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。     

HPLC法测定核桃仁不同品种、不同部位中的鞣花酸含量

建立高效液相色谱(high performance liquid chromatography,HPLC)法测定核桃仁中鞣花酸含量,对比分析不同品种及温185核桃仁不同部位中的鞣花酸含量.确定检测鞣花酸的色谱条件为Phenomenex Synergi Hydro-RP C18色谱柱(4.6 mm×250 mm,4μm)...     

高效液相色谱法测定成熟龙眼各部位游离鞣花酸含量

采用高效液相色谱法测定石峡、储量两个品种成熟龙眼各部位游离鞣花酸含量。鞣花酸提取条件为80%甲醇溶液,70℃回流2h。色谱条件为:C18色谱柱(250mm×4.5mm,5μm)、柱温35℃、进样体积10μL、检测波长254nm、流速1mL/min,流动相为甲醇-水-冰乙酸(50:49.7:0.3,V/V)。鞣花酸在1～200μg/mL范围内与峰面积呈线性关系,相关系数为0.999。取5个样品瓶分别进样,得到样品保留时间相对标准偏差(RSD)值为1.0%,峰面积RSD值为0.8%。对同一瓶样品连续进样10次,保留时间和峰面积的RSD值分别为0.15%和0.2%。加标回收结果表明平均加标回收率为95.97%～101.9%。测定结果表明石峡和储量各部位鞣花酸的含量大小顺序均为果核>果皮>果肉,其中果皮和果核中鞣花酸远大于已报道的其他水果中的含量。     

反相高效液相色谱法测定石榴籽中鞣花酸的含量

目的：建立检测石榴籽中鞣花酸的含量测定方法。方法：采用反相高效液相色谱法,Arcus EP-C18(250mm×4.6mm,5μm)色谱柱,以甲醇-0.1% TFA溶液为流动相梯度洗脱,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100.49%。结论：本方法精密可靠,可作为检测石榴籽中鞣花酸含量的方法。     

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。     

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。     

高效液相色谱法测定番石榴叶中的游离鞣花酸

研究高效液相色谱法测定番石榴叶中鞣花酸的方法。以甲醇(含0.01% HCl)作为提取液,70℃加热回流;色谱条件为：Intersil ODS-SP色谱柱(250mm×4.5mm,5μm),柱温35℃,进样体积10μL,紫外检测波长254nm;流动相为3%冰乙酸(A)-纯甲醇(B);梯度洗脱程序：0～4min B由20%至45%,保持6min,10～15min由45%至70%,15～20min由70%至100%,保持1min。结果表明：本方法的相对标准偏差为0.6%,鞣花酸在0.5～200mg/L范围内与峰面积呈良好的线性关系,相关系数为0.9998,加标回收率为95.86%～98.31%。本方法用于测定鞣花酸是切实可行的;番石榴叶片中鞣花酸含量高低与叶片的成熟度有关,其中幼嫩叶含量为0.814mg/g、成熟叶含量为0.023mg/g,而老叶中未检出。     

Caco-2细胞研究石榴皮中活性成分鞣花酸的摄取特性

目的:研究鞣花酸在Caco-2细胞的摄取特性。方法:采用Caco-2细胞单层模型研究鞣花酸在不同给药时间、药物浓度、p H介质、培养温度、有无抑制剂条件下对细胞摄取的影响。采用高效液相色谱法(HPLC)测定鞣花酸在细胞中的含量。结果:鞣花酸在0.0254~1.016μg范围内呈现良好的线性关系(r=0.9997)。鞣花酸在Caco-2细胞上的摄取与给药时间、药物浓度、p H介质、培养温度、抑制剂密切相关。结论:建立的HPLC检测方法简便、灵敏、准确,鞣花酸在Caco-2细胞上的摄取主要是主动转运,抑制剂不同程度参与鞣花酸的转运过程。     

黑莓、树莓不同发育时期叶片鞣花酸与抗氧化、糖类物质的相关性分析

本研究拟在初步揭示悬钩子属浆果不同发育时期叶片中鞣花酸的变化规律。以黑莓、树莓及杂交类型共21个品种不同时期叶片为试材,用超声辅助溶剂提取法提取叶片中鞣花酸,用紫外分光光度法快速检测鞣花酸含量。在此基础上,选取其中5个品种分析评价了叶片抗氧化能力、抗氧化物质含量及糖类物质含量与鞣花酸含量的关系。结果表明,黑莓、树莓鞣花酸在嫩叶中含量较高,随着叶片的发育和衰老呈下降趋势,以黑莓‘宝森’嫩叶鞣花酸含量最高达39.622 mg/g,以树莓‘哈瑞泰兹’老叶中鞣花酸含量最低为7.649 mg/g。进而分析5个品种叶片的抗氧化能力及抗氧化物质含量,发现其与鞣花酸含量变化趋势相似,而叶片中糖类物质含量与鞣花酸含量变化呈负相关。研究结果为黑莓、树莓叶片中鞣花酸的功能研究及进一步开发利用提供部分基础理论依据。     

