高效液柑色谱-串联质谱测定砀山酥梨中的酚类物质

本文研究了高效液相色谱．电喷雾串联质谱定性、定量检测砀山酥梨及其制品中的酚类物质。样品中的酚类物质在70min内得到分离，酚类物质由色谱的保留时间、质谱信息和参考文献确定。质谱检测在负离子模式下，利用全扫描模式、多反应检测模式和中性丢失模式进行。本研究在砀山酥梨中共检出酚类27种，主要酚类物质是绿原酸，含量为180mg／L，其次为熊果苷、没食子酸、咖啡酸、咖啡奎宁酸、表儿茶素、儿茶素、柚（皮）苷、芦丁、槲皮素．葡萄糖苷、槲皮素-半乳糖苷、槲皮素-鼠李糖苷、香豆酸-葡萄糖苷、槲皮紊-戊糖苷、鼠李糖素-葡萄糖苷、槲皮索等。     

高效液相色谱-串联质谱测定黄冠梨果皮中主要酚类物质

为研究黄冠梨果皮中的主要酚类物质,采用了高效液相色谱—串联质谱方法定性、定量检测黄冠梨果皮中主要酚类物质,并评价该方法定量分析的准确性、线性和检出限。以甲醇—1%乙酸溶液为流动相做梯度洗脱,柱温30℃,检测波长280 nm,流速为0.6 mL/min;质谱检测在负离子模式下,利用全扫描模式、多反应检测模式和中性丢失模式进行。在此方法下,样品中的酚类物质在50 min内得到分离并由色谱的保留时间和质谱信息确定。结果表明:黄冠梨果皮中共检测出15种酚类物质,主要以绿原酸含量最多,含量为65.765 2 mg/100gDW,其次是熊果苷、儿茶素、香豆酸、咖啡酸、槲皮素等。     

不同整形方式对‘赤霞珠’葡萄果皮非花色苷酚的影响

以欧亚种酿酒葡萄‘赤霞珠’为试材,通过高效液气相色谱-质谱联用法研究单干单臂、单干双臂和单干双层双臂3 种整形方式对葡萄果皮非花色苷酚类物质的影响,并采用聚类分析和偏最小二乘判别分析（partial least squares-discrimination analysis,PLS-DA）筛选不同整形方式间单体非花色苷酚的差异组分。结果表明：与单干单臂和单干双层双臂整形相比,单干双臂整形可以提高葡萄果皮中的槲皮素-3-O-葡萄糖醛酸苷、槲皮素-3-O-葡萄糖苷、山柰酚-3-O-半乳糖苷、槲皮素-3-O-鼠李糖苷类黄酮醇物质的含量;单干单臂和单干双臂整形葡萄果皮中杨梅酮-3-O-葡萄糖苷、异鼠李亭-3-O-葡萄糖苷、丁香亭-3-O-葡萄糖苷类黄酮醇类物质和表没食子酸儿茶素、儿茶素类黄烷醇类物质及原儿茶酸类、香草酸酚酸类物质的含量均高于或显著高于单干双层双臂整形,而单干双层双臂整形葡萄果皮中仅原花色素C1的含量较高。聚类分析结果显示,部分单体非花色苷酚类物质受年份的影响较大,且单干单臂和单干双臂整形对非花色苷酚类物质的影响模式类似,而两者与单干双层双臂整形区别较大。PLS-DA结果表明,不同整形方式的葡萄果皮中单体非花色苷酚类物质组分差异较大。槲皮素-3-O-葡萄糖苷、杨梅酮-3-O-葡萄糖苷等黄酮醇类物质是3 类整形方式所处理的葡萄果皮的主要差异组分,且对单干双臂整形处理组的贡献值较大,单干双臂整形的葡萄果皮中含有较高的黄酮醇类物质含量。综上,单干双臂整形方式有助于葡萄果皮非花色苷酚类物质的积累,可被应用于陕西关中地区酿酒葡萄的栽培生产中。     

