椴树蜜、苕子蜜和刺槐蜜红外指纹图谱及抗氧化研究
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摘要
椴树蜜,主要以椴树科,紫椴(Tilia amurensis)和糠椴(Tilia mandschurica)为蜜源植物,是我国东北特有蜜种。蜜色为特浅琥珀色、浅琥珀色,具有浓郁的香味,容易结晶,为一等蜜。具有养血补虚、清热、补中益气、解毒、润燥、镇静的功效。较受消费者喜欢,是难得的森林蜜种。
苕子蜜,主要以豆科,广布野豌豆(Vicia cracca L.)和长绒毛野豌豆(Vicia villosa rothL.)为蜜源植物。苕子蜜既具有普通蜂蜜的解毒润燥作用,又有苕子清凉养颜、滋润肌肤、增强皮肤弹性、保持皮肤柔软等独特功效,颇受女士的青睐。苕子蜜呈浅琥珀色,结晶颗粒较细,气味清香,味道甘甜,为一等蜜。
刺槐蜜,主要以豆科,刺槐属植物洋槐(Robinia pseudoacacia L.),又名刺槐,为蜜源植物。洋槐蜜白色、水白色,透明,粘稠状液体,含有独特的刺槐花香,为一等蜜。
针对市场上,蜂蜜传统鉴定方法的不确定性和人为差异等因素,目前急需寻找科学的蜂蜜标准化检验体系的情况下,本实验以椴树蜜、苕子蜜和刺槐蜜为研究对象,深入研究苕子蜜、椴树蜜和刺槐蜜的质量评价方法,开发蜂蜜抗氧化作用的药用价值为目的,采用较先进的研究手段——高效液相色谱仪,紫外-可见分光光谱仪和傅里叶变换红外光谱仪三种精密实验仪器,从蜂蜜来源入手,寻找椴树蜜、苕子蜜和刺槐蜜质量评价的方法。同时,通过比较不同品种、地域的蜂蜜的还原能力和清除DPPH自由基,抑制羟自由基(·OH)、超氧阴离子自由基(O2-.)和亚硝基自由基能力的差异,评价了16种单花蜂蜜36个样本的抗氧化活性,为开发蜂蜜的药用价值和确保蜂蜜质量提供参考。
本文建立了苕子蜜、椴树蜜和刺槐蜜的傅里叶变换红外光谱(FTIR)指纹图谱及二阶导数红外光谱指纹图谱,二阶导数红外光谱指纹图谱能有效地区分苕子蜜、椴树蜜和刺槐蜜与其它品种蜂蜜,并结合聚类分析方法,基本可以把苕子蜜、椴树蜜和刺槐蜜的品种不同、产地不同和质量差异区分开,为市场上评价蜂蜜质量的方法提供一些技术。
建立了异硫氰酸苯酯柱前衍生反相高效液相色谱同时测定蜂蜜中17种游离氨基酸的实验方法。以正亮氨酸为内标,异硫氰酸苯酯为柱前衍生试剂,Venusil-AA氨基酸分析专用柱(5μm,4.6mm×250mm),醋酸钠缓冲溶液(pH6.5)-乙腈溶液和80%乙腈水溶液为流动相,采用梯度洗脱,流速：1ml/min,柱温：40℃,紫外检测器在254 nm波长处检测。氨基酸浓度在0.5μmol～0.0125μmol范围内,线性相关系数均大于0.99。17种氨基酸的平均回收率在91.3%～103.1%。该方法灵敏、准确,可作为测定蜂蜜中氨基酸的有效方法。测定结果表明：16种蜂蜜(桉树蜜、枣花蜜、黄连蜜、椴树蜜、党参蜜、龙眼蜜、紫云英蜜、荔枝蜜、荆条蜜、益母草蜜、蒲公英蜜、山花蜜、五味子蜜、枇杷蜜、苕子蜜、刺槐蜜)(深色和浅色)不同地域的36个样品的游离氨基酸含量,得到蜂蜜中的游离氨基酸主要以脯氨酸为主,占游离氨基酸总量69.82%,16种蜂蜜中平均含游离氨基酸543mg/kg左右,这一结果为进一步研究蜂蜜的营养价值提供了前提条件。
通过体外抗氧化实验(抑制DPPH自由基、超氧阴离子(02-.)自由基、羟自由基和亚硝基)的研究表明,蜂蜜对四种自由基均具有一定的清除能力,其中蜂蜜对DPPH自由基、超氧阴离子(02-.)自由基的清除能力强于对羟自由基和亚硝基的清除。采用Duncan's multiple range test统计学方法分析比较了16种蜂蜜,不同地域的36个蜂蜜样品之间对DPPH自由基、羟自由基、超氧阴离子自由基和亚硝基抑制能力的显著性差异,证明不同品种的蜂蜜,甚至同一品种而产地不同的蜂蜜,抗氧化能力有很大差异。