石榴汁中部分多酚类物质的分离鉴定

目的:分离鉴定天然石榴汁中主要多酚类物质.方法:采用乙酸乙酯、正丁醇萃取多酚类物质,并应用液质联用分析方法,对其有关成分进行鉴定.结果:初步分析结果表明,石榴汁中含有鞣花酸、石榴素、芹菜配基、芹菜配基-7-O-β-D-吡喃葡萄糖苷、芦丁、3,3\,4',5,7-五羟基黄烷酮、3,3',4',5,7-五羟基黄烷酮-6-D-吡喃葡萄糖苷.结论:石榴汁中多酚类物质含量丰富,有必要进行深入研究,以明确其主要功能成分.     

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.     

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.     

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。     

Comparison of the polyphenolic composition and antioxidant activity of European commercial fruit juices

Thirty six commercial European fruit juices were tested to ascertain their antioxidant capacity and polyphenolic composition. Six of the products were labelled 100% pomegranate juice, the others included 20 brands of diluted pomegranate juice or pomegranate blended with other fruit juices and 10 different non-pomegranate fruit juices. The antioxidant capacity of all the juices was determined while anthocyanin, ellagitannin and ellagic acid profiles of the 26 pomegranate juices and pomegranate juice blends were obtained using HPLC-PDA-MS(2). Additional analysis was conducted on seven of the juices using HPLC with an on-line antioxidant detection system. Three of the "pure" pomegranate juices had the highest ellagitannin content and the highest antioxidant capacity. Only one of these three juices was rich in anthocyanins. The other "pure juices" had differences in their HPLC "pomegranate" fingerprint and also had a lower antioxidant capacity, in some cases lower than that of some of the blended juices. Vitamin C rather than phenolic compounds was the major contributor to the antioxidant capacity for some of the juices. Statistical analysis of both the antioxidant assay and the HPLC on-line antioxidant data demonstrated that the ellagitannins were the major antioxidants in the pomegranate juices. The complexity of the polyphenolic profile of pomegranates necessitates the use of HPLC-PDA-MS(2) for a thorough evaluation of juice composition and authenticity.     

气相色谱法测定石榴葡萄酒中的甲醇和杂醇油

以石榴和葡萄为原料榨汁,混合果汁经酸度和糖度调整后(添加蔗糖调整糖度,添加酒石酸和碳酸钙调整酸度),添加酵母,通过前发酵、后发酵,酿制出石榴葡萄酒.再以4-甲基-2-戊醇为内标物,建立了一种可同时测定石榴葡萄酒中甲醇和杂醇油含量的气相色谱测定方法.该方法相对标准偏差为2.1797％～3.2125％,加标回收率为95.8143％～101.9143％,具有较高的精密度和准确度.最后以该方法测定石榴葡萄酒中的甲醇和杂醇油含量,检测结果甲醇未检出,杂醇油(以异丁醇和异戊醇计)0.0228g/100mL.     

浓缩石榴汁中熊果苷的超高效液相-串联质谱检测方法研究

建立了浓缩石榴汁样品中熊果苷含量测定的固相萃取-超高效液相-串联质谱测定方法。样品用水稀释,HLB和氨基固相萃取柱净化,采用BEHAmide色谱柱(100mm×2.1mm,1.7μm)分离,以乙腈和水梯度洗脱,多反应监测模式测定,定量离子对为m/z271.2>108.0,定性离子对为m/z271.2>160.9,外标法定量。熊果苷的检测限为0.006mg/kg,在0.004～0.2mg/L浓度范围内,熊果苷的线性相关系数为0.9994,熊果苷的加标回收率均在80.6%～108.1%范围内,相对标准偏差均低于8.3%。该方法样品净化效果良好,检测简便、快速、准确,能够满足浓缩石榴汁中熊果苷含量测定和定性确证的要求。     

Identification of Ellagic Acid in Blackberry Juice Sediment

ABSTRACT: The sediment in a commercial reconstituted 'Evergreen' blackberry ( Rubus laciniatus L.) juice concentrate was found to be composed of ellagic acid, protein, and other unidentified compounds. The qualitative tannin and protein-tannin haze test indicated that the sediment was predominantly tannin or protein-tannin complexes. Nitrogen determination showed the sediment to be 6.69%± 2.21% protein on a dry-weight basis. Almost all of the extractable material was identified as ellagic acid by high-performance liquid chromatography and liquid chromatography-mass spectrometry. The ellagic acid content of the wet sediment was 0.05 g/100 g, whereas it was 7.41 g/100 g in freeze-dried sediment. Tannase enzyme did not significantly decrease the concentration of ellagitannins in Marion blackberry ( Rubus spp. Hyb.) juice in this study.