滇产红花的高效液相指纹图谱研究

采用高效液相色谱法建立中药材滇产红花的高效液相特征指纹图谱。使用美国安捷伦1100紫外检测高效液相色谱仪进行试验,采用“中药色谱指纹图谱相似度评价系统2004A版”电脑软件处理试验数据,建立滇产红花的HPLC指纹图谱,并对25批滇产红花指纹图谱进行相似度评价。从云南大理和丽江等不同产地采集的25批样品共得到14个色谱共有峰,其基本特征一致,云南不同产地红花指纹图谱相似度有一定的差异。羟基红花黄色素A是红花中的黄酮类化合物,性质稳定,峰比较突出明显,且在色谱峰中间位置出峰,可以作为参照物。建立的高效液相(high performance liquid chromatography,HPLC)指纹图谱能够标示红花化学成分特征色谱图,为红花中药材的鉴别和研究提供了比较完善的信息。所建立的方法简便准确,经济便捷,重复性好,为红花药材的质量控制提供了一定的科学依据。     

不同品种橘皮中的黄酮分析

目的：检测六种柑橘皮中黄酮类化合物的含量,为天然产物开发提供数据参考。方法：采用紫外-可见分光光度法和高效液相色谱法分别建立标准品的标准曲线方程和检测方法。结果：不同品种柑橘皮不仅总黄酮含量相差悬殊,含有黄酮的种类和各自含量也差别很大。六种柑橘皮苦柑中总黄酮的含量最高,达到干重的9.26%,新橙皮苷是其黄酮的主要组成部分。结论：进行天然产物的开发,根据目标物质含量对优良品种筛选是必要的。     

柑橘醋中黄酮类化合物含量及其与总黄酮含量关系研究

采用柱层析、薄层色谱、高效液相色谱法等技术,分离鉴定出柑橘醋中的黄酮类化合物柚皮苷、柚皮素、橙皮素,通过高效液相色谱法对这3种化合物在9个批次柑橘醋产品中的绝对含量进行了测定。同时,采用亚硝酸钠-硝酸铝-氢氧化钠显色法对9个批次柑橘醋中总黄酮含量进行了测定。通过对9个批次样品中柚皮苷、柚皮素、橙皮素的含量及比例分配与其总黄酮含量之间的关系发现,样品总黄酮含量的高低与样品中柚皮苷、柚皮素、橙皮素的含量及比例分配存在一定关系,总黄酮含量的高低与柚皮苷含量呈正比例关系,与柚皮素、橙皮素的含量呈反比例关系。     

利用糖类成分评价我国油菜原蜜的品质

目的以油菜原蜜为研究对象,通过测定其中果糖、葡萄糖、蔗糖和麦芽糖的含量评价我国大宗原蜜的品质。方法采用高效液相色谱示差折光法测定我国10个省份共36种油菜原蜜样品中果糖、葡萄糖、蔗糖和麦芽糖的含量。结果用液相色谱方法测定蜂蜜中的果糖、葡萄糖、蔗糖和麦芽糖,最低检测限分别为0.079 g/100 g、0.018 g/100 g、0.037 g/100 g及0.094 g/100 g,定量检测限分别为0.263 g/100 g、0.061 g/100 g、0.124 g/100 g及0.314 g/100 g。36种油菜原蜜中果糖、葡萄糖、蔗糖和麦芽糖的含量范围分别为26.00~38.16g/100 g、23.65~39.68 g/100 g、0.14~0.62 g/100 g及1.09~3.10 g/100 g。结论油菜原蜜样品中的蔗糖含量都符合我国蜂蜜国家标准,但其中31.8%的样品中葡萄糖和果糖总量低于国家标准。目前,我国大宗蜂蜜不存在人为掺入蔗糖的问题。     

