| China is a country which is rich in the part of Chinese herbal medicine plant resource particularly while many kinds of chemical compositions are very complex and with strong physiological activities and higher clinical values. Such as alkaloids, which is a kind of alkaline nitrogen compounds existing in organism. Meanwhile they are urgent to be developed to provide rich material resources and infrastructures for plant medicine, functional food, health care products and cosmetics. However the contents of alkaloids in most plant resources are lower and the structures of alkaloids are complicated, therefore, the separations and purifications of alkaloids are quietly difficult. In this paper, in view of the technical problems in the separation and purification of different types of alkaloids compounds, we should investigate the feasibility which is exploring a new type of liquid-liquid chromatographic separation technology-- PZRCCC to separate and purify the different types of alkaloids, then we should study the characteristics of the solvent systems of different types of alkaloids separation and purification. In the end, set up a key separation technology basing on PZRCCC and provide a new method and a new thinking for the development and utilization for the rapid extraction, concentration, separation, purification and structural identification of different types of alkaloids in natural profucts. The main results and discussions are as follows:1. The study established a method to separate and purify ndole alkaloids by PZRCCC.The research applied the method of PZRCCC, selected evodia and Rauwolfia as experimental materials and investigated the effect of parameters in the separation of PZRCCC about the solvent system compositions, proportions, concentrations of acid and alkali and so on.The results showed: Choosing Chloroform- ethyl acetate- methanol- water(3:1:3:3, v/v) as a two-phase solvent system which can obviously seperate the tryptamine alkaloids, evodiamine and dehydroevodiamine. From 2 g crude extract of evodia disposable isolated tryptamine indole alkaloids-- 101 mg evodiamine and 304 mg dehydroepiandrosterone evodiamine in a single run, which purity are 90.0 % and 93.1 %. At the same time, choosing petroleum ether- ethyl acetate- methanol- water(5:5:2:8, v/v) as a two-phase solvent system. As a result, monoterpenoid indole alkaloids-- 112 mg of reserpine and 21 mg yohimbine were obtained from 3 g of crude extract in a single run. The analysis of the isolated compounds were determined with purities of 90.0 % and were carried out by electrospray ionization-mass spectrometry(ESI-MS), 1H-NMR, and 13C-NMR.2. The study established a method to separate and purify benzophenanthridines alkaloids by PZRCCC.The research applied the method of PZRCCC, selected Macleaya cordata. and Tinospora Capillipes Gagnep. as experimental materials and investigated the effect of parameters in the separation of PZRCCC about the solvent system compositions, proportions, concentrations of acid and alkali and so on.The results showed: Choosing the two-phase solvent system chloroform–ethyl acetate–methanol–water(3:1:3:2, v/v). From 2.0 g of crude extract, chelerythrine(600 mg) and sanguinarine(750 mg) were obtained in a single run with the purity of both over 95.0% as determined. At the same time, choosing Chloroform- methanol- water(4:3:2, v/v) as a two-phase solvent system. From 2 g of crude extract, 512 mg benzophenanthridines alkaloid-- palmatine and 421 mg protoberberine alkaloid-- jatrorrhizine with purities of 95.1 % and 95.3 % were obtained. All were carried out by electrospray ionization-mass spectrometry(ESI-MS), 1H-NMR, and 13C-NMR.3. The study established a method to separate and purify piperidine alkaloids by twotwo extraction and PZRCCC.The research applied the method of PZRCCC, selected Areca catechu L.as experimental materials and combinated of the two extraction, according to the physical and chemical properties of these alkaloids, then investigating the effect of parameters in the separation of PZRCCC about the solvent system compositions, proportions, concentrations of acid and alkali and so on.The results showed: Choosing petroleum ether- ethyl acetate- methanol- water(3:7:3:7, v/v) as a two-phase solvent system. As a result, piperidine alkaloid-- 75 mg of arecoline with purities of 91 % was obtained from 1 g of crude extract in a single run which was carried out by electrospray ionization-mass spectrometry(ESI-MS), 1H-NMR, and 13C-NMR.4. The study established a method to separate and purify macrocyclic alkaloids by MCI column elution and PZRCCCThe research applied the method of PZRCCC, selected Carica papaya L. Leaves as experimental materials and combined with MCI column elution according to the nature of the macrocyclic alkaloids, then investigating the effect of parameters in the separation of PZRCCC about the solvent system compositions, proportions, concentrations of acid and alkali and so on.The results showed: The MCI column can remove pigment matrix well and the macrocyclic alkaloids have a preliminary enrichment. Choosing petroleum ether- ethyl acetate- methanol- water(5:5:2:8, v/v) and petroleum ether- ethyl acetate- methanol- water(5:5:1:9, v/v)as two-phase solvent systems used in tandem. As a result, macrocyclic alkaloids-- 112 mg of carpaine and 21 mg pseudocarpaine were obtained from 2 g of crude extract. The analysis of the isolated compounds were determined with purities of 90.0 % which were carried out by electrospray ionization-mass spectrometry(ESI-MS), 1H-NMR, and 13C-NMR.5. The study established a method to separate and purify aporphine alkaloids by ultrasound extraction and PZRCCCThe research applied the method of PZRCCC, selected Lindera aggregata(Sims) Kosterm. as experimental materials and combined the ultrasonic extraction, then investigating the effect of parameters in the separation of PZRCCC about the solvent system compositions, proportions, concentrations of acid and alkali and so on.The results showed: Choose Chloroform- methanol- water(4:3:3, v/v) as a two-phase solvent system. From 1 g of crude extract, 150 mg aporphine alkaloid-- boldine was obtained with purity of 95 % which was carried out by electrospray ionization-mass spectrometry(ESI-MS), 1H-NMR, and 13C-NMR.Through the separation and purification of different types of alkaloids by PZRCCC from a few kinds of medicinal materials in above, the general steps of separation of alkaloids are determined--choose suitable solvent system, adjust percentage of solvent and concentration of acid-akkali or change the instrument parameters and power, at the same time, it can combine other techniques(MCI column chromatography, ultrasonic ectraction) in order to achieving the different types of alkaloids successfully. All in all, the study provide a new technology and a new idea for separation and purification of different types alkaloids. |
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