甜叶菊花和叶中酚类成分分析及含量测定

以高效液相色谱-串联四极杆质谱分析仪,结合质谱分析及比对对照品或文献报道化合物的保留时间和质谱参数,在甜叶菊花和叶中鉴定了16种酚类化合物。建立一种利用高效液相色谱法测定甜叶菊中16种酚类化合物含量的方法,并对甜叶菊一新品种盛花期花和叶中酚类的含量进行测定。结果表明,甜叶菊花和叶中酚酸化合物以新绿原酸、绿原酸、隐绿原酸、异绿原酸B、异绿原酸A、异绿原酸C为主,异绿原酸A含量最高,绿原酸含量次之;花和叶中类黄酮化合物主要为芦丁、槲皮素-7-O-葡萄糖苷、木犀草素-7-O-葡萄糖苷、木犀草素-7-O-芸香糖苷、槲皮素-3-O-木糖苷、槲皮素-3-O-鼠李糖苷、芹菜素-7-O-葡萄糖苷、山奈酚-3-O-阿拉伯糖苷、山奈酚-3-O-鼠李糖苷,槲皮素-3-O-鼠李糖苷含量最高;盛花期甜叶菊花和叶中酚类总量达（22.31±0.51）mg/g和（91.11±2.32）mg/g。     

杜鹃花色素的分离与鉴定分析

运用特征性颜色反应、高效液相色谱(HPLC)和质谱(MS)等技术对浅红色毛叶杜鹃(R. championae Hook)花瓣花色素提取液进行初步鉴定和含量测定。结果表明：该杜鹃品种的花色素含有非红色的黄酮类化合物,主要有黄酮、黄酮醇、二氢黄酮等色素,其中还有部分酚类色素。高效液相色谱(HPLC)显示杜鹃花色素提取液至少包括39种物质,其中有7种可以确定,分别为杨梅黄素3-半乳糖苷、槲皮素3-半乳糖苷、槲皮素-鼠李糖苷、槲皮素3-葡萄糖苷、锦葵素3-阿拉伯糖苷-5葡萄糖苷、杨梅黄素3-鼠李糖苷、绿原酸。利用紫外分光光度法测得杜鹃花瓣中花色苷的含量为131.025mg/100g,总黄酮含量为10.68mg/g。     

UHPLC-DAD-ESI-MS/MS法分析紫山药块根和茎叶中酚类物质

本文采用微波辅助法,以80%甲醇作为溶剂提取经冻干后紫山药块根和地上茎叶中的多酚类物质,提取物经超高效液相色谱分离、电喷雾串联三重四级杆质谱正和负离子方式检测分析。根据液相保留时间、紫外吸收、相对分子质量和二级碎片离子等信息,结果表明,紫山药块根和茎叶中酚类物质主要为酚酸、花色苷及黄酮类化合物。分析得到的29种酚类化合物中,5-O-咖啡酰奎宁酸、阿魏酰奎宁酸(tR:12.289)、迷迭香酸、芦丁和槲皮素为块根和茎叶共有组分,块根中另含有芥子酸、芥子酸葡萄糖苷、丁香酸衍生物、香豆酸衍生物、阿魏酸衍生物、芥子酸二葡萄糖苷和花色苷等11种酚类物质。茎叶中除5种共有组分,还含有咖啡酸、咖啡酰奎宁酸甲酯、咖啡酰莽草酸、对香豆酰奎宁酸、香豆酰奎宁酸甲酯、山奈酚-3-O-芦丁糖苷与迷迭香酸甲酯等13种酚类化合物。     

高效液相色谱-串联质谱法测定各生长期紫苏中酚类物质的含量

目的：建立一种简单、快速、准确的高效液相色谱-串联质谱法,同时分离与检测紫苏叶、茎和根中酚类化合物含量,探讨紫苏各部位酚类物质的累积规律,为确定紫苏各部位最佳采收时期提供依据。方法：对紫苏进行人工栽培,样品分期采集,采集后的样品经乙醇提取,采用高效液相色谱-串联质谱法进行分析,对影响色谱、质谱的重要参数进行优化,通过对照标准物质的保留时间及质谱分析,确定相应的化合物;通过标准曲线法定量,以其中13 种酚类化合物为主要参考指标,观察它们在各部位的动态含量变化。结果：鉴定出化合物：咖啡酸、咖啡酸-3-O-葡萄糖苷、迷迭香酸、迷迭香酸-3-O-葡萄糖苷、木犀草素、木犀草素-7-O-葡萄糖苷、木犀草素-7-O-葡萄糖苷酸、木犀草素-7-O-二葡萄糖苷酸、芹菜素、芹菜素-7-咖啡酰基葡萄糖苷、芹菜素-7-O-二葡萄糖苷、野黄岑素-7-O-二葡萄糖苷、迷迭香酸甲酯,以上化合物中酚酸及其糖苷化合物含量呈先降低,至结果期急剧上升达到最大,黄酮及其糖苷化合物含量则递增至结果期达到最大。其中鉴定出迷迭香酸和木犀草素-7-O-葡萄糖苷酸分别在结果期达到最大,含量分别为21.41 mg/g和19.89 mg/g,结果期叶部位总酚类含量达到最大,含量为47.50 mg/g。结论：通过对紫苏各个部位不同采收时期黄酮、黄酮糖基化、酚酸的动态变化、总含量变化和部位成分分布的研究,对紫苏酚类资源的利用提供参考依据。     

