Extraction And Purification Of Alkaloids From Lotus Leaf

A traditional Chinese herb called lotus leaf is approved as'material that can be used in healthy foods'by Ministry of Health, P. R. China.In the present study, there were few paper about the separation and preparation of lotus leaf alkaloids. In this paper, the technology for extraction of alkaloid from lotus leaf was studied. Quantitative analysis of nuciferine by HPLC was established here. Two methods-high speed countercurrent chromatography and high performance liquid chromatography for separation and purification of alkaloids were applied. The main results were described as follows:1. The technology for extraction of alkaloidThe technology for extraction of alkaloid from lotus leaf was studied. Through combination of artificial neural network (ANN) with traditional orthogonal design, a new method for analyzing, processing and optimizing the test data was put forward. The optimum condition for extraction was summarized, namely, pH value 2.5,extracting temp. 85℃, duration of extraction 1.75h and solid to liquid ratio 1:35. The satisfactory results confirmed the reasonableness and practicability.2. Alkaloids of lotus leaf were identified by TLCUsing thin layer chromatography(TLC) for identifying the alkaloid in the extract demonstrated that the kinds of alkaloid changed with the extract solvent. Chloroform-methanol- lignocaine amine (90:9:1)mixture as a mobile phase. The nuciferine in the lotus leaf were found.3. Separation and purification of alkaloids from Lotus Leaf by high speed countercurrent chromatographyHigh speed countercurrent chromatography (HSCCC) was applied to the separation of alkaloids from Lotus Leaf the first time. High speed countercurrent chromatography with a two-phase solvent system composed of Chloroform:methanol:pH4.0 water(4:3:2,volume ratio). Five collections were separated. The results were verified by TLC, in which three components exhibited orange-red colour with bismuth potassium iodide reagent. HSCCC is a convenient, feasible and effective technique for the separation and purification of active components in Chinese herbal medicines.4. Separation and purification alkaloids from Lotus Leaf by high performance liquid chromatography. And found the method of HPLC analytical of nuciferine.A method using preparative high performance liquid chromatography (HPLC) to isolate four individual alkaloids from lotus leaf was investigated. A preparative C18 column was employed in the procedure. First, by isocratic elution with methanol:water:lignocaine amine (75:25:0.0125,V/V) as mobile phase at a flow rate of 8 ml/min, two mixture (part1, part2) were obtained. Then, by isocratic elution with acetonitrile:water:lignocaine amin(e55:45:0.0125,60:60:0.0125)as mobile phase at a flow rate of 8 ml/min for part1 and part2, respectively, three substance were obtained. HPLC analytical results showed that the purity of each alkaloid was over 97%. Under the chromatographic condition, every peak was completely baseline separated.In a word, the technology for extraction of alkaloid from lotus leaf was founded. High speed countercurrent chromatography (HSCCC) and performance liquid chromatography (HPLC) are two efficient methods for separation and preparation of alkaloid.