| At present,the use of antibiotic additives in feed is banned in China,and the study of antibiotic substitutes has been paid more and more attention.Studies have shown that honeysuckle contains a large amount of chlorogenic acid,chlorogenic acid has superior antibacterial,antiviral and lipid regulation performance,making it has a great application prospect in the function of replacing antibiotics.With the rise of the pharmaceutical industry and the expansion of the health products and food market,the demand for the green health additive chlorogenic acid is also increasing year by year,but the supply of chlorogenic acid in the current market is in short supply,which is mainly due to the insufficient technological level of extracting chlorogenic acid from honeysuckle at present.This thesis mainly in sichuan bazhong nanjiang characteristic economy crops "south silver 1" new honeysuckle flower,stem and leaf of the research for comprehensive utilization of high efficiency,high quality development,research and development of high and new technology,developed the high economic efficiency and market competitiveness for luyuan acid extraction of new technology,new preemption honeysuckle industry development.Implementation from market demand and actual situation,the paper adopts the single factor method and orthogonal experimental method to determine the "south silver 1" new honeysuckle flower,stem and leaf of luyuan acid of high efficient and environmentally friendly new extraction technology and the purification technology related to it,and respectively by high performance liquid chromatography(HPLC)method to measure its luyuan acid content in flowers,stems,leaves,In order to effectively extract planting in the rest of the hundreds of millions of tons of stem leaves after picking honeysuckle cane luyuan acid,rich in ease market demand for luyuan acid,and reduce the cost of traditional extraction technology,to promote the characteristic economy crops of flos lonicerae in sichuan province industry technologies provide a good demonstration effect,the drive to produce more significant social and economic benefits,It has practical value and practical significance.In this experiment,the optimal process parameters for extraction,decolorization,separation and purification of chlorogenic acid from the flowers,stems and leaves of "Nanyin No.1" new strain of Flos Lonicerae were determined,and the following studies were carried out respectively:(1)Feasibility analysis of determination of chlorogenic acid content by HPLCIn this paper,the feasibility of determination of chlorogenic acid by HPLC was confirmed from the aspects of precision,stability,reproducibility and recovery.The experimental results show that the system has good precision,high stability of chlorