高效液相色谱-电喷雾质谱法分析牡丹花中花色苷类化合物

利用高效液相色谱与二极管阵列检测器/电喷雾质谱联用技术研究了牡丹花中的花色苷类化合物,分离检测了五种花色苷,结合紫外吸收光谱和质谱信息分别鉴定为矢车菊-3,5-O-二葡萄糖苷,矢车菊-3-O-葡萄糖苷、芍药-3,5-O-二葡萄糖苷、芍药-3-O-葡萄糖苷和天竺葵-3,5-O-二葡萄糖苷;比较了六个牡丹品种的花色苷组成。     

紫皮豇豆(Vigna unguiculata L.)花色苷的研究

首次应用高效液相色谱-二极管阵列检测器-质谱联用技术(HPLC-DAD-MS)鉴定出紫皮豇豆主要含有三种花色苷,分别为飞燕草色素、矢车菊色素、天竺葵色素的葡萄糖苷,其中矢车菊葡萄糖苷相对百分含量达到93.4%;应用pH示差法测定出其单体花色苷的含量为57.15mg/100g鲜重;应用DPPH法测定结果显示,花色苷粗提液有较强的抗氧化能力。     

野生桑葚中花色苷成分分析

运用固相萃取纯化技术与高效液相色谱/二极管阵列检测器/电喷雾质谱联用技术,采用ZorbaxSB-C18色谱柱(250mm×4.6mm,5μm),甲醇-5%甲酸水溶液为流动相,流速为1.0mL/min,检测波长为520nm,以矢车菊素3-葡萄糖苷为对照,外标法测定了野生桑葚花色苷含量,并通过紫外扫描光谱和电喷雾质谱正离子碎片信息确定了花色苷的成分组成。结果表明:野生桑葚总花色苷含量为154.27mg/100g,含有的三种花色苷成分分别为矢车菊素3-葡萄糖苷、矢车菊素3-芸香糖苷和天竺葵素3-葡萄糖苷,其相对含量为67.52%、31.29%和1.06%。     

咖啡酸对紫玉米花色苷的辅色作用研究

研究咖啡酸对紫玉米花色苷溶液的吸收光谱、颜色和花色苷组分的影响。结果表明,添加咖啡酸后,花色苷溶液的颜色加深,最大吸收峰发生蓝移,这表明咖啡酸对紫玉米花色苷具有辅色效应。热稳定性实验结果表明,添加咖啡酸能抑制花色苷因热褪色的速率,而增加花色苷褪色反应的半衰期和活化能。经高效液相色谱/ 电喷雾质谱联用技术检测结果表明,咖啡酸对紫玉米花色苷的辅色作用是通过咖啡酸与不同的花色苷发生加成形成矢车菊素-3- 葡萄糖苷- 乙烯基儿茶酚、天竺葵素-3- 葡萄糖苷- 乙烯基儿茶酚和芍药素-3- 葡萄糖苷- 乙烯基儿茶酚等花色苷衍生物而实现的。     

黑豆皮花色苷的中压液相色谱制备及结构鉴定

以黑豆皮为实验材料,用乙醇浸提法对黑豆皮中的花色苷进行提取,用大孔吸附树脂对花色苷进行纯化,经冷冻干燥得到黑豆皮花色苷粗品。利用中压制备色谱对花色苷组分进行分离,通过质谱分析鉴定经中压制备色谱分离后的花色苷组分。结果表明:黑豆皮花色苷粗品中的总花色苷含量为26.9%,经中压制备色谱对花色苷粗品进行分离后的2峰中矢车菊素-3-葡萄糖苷纯度达到91.46%。黑豆皮中的主要花色苷为天竺葵素-3-O-芸香糖苷、芍药色素-3-O-葡萄糖苷、矢车菊素-3-O-葡萄糖苷和锦葵素-3-葡萄糖苷-4-乙醛。     