超高效液相色谱-串联质谱法检测蜂蜜中的内源性酚酸和黄酮类物质

本研究建立了超高效液相色谱-串联质谱法测定蜂蜜中18种内源性酚酸、黄酮类物质检测方法。样品经盐酸水溶液提取,HLB固相萃取柱净化,采用Acquity UPLC HSS T3(2.1*100 mm,1.8μm)色谱柱进行分离,以乙腈-10 mmol/L醋酸铵溶液为流动相进行梯度洗脱,电喷雾电离,负离子多反应监测模式检测,外标法定量。实验结果表明,所建立的方法线性范围宽,能适用于各类蜂蜜中酚酸、黄酮类物质的定量分析。各物质线性关系良好,相关系数(r2)≥0.991;各分析物检出限在20~200μg/kg之间,加标回收率范围为69.2%~94.1%,RSD为0.9%~9.5%。通过对150批次天然成熟蜂蜜、市售洋槐蜜及以不同比例(20%、40%、60%、80%)糖浆混合的洋槐蜜进行检测,发现不同蜜源蜂蜜中,酚酸、黄酮的种类与含量差异较大。其中,枣花蜜中阿魏酸平均含量最高,为550μg/kg;洋槐蜜中芹菜素平均含量最高,含量为3910μg/kg;咖啡酸在荆条蜜含量最高,为1721μg/kg;糖浆掺假蜂蜜中目标物含量明显低于真正蜂蜜。该方法前处理操作简便、分析速度快、准确度高,可用于蜂蜜中多种内源性酚酸、黄酮类物质的同时测定。该方法为通过检测不同蜜源中酚酸及黄酮类物质含量建立蜂蜜指纹图谱,提供技术参考。     

糖源蜜、花源蜜和市售蜂蜜品质的比较研究

该文对糖源蜜、花源蜜以及市售蜂蜜(黑蜂雪蜜、洋槐蜂蜜、天然蜂蜜、紫云英蜂蜜)的水分含量、电导率值、色度、果糖、葡萄糖、蔗糖、pH值、游离酸、总酸和蛋白质含量等基本理化成分以及矿物质含量、淀粉酶活性、总酚酸含量、总黄酮含量等活性成分进行测定。结果表明:糖源蜜、花源蜜以及市售4种蜂蜜的基本理化成分含量均符合我国国家标准,表明这6种蜂蜜均为新鲜蜂蜜以及来自不同的蜜源;活性成分含量测定结果表明,除紫云英蜂蜜的淀粉酶活性没有达到8 mL/(g·h),不符合标准外,其他蜂蜜均符合,且糖源蜜的酶活性接近于天然蜂蜜,花源蜜的酶活性远远高于黑蜂雪蜜。此外,蜂蜜矿物质、总酚酸、总黄酮的含量因蜜源不同存在很大的差异。     

不同蜜源蜂蜜挥发性成分差异分析

为考察不同蜜源蜂蜜挥发性成分差异,采用固相微萃取结合气相色谱与质谱联用技术对8种不同蜜源蜂蜜成分进行鉴定分析。结果表明:8种蜂蜜的挥发物中共鉴定59种挥发性化合物,椴树蜜中有30种,荆花蜜中有21种,荆条蜜中有16种,苕子蜜中有18种,洋槐蜜中有24种,野山花蜜中有27种,野酸枣蜜中有24种,枣花蜜中有32种。聚类分析结果表明,不同蜜源的蜂蜜相似度不大,通过主成分分析(principal components analysis,PCA)结果表明,壬醛和生物碱乙酸甲酯作为潜在特征标记物,可以区分不同蜜源的蜂蜜,为鉴定不同蜜源蜂蜜奠定基础。     

高效液相色谱-电化学检测指纹图谱鉴别3种单花种蜂蜜花源

基于高效液相色谱-电化学检测器(high performance liquid chromatography-electrochemical detection,HPLC-ECD)技术建立一种新的蜂蜜花源鉴别方法。以采自中国不同地区的3种单花种蜂蜜为研究对象,构建了3种单花种蜂蜜的HPLC-ECD指纹图谱,提取HPLC-ECD图谱共有峰面积信息并应用主成分分析和系统聚类分析进行蜂蜜花源分类,并对完全未参与建模的蜂蜜样品进行验证。结果表明,45个蜂蜜样品(枸杞蜜、荆条蜜、荔枝蜜各15个),均可通过主成分分析和系统聚类分析按照其花源正确分类,正确率达到100%。该蜂蜜花源鉴别的模型对完全未参与建模的枸杞蜜、荆条蜜和荔枝蜜样品的正确预判率可达到100%、80%和100%。研究表明,HPLCECD指纹图谱技术应用主成分分析和系统聚类分析可以作为一种快速、准确、绿色的判别蜂蜜花源的方法。     

单花蜂蜜多酚类物质的抗氧化活性

以5种中国单花原蜜,不同产地的42个样本通过Amberlite XAD-2吸附树脂得到的多酚提取物为原料,采用分光光度法,测定总酚酸和总黄酮含量、还原能力、抗氧化活性(清除DPPH自由基、O2－ ·能力),评估蜂蜜多酚类物质抗氧化活性及蜂蜜品种、颜色和地域差异对清除自由基的影响。结果表明：样本总黄酮含量为(9.41±0.18)～(92.76±0.13)mg/kg;总酚酸含量为(9.10±0.05)～(149.57±0.14)mg/100g;深色枣花蜜抗氧化能力最强,对DPPH自由基、O2－ ·清除作用分别为EC50＝(0.042±0.014)g/mL和EC50＝(0.038±0.009)g/mL。提示蜂蜜的多酚类物质影响抗氧化活性,酚酸、黄酮含量与抗氧化能力呈现正相关系;色深原蜜的酚酸、黄酮含量高于色浅原蜜,抗氧化能力也强于浅色原蜜。     

