反相高效液相色谱法测定东北地区6 种红树莓果实中鞣花酸含量

建立反相高效液相色谱法测定东北地区6 种红树莓果实中鞣花酸含量,同时与蓝莓进行比较分析。色谱条件：色谱柱SinoChrom DS-BP C18（150 mm×4.6 mm,5 μm）;流动相为CH3OH-0.3%磷酸（40∶60,V/V）;流速1 mL/min,柱温30 ℃,检测波长254 nm。结果表明：鞣花酸得到很好地分离,鞣花酸含量在0.1～1 μg范围内与峰面积呈良好的线性关系,R2为0.999 3。精密度实验相对标准偏差为0.06%,重复性实验相对标准偏差为0.09%,稳定性实验相对标准偏差为0.39%,平均加样回收率为93.00%～98.62%,相对标准偏差为0.21%～0.42%,该方法简单、准确、重复性好。测得不同品种红树莓果实中总鞣花酸含量为39.10～155.98 mg/100 g;游离鞣花酸含量为1.24～8.26 mg/100 g,红树莓中含有丰富的鞣花酸,具有潜在的开发利用价值。     

RP-HPLC法测定石榴汁中的4种多酚类成分

目的:建立检测石榴汁中没食子酸、石榴皮鞣素、安石榴苷和鞣花酸的测定方法。方法:采用高效液相色谱法,ArcusEP-C18(250mm×4.6mm,5μm)色谱柱,以甲醇-0.1%TFA为流动相梯度洗脱,流速1.0mL/min,没食子酸的检测波长254nm,石榴皮鞣素、安石榴苷和鞣花酸的检测波长377nm。结果:没食子酸、石榴皮鞣素、安石榴苷和鞣花酸分别在0.020~0.320μg,0.038~0.608μg,0.074~1.184μg,0.039~0.624μg之间线性关系良好,平均加样回收率分别为97.05%、96.21%、95.30%、98.14%。结论:本方法精密可靠,可作为石榴汁中没食子酸、石榴皮鞣素、安石榴苷和鞣花酸4种多酚成分的检测方法。     

高效液相色谱法同时测定保健食品中的安石榴甙和鞣花酸

目的建立同时测定保健食品中安石榴甙、鞣花酸的高效液相色谱法(HPLC)方法。方法采用迪马钻石Ⅱ(4.6 mm×250 mm,5μm)色谱柱,乙腈+0.2%磷酸水溶液为流动相梯度洗脱,在波长256 nm下进行检测。结果方法的线性范围:α安石榴甙24～240μg/ml;β安石榴甙56.0～560μg/ml;鞣花酸60.0～600μg/ml。相关系数r为0.999 7～0.999 9;检出限:α安石榴甙、β安石榴甙、鞣花酸分别为2.36、2.78、2.67μg/g;定量限:7.08、8.34、8.01μg/g。高低两个浓度水平加标回收率97.5%～104%;相对标准偏差(RSD)均<2.5%。结论本方法简便、快速、准确、重现性好,适用于保健食品中安石榴甙和鞣花酸的测定。     

石榴功能物质鞣花酸研究进展

鞣花酸是石榴中重要的酚类物质之一,具有抗氧化、抗癌、抗炎、抗菌等生物学效应,与石榴保健功能密切相关。笔者概括了鞣花酸的结构、理化性质、石榴中鞣花酸提取工艺和含量测定、代谢途径和保健功能等,以期为石榴功能品质的栽培调控和加工产品的开发利用提供参考。     

超临界CO2萃取石榴籽油及石榴酸甲酯富集

采用超临界CO2 萃取技术从石榴籽中提取石榴籽油,对其脂肪酸和甘三酯组成和含量进行分析;通过单因素和正交优化试验研究尿素包合法富集石榴酸甲酯。超临界CO2 萃取技术与有机溶剂萃取相比更有利于获得高石榴酸含量的石榴籽油,石榴籽油中检测出14 种甘三酯。尿素包合法富集石榴酸甲酯的最佳工艺为尿素包合温度5 ℃、包合时间20 h、尿素与脂肪酸甲酯的质量比1∶1、尿素和95%乙醇的质量比1∶5,此最佳包合条件下获得的石榴酸甲酯的含量为78.82%。     