新疆药桑椹中花色苷的分离与鉴定

采用高速逆流色谱分离制备药桑花色苷。以甲基叔丁基醚-正丁醇-乙腈-水-三氟乙酸(2:2:1:5:0.01,V/V)为溶剂体系,进样量50mg,分离得到纯度分别为99.24%、88.5%、99.9%和96%的4个花色苷单体。通过高速逆流色谱(HSCCC)、紫外-可见光谱、质谱进行结构鉴定,初步确定馏分1为矢车菊-3-O-芸香糖苷,馏分2为天竺葵-3-O-芸香糖苷,馏分3为矢车菊-3-O-葡萄糖苷,馏分4为天竺葵-3-O-葡萄糖苷。此法高效、稳定、简捷易行。     

利用中压制备液相色谱从桑葚中快速制备矢车菊素-3-葡萄糖苷单体

单体花色苷的快速大量制备长久以来是花色苷产业化中的难题,而中压制备液相色谱在产业应用中有着很大的开发空间。选取花色苷组分较单一的桑葚为实验原料,经提取分离总花色苷后使用填装有反相C18填料的耐压玻璃柱作为中压制备液相色谱柱,纯化制备矢车菊素-3-葡萄糖苷单体。结果显示：3 个色谱分离峰中目的峰（峰2）经高效液相色谱和质谱确证为由矢车菊素-3-葡萄糖苷（cyanidin-3-glucoside,C3G）和矢车菊素-3-芸香糖苷（cyanidin-3-rutinoside,C3R）组成,采用峰面积归一化法计算得到C3G纯度为73.56%;通过对峰2采用切割方式进行收集,C3G纯度达到98%以上,单次收集到C3G单体溶液650?mL。中压制备液相色谱法单次上样量大、步骤简洁、成本低廉,可为矢车菊素-3-葡萄糖苷单体的规模化生产提供一定的参考。     

超声对花色苷稳定性的影响及其降解动力学

为研究超声对花色苷稳定性影响规律及其降解动力学,采用单因素试验在超声提取模拟体系中研究4 种因 素（功率密度、温度、溶剂、pH值）对5 种常见花色苷稳定性的影响规律,采用试错法分析花色苷超声降解动力 学,并结合液相色谱-质谱联用技术分析超声处理后的降解产物。结果表明：随着温度的升高,花色苷在超声处理 下的降解率下降;5 种花色苷在体积分数70%乙醇溶液中降解最少;pH 1～5范围内,花色苷降解率呈先增加后减 少趋势。飞燕草素-3-葡萄糖苷的超声降解符合一级动力学模型,矢车菊素-3-葡萄糖苷、天竺葵素-3-葡萄糖苷、锦 葵素-3-葡萄糖苷、芍药素-3-葡萄糖苷的超声降解符合零级动力学模型。在天竺葵素-3-葡萄糖苷、锦葵素-3-葡萄糖 苷、芍药素-3-葡萄糖苷、矢车菊素-3-葡萄糖苷的降解产物中都检测到8-β-D-吡喃葡萄糖基-2,4-二羟基-6-氧代环己 基-2,4-二烯基乙酸,同时也检测到山柰酚、槲皮素、丁香酸、阿魏酸、2,6-二甲氧基苯酚等。     

大孔树脂-中压柱层析联用分离纯化蓝莓花色苷

以蓝莓果实为原料,采用大孔树脂-中压柱层析联用分离纯化蓝莓花色苷。分别比较6 种不同类型树脂 对蓝莓花色苷静态吸附-解吸效果,优化大孔树脂分离纯化蓝莓花色苷的工艺。结果表明：D101大孔树脂对蓝莓 花色苷的分离效果最佳,对花色苷的吸附属于多分子层吸附。在柱压力为1 MPa、温度25 ℃、上样液质量浓度为 0.073 mg/mL、洗脱剂乙醇体积分数为80%、流速5 mL/min条件下,经D101大孔树脂柱分离后,花色苷纯度从5.53% 增加到75.58%,提高了12.67 倍。采用Sephadex LH-20中压柱层析对蓝莓花色苷进一步分离纯化,主要得到1 种花 色苷组分,通过高效液相色谱和高效液相色谱-电喷雾质谱联用对蓝莓花色苷进行定性和定量分析,确定该组分为 矢车菊-3-O-葡萄糖苷,纯度达到90.88%。     