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。     

苹果酵素自然发酵工艺优化及品质分析

为了优化苹果酵素自然发酵工艺,通过Box-Behnken试验设计,研究初始pH值、糖添加量、苹果片质量浓度对苹果酵素感官评分和超氧化物歧化酶（SOD）活性的影响,并采用超高效液相色谱（UPLC）及液相色谱-质谱联用技术（LC-MS）分析了酚类物质的组成和含量。结果表明,优化出自然发酵工艺条件为初始pH值4.6,糖添加量15%,苹果片含量200 g/L。在此优化条件下,苹果酵素感官评分为（8.45±0.24）分、SOD活性为（40.26±1.03） U/mL。苹果酵素共检测出15种酚类物质,其中绿原酸含量最高,原花青素B2、表儿茶素、原花青素B1、槲皮素-半乳糖苷、儿茶素以及根皮苷次之。     

白首乌酒对癫痫大鼠神经元保护作用及其机制研究

为了优化苹果酵素自然发酵工艺，通过Box-Behnken试验设计，研究初始pH值、糖添加量、苹果片质量浓度对苹果酵素感官评分和超氧化物歧化酶（SOD）活性的影响，并采用超高效液相色谱（UPLC）及液相色谱-质谱联用技术（LC-MS）分析了酚类物质的组成和含量。结果表明，优化出自然发酵工艺条件为初始pH值4.6，糖添加量15%，苹果片含量200 g/L。在此优化条件下，苹果酵素感官评分为（8.45±0.24）分、SOD活性为（40.26±1.03） U/mL。苹果酵素共检测出15种酚类物质，其中绿原酸含量最高，原花青素B2、表儿茶素、原花青素B1、槲皮素-半乳糖苷、儿茶素以及根皮苷次之。     

Multiclass Determination of Phenolic Compounds in Different Varieties of Tomato and Lettuce by Ultra High Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry

Tomato and lettuce are main components of the Mediterranean Diet and it is important to know their content in phenolic compounds, considering they have beneficial effects on human health. In this work, more than 25 phenolic compounds were simultaneously analyzed by ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry, including phenolic acids, flavonols, and flavones. Several varieties of lettuce and tomato were evaluated and total phenolic content ranged from 68 to 212 mg/kg dry weight for lettuce, and from 341 to 1296 mg/kg dry weight for tomato, being the chlorogenic acid the compound detected at the highest concentration in both types of vegetables (lettuce from 9 to 126 mg/kg dry weight, tomato from 96 to 538 mg/kg dry weight), although in tomato the content of quercetin-3-O-rutinoside (from 29 to 460 mg/kg dry weight) was also important. On the other hand, the minor compounds detected were quercetin-3-O-derivate (up to 1 mg/kg dry weight) in tomato, and apigenin O derivate, luteolin C glucoside, and isorhamnetin 3-O-glucoside in lettuce. It was observed that the total phenolic content in the plum cherry tomato was significantly higher than in the other varieties, whereas in lettuce, the differences were not statistically different.     

Characterization of phenolic compounds in Chinese hawthorn (Crataegus pinnatifida Bge. var. major) fruit by high performance liquid chromatography–electrospray ionization mass spectrometry