超高效液相色谱法测定不同产地槐花中4种黄酮含量

目的 建立槐花中4种黄酮类成分的超高效液相(UPLC)含量测定方法,并比较不同产地槐花中4种黄酮类成分的含量差异及聚类分析。方法 色谱柱为ACQUITY UPLC BEH C18色谱柱(2.1mm×50mm,1.7μm);流动相A为甲醇溶液,B为0.1%乙酸水溶液,梯度洗脱。柱温为30 ℃,检测波长为360 nm;通过TB tools 热图绘制选项以及PCA主成分分析图绘制功能进行各产地样品的聚类分析;结果 四种黄酮对照品在相应浓度范围内与峰面积均呈现良好线性关系;回收率均在95%- 105%之间;不同产地槐花中的4种黄酮类成分含量差异较大,基于4种黄酮类成分含量对不同产地的槐花的PCA主成分分析及聚类热图分析结果表明,槐花具有明显的产地聚类特性。结论 所建立的UPLC含量测定方法操作简单,结果稳定准确,专属性强,并且分析时间短,能够降低分析成本,适合作为槐花的含量测定方法。不同产地槐花质量差异明显,可通过优选产地从而保证其来源的质量稳定性以及用药安全性。     

中蜂与意蜂荔枝蜜挥发性成分的测定与对比分析

采用顶空固相微萃取(headspace solid phase microextraction,HS-SPME)-气相色谱质谱联用(gas chromatography-mass spectrometry,GC-MS)技术对我国不同产地采集的中蜂和意蜂荔枝蜜的挥发性成分进行检测,并分析和评价中蜂和意蜂荔枝蜜挥发性成分的差异。在优化后的试验条件下,荔枝蜜样品中共检测出98种挥发性物质,主要包括萜烯类、醇类、醛类、芳香类等,其中含量最高的是芳樟醇氧化物。对比研究发现,正庚醇、正壬醇、乙酸苯乙酯、丁香醇仅在中蜂荔枝蜜中被检出,而癸醛、环氧芳樟醇氧化物和某未知色谱峰(RT:27.98)仅在意蜂荔枝蜜中被检出,这几种差异性成分可以有效区分中蜂和意蜂荔枝蜜。     

贵州黄花梨蜂蜜的品质分析

首次对贵州中华蜜蜂黄花梨蜂蜜的感官指标、理化指标、卫生指标、营养成分、活性成分和安全性指标等进行检测分析,探讨其品质特性和安全性。根据相关标准规定的方法对上述各项指标进行测定,结果显示贵州黄花梨蜂蜜呈浅琥珀色,口感甘甜,有黄花梨花香味,常温下呈粘稠状,低温易结晶;水分含量19.43%,果糖和葡萄糖含量66.47%,蔗糖含量0.25%,酸度37.44 mL/kg,羟甲基糠醛含量0.91 mg/kg,淀粉酶活性28.20 mL·g-1·h-1;微生物含量符合标准规定;铁元素含量3.1767 mg/kg,锌元素含量1.6870 mg/kg,蛋白质含量0.36 g/100g,维生素C含量1.38 mg/100g,在检测的16种氨基酸中,天冬氨酸的含量最高（0.0400 g/100g）,谷氨酸（0.0373 g/100g）、异亮氨酸（0.0303 g/100g）、脯氨酸（0.0280 g/100g）和苯丙氨酸（0.0277 g/100g）次之,酪氨酸和蛋氨酸则未检出;超氧化物歧化酶活性4.29 U/g,总三萜含量65.97 mg/100g,总黄酮含量和兽药残留量未检出,重金属含量极低。本研究结果表明了产自贵州的黄花梨蜂蜜为健康安全的蜂蜜食品,并达到了蜂蜜标准的一级品水平要求。     

Biomedical Activity and Related Volatile Compounds of Thai Honeys from 3 Different Honeybee Species