结球红菊苣(Cichoricum intybus var.foliosum Hegi.)花色苷初步研究

采用pH示差法测定结球红菊苣中花色苷的含量,并采用高效液相色谱-光电二极管阵列检测器-离子阱/飞行时间质谱(HPLC-DAD-IT/TOFMS)技术对结球红菊苣花色苷溶液进行了检测和鉴定。结球红菊苣花色苷溶液是从结球红菊苣中用0.1%盐酸-甲醇溶液提取,并经C18固相萃取小柱纯化制备而成。结果表明,结球红菊苣中花色苷含量为37.3±12.25mg/100gFW,主要由四种矢车菊花色素的糖苷衍生物组成,分别是矢车菊-3-O-葡萄糖苷、矢车菊-3-丙二酰半乳糖苷、矢车菊-3-丙二酰葡萄糖苷、矢车菊-3-琥珀酰葡萄糖苷,其中矢车菊-3-丙二酰葡萄糖苷含量最高,占结球红菊苣叶子中花色苷总量的82.01%。     

Direct Acylation of Cyanidin-3-Glucoside with Lauric Acid in Blueberry and Its Stability Analysis

Cyanidin-3-glucoside is an anthocyanin that is abundant in blueberry. Lauric acid was used as the acyl donor in the acylation of cyanidin-3-glucoside. Preparative high performance liquid chromatography was used to separate and purify the acylated cyanidin-3-glucoside, the acylated rate was 30.78%. Fourier transform infrared spectroscopy and liquid chromatography mass spectrometry were used to confirm the basic structure of cyanidin-3-glucoside. Lauric acid was combined with cyanidin-3-glucoside. Lauric acid reacted with the glucoside’s primary hydroxyl group and removed a molecule of water, thereby resulting in the obtained structure. The compositions of the unacylated cyanidin-3-glucoside and acylated cyanidin-3-glucoside were compared under different temperatures and under illumination. The effects of different concentrations of additive compounds on the stability of acylated cyanidin-3-glucoside were compared. Stability of acylated cyanidin-3-glucoside was obviously higher than that of the unacylated cyanidin-3-glucoside because of the higher stability of the ester group than the hydroxyl group. The primary hydroxyl esterification of glucoside improved cyanidin-3-glucoside.     

云南丽江和吉林靖宇11种越橘果实花色苷组分的分析

采用高效液相色谱法对云南丽江、吉林靖宇11种越橘果实花色苷组分进行测定。通过标准品的分析,建立了飞燕草素-3-半乳糖苷、飞燕草素-3-葡萄糖苷、矢车菊素-3-半乳糖苷、矢车菊素-3-葡萄糖苷、锦葵色素-3-半乳糖苷、锦葵色素-3-葡萄糖苷的回归方程,相关系数为0.9939~0.9968,精密度RSD为2.00%~3.73%,回收率在98.90%~100.99%,方法准确可行。通过6种标准品的对比,供试的15个样品中,所有越橘品种均有飞燕草素-3-半乳糖苷、矢车菊素-3-半乳糖苷、锦葵色素-3-半乳糖苷3种花色苷,飞燕草素-3-半乳糖苷平均含量为223.99 μg/g、锦葵色素-3-半乳糖苷平均含量为153.34 μg/g,二者占总花色苷的65%。采用欧氏距离聚类分析表明,集群1为飞燕草素-3-半乳糖苷含量较高的三种越橘,代表品种为丽江雷戈西、丽江奥尼尔、靖宇杜克;集群2为6种花色苷总含量较高的越橘品种,代表品种为丽江北陆与靖宇早蓝,说明飞燕草素-3-半乳糖苷含量与花色苷总含量是评价越橘花色苷特点的重要因子。     