Hawthorn (Crataegus spp.) fruits are increasingly popular as raw materials for nutraceuticals and functional foods. As a major group of bio-active components of hawthorn, the phenolic compounds of the fruits have not been well characterized so far. After extraction with 80% aqueous ethanol, the phenolics of the fruits of a major Chinese hawthorn variety, Crataegus pinnatifida Bge. var. major, were separated by polyamide column chromatography, followed by analyses by high performance liquid chromatography combined with diode array UV spectrometry and electrospray ionization mass spectrometry. Ideain (cyanidin-3-O-galactoside), chlorogenic acid, procyanidin B2 [epicatechin-(4β → 8)-epicatechin], epicatechin, hyperoside (quercetin-3-O-galactoside) and isoquercitrin (quercetin-3-O-glucoside) were identified with UV spectra, mass spectra and reference compounds. In addition, 35 compounds were tentatively identified based on UV and mass spectra. These compounds were mostly B-type procyanidins (PA) and their glycosides including aglycons of 3 dimers, 3 trimers, 8 tetramers, 4 pentamers, 2 hexamers and 2 glycosides of PA monomers, 7 glycosides of PA dimers, 1 glycoside of a PA trimer, 2 glycosides of PA tetramers, 1 glycoside of a PA pentamer, and 2 glycosides of quercetin. This is the first systematic study of phenolic compounds in Chinese hawthorn fruits and the first report of the presence of glycosylated procyanidins in hawthorn.     

基于非靶向代谢组学评价传统发酵对客家酸芥菜酚类化合物组成的影响

芥菜的酚类物质组成及乳酸发酵对其影响仍未完全探明。该文采用高效液相色谱串联质谱的非靶向代谢组学,结合主成分分析、相关性分析和偏最小二乘判别分析等,对广西传统客家酸芥菜(客家擦菜)的原料和成品进行了酚类物质组成分析与对比。结果表明,新鲜芥菜和客家擦菜共初步定性检出136种酚类物质,其中119种(包括25种多酚类物质、34种酚酸物质、60种黄酮类物质)为芥菜产品中首次报道,例如大豆苷元、雌马酚。发酵改变了芥菜的酚类物质组成,其中发酵后原儿茶酸等7种酚酸、槲皮素等9种黄酮类物质和血竭素的含量显著升高,水杨酸等7种酚酸和槲皮素-3-O-葡萄糖醛酸苷等5种黄酮类物质的含量显著下降。通过KEGG数据库的比对分析可知,发酵后的酚类差异代谢物涉及发酵过程的7条代谢通路。基于非靶向代谢组学分析,该文首次较系统报道了芥菜及其乳酸发酵产品的酚类物质组成和它们的差异,为探明多酚物质在蔬菜乳酸发酵过程中的变化机制提供理论参考。     

Phenolic Compounds from the Leaves of Vitis labrusca and Vitis vinifera L. as a Source of Waste Byproducts: Development and Validation of LC Method and Antichemotactic Activity

The cultivation of Vitis (Vitaceae) grape varieties is one of the most important economic activities in agribusiness in southern Brazil. Vitis varieties are rich in polyphenolic compounds with several pharmacological and biological activities, such as antioxidant action. In this context, this study analyzed qualitatively and quantitatively the anthocyans and flavonoids found in the leaves of grape varieties Vitis vinifera and Vitis labrusca. For this purpose, vine leaf extracts were prepared and the chemical profile of each was characterized by LC/MS-MS. Two high performance liquid chromatography-validated methods were performed using UV/VIS-LC-DAD detector to quantify phenolic compounds. The main anthocyanins isolated from vine leaves were cyanidin-3-O-glucoside and peonidin-3-O-glucoside. The flavonoids identified were rutin, quercetin-3-O-galactoside, quercetin-3-O-glucoside, and quercetin-3-O-glucuronide, which was the predominant compound. The Waters X-Terra® RP18 column allowed the effective separation of quercetin-3-O-glucuronide from the other flavonoids for the first time, besides the partial separation of quercetin-3-O-galactoside from quercetin-3-O-glucoside. Furthermore, another phenolic compound was confirmed by MS spectrometry, using direct infusion, as being trans-caftaric acid. The present study also investigates the antichemotactic activity in vitro of grape crude extracts, fractions, and isolated compounds. It was demonstrated that almost all fractions and isolated compounds showed increased antichemotactic effect in response to LPS with a more pronounced values of IC₅₀ for anthocyanins fraction, rutin, quercetin-3-O-galactoside, and trans-caftaric acid (0.9, 1.6, 3.7, and 5.1 ng/mL, respectively).     