采用Folin-Ciocalteu比色法测定36个蜂蜜样品总酚酸含量,蜂蜜的总酚酸含量：由10.43±0.69mg/100g到168.9±0.04 mg/100g,刺槐蜜(昆明)总酚酸含量最低,桉树蜜(广东)总酚酸含量最高,说明蜂蜜品种差异、颜色差异、地域差异影响蜂蜜中的酚酸含量。利用紫外-可见分光光度法测定36个蜂蜜样品总黄酮含量和还原能力,36个样本的总黄酮含量：由9.410±0.06 mg/kg到99.63±0.04 mg/kg,色浅的刺槐蜜(昆明)含量最低,色深的桉树蜜(广西)含量最高,含量相差近10倍。采用Duncan's multiple range test统计学方法分析比较了36个蜂蜜样品之间总酚酸含量和总黄酮含量的显著性差异。同时测得蜂蜜的还原能力与总酚酸、黄酮含量的相关系数分别为：R=0.940368和R=0.79281,说明蜂蜜的还原能力与总酚酸、黄酮含量呈线性关系变化,同时蜂蜜的颜色越深,还原能力与总酚酸、黄酮含量越高,反之,还原能力与总酚酸、黄酮含量越低。
通过实验还测得蜂蜜的总酚酸含量与清除DPPH自由基和超氧阴离子自由基作用的相关系数分别为：R=0.85074和R=0.83493；总黄酮含量与抑制DPPH自由基和超氧阴离子自由基作用的相关系数分别为：R=0.62692和R=0.66133,说明蜂蜜的抗氧化能力受其多酚类(类黄酮和酚酸)成分的影响,不同品种的蜂蜜,甚至同一品种而产地不同,蜂蜜的抗氧化能力差异很大。验证了蜂蜜的总酚酸、黄酮含量、还原能力与蜂蜜体外抗氧化能力呈现正相关系；原蜜的颜色也影响抗氧化能力,色深原蜜的酚酸和黄酮含量高于色浅的原蜜,深色蜂蜜比浅色蜂蜜有更好的抗氧化活性。
蜂蜜作为健康营养物质,文章中展示了如何利用显微镜技术分析花粉形态,确定蜜源；利用评价蜂蜜样本的植物来源、色泽、结晶状态、气味或滋味等的感官特征来判断蜂蜜真伪；利用傅里叶变换红外光谱仪,建立椴树蜜、苕子蜜和刺槐蜜的普通红外光谱(FTIR)指纹图谱及二阶导数红外光谱指纹图谱,作为蜂蜜的质量评价方法,为今后建立科学的蜂蜜标准化检验体系提供实验依据。同时还比较了了苕子蜜、椴树蜜和刺槐蜜等16种蜂蜜36个不同地域样品的抗氧化能力差异。实验证明,蜂蜜的质量取决于它们自身的化学成分和蜜源植物属性,受到植物来源、地理来源等因素的影响,因而鉴定和量化蜂蜜中的微量化学物质意义重大。
Linden, mainly Tilia amurensis and Tilia mandschurica, is a kind of plants from Tilia, one of the peculiar nectariferous plants in Northeast China. Linden honey ranks as first class honey, for its properties of light amber or pale light amber color, strong fragrance, and easily crystallized. As its popularity among consumers and rare collection in the forestry, it is cherished for the functions of nurturing blood and improving weakness, reducing body heat, neutralizing poison, relieving dryness, and tranquilizing spirit. The forests in Heilongjiang and Jilin Provinces serve as the major Linden honey sources.