Flavonoid Compounds in the Strawberry Fruit

SUMMARY— The flavonoid compounds of the strawberry fruit were studied by paper chromatography, spectrophotometry and color reactions, Catechin, quercetin–3-glucoside, kaempferol–3–glucoside and leucocyanidins of varying degrees of polymerization were found besides the two anthocyanins, pelargonidin-3-glucoside and cyanidin-3-glucoside.
When the leucocyanidins were fractionated into water and methanol insoluble, ethyl acetate–soluble and water–soluble fractions, the latter fraction, generally representing glycosylated leucoanthocyanidins, was found to be the largest of the three in both ripe and unripe strawberries.     

Phenolic profiles of Andean purple corn (Zea mays L.)

Qualitative and quantitative high performance liquid chromatography with diode array detection (HPLC-DAD) was performed to characterize the presence of phenolic compounds in Andean purple corn. Phenolic compounds were analyzed by separating them in two main fractions: a water fraction (WF) and an ethyl acetate fraction (EAF). The WF rich in anthocyanins revealed the presence of cyanidin-3-glucoside, pelargonidin-3-glucoside and peonidin-3-glucoside. The respective acylated anthocyanin glucoside forms of these compounds were also detected following alkaline hydrolysis. The EAF was composed of phenolic acids such as p-coumaric, vanillic acid, protocatechuic acid, flavonoids such quercetin derivatives and a hesperitin derivative. Alkaline and acid hydrolysis of the EAF revealed the presence of p-coumaric and ferulic acid as main components in four bound hydroxycinnamic acid forms present in the ethyl acetate fraction.     

Characterisation of phenolic phytochemicals and quality changes related to the harvest times from the leaves of Korean purple perilla (Perilla frutescens)

The objective of this study was to investigate the profiles of phenolic phytochemicals in the leaves of Korean purple perilla (cv. Bora, Perilla fructescens) using reversed-phase C₁₈ column chromatography and HPLC with DAD-ESI/MS analysis. Changes in their contents were also the first reported through eight different harvest times during two months. They were characterised as five anthocyanins and three phenolic acids including cyanidin-3,5-di-O-glucoside (1), cyanidin-3-O-glucoside (2), cyanidin-3-O-(6-O-caffeoyl)glucoside-5-O-glucoside (3), cyanidin-3-O-(6-O-coumaroyl)glucoside-5-O-glucoside (4), cyanidin-3-O-(6-O-coumaroyl)glucoside (5), caffeic acid (6), rosmarinic acid (7), and rosmarinic acid methylester (8). Significant differences were observed between individual and total phytochemical contents, especially, cyanidin-3-O-(6-O-coumaroyl)glucoside-5-O-glucoside (4) and rosmarinic acid (7) exhibited the predominant constituents. Among different harvest times, the highest content was found with 82.473 mg/g on 21st September, while the lowest was 39.000 mg/g on 17th August. These results may be useful in determining the optimal harvest time at which phenolic phytochemicals reaches a maximum level in mid-September.     

HPLC法测定桃不同品种果实中花色苷组成和含量

目的： 用高效液相色谱法测定桃不同品种果实中花色苷组成和含量。方法： 色谱柱为C1 8柱(150mm×4.6mm,5μm),流动相为乙腈：4%磷酸溶液(V/V=12:88,pH2),检测波长520nm,流速0.8mL/min,进样量20μL,柱温30℃。结果：桃3个品种果实中主要含有1种花色苷：花青素-3-葡萄糖苷,花青素-3-葡萄糖苷在0.4～100μg/mL内线性关系良好(R=0.9992),低、中、高质量浓度的平均回收率为85.7%、89.4%、91.2%。红、黄、白肉桃果肉中的花青素-3-葡萄糖含量分别为103.7、30.6、21.5mg/kg。结论：本方法操作简单,结果准确可靠,重复性好,可作为桃果实中花青素-3-葡萄糖苷的定量分析方法。     

Flavonoids in three olive (Olea europaea) fruit varieties during maturation

The objective of the present paper was the characterisation of olive fruit varieties through the formation and variation during maturation of their flavonols, flavones and anthocyanins. An olive variety with a constant cyanidin-3-rutinoside:cyanidin-3-glucoside ratio of 1 exhibited lower contents of luteolin-7-glucoside and quercetin-3-rutinoside than varieties in which the only anthocyanin detected was cyanidin-3-rutinoside.     