鞣花酸微球的制备及其对前脂肪细胞生长和成脂分化的影响

目的：采用海藻酸钠/壳聚糖对石榴皮鞣花酸进行包裹,制备成鞣花酸微球,并观察其对前脂肪细胞的生 长及成脂分化的影响。方法：利用扫描电子显微镜、傅里叶变换红外光谱仪和释放率分析仪鉴定制备的鞣花酸微 球;噻唑蓝法检测不同质量浓度的鞣花酸微球在6、12、24 h和36 h时对前脂肪细胞生长的影响,苏木精-伊红染色 观察细胞形态及数量的变化;油红O染色观察鞣花酸微球对前脂肪细胞成脂分化的影响,异丙醇萃取测定脂肪生成 量。结果：成功制备了海藻酸钠/壳聚糖包裹的鞣花酸微球,0.1、0.3 g/L和0.5 g/L的鞣花酸微球能够抑制前脂肪细 胞的生长及成脂分化,且具有剂量-效应关系,对脂滴生成的抑制率可达80%。结论：以海藻酸钠/壳聚糖作为石榴 皮鞣花酸的包裹材料制备鞣花酸微球是可行的,鞣花酸微球具备潜在的保健价值。     

RP-HPLC法测定石榴汁中鞣花酸的含量

目的:建立检测石榴汁中鞣花酸的含量测定方法。方法;采用高效液相色谱法,ArcusEP-C18(250mm×4.6mm,5μm)色谱柱,以甲醇-0.1%TFA为流动相梯度洗脱,流速1.0mL/min,检测波长254nm。结果:鞣花酸0.004～0.064μg之间线性关系良好,平均加样回收率为99.32%。结论:方法精密可靠,可作为检测石榴汁中鞣花酸含量的方法。     

高效液相色谱分析蓝莓发酵液中鞣花酸和没食子酸

建立了高效液相色谱法测定蓝莓发酵液中鞣花酸含量和没食子酸含量的两种方法。鞣花酸的测定条件：Waters symmetry C18色谱柱,柱温25℃,流动相为46∶2∶52的甲醇—乙酸乙酯—KH2PO4/H3PO4溶液,流速0.8m L/min,检测波长254nm,检测蓝莓发酵液中鞣花酸平均回收率为92.68%,RSD为0.97%。没食子酸的测定条件：Waters symmetry C18色谱柱（250×4.6mm,5μm）,柱温35℃,流动相为甲醇：水（0.1%磷酸）=10∶90,流速为1.0m L/min,检测波长273nm,该方法检测蓝莓发酵液中没食子酸平均回收率为99.0%,RSD为0.73%。两种方法均操作简单快速,重现性好,准确度高,可分别用于香菇等食用菌发酵蓝莓制品中鞣花酸和没食子酸含量的测定。     

石榴皮多酚的提取、制备及其应用研究进展

石榴皮作为一种可再生资源,其含有丰富的多酚,在食品中具有广阔的应用前景。该文对石榴皮多酚的提取、纯化、检测和应用研究现状进行综述;并介绍石榴皮中最重要的两种多酚——安石榴苷和鞣花酸的研究现状,特别是对安石榴苷的水解、鞣花酸的提取及制备等进行详细阐述;并以大白鼠为模型,阐明安石榴苷体内代谢的一般途径。     

响应面试验优化超声波辅助提取圆叶葡萄鞣花酸和总酚工艺

以圆叶葡萄Noble皮和籽为原料,使用丙酮提取溶剂（丙酮-水-盐酸体积比70∶29∶1）,采用超声波辅助提取圆叶葡萄中以鞣花酸为主的多酚类化合物。在单因素试验的基础上,利用响应面试验优化圆叶葡萄皮中鞣花酸和总酚提取工艺,建立数学回归模型,并分析双因素间的交互作用。结果表明,影响圆叶葡萄皮中鞣花酸和总酚含量的显著因素为料液比、提取时间和超声功率,得到的最佳提取工艺条件为料液比1∶30（g/mL）、提取时间28 min、超声功率616 W,此条件下鞣花酸和总酚含量分别为616.21 μg/g和15.06 mg/g（以没食子酸当量计,鲜质量）。用响应面试验对圆叶葡萄中鞣花酸和总酚提取进行优化,可得到最佳工艺条件,验证结果与理论预测拟合度高,可为实际生产提供理论依据。     

