葛根、大黃、地黃之定性、定量分析與數據化指紋圖譜研究

林靖哲(Lin Ching-Che)

指導教授 ： 許順吉

國立臺灣師範大學/理學院/化學系/博士(2004年)

摘要

無資料

關鍵字

葛根 ； 大黃 ； 地黃 ； LC-MS ； ELSD

並列摘要

High-performance liquid chromatography (HPLC) is the most commonly used instrument for analyzing the components in Chinese herb medicine. As for compounds which do not have any UV absorption group, ELSD or mass spectrometer coupled HPLC is used for analysis. Advantages of each instrument are combined to expand the chemical evaluation of Chinese herb medicine.

There are five parts in this study. The first part is the LC-MS analysis and plant origin identification of Puerariae Radix, which is dry roots from Leguminosae Pueraria lobata Ohwi. Puerariae Radix is an important Chinese herbal medicine to expel superficial evils. Its major components are isoflavones. LC-MS was used in this study to separate and identify 12 marker compounds simultaneously. From the fragmentation patterns of these marker components, a rule for identifying isoflavones can be established, and structure of unknown compounds can be predicted. Two kinds of Puerariae Radix are available on the market, P. lobata Ohwi and P. thomsonii Benth. In this study, fingerprint analysis of Puerariae Radix was accomplished using the statistical software with other data processing methods.

The second part is the LC-MS analysis and plant origin identification of Rhei Rhizoma, which is dry roots and stems from Polygonaceae plants. Rhei Rhizoma is an important purgastive herbal medicine. Its components include anthraquinones, dianthraquinones, stilbenes, and galloylglucoses. LC-MS was used in this study to isolate and identify 19 index components simultaneously. Comparing the fragments of MSn mass spectra in the TIC diagram from unknown peaks with previous reports, their structure can be predicted. Three kinds of Rhei Rhizoma are available on the market, Rheum tanguticum Maxim, Rheum palmatum Linne, and R. officinale Baillon. Plant origin identification of three species was accomplished using graphic identification technology and HPLC fingerprint analysis.

The third part is the development of analytical method for Rehamnnia with HPLC-UV-ELSD and chemical identification of Rehmanniae Radix and Rehmanniae Preparata Radix. Pharmacological significance of processing and different results from various processing methods for Rehamnnia were discussed in this study as well. The major components of Rehamnnia are saccharides. With a mobile phase of 0.1% formic acid and methanol, a Hypercarb porous graphitic column, and an ELSD detector, saccharides in Rehmanniae were isolated and quantitatively analyzed. Results showed that carbohydrates in Rehmanniae are much different after processing. Therefore, the peak area ratio of mannintriose and stachyose, which are the two major components of Rehmanniae, was used to differentiate Rehmanniae Radix and Rehmanniae Preparata Radix.

Catalpol is the active ingredient to decrease blood sugar. It is destroyed during the processing. Therefore, the pharmacological reactions of Rehmanniae Preparata Radix are different from Rehmanniae Radix. It is also found that Maillard reaction occurs during the processing of Rehmanniae. The intermediate product, 5-HMF, has the activity of Aldose reductase inhibiotor (ARI). The end products, melanoidins, and oligosaccharides have the functions to activate intestinal probiotics, decrease cholesterol, regulate immunity, increase anti-oxidation, and increase the absorption of minerals. This also shows that clinical effects of Rehmanniae Preparata Radix are different from Rehmanniae Radix. In this study, several groups of samples from Rehmanniae were prepared to study different results due to various time of processing, frequency of processing, and incubation solutions. It was found that sample which was steamed 3 hours and dried for 9 cycles is the best. Addition of acidic substance during processing increased the conversion of 5-HMF from saccharides.

The fourth part is the analysis of trace elements. The content of trace elements in P. lobata Ohwi and P. thomsonii Benth, and the contents of trace elements in different stages of processing of Rehamnnia were analyzed. Results showed that the elemental profile of P. lobata Ohwi and P. thomsonii Benth are somewhat different. P. thomsonii Benth contains higher amount of important elements than P. lobata Ohwi. As to Rehamnnia, most elements decrease with number of processing cycles increase. However, some elements such as Fe, Zn, Mn, Co, Ga, Sr, Ba, Ni increases.

The last part is the analysis of amino acids in almond seeds and Rehamnnia. Amino acids do not have characteristics of UV absorption, and they are very polar molecules. In this study, AccQ-Tag reagent, which is a newly developed derivative reagent by Waters, was used to perform a pre-column derivative reaction. The stable derivative products were then analyzed quantitatively by reverse phase HPLC. It is found that there are 18 kinds of amino acids in seed of P. armeniaca L.var. ansu Maxim sample. The content in P. armeniaca L.var. ansu Maxim is much higher than Prunus armeniaca L. There are 17 kinds of amino acids found in Rehamnnia. The content in Rehmanniae Radix is much higher than Rehmanniae Preparata Radix.

並列關鍵字

pueraria ； rhubarb ； rehmanniae ； LC-MS ； ELSD