拮抗砀山梨炭疽菌的内生菌筛选及抑菌效果分析

从健康砀山梨中分离出15 株内生菌,筛选出1 株对砀山梨胶孢炭疽菌抑菌活性较高的菌株,命名为DSL-9,初步鉴定为枯草芽孢杆菌（Bacillus subtilis）。DSL-9菌株对炭疽菌的抑制主要是通过胞外分泌物作用的结果。采用硫酸铵沉淀法获得DSL-9的粗蛋白提取液,蛋白提取液在30～60?℃抑菌率无明显变化,大于60?℃抑菌率下降;在pH?7～10区间抑菌活性较高,且无明显变化;紫外线对抑菌物质无明显影响;蛋白提取液经蛋白酶处理后,抑菌活性下降。研究内生菌DSL-9对感染炭疽菌砀山梨防御酶活性的影响,发现内生菌DSL-9可提高砀山梨果实对病菌的抗性。从砀山梨中分离筛选的枯草芽孢杆菌DSL-9?对砀山梨炭疽病害的生物防治具有潜在的应用价值。     

Selection of elderberry (<Emphasis Type="Italic">Sambucus nigra</Emphasis> L.) genotypes best suited for the preparation of juice

Experiments with elderberry genotypes, ripening stage and pectinolysis were carried out in order to investigate the possibility to improve different quality characteristics of elderberry juice. Raw juice was processed using either an optimised method with respect to enzyme concentration and treatment time or produced directly without any enzymatic treatment. Neochlorogenic acid, chlorogenic acid, cyanidin-3-sambubioside-5-glucoside, cyanidin-3,5-diglucoside, cyanidin-3-sambubioside, cyanidin-3-glucoside, quercetin-3-rutinoside (rutin) and quercetin-3-glucoside (isoquercitrin) were identified in the raw juice by liquid chromatography-mass spectrometry (LC-MS) and quantified by high-performance liquid chromatography-diode array detection (HPLC-DAD). The content of phenolic acids and flavonoids in elderberries were shown to be affected by berry development stage at harvest (ripening stage). Furthermore, a considerable variation in juice yield and physicochemical properties such as turbidity, soluble solids, titratable acidity, and content of phenolic acids and flavonoids was found between juices processed from elderberries by pectinolysis of different elderberry genotypes. Pectinolysis appeared not to have any major effect on juice yield and the content of phenolic acids and flavonoids in the juice compared to non-enzymatic processed juice. Factor analysis revealed correlations among quality characteristics such as soluble solids, titratable acidity, turbidity, and content of phenolic acids and flavonoids and it is shown that these correlations may be useful for the selection of elderberry genotypes best suited for the preparation of juice.     

Changes in the volatile compounds and chemical and physical properties of Yali pear (Pyrus bertschneideri Reld) during storage

Volatiles from stored Yali pear (Pyrus bertschneideri Reld) were studied using high-resolution gas chromatography and the solid-phase microextraction (SPME) method of gas chromatography/mass spectrometry (GC/MS). The dominant components were ethyl butanoate, ethyl hexanoate, α-farnesene, hexanal, ethyl acetate, hexyl acetate, ethanol and so on. By using GC-olfactometry, it was demonstrated that the volatile compounds from SPME were responsible for the aroma of Yali pear. The levels of sugars, organic acids, and phenolic acids in Yali pear were investigated using high-performance liquid chromatography. Fructose was the dominant sugar, followed by glucose and sucrose. With increasing storage time, sucrose levels decreased, however fructose and glucose levels did not changed remarkably. There was a slight decrease in flesh firmness during storage. The general soluble solids concentration, slightly decreased after 5 months storage. Some aroma volatile components increased during storage, while others decreased, especially the esters. The organic acids and phenolic acids also changed. Yali pear flavor was affected by changes in the levels of volatile compounds, and chemical and physical properties.     

Comparative study of six pear cultivars in terms of their phenolic and vitamin C contents and antioxidant capacity

The main phenolic compounds in six pear cultivars were identified and quantified using high‐performance liquid chromatography/diode array detection (HPLC/DAD) and HPLC/electrospray ionisation mass spectrometry (HPLC/ESIMS). Major quantitative differences were found in the phenolic profiles. The peel contained higher concentrations of chlorogenic acid, flavonols and arbutin than the flesh, where only chlorogenic acid was detected. Total phenolics ranged from 1235 to 2005 mg kg^?1 in the peel and from 28 to 81 mg k g^?1 in the flesh. Ascorbic acid and dehydroascorbic acid were detected in the peel, whereas only dehydroascorbic acid was present in the flesh. The ranges of vitamin C content were from 116 to 228 mg kg^?1 in the peel and from 28 to 53 mg kg^?1 in the flesh. The antioxidant capacity was correlated with the content of chlorogenic acid (r = 0.46), while ascorbic acid made only a small contribution to the total antioxidant capacity of the fruit. Copyright © 2003 Society of Chemical Industry