信阳毛尖高效液相色谱指纹图谱的初步研究

发布时间：2018-01-05 08:13

本文关键词：信阳毛尖高效液相色谱指纹图谱的初步研究　出处：《河南科技学院》2017年硕士论文　论文类型：学位论文

【摘要】：茶叶品质与产地、自然环境及加工工艺关系十分密切,产地往往成为茶叶品质的代名词。信阳毛尖产于河南信阳,历史比较悠久,生态环境优越,茶叶品质较佳,属于绿茶类。mI河区、平桥区、光山县等茶区域是其主要产地。近年来,由于“信阳毛尖”地理标志产品的品牌响、经济效益高、影响力大,致使江苏、湖北、浙江等地方有些茶企用本地绿茶原料,模仿信阳毛尖的加工工艺制作信阳毛尖茶,导致市场上“信阳毛尖”标价过高而往往质量低的掺假现象时有发生,成为长期困扰信阳毛尖原产地保护的难题。高效液相色谱指纹图谱技术是对茶叶进行产区鉴别的分析方法之一,当前茶叶高效液相色谱指纹图谱多是基于茶叶中多酚类等特征成分的含量,根据茶叶成分的特殊性,鉴别产地和品种。本文收集了来自信阳地区及外地其他地区的62个茶样,进行高效液相色谱测定,从高效液相色谱指纹图谱的研究出发,借鉴中药指纹图谱技术并结合主成分分析及聚类分析法来鉴别信阳毛尖茶的产地和品质。其研究内容如下:1、应用高效液相色谱方法建立信阳毛尖茶指纹图谱。经方法学考察,该方法精密度、稳定性和重复性都较好,适合对信阳毛尖茶进行分析研究。通过pH、流动相的梯度洗脱方式及流速的改变,确定的最优色谱条件为:ZORBAX SB-C18反相柱(4.6×150 mm,5μm);检测波长为280 nm;柱温35℃;A相为2%乙酸;B相为乙腈;梯度洗脱方式为:96%A(0 min)→88.5%A(15 min)→87.6%A(20 min)→86.3%A(30 min)→81.3%A(35 min)→96%A(40 min);检测器:紫外检测器;流速0.8 mL/min;洗脱时间为40 min;进样量为10μL。通过各个组分的保留时间来进行定性分析,并根据峰面积进行了定量测定。2、按照1中优化的色谱条件进行样品的测定分析,以咖啡碱(CAF)峰为参照峰,最终选择了19个色谱共有峰作为主要的特征峰来构建信阳毛尖多元化学指纹图谱的组成因子。从图谱的整体性出发,运用中药色谱指纹图谱相似度评价系统软件进行相似度分析,并结合主成分分析、聚类分析等方法对色谱图进行产地识别的初步分析。主成分分析结果表明提取的4个主成分累计方差贡献率为82.647%,根据前两个主成分得分图发现主成分分析方法不能完全区分信阳地区的茶样与外地茶,但还是能够显示他们之间的差异。聚类分析发现同一产地不同品种的信阳毛尖茶可以很好地区分开,而同一品种不同产地的信阳毛尖茶区分不开。3、氨基酸的含量与茶叶品质密切相关,茶叶中氨基酸含量的差异更能直接地反映品种的特性,通过不同茶叶游离氨基酸、茶氨酸、呈味氨基酸的比较以及从绘制的氨基酸指纹图谱分析不难发现,同一采摘时期下信阳群体种的信阳毛尖游离氨基酸总量明显低于安吉白茶种;不同品种信阳毛尖及同一品种不同采摘时期制作的信阳毛尖茶样的必需氨基酸平均含量均存在显著差异。以缬氨酸为参照氨基酸绘制的必需氨基酸指纹图谱中,光山县信阳毛尖与其他地区之间必需氨基酸含量差异较大,尤其是苏氨酸的含量。总游离氨基酸指纹图谱中,茶氨酸的含量相对最高,其次是谷氨酸。不同产地茶叶样品必需氨基酸的含量中基本上呈现的规律是:光山县的茶氨酸和谷氨酸含量相对较高,其次是黑龙潭、白龙潭,最低的是土门地区。总之,我们可以从绘制茶叶氨基酸指纹图谱中,根据茶氨酸、谷氨酸和苏氨酸相对含量的高低来可以作为茶叶产地识别的一个依据。
[Abstract]:The tea quality and origin, the natural environment and the processing technology of the close relationship between the origin often become synonymous with tea quality. Xinyang Maojian tea produced in Henan Xinyang, has a long history, excellent ecological environment, the quality of tea is good, which belongs to the class of.MI Green Tea District, Pingqiao District, Guangshan county tea area is the main producing areas in recent years. Because, "Xinyang Maojian tea" geographical indication brand ring, high economic efficiency, high impact, resulting in Jiangsu, Hubei, Zhejiang and other places some tea enterprises with local Green Tea raw materials, processing technology to imitate Xinyang Maojian tea production Xinyang Maojian Tea, causing adulteration on the market "Xinyang Maojian tea" overpriced and often low quality when had become a long-term problem of Xinyang Maojian tea origin protection. The HPLC fingerprint analysis technology is one of the areas for the identification of tea, the tea liquid Chromatographic fingerprint is content of polyphenols in tea based on characteristic components, according to the particularity of tea ingredients, origin and variety identification. This paper collected 62 tea samples from Xinyang area and other areas of the field, HPLC fingerprint spectrum of the high performance liquid chromatography of from the traditional Chinese medicine fingerprint technology and production and quality combined with principal component analysis and cluster analysis method to identify Xinyang Maojian Tea. The research contents are as follows: 1, Xinyang Maojian Tea fingerprint by high performance liquid chromatography method is established. The methodology study, the precision, stability and repeatability are good for study the Xinyang Maojian