超高效液相色谱-串联四极杆质谱联用法同时测定蒙药全石榴、中药石榴皮及新鲜石榴中7 种成分

采用超高效液相色谱-串联四极杆质谱联用法,同时检测蒙药全石榴皮及籽、中药石榴皮、鲜石榴皮及籽中诃子鞣质A、诃子鞣质B、诃子裂酸、五没食子酰葡萄糖、鞣料云实素、诃黎勒酸和没食子酸7 种成分。选用表儿茶素-4-乙酰半胱氨酸作为内标。石榴样品经粉碎用甲醇（含内标液）溶液超声提取,采用ZORBAX Eclipse XDB-C18柱（2.1 mm×50 mm,1.8 μm）,以甲醇-0.1%甲酸溶液为流动相,梯度洗脱,以电喷雾离子源,采用超高效液相色谱-串联四极杆质谱联用动态多反应监测模式测定。结果表明,7 种成分检出限均小于0.013 5 μg/mL,定量限均小于0.054 2 μg/mL,在0.003 4～13.875 0、0.013 5～13.875 0、0.003 4～13.875 0、0.216 8～55.500 0、0.867 2～55.500 0、0.216 8～55.500 0、0.003 4～13.875 0 μg/mL范围内,峰面积与质量浓度呈良好线性关系,相关系数均大于0.99;方法回收率为99.44%～105.54%;其日内及日间精密度实验的相对标准偏差分别为0.11%～0.95%和0.14%～1.21%。该方法灵敏度高、稳定性强、操作简便、快捷、准确,可用于蒙药全石榴、中药石榴皮、鲜石榴及其相关食品药品的质量控制。     

A NEW METHOD FOR ELLAGIC ACID PRODUCTION FROM POMEGRANATE HUSK

Pomegranate ( Punica granatum L.) husk, a by-product of the pomegranate juice industry, is an inexpensive and abundant source of ellagic acid. Ellagic acid is widely used as functional food for its physiological functions. It is the breakdown product of ellagitannins. To date, the preparation of ellagic acid from pomegranate husk has not been reported. This article reports a new process for ellagic acid production from pomegranate husk by extraction of tannins followed by acid hydrolysis and purification by extraction and recrystallization. Several tests were conducted to obtain optimum conditions including extraction of tannins by varying solvents, acid concentration and reaction time for acid hydrolysis and the volume of methanol used for purification. Ellagic acid (3.5 g) with 90% purity from 100 g pomegranate husk was obtained. This new method is easy to scale up. All equipment used in this production process is widely used in food processing industry. The cost of production is low. It is suitable for industrial applications.

PRACTICAL APPLICATIONS

The production of ellagic acid is easier and the yield and purity of ellagic acid produced this way are higher than before. This method can be used not only for experiment in laboratory but also for industrial applications. The material – pomegranate husk – is a by-product of the pomegranate juice industry, so it is very cheap and easy to get. High-purity ellagic acid produced this way is sold to many companies back home and abroad. It is used as food additive and cosmetic material because of its antioxidant activity and whiteningfunction. The toxicity of pomegranate husk is lower than that of gallnut, which has been the main material of ellagic acid production in the past. Reagents are common and inexpensive; some of them are reusable.     

HPLC/DAD/ESI-MS analyses and anti-radical activity of hydrolyzable tannins from different vegetal species

In the present work, several tannin extracts from myrtle and pomegranate were analyzed and characterized using HPLC/DAD/ESI-MS methods. Both aqueous and hydroalcoholic myrtle leaf extracts were found to be rich in galloyl-glucosides, galloyl-quinic acids, ellagitannins and flavonoids. In these extracts we observed a predominance of galloyl-glucosides and galloyl-quinic derivatives with respect to ellagic derivatives; 87.14% and 12.86%, respectively, average gallic and ellagic derivatives with respect to total tannins content. In pomegranate extracts, substantial differences were found in polyphenol contents between peel and seed extracts with regard to the relative abundance of gallic and ellagic acid derivatives: 28.81% and 71.19%, respectively, as average total gallic and ellagic derivatives in peel extracts; 61.30% and 38.70%, in seed extracts. The antiradical properties were evaluated and compared to those of two commercial extracts of chestnut bark and grape seeds. The collected data could suggest the use of myrtle and pomegranate extracts as nutraceuticals and functional foods for their important antioxidant properties.     

