类化合物是其主要的化学成分,且化合物2,3-(s)-六羟基联苯二甲酰基-D-葡萄糖、没食子苷、木鞣质C为首次从黄刺玫果中发现。本分析结果可为黄刺玫果资源在功能性食品开发利用、质量控制及阐明其药效物质基础提供理论参考。     

Determination of Omega Fatty Acid Profiles in Egg Yolk by HILIC-LC-MS and GC-MS

The paper presents the analysis of the profile composition of fatty acids in the molecules of phosphatidylcholine and phosphatidylethanolamine, by using hydrophilic interaction liquid chromatography and gas chromatography coupled with mass spectrometry. The profiles of 15 phosphatidylcholine and 8 phosphatidylethanolamine species were analyzed with a newly developed hydrophilic interaction liquid chromatography (HILIC)–electrospray ionization (ESI)–tandem mass spectroscopy (MS/MS) method, by using a new stationary bonded phase. The application of the new method in control and experimental groups of egg yolk revealed significant differences in the composition of phospholipid species containing mainly polyunsaturated fatty acids. Additionally, using GC-MS, the profile of fatty acids in four groups with different dietary supplementation of hens was analyzed and 20 fatty acids in egg yolks were determined. Monounsaturated fatty acids were found in higher amounts than saturated fatty acids and polyunsaturated fatty acids in egg yolks. Oleic acid (18:1) was the major monounsaturated fatty acid in egg yolk while palmitic acid (16:0) was the major saturated fatty acid. Linoleic acid (18:2), arachidonic acid (20:4), and docosahexaenoic acid (22:6) reached the highest levels among the polyunsaturated fatty acids.