维药刺糖化学成分的基础研究
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摘要
目的:研究维药刺糖挥发油及非挥发油部分的化学成分,并对刺糖中糖类物质和总黄酮的含量进行测定。方法:1.采用系统预试验的方法对刺糖化学成分进行初步分析。2.用3、5-二硝基水杨酸法测定刺糖中总糖及还原糖的含量,用硫酸-苯酚法测定刺糖中总糖的含量。3.采用荧光分光光度法测定刺糖中总黄酮的含量。4.采用水蒸气蒸馏的方法对刺糖挥发油部分进行提取,并通过气相色谱-质谱连用技术分析,以面积归一化法确定各组分的相对百分含量,经质谱数据系统(Nist147谱库及Wiley7谱库)检索,对挥发油中的化学成分进行了鉴定。5.非挥发油部分采用回流提取法进行提取,提取物通过大孔吸附树脂色谱法、凝胶色谱法、硅胶色谱法和重结晶等方法进行化学成分的分离,分离所得的化合物借助~1H-NMR、~(13)C-NMR等波谱法和化学法鉴定结构。结果:1.刺糖中可能含有的成分蛋白质、氨基酸、酚类、糖及其苷类、黄酮、醌类、香豆素、强心苷、萜及挥发油等。2.维药刺糖中总糖含量较高,约75%左右,其中还原糖是9.29%。3.在70%的乙醇中加入pH7的三酸缓冲溶液,芦丁荧光强度最大。该法检测限为6.96×10~(-8) mol/L,线性回归方程为y=10.485x+1.3728 ,相关系数为r=0.9995 ,荧光强度与浓度的线性范围在2.7×10~(-7)mol/L～1.35×10~(-5)mol/L,RSD为1.8%。4.从刺糖的挥发油部分分离鉴定出62种化合物,已鉴定的化合物组分占总流出组分的93.9%。6种含量较高的成分为:2,3-二甲基丁烷、甲基环戊烷、5-乙酰基二氢-2(3H)-呋喃酮、3-己酮、2-乙基-1-十二烯、庚烷等。5.从刺糖中分离得到了5个单体化合物,运用现代波谱技术结合化学方法鉴定了其中4个化合物的结构:12-烯-乌苏烷醇,邻苯二甲酸丙二酯, D-Glu-1β-6α-D-Glu,D-Glu-1β-6α-D-Glu-1α-6α-D-Glu。结论:1.硫酸-苯酚法和3,5-二硝基水杨酸法测定的总糖结果相似,均为稳定可靠、简便易行的糖含量测定方法。2.用荧光分析法测定维药刺糖中总黄酮的含量灵敏度高,其准确性、重现性、线性关系、稳定性均能达到科研和生产的要求,操作简便,是药用植物中总黄酮含量测定的较佳方法。3.挥发油成分分析结果可为刺糖的质量控制提供依据。4. 4种化合物均为首次从维药刺糖中分离得到。
Objective: To search for the constituents of Saccharum Alhagi’s essential oils and fixed oils, To determine the content of total sugar, reducing sugar and flavone. Methods: 1. Qualitative test of the chemical ingredients contained in Saccharum Alhagi has been conducted with systemic preparative test method. 2. To determine the content of total sugar,reducing sugar in radix angelicae sinensis by pectrophotometry and 3 ,5-dinitrosalicylic acid (DNS) were used to colour reaction and use the phenol -sulfuric acid show a color of purple about sugar contents of Saccharum Alhagi. 3. The flavone contents in saccharum alhagi were determined using rutin as standard sample, then researches the effects of solvent pH value and time.4. Essential oils of Saccharum Alhagi were extracted by Steam distillation (SD), and analyzed by gas chromatography-mass spectrometry (GC/MS). The volatile components from the plants were identified with the help of the normal mass spectral data (NIST and WILLY database).The relative contents of volatile components were calculated by the peak area normalization method.5. The fixed oils were extracted by back streaming method,isolated by means of column chromatography over normal phase silica gel and SephadexLH-20, recrystallization,structures were identified by spectroscopic method including ~1H-NMR, ~(13)C-NMR and chemical method.Results: 1.The result showe that Saccharum Alhagi possibly contain amino acid,protein,organic acid,saccharides,saccharides glycolsides,flavonoids,anthraquinone,coumarin,cardiac glycolsides,essential oils,etc.2. The content of total suga was 75% and the content of reducing sugar was 9.29%. 3. The results indicated that fluorescence is the strongest while solvent is 70% ethanol and put in moderator (pH=7.0). The detection limit was 6.96×10~(-8) mol/L, regression equation was y=10.485x+1.3728, the correlation coefficient was 0.9995, the range of linearity relationship was 2.7×10~(-7)mol/L～1.35×10~(-5)mol/L , and RSD was 1.8%. 4. Sixty-two components have been identified from the essential oil and their amount accounted to be 93.93%.The main components were 2,3-Dimethylbutane , Methylcyclopentane ,5-Acetyldihydro-2(3H)-furanone,3-Hexanone,2-Ethyl-1-dodecene,Heptane. 5. We have isolated four compounds, them were determined. Conclusions:1. DNS method and phenol -sulfuric acid method is convenient, rap id and reliable. It can be used for the quality control about Saccharum Alhagi. 2. The method is a simple, rapid and accurate approach to detect flavone content in saccharum alhagi and has a good application prospect.3. This result of the analysis of the chemical components of essential oil from Saccharum Alhagi can be applied to analyze the essential oil components extracted from Saccharum Alhagi. 4. Four compounds were isolated from Saccharum Alhagi for the first time.