Anthocyanin Pigment Composition of Blackberries

ABSTRACT: The anthocyanin pigments in 51 blackberry samples were isolated and characterized by highperformance liquid chromatography (HPLC) with UV-visible detection, electrospray mass spectroscopy (ESMS), and hydrolysis and saponification reactions. Five pigments were detected: cyanidin-3-glucoside, cyanidin-3-rutinoside, cyanidin-3-xyloside, cyanidin-3-glucoside acylated with malonic acid, and an unidentified acylated derivative of cyanidin-3-glucoside. Four different quantitative patterns were evident among the cultivars and selections. Total anthocyanin content ranged from 70.3 to 201 mg/100 g with a mean of 137 mg/100 g. Ten commercial blackberry juice concentrate samples were analyzed. Total anthocyanin ranged from 12.3 to 107 mg/100 g on a single-strength juice basis, and 1 sample had an atypical profile.     

超声辅助果胶酶法提取红树莓花色苷及其成分测定

本研究利用超声辅助果胶酶法来提取制备红树莓花色苷,通过单因素实验,研究花色苷提取工艺中果胶酶浓度、料液比、酶解pH、酶解温度、超声时间和超声功率对提取液中花色苷含量的影响,结合响应面实验对提取工艺进行了优化,对比超声辅助果胶酶法和单一提取法所得的花色苷含量,并利用超高效液相色谱仪串联四级杆/飞行时间质谱(UPLC-Q/TOF)对树莓花色苷进行结构鉴定。结果表明:红树莓花色苷最佳提取条件为:果胶酶浓度5 mg/g、料液比1:15(g/mL)、酶解pH3、酶解温度50℃、酶解时间60 min、超声时间20 min和超声功率450 W,此时所得花色苷含量为127.51 mg/100 g。超声辅助酶法所得到花色苷含量较酶法提高了24.58 mg/100 g,较超声法提高40.40 mg/100 g,较单一提取法,超声辅助酶法具有更好的提取效果。经过超高效液相色谱三重四级杆飞行质谱分离中主要花色苷为:芍药素-3-葡萄糖苷、矢车菊素-3-葡萄糖苷、矢车菊素-3-阿拉伯糖苷和矢车菊素-3-芸香糖苷。     

Mechanisms of anthocyanin degradation in grape must-like model solutions

Oxidative degradation of o-diphenolic (eg cyanidin-3-glucoside) and non-o-diphenolic (eg malvidin-3-glucoside) anthocyanins in the presence of caffeoyltartaric acid and grape polyphenoloxidase was studied in model solutions. Both anthocyanins reacted with the enzymically generated caffeoyltartaric acid o-quinone. Kinetic studies indicated that cyanidin-3-glucoside was degraded mostly by coupled oxidation whereas malvidin-3-glucoside formed adducts with caffeoyltartaric acid quinone. In solutions containing equimolar amounts of the anthocyanins, both reactions took place but coupled oxidation of cyanidin-3-glucoside occurred at a faster rate, partly protecting malvidin-3-glucoside. The occurrence of coupled oxidation resulted in sparing of caffeoyltartaric acid. Several oxidation products, including red pigments, were detected by HPLC. The UV-Vis spectra of the coloured condensation products suggested that they contained both caffeoyltartaric acid and anthocyanin moieties. All these pigments were gradually degraded to colourless compounds either by enzymatic oxidation or by further reaction with quinones.