Vetch honey, belonging to Leguminosae, includes Vicia cracca L. and Vicia Villosa Roth L., which are defined as nectariferous plants of Vicia spp. honey. Rare produced in Yunnan-Kweichow Plateau, Vicia spp. honey bears not only the property of common honeys, such as poison neutralization and dryness-syndrome relief, but also the peculiar function of muscle and skin moisturizing, and reinforcement elasticity of skin as well. In light of its property, Vetch honey is especially popular with women consumers.
Robinia pseudoacacia L., a kind of plants from Leguminosae, also named as locust, is one of the main nectariferous plants in China. With the features of white, or water-like white color, transparent, creamy liquid and unique flower fragrance of Robinia pseudoacacia L.. Acacia honey boasts as the first class honey.
In this experiment, Linden honey, Vetch honey, and Acacia honey are chosen as subjects to further the research on their quality appraisal measures. The article focused on the investigation of three kinds honey and the development of the antioxidant and sunscreen medicine. Meanwhile, the relationship of honey and nectariferous plants was studied with high performance liquid chromatograph (HPLC), ultraviolet spectra (UV) and infrared spectra (IR). In light of honey sources, quality appraisal measures were ascertained for the three kinds of honeys. Then, the antioxidant activities of three honeys were appraised by comparing their reducing power and the diversity on scavenging effects on DPPH-free radical, hydroxyl radical, NO2 radical and superoxide radical. The study may serve as reference for honey's wellbeing function and quality security.
The study established the common infrared spectrum (FTIR) fingerprint and second derivative infrared spectrum fingerprint of Linden honey, Vetch honey, and Acacia honey. It proved that second derivative infrared spectrum fingerprint efficiently distinguished Linden honey, Vetch honey, and Acacia honey from other kinds of honeys of different sources and types in the market. Cluster analysis method was used along with IR fingerprint to analyze original honey against the honey sold in market, which can separate original honey and the processed ones. By cluster analysis, colour, botanic or geographic differences of three kinds honeys were also clarified to give light to quality evaluation measures for honeys.
A reversed phase high performance liquid chromatographic method was utilized for simultaneous determination of 17 free amino acids in Panax quinquefolium L.. The amino acid samples were mixed with norleucine as internal standard, derivatized with Phenylisothiocyanate (PITC), and then were separated on a Venusil-AA (5μm,4.6 mm X 250 mm) column specially for analysis amino acid using NaAC buffer solution (PH6.5)-acetonitrile solution and 80% acetonitrile solution as a mobile phase with gradient elution technique. Flow rate:1 ml/min, column temperature:40℃, The UV detector at 254 nm was utilized for amino acid quantification. All 17 amino acid concentration was set at the range of 0.5μmol-0.0125μmol, the linear correlation coefficients of the amino acids were above 0.99. The average recoveries of amino acids were 91.3%～103.1%. This technique was simple, accurate, and eligible for qualitative analysis of amino acid in honey. The research targeted 36 honey samples of 16 geographic sources of different colors and bionic sources. They were:Acacia, Codonopsis, Astragalus, Eriobotrya, Vetch, Red date, Coptidis and Eucalyptus honey, and so on. The result indicated that proline was the major element in free amino acid and constituted over 69.82%, averaging about 543 mg/kg in the honey samples. The finding may serve as the presupposition for further study of honey nutrition.
Scavenging effects on DPPH free radical, hydroxyl radical, NO2- radical and superoxide radical have indicated that there exists certain scavenging effects in honey, and the effects of scavenging DPPH free radical and superoxide radical are better than scavenging hydroxyl radical and NO2- radical. Duncan's multiple range test was applied to analyze 36 honey samples of 16 geographic sources on their diversity of scavenging effect on DPPH free radical, hydroxyl radical, NO2- radical and superoxide radical. The result of the experiment indicated that there existed large difference of antioxidant activities within honeys of bionic sources, and even within the honeys of same bionic source but different geographic source.