Objective: To search for the constituents of Saccharum Alhagi’s essential oils and fixed oils, To determine the content of total sugar, reducing sugar and flavone. Methods: 1. Qualitative test of the chemical ingredients contained in Saccharum Alhagi has been conducted with systemic preparative test method. 2. To determine the content of total sugar,reducing sugar in radix angelicae sinensis by pectrophotometry and 3 ,5-dinitrosalicylic acid (DNS) were used to colour reaction and use the phenol -sulfuric acid show a color of purple about sugar contents of Saccharum Alhagi. 3. The flavone contents in saccharum alhagi were determined using rutin as standard sample, then researches the effects of solvent pH value and time.4. Essential oils of Saccharum Alhagi were extracted by Steam distillation (SD), and analyzed by gas chromatography-mass spectrometry (GC/MS). The volatile components from the plants were identified with the help of the normal mass spectral data (NIST and WILLY database).The relative contents of volatile components were calculated by the peak area normalization method.5. The fixed oils were extracted by back streaming method,isolated by means of column chromatography over normal phase silica gel and SephadexLH-20, recrystallization,structures were identified by spectroscopic method including ~1H-NMR, ~(13)C-NMR and chemical method.Results: 1.The result showe that Saccharum Alhagi possibly contain amino acid,protein,organic acid,saccharides,saccharides glycolsides,flavonoids,anthraquinone,coumarin,cardiac glycolsides,essential oils,etc.2. The content of total suga was 75% and the content of reducing sugar was 9.29%. 3. The results indicated that fluorescence is the strongest while solvent is 70% ethanol and put in moderator (pH=7.0). The detection limit was 6.96×10~(-8) mol/L, regression equation was y=10.485x+1.3728, the correlation coefficient was 0.9995, the range of linearity relationship was 2.7×10~(-7)mol/L～1.35×10~(-5)mol/L , and RSD was 1.8%. 4. Sixty-two components have been identified from the essential oil and their amount accounted to be 93.93%.The main components were 2,3-Dimethylbutane , Methylcyclopentane ,5-Acetyldihydro-2(3H)-furanone,3-Hexanone,2-Ethyl-1-dodecene,Heptane. 5. We have isolated four compounds, them were determined. Conclusions:1. DNS method and phenol -sulfuric acid method is convenient, rap id and reliable. It can be used for the quality control about Saccharum Alhagi. 2. The method is a simple, rapid and accurate approach to detect flavone content in saccharum alhagi and has a good application prospect.3. This result of the analysis of the chemical components of essential oil from Saccharum Alhagi can be applied to analyze the essential oil components extracted from Saccharum Alhagi. 4. Four compounds were isolated from Saccharum Alhagi for the first time.
引文
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