Tea. By pH, flow velocity and the change of phase gradient elution, the optimal chromatographic conditions were determined: ZORBAX SB-C18 reversed-phase column (4.6 x 150 mm, 5 m); the detection wavelength was 280 nm; column The temperature of 35 DEG C; A phase of 2% acetic acid; B phase acetonitrile; gradient elution method is as follows: 96%A (0 min), 88.5%A (15 min), 87.6%A (20 min), 86.3%A (30 min), 81.3%A (35 min), 96%A (40 min); the UV detection rate of 0.8; mL/min; elution time was 40 min; the retention time of the injection volume was 10 L. by each component to the qualitative analysis, and quantitative determination of.2 according to the peak area, were analyzed according to the chromatographic conditions in 1 samples of optimization, caffeine (CAF) peak as the reference peak, the final choice of 19 a common factor chromatography peak as the main characteristic peaks to build Xinyang Maojian multiple chemical fingerprint. From the overall view of the similarity analysis using chromatographic fingerprint similarity evaluation system software, combined with principal component analysis, cluster analysis and other methods were analyzed to identify the origin of the chromatogram. Principal component analysis showed that 4 principal components are extracted from the cumulative variance contribution rate of 82.647%, according to the principal component analysis method can not fully differentiate tea samples with the field of tea in Xinyang before the two principal component scores charts to find, but still able to show the difference between them. The clustering analysis found the same origin of different varieties of Xinyang Maojian tea tea can well separate areas, and the same species from different habitats Xinyang Maojian Tea can't distinguish between.3, and the content of amino acid of tea quality is closely related to the differences of contents of amino acids in tea can directly reflect the variety characteristics, the free amino acids in different tea, theanine, a comparison of amino acids and amino acids from the taste fingerprint analysis of drawing is not difficult to find, with a total time of picking Xinyang Maojian free amino groups of Xinyang is significantly lower than that of Anji white tea varieties in Xinyang hair tip; And the same species of different average content of essential amino acid picking time making Xinyang Maojian Tea samples had significant difference. The fingerprint of essential amino acids in the amino acid reference drawing for valine in Guangshan County, Xinyang Maojian and other areas essential amino acid content is different, especially threonine. The contents of total free amino acid content in the fingerprint. Theanine is the highest, followed by glutamic acid. Basically the rules of the content of essential amino acids in tea samples: theanine and glutamic acid content in Guangshan county is relatively high, followed by the Black dragon Pool, white dragon pool, is the lowest in Tumen area. In a word, we can draw from the fingerprint of amino acids in tea and according to the relative content of theanine, glutamic acid and threonine level can be used as a basis for identifying the origin of tea.

【学位授予单位】：河南科技学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S571.1

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