Folin-Ciocalteu method was used to determine the content of total phenolic acid in 36 honey samples of 16 geographic sources. Contents of total phenolic acid ranged from 10.43±0.69mg/100g to 168.9±0.04 mg/100g, Eucalyptus honey (Canton) ranking the highest and Acacia honey (Kunming) the lowest. It showed the difference of total phenolic acid contents in honeys was due to the effects of color, bionic and geographic aspects. Total flavonoid contents and reducing power were determined by ultraviolet spectrophotometry and the results showed that 36 samples ranged from 9.410±0.06 mg/kg to 99.63±0.04 mg/kg, the dark colored Eucalyptus honey (Guangxi) ranking the highest and the light colored Acacia honey (Kunming) the lowest. Significant differences total phenolic acid and total flavonoid contents within 36 honey samples were made by using Duncan's multiple range test. The relevance between reducing power and total phenolic acid contents, total flavonoid contents respectively were R=0.940368 and R=0.79281. It indicated that there existed linear relations between reducing power and total phenolic acid contents, total flavonoid contents respectively. The darker the honey appeared, the higher reducing power, total phenolic acid contents, and total flavonoid contents were, and vice versa. The results also indicateed that the relevance between total phenolic acid contents and scavenging effects on DPPH-free radical and superoxide radical respectively were R=0.88783 and R=0.86443. Meanwhile, the relevance between total flavonoid contents and scavenging effects on DPPH-free radical and superoxide radical respectively were R=0.71286 and R=0.73946. The findings indicate that the antioxidant capacities of honey rely on their contents of phenolic compounds. The antioxidant capacities differ in honeys of various bionic sources, and are also affected by geographic differences. Positive correlations are found in total phenolic acid contents and total flavonoid contents to reducing power and antioxidant capacities respectively. Darker colored honeys have more effects on its antioxidant capacities, and higher total phenolic acid contents and total flavonoid contents than their lighter colored counterparts.
Microscopical techniques were applied to investigate and compare the morphological properties of pollen, an obvious indicator of honey. In light of sensory features of honey in honey samples, i.e., floral origin, color, aroma and taste, the quality of honey was appraised. In use of infrared spectra device, it set up the common IR infrared spectrum fingerprint and second derivative infrared spectrum fingerprint of Linden honey, Vetch honey, and Acacia honey. The quality evaluation measures for honeys were established as the touchstone for honey sold in market.36 honey samples of 16 geographic sources were compared in their antioxidant performance to prove that the antioxidant activities of phenolic compounds in dark color honeys were better than that in the light color ones. The antioxidant performance functioned in reducing power and scavenging effect. It has been proved the quality of honey relies on its chemical properties and bionic sources, and is also affected by geographic differences. Therefore, it is significant to appraise and measure the chemical matters in honey.