鞣花酸磷脂复合物制备及其对植物油抗氧化作用

以鞣花酸与磷脂的复合率为评价指标,以石榴皮鞣花酸和大豆卵磷脂为原料,采用超声-微波协同技术,通过正交实验得到制备鞣花酸磷脂复合物的最优条件,并研究了鞣花酸磷脂复合物的理化性质及其对植物油氧化稳定性的影响。结果表明,鞣花酸磷脂复合物的最佳制备工艺为：以乙醇为反应溶剂,鞣花酸质量浓度为 3 mg /mL,投料比（磷脂:石榴皮鞣花酸质量比）为 3:1,微波功率为300 W,反应时间为 35 min,鞣花酸磷脂复合物复合率为91.80%。光谱分析表明,鞣花酸与磷脂未形成新的化合物而是通过分子间作用力形成的复合物。与鞣花酸相比,复合物在水和在正辛醇中溶解性均稍有改善。添加鞣花酸及其磷脂复合物可以延缓葡萄籽油和核桃油的氧化反应。石榴皮鞣花酸及其磷脂复合物对核桃油的抗油脂氧化效果更好。本实验为石榴皮鞣花酸的进一步资源化开发利用提供了一定的技术依据。     

石榴皮中鞣花酸的抗衰老性能及机理研究

利用光谱法研究了石榴皮中鞣花酸的抗衰老性能和机理。首先,通过分光光度法考察了鞣花酸对DPPH·、·OH和·O2-等3种自由基的清除率和弹性蛋白酶抑制率,结果证明了鞣花酸抗衰老能力强。其次,以弹性蛋白酶为研究对象,利用光谱法探讨了鞣花酸对弹性蛋白酶的作用机理。结果表明,鞣花酸与弹性蛋白酶发生作用并形成络合物,当鞣花酸的质量浓度为4.57 mg/m L时,鞣花酸对弹性蛋白酶的抑制率高达88.26%。当鞣花酸浓度低于4.0×10-5mol/L时,其荧光猝灭机理主要是静态猝灭,鞣花酸浓度较高时动态猝灭所占比例增加。因此,鞣花酸可以作为天然自由基清除剂与弹性蛋白酶抑制剂应用于抗衰老化妆品中。     

Quantitative Determination of Punicalagin and Related Substances in Different Parts of Pomegranate

Three simple and efficient high-performance liquid chromatography (HPLC) methods for punicalagin and related substances in different parts of pomegranate were established. The mobile phases for α-punicalagin, β-punicalagin, gallic acid, and ellagic acid consisted of acetonitrile and 2% glacial acetic acid solutions, and that for pentagalloylglucose consisted of acetonitrile and 0.5% phosphoric acid. The mobile phase of shikimic and 3-dehydroshikimic acids consisted of methanol and 1% phosphoric acid. The limit of detection values of seven substances ranged from 0.15 to 0.24 mg/g. The HPLC method of these substances showed high precision, good stability, and repeatability. Their recovery rates were in the range of 98.9–102.5%, and their relative standard deviation values were less than 2.2%. Punicalagin and related substances were determined in the pericarp, juice, seed, and leaf of “Taishanhong” and “Taishansanbaitian” cultivars. The highest punicalagin content was 138.232 mg/g in “Taishanhong” pericarp. Pentagalloylglucose mainly existed in leaf, and the six other substances were mainly present in pericarp. HPLC method can be used to further research on punicalagin metabolism in the pericarp, seeds, juices, and leaves of pomegranate.     

Quantitative determination of major polyphenol constituents in pomegranate products

The anti-oxidant content and potential health benefits associated with consuming pomegranate and pomegranate-containing products has lead to increased consumer demand for this crop resulting in it becoming a high value crop. The potential health benefits and high anti-oxidant content of this fruit is attributed to the polyphenolic compounds it contains, including the ubiquitous phenolic acids, gallic acid and ellagic acid, and punicalagin A and punicalagin B, two polyphenolics unique to this fruit. A rapid HPLC–UV method targeting these four metabolites requiring minimal sample cleanup and offering runtimes half as long as existing methods was established. Within day and inter-day run-to-run variability for the four metabolites ranged from 1.9% to 6.6% and 5.3% to 11.4%, respectively. Spike recovery percentages for gallic acid, punicalagin A, punicalagin B and ellagic acid were found to be 98.5%, 92.4%, 95.5%, and 96.5%, respectively. This method was applied to the evaluation of various pomegranate products, including commercial drinks, handmade juice, and marc extracts. This method may be readily used to verify the presence of pomegranate metabolites in juices, extracts, and other products.