| Citrus aurantium L. is mainly distribution in Sichuan, Chongqing, Hunan, Jiangxi. Fructus Aurantii Immaturus and Fructus Aurantii are from Citrus aurantium L. at different harvest periods. Fructus Aurantii Immaturus is collected in May and June. Fructus Aurantii is collected in July when its peel still green. Fructus Aurantii Immaturus and Fructus Aurantii from Sichuan are good quality. Hunan is the production base of Fructus Aurantii Immaturus and Fructus Aurantii. Fructus Aurantii Immaturus and Fructus Aurantii from Jiangxi are the famous-region drugs. Both from the same source, but the action difference because of differert harvesting periods. Fructus Aurantii Immaturus is only used synephrine as quality control indicator and Fructus Aurantii is used naringin and neohesperidin for quality control indicators in current pharmacopoeia standard. It does not reflect both common and characteristic.Due to quality assessment system imperfect and lack of system analysis and comparision, so this paper engaged to harvesting periods and used main pharmacological compounds of flavonoids, alkaloids, phenolic acids and limonin as indicators. Systematically study dynamic changes of effective components in the entire growth cycle. Focus on the following research:1. Studies on composition analysis and comparision of fruit from Citrus aurantium L. at different harvest periodsOn the basis of Pharmacopoeia of the People’s Republic of China (2010edition), increasing assay of total flavonoids, total alkaloids, total phenolic acids, narirutin, hesperidin and limonin.1.1FlavonoidsFlavonoids in Fructus Aurantii Immaturus and Fructus Aurantii are mainly narirutin, naringin, hesperidin and neohesperidin. This study used HPLC and UV methods for determination of the four kinds of flavonoid and total flavonoids. The results showed that total flavonoids decreased from41.13%to8.28%with the increase of fruit maturity. Hesperidin and neohesperidin were significantly degressive trend.But the contents of naringin and narirutin first increased and then decreased. Transforming relationship among flavonoids needs further study.1.2AlkaloidsUsing HPLC and UV methods for determination of total alkaloids and synephrine. The results showed that total alkaloids decreased from10.35%to1.2%and synephrine decreased from0.79%to0.22%with the increase of fruit maturity.1.3Total phenolic acidsReported in the literature, phenolic acids not only have antioxidant, anti-free radicals, inhibition of LDL oxidation and prevention of cardiovascular disease, but also have the function of anti-cancer, anti-inflammatory and anti-platelet aggregation. At present, phenolic acids in Fructus Aurantii Immaturus and Fructus Aurantii are studied less, this is the first comparative analysis of the variation of total phenolic acids content in the fruit from Citrus aurantium L. at different harvest periods. Results, with the increase of fruit maturity, the total phenolic acids content decreasing from8.73%to0.55%. The results provided experimental basis for the raw material selection on phenolic acids extration.1.4LimoninLimonin as a bitter substance, widespread in the citrus peel. Reported in the literature, it has wide range of pharmacological activity on the anti-inflammatory, analgesic, anti-cancer. This study used HPLC method for the first time to analyse variation of the limonin in the fruit from Citrus aurantium L. at different harvest periods. The results showed that the content of limonin was highest (0.5%) in mid-June with the increase of fruit maturity. Then its content decreased, increased, decreased by the change. In early September, the content of limonin was the lowest (0.26%). The results provided experimental basis for the raw material selection on limonin extration.These results showed that the contents of total alkaloids, total flavonoids, total phenolic acids, synephrine,hesperidin and neohesperidin were significantly degressive trend with the increase of fruit maturity. But the contents of naringin, narirutin, limonin showed a downward trend overall. The results provided an experimental basis for formulating harvest periods.2. Comparative study on ingredients in different parts of Fructus AurantiiDue to core flesh easily moldy, degenerative and vermiculate. In the traditional medication, only using the peel of the Fructus Aurantii and core flesh to discard. Previously reported in the literature, there is no comparative study on active ingredients in peel and core flesh of Fructus Aurantii. In order to better comprehensive utilization of Fructus Aurantii, to compare with the contents of flavonoids and alkaloids in peel and core flesh of Fructus Aurantii. The results showed there were five active ingredients of narirutin, naringin, hesperidin, neohesperidin and synephrine. And the content of active ingredients in peel was higher than core flesh. This study provides the basis for the comprehensive utilization on resources of Fructus Aurantii.3. Comparative study on essential oil