苕子蜜,主要以豆科,广布野豌豆(Vicia cracca L.)和长绒毛野豌豆(Vicia villosa rothL.)为蜜源植物。苕子蜜既具有普通蜂蜜的解毒润燥作用,又有苕子清凉养颜、滋润肌肤、增强皮肤弹性、保持皮肤柔软等独特功效,颇受女士的青睐。苕子蜜呈浅琥珀色,结晶颗粒较细,气味清香,味道甘甜,为一等蜜。
刺槐蜜,主要以豆科,刺槐属植物洋槐(Robinia pseudoacacia L.),又名刺槐,为蜜源植物。洋槐蜜白色、水白色,透明,粘稠状液体,含有独特的刺槐花香,为一等蜜。
针对市场上,蜂蜜传统鉴定方法的不确定性和人为差异等因素,目前急需寻找科学的蜂蜜标准化检验体系的情况下,本实验以椴树蜜、苕子蜜和刺槐蜜为研究对象,深入研究苕子蜜、椴树蜜和刺槐蜜的质量评价方法,开发蜂蜜抗氧化作用的药用价值为目的,采用较先进的研究手段——高效液相色谱仪,紫外-可见分光光谱仪和傅里叶变换红外光谱仪三种精密实验仪器,从蜂蜜来源入手,寻找椴树蜜、苕子蜜和刺槐蜜质量评价的方法。同时,通过比较不同品种、地域的蜂蜜的还原能力和清除DPPH自由基,抑制羟自由基(·OH)、超氧阴离子自由基(O2-.)和亚硝基自由基能力的差异,评价了16种单花蜂蜜36个样本的抗氧化活性,为开发蜂蜜的药用价值和确保蜂蜜质量提供参考。
本文建立了苕子蜜、椴树蜜和刺槐蜜的傅里叶变换红外光谱(FTIR)指纹图谱及二阶导数红外光谱指纹图谱,二阶导数红外光谱指纹图谱能有效地区分苕子蜜、椴树蜜和刺槐蜜与其它品种蜂蜜,并结合聚类分析方法,基本可以把苕子蜜、椴树蜜和刺槐蜜的品种不同、产地不同和质量差异区分开,为市场上评价蜂蜜质量的方法提供一些技术。
建立了异硫氰酸苯酯柱前衍生反相高效液相色谱同时测定蜂蜜中17种游离氨基酸的实验方法。以正亮氨酸为内标,异硫氰酸苯酯为柱前衍生试剂,Venusil-AA氨基酸分析专用柱(5μm,4.6mm×250mm),醋酸钠缓冲溶液(pH6.5)-乙腈溶液和80%乙腈水溶液为流动相,采用梯度洗脱,流速：1ml/min,柱温：40℃,紫外检测器在254 nm波长处检测。氨基酸浓度在0.5μmol～0.0125μmol范围内,线性相关系数均大于0.99。17种氨基酸的平均回收率在91.3%～103.1%。该方法灵敏、准确,可作为测定蜂蜜中氨基酸的有效方法。测定结果表明：16种蜂蜜(桉树蜜、枣花蜜、黄连蜜、椴树蜜、党参蜜、龙眼蜜、紫云英蜜、荔枝蜜、荆条蜜、益母草蜜、蒲公英蜜、山花蜜、五味子蜜、枇杷蜜、苕子蜜、刺槐蜜)(深色和浅色)不同地域的36个样品的游离氨基酸含量,得到蜂蜜中的游离氨基酸主要以脯氨酸为主,占游离氨基酸总量69.82%,16种蜂蜜中平均含游离氨基酸543mg/kg左右,这一结果为进一步研究蜂蜜的营养价值提供了前提条件。
通过体外抗氧化实验(抑制DPPH自由基、超氧阴离子(02-.)自由基、羟自由基和亚硝基)的研究表明,蜂蜜对四种自由基均具有一定的清除能力,其中蜂蜜对DPPH自由基、超氧阴离子(02-.)自由基的清除能力强于对羟自由基和亚硝基的清除。采用Duncan's multiple range test统计学方法分析比较了16种蜂蜜,不同地域的36个蜂蜜样品之间对DPPH自由基、羟自由基、超氧阴离子自由基和亚硝基抑制能力的显著性差异,证明不同品种的蜂蜜,甚至同一品种而产地不同的蜂蜜,抗氧化能力有很大差异。
采用Folin-Ciocalteu比色法测定36个蜂蜜样品总酚酸含量,蜂蜜的总酚酸含量：由10.43±0.69mg/100g到168.9±0.04 mg/100g,刺槐蜜(昆明)总酚酸含量最低,桉树蜜(广东)总酚酸含量最高,说明蜂蜜品种差异、颜色差异、地域差异影响蜂蜜中的酚酸含量。利用紫外-可见分光光度法测定36个蜂蜜样品总黄酮含量和还原能力,36个样本的总黄酮含量：由9.410±0.06 mg/kg到99.63±0.04 mg/kg,色浅的刺槐蜜(昆明)含量最低,色深的桉树蜜(广西)含量最高,含量相差近10倍。采用Duncan's multiple range test统计学方法分析比较了36个蜂蜜样品之间总酚酸含量和总黄酮含量的显著性差异。同时测得蜂蜜的还原能力与总酚酸、黄酮含量的相关系数分别为：R=0.940368和R=0.79281,说明蜂蜜的还原能力与总酚酸、黄酮含量呈线性关系变化,同时蜂蜜的颜色越深,还原能力与总酚酸、黄酮含量越高,反之,还原能力与总酚酸、黄酮含量越低。
通过实验还测得蜂蜜的总酚酸含量与清除DPPH自由基和超氧阴离子自由基作用的相关系数分别为：R=0.85074和R=0.83493；总黄酮含量与抑制DPPH自由基和超氧阴离子自由基作用的相关系数分别为：R=0.62692和R=0.66133,说明蜂蜜的抗氧化能力受其多酚类(类黄酮和酚酸)成分的影响,不同品种的蜂蜜,甚至同一品种而产地不同,蜂蜜的抗氧化能力差异很大。验证了蜂蜜的总酚酸、黄酮含量、还原能力与蜂蜜体外抗氧化能力呈现正相关系；原蜜的颜色也影响抗氧化能力,色深原蜜的酚酸和黄酮含量高于色浅的原蜜,深色蜂蜜比浅色蜂蜜有更好的抗氧化活性。
蜂蜜作为健康营养物质,文章中展示了如何利用显微镜技术分析花粉形态,确定蜜源；利用评价蜂蜜样本的植物来源、色泽、结晶状态、气味或滋味等的感官特征来判断蜂蜜真伪；利用傅里叶变换红外光谱仪,建立椴树蜜、苕子蜜和刺槐蜜的普通红外光谱(FTIR)指纹图谱及二阶导数红外光谱指纹图谱,作为蜂蜜的质量评价方法,为今后建立科学的蜂蜜标准化检验体系提供实验依据。同时还比较了了苕子蜜、椴树蜜和刺槐蜜等16种蜂蜜36个不同地域样品的抗氧化能力差异。实验证明,蜂蜜的质量取决于它们自身的化学成分和蜜源植物属性,受到植物来源、地理来源等因素的影响,因而鉴定和量化蜂蜜中的微量化学物质意义重大。