This study investigated the effect of 3 factors (floral source, honeybee species, and postcollection processing) that influence the antibacterial activity, free radical reduction, and other biochemical compositions of different honey types typical of Thailand. Honey samples from 3 honeybee species (Apis mellifera, Apis cerana, and Apis dorsata) were obtained from 9 floral sources (longan, wild flower, lychee, coffee, sunflower, sesame, bitter bush, para‐rubber, and manuka as a control) in different regions of Thailand. These samples were evaluated for both their total and nonperoxide antibacterial activity against 10 human pathogens by agar incorporation technique. Honey samples were further analyzed to evaluate the capacity for free radical‐scavenging activity, total phenolic content, and the total flavonoid contents by the 2,2‐diphenyl‐1‐picrylhydrazyl assay, Folin–Ciocalteu method, and aluminum chloride colorimetric assay, respectively. Furthermore, the volatile organic compounds (VOCs) of Thai honey samples were investigated by headspace solid‐phase microextraction and gas chromatography‐mass spectrometry analysis. Findings of this study suggest a strong correlation between floral origin and honeybee species on one hand, and differences in %Brix, total acidity, protein content, antimicrobial activities, free radical reduction, phenolic, and flavonoid contents on the other hand. Moreover, VOCs of wild and coffee honey types were remarkably different, depending on the floral source. Both honeys contained characteristics of VOCs, some of which are involved in antibacterial and antioxidant activities.     

中蜂枇杷蜜与意蜂枇杷蜜中的理化指标及抗氧化活性分析

目的分析中蜂枇杷蜜与意蜂枇杷蜜的理化指标及其抗氧化活性。方法以江苏中蜂枇杷蜜与意蜂枇杷蜜为研究对象,对其理化指标和总酚酸、总黄酮含量进行测定,采用DPPH·和ABTS~+自由基清除试验测定其抗氧化活性。结果中蜂枇杷蜜和意蜂枇杷蜜的理化指标均符合GH/T 18796—2012《蜂蜜》,中蜂枇杷蜜和意蜂枇杷蜜之间的水分、淀粉酶值、葡萄糖、果糖、蔗糖、麦芽糖、乳糖、灰分均存在差异性;中蜂枇杷蜜的总酚、总黄酮、清除DPPH·自由基能力和ABTS~+自由基能力均显著优于意蜂枇杷蜜;枇杷蜜中总黄酮与清除ABTS~+自由基能力均存在良好线性关系,枇杷蜜中总酚与清除DPPH·自由基能力和ABTS~+自由基能力均存在良好线性关系。结论通过对中蜂枇杷蜜与意蜂枇杷蜜的理化指标及其抗氧化活性的检测,为中蜂枇杷蜜与意蜂枇杷蜜鉴定与检测提供理论依据,为枇杷蜜产品的开发提供理论支撑。     

Hesperetin: A marker of the floral origin of citrus honey

Seventeen flavonoid aglycones were identified in various experimental and commercial citrus honey samples by HPLC analysis. The flavanone hesperetin was detected in all samples. This flavanone was not detected in any of the honey samples, from diverse floral origin (including rosemary, lavender, sunflower, almond, sweet chestnut, white clover, Erisarum, Robinia, Rhododendron, Tilia, Prosopis, Eucalyptus and Calluna honeys) previously investigated. The analysis of the flavonoids present in orange nectar revealed that the flavanone hesperidin (hesperetin-7-rutinoside) was the major flavonoid detected and, therefore, this should be the main source of the hesperetin found in citrus honey. Hesperetin should be produced by hydrolysis of hesperidin by the bee enzymes present in honey. These results suggest that hesperetin could be used as a marker for the botanical origin of citrus honey.     

Honey major protein characterization and its application to adulteration detection

The major proteins in honey have different molecular weights depending upon the honeybee species. To confirm the origin of major honey proteins, honey protein produced by Apis cerana or Apis mellifera were purified and analyzed by MALDI-TOF. Two major proteins were identified as a major royal jelly protein 1. Although two major proteins shared primary structure, they showed different molecular weights of 56 and 59 kDa, respectively. To discriminate the honeybee species producing honey using SDS–PAGE, artificial marker proteins, 56 and 59 kDa, were produced from Escherichia coli. Two artificial marker proteins were co-electrophoresed with honey samples and the difference in molecular weight was readily distinguished by SDS–PAGE. Therefore, the measurement of major proteins in honey is a useful method to discriminate the honey that produced from different honeybee species.     