in Fructus Aurantu Immaturus and Fructus Aurantu3.1Study on extraction methods of essential oilThe essential oil as one of the two drugs main effective ingredients, the traditional steam distillation usually be used to extract essential oil. While other methods studied less, such as solvent refluxing, ultrasonic extraction. In this study, GC-MS was used to analyze essential oil extracted by petroleum ether ultrasonic, ether ultrasonic, N-hexane ultrasonic, petroleum ether reflux, ether reflux, N-hexane reflux, steam distillation, pharmacopoeia A method. To analyze the different extraction methods on chemical composition of volatile oil. The results showed different extraction methods had important influence on chemical constituents of the volatile oil. Lots of macromolecular in solvent refluxing extraction. More of micromolecule in steam distillation extraction and pharmacopoeia A method extraction. Components of volatile oil extracted by2010Chinese Pharmacopoeia Appendix XD A method were more rich, and identified more. The next was steam distillation. The research of essential oil should consider a variety of extraction methods.3.2Comparative study on essential oil in Fructus Aurantu Immaturus and Fructus Aurantii from different originsLimonene and linalool were the main components of essential oil in Fructus Aurantii Immaturus and Fructus Aurantii. The essential oil had significant difference at different origins, different growth cycle. With the extension of the growth cycle, linalool, a-terpineol, carvacrol and geraniol showed a certain regular changes. The content of linalool and carvacrol showed declined with the extension of the growth cycle. But, the content of a-terpineol and geraniol showed increased.3.3Comparative study on essential oil in processed products of Fructus Aurantii Immaturus and Fructus AurantiiDifferences were found by comparing with essential oil before and after processing. After processing, Limonene was increased which was consistent with the reports. Meanwhile, the ratio of various components changed and there were some of the new components to emerge. Fructus Aurantii Immaturus from Jiangxi after processing, the total compounds reduced from38to29. The ratio of linalool and limonene decreased from the raw product of0.57to0.29; Fructus Aurantii from Jiangxi after processing, the ratio of linalool and limonene decreased from the raw product of1.06to0.25. The content of some compounds were increased or decreased. The results provided an experimental basis for further study of the processing mechanism.4. Optimizing extraction process of the total phenolic acids for the first timeAt present, extraction methods study were focused on flavonoids, synephrine and essential oil. The phenolic acids was studied less. So using ultrasonic wave and Box-Behnken statistical method to optimize extraction process of the total phenolic acids. Using gallic acid for the contrast and colorimetric method to determine. The Optimal ultrasonic extraction process is as follows:the ethanol concentration is56%,the ratio of solvent to material is28:1, the temperature is30℃, the ultrasonic power is400W, extraction times are2and each time20min.The study provided basis for development and utilization of phenolic acids in Fructus Aurantii Immaturus and Fructus Aurantii.5. Study on quality evaluation of Fructus Aurantii Immaturus and Fructus Aurantii.At present, the chemical evaluation method is widely used in quality evaluation. But there are limitations. For example, pharmacopoeia standards to evaluate the same source of herbs by the different indicators.So this paper used multi-index to study collection period. With the extension of the growth cycle, the contents of total alkaloids, total flavonoids, synephrine, hesperidin and neohesperidin were significantly degressive trend.But the contents of naringin, narirutin, limonin showed a downward trend overall. Therefore, when evaluating quality of medical material, use naringin, neohesperidin and synephrine to formulate the content limits of Fructus Aurantii Immaturus and Fructus Aurantii. It can comprehensively reflect the character and individuality of the same source of herbs.Because the complexity and instability of essential oil, the total amount of essential oil reduced with the extension of the growth cycle, the content of a-terpineol and geraniol increased, and the content of linalool and carvacrol declined with the extension of the growth cycle. So, the quality evaluation of essential oil should focus on the total amount of essential oil and the contents of a-terpineol, geraniol, linalool, carvacrol.The study provided exemplary for the quality evaluation of medical material with the same sources. Due to the limit of time and research funds, this paper only studied the herbs in Rongchang from Chongqing. Other areas of herbs are further to study. On this basis, to improve the harvesting standards and quality evaluation system. |
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