Linden, mainly Tilia amurensis and Tilia mandschurica, is a kind of plants from Tilia, one of the peculiar nectariferous plants in Northeast China. Linden honey ranks as first class honey, for its properties of light amber or pale light amber color, strong fragrance, and easily crystallized. As its popularity among consumers and rare collection in the forestry, it is cherished for the functions of nurturing blood and improving weakness, reducing body heat, neutralizing poison, relieving dryness, and tranquilizing spirit. The forests in Heilongjiang and Jilin Provinces serve as the major Linden honey sources.
Vetch honey, belonging to Leguminosae, includes Vicia cracca L. and Vicia Villosa Roth L., which are defined as nectariferous plants of Vicia spp. honey. Rare produced in Yunnan-Kweichow Plateau, Vicia spp. honey bears not only the property of common honeys, such as poison neutralization and dryness-syndrome relief, but also the peculiar function of muscle and skin moisturizing, and reinforcement elasticity of skin as well. In light of its property, Vetch honey is especially popular with women consumers.
Robinia pseudoacacia L., a kind of plants from Leguminosae, also named as locust, is one of the main nectariferous plants in China. With the features of white, or water-like white color, transparent, creamy liquid and unique flower fragrance of Robinia pseudoacacia L.. Acacia honey boasts as the first class honey.
In this experiment, Linden honey, Vetch honey, and Acacia honey are chosen as subjects to further the research on their quality appraisal measures. The article focused on the investigation of three kinds honey and the development of the antioxidant and sunscreen medicine. Meanwhile, the relationship of honey and nectariferous plants was studied with high performance liquid chromatograph (HPLC), ultraviolet spectra (UV) and infrared spectra (IR). In light of honey sources, quality appraisal measures were ascertained for the three kinds of honeys. Then, the antioxidant activities of three honeys were appraised by comparing their reducing power and the diversity on scavenging effects on DPPH-free radical, hydroxyl radical, NO2 radical and superoxide radical. The study may serve as reference for honey's wellbeing function and quality security.
The study established the common infrared spectrum (FTIR) fingerprint and second derivative infrared spectrum fingerprint of Linden honey, Vetch honey, and Acacia honey. It proved that second derivative infrared spectrum fingerprint efficiently distinguished Linden honey, Vetch honey, and Acacia honey from other kinds of honeys of different sources and types in the market. Cluster analysis method was used along with IR fingerprint to analyze original honey against the honey sold in market, which can separate original honey and the processed ones. By cluster analysis, colour, botanic or geographic differences of three kinds honeys were also clarified to give light to quality evaluation measures for honeys.