基于GC-MS非靶向代谢组学分析贵州特色蜂蜜的代谢物差异

该研究以中华蜜蜂（Apis cerana,简称中蜂）特色蜂蜜蓝莓蜜、野蔷薇蜜为研究对象,以荆条蜜、野桂花蜜为对照,采用GC-MS非靶向代谢组学,对特色蜂蜜代谢物聚类、功能富集分析,并筛选差异代谢物及功能注释。结果表明,4种蜂蜜共检测到90种代谢物,富集到210条通路上,其中富集前20的通路在碳水化合代谢（8条）、总览（9条）、膜运输（3条）三个模块;4种蜂蜜中都检测出去甲肾上腺素（Norepinephrine,NE）,它富集到了唾液分泌、胃酸分泌等通路。根据VIP>1、log2FC>1、p<0.05筛选出2-酮丁酸、硬脂酸、肌醇等12种显著差异代谢物;2-酮丁酸和3-己烯二酸分别是蓝莓蜜和野蔷薇蜜的特征性物质。在改善人体消化吸收、促进蛋白质合成效果顺序为：野桂花蜜>荆条蜜>蓝莓蜜>野蔷薇蜜。该研究从代谢组学角度分析了不同蜂蜜营养成分差异,初步确定蓝莓蜜、野蔷薇蜜的特征性物质,其结果为鉴别蜂蜜真假、蜂蜜溯源识别提供相应参考。     

Metal content of southern Italy honey of different botanical origins and its correlation with polyphenol content and antioxidant activity

Seventy‐eight samples of southern Italy honey from five different floral origins (chestnut, eucalyptus, citrus, multifloral and sulla) were screened to quantify the polyphenol and metal contents, evaluate the antioxidant activity and determine the correlations between the parameters analysed. The average polyphenol content was 12.06 mg gallic acid equivalent per 100 g honey and 7.92 mg quercetin equivalent per 100 g honey, for total phenolic and flavonoid contents, respectively. The antioxidant activity ranged from 58.40% (eucalyptus honey) to 60.42% (chestnut honey) in the ABTS assay, from 152.65 μm Fe (II) (citrus honey) to 881.34 μm Fe (II) (chestnut honey) in the FRAP assay, and from 54.29% (citrus honey) to 78.73% (chestnut honey) in the DPPH assay. Fe and Zn were the most abundant among the tested metals, while Cd, Co and Mo were those less present. Chestnut honey presented the highest polyphenol content, antioxidant activity and metal content. The correlations between the analysed parameters were statistically significant (P < 0.05). The correlations between metal content and both total phenolic and antioxidant activities were particularly interesting, suggesting a relationship between metal and polyphenol contents in honey.     

Characterisation of the phenolic and flavonoid fractions and antioxidant power of Italian honeys of different botanical origin

BACKGROUND: Twenty‐seven Italian honey samples of different floral origin were analysed for total phenolic and flavonoid contents by a spectrophotometric method and for antioxidant power and radical‐scavenging activity by the ferric‐reducing/antioxidant power (FRAP) and 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) assays respectively. In addition, the phenolic and flavonoid profiles were analysed using high‐performance liquid chromatography with UV detection (HPLC‐UV). RESULTS: The results of this study showed that honey contains copious amounts of phenolics and flavonoids. HPLC‐UV analysis showed a similar qualitative polyphenolic profile for all honey samples analysed. The main difference among samples was in the contribution of individual analytes, which was affected by floral origin. Total phenolic and flavonoid contents varied from 60.50 to 276.04 mg gallic acid equivalent kg^?1 and from 41.88 to 211.68 mg quercetin equivalent kg^?1 respectively. The antioxidant capacity was high and differed widely among samples. The FRAP value varied from 1.265 to 4.396 mmol Fe²⁺ kg^?1, while the radical‐scavenging activity expressed as DPPH‐IC₅₀ varied from 7.08 to 64.09 mg mL^?1. Correlations between the parameters analysed were found to be statistically significant (P < 0.05). CONCLUSION: The present study shows that honey contains high levels of phenolics and flavonoids and that the distribution of these compounds is influenced by the honey's floral origin. Copyright © 2009 Society of Chemical Industry     

Immunological characterization of honey major protein and its application

Honey contains a small amount of proteins and we purified the major protein in honey produced by Apis cerana and by Apis mellifera, respectively. Each purified major protein was injected in rats and after repeated immunization; antibodies were obtained and verified with Western-blot. Although honey samples used in this study differ due to geographical variations, storage periods and botanical origins, each antibody was reacted specifically to its antigen with high affinity. These results showed that the major proteins in honey derived from A. cerana and from A. mellifera have different surface structures. This feature can be applied for discrimination of honey from different honeybee species and for detection of adulterated honey by ELISA with high sensitivity.