A reversed phase high performance liquid chromatographic method was utilized for simultaneous determination of 17 free amino acids in Panax quinquefolium L.. The amino acid samples were mixed with norleucine as internal standard, derivatized with Phenylisothiocyanate (PITC), and then were separated on a Venusil-AA (5μm,4.6 mm X 250 mm) column specially for analysis amino acid using NaAC buffer solution (PH6.5)-acetonitrile solution and 80% acetonitrile solution as a mobile phase with gradient elution technique. Flow rate:1 ml/min, column temperature:40℃, The UV detector at 254 nm was utilized for amino acid quantification. All 17 amino acid concentration was set at the range of 0.5μmol-0.0125μmol, the linear correlation coefficients of the amino acids were above 0.99. The average recoveries of amino acids were 91.3%～103.1%. This technique was simple, accurate, and eligible for qualitative analysis of amino acid in honey. The research targeted 36 honey samples of 16 geographic sources of different colors and bionic sources. They were:Acacia, Codonopsis, Astragalus, Eriobotrya, Vetch, Red date, Coptidis and Eucalyptus honey, and so on. The result indicated that proline was the major element in free amino acid and constituted over 69.82%, averaging about 543 mg/kg in the honey samples. The finding may serve as the presupposition for further study of honey nutrition.
Scavenging effects on DPPH free radical, hydroxyl radical, NO2- radical and superoxide radical have indicated that there exists certain scavenging effects in honey, and the effects of scavenging DPPH free radical and superoxide radical are better than scavenging hydroxyl radical and NO2- radical. Duncan's multiple range test was applied to analyze 36 honey samples of 16 geographic sources on their diversity of scavenging effect on DPPH free radical, hydroxyl radical, NO2- radical and superoxide radical. The result of the experiment indicated that there existed large difference of antioxidant activities within honeys of bionic sources, and even within the honeys of same bionic source but different geographic source.
Folin-Ciocalteu method was used to determine the content of total phenolic acid in 36 honey samples of 16 geographic sources. Contents of total phenolic acid ranged from 10.43±0.69mg/100g to 168.9±0.04 mg/100g, Eucalyptus honey (Canton) ranking the highest and Acacia honey (Kunming) the lowest. It showed the difference of total phenolic acid contents in honeys was due to the effects of color, bionic and geographic aspects. Total flavonoid contents and reducing power were determined by ultraviolet spectrophotometry and the results showed that 36 samples ranged from 9.410±0.06 mg/kg to 99.63±0.04 mg/kg, the dark colored Eucalyptus honey (Guangxi) ranking the highest and the light colored Acacia honey (Kunming) the lowest. Significant differences total phenolic acid and total flavonoid contents within 36 honey samples were made by using Duncan's multiple range test. The relevance between reducing power and total phenolic acid contents, total flavonoid contents respectively were R=0.940368 and R=0.79281. It indicated that there existed linear relations between reducing power and total phenolic acid contents, total flavonoid contents respectively. The darker the honey appeared, the higher reducing power, total phenolic acid contents, and total flavonoid contents were, and vice versa. The results also indicateed that the relevance between total phenolic acid contents and scavenging effects on DPPH-free radical and superoxide radical respectively were R=0.88783 and R=0.86443. Meanwhile, the relevance between total flavonoid contents and scavenging effects on DPPH-free radical and superoxide radical respectively were R=0.71286 and R=0.73946. The findings indicate that the antioxidant capacities of honey rely on their contents of phenolic compounds. The antioxidant capacities differ in honeys of various bionic sources, and are also affected by geographic differences. Positive correlations are found in total phenolic acid contents and total flavonoid contents to reducing power and antioxidant capacities respectively. Darker colored honeys have more effects on its antioxidant capacities, and higher total phenolic acid contents and total flavonoid contents than their lighter colored counterparts.
Microscopical techniques were applied to investigate and compare the morphological properties of pollen, an obvious indicator of honey. In light of sensory features of honey in honey samples, i.e., floral origin, color, aroma and taste, the quality of honey was appraised. In use of infrared spectra device, it set up the common IR infrared spectrum fingerprint and second derivative infrared spectrum fingerprint of Linden honey, Vetch honey, and Acacia honey. The quality evaluation measures for honeys were established as the touchstone for honey sold in market.36 honey samples of 16 geographic sources were compared in their antioxidant performance to prove that the antioxidant activities of phenolic compounds in dark color honeys were better than that in the light color ones. The antioxidant performance functioned in reducing power and scavenging effect. It has been proved the quality of honey relies on its chemical properties and bionic sources, and is also affected by geographic differences. Therefore, it is significant to appraise and measure the chemical matters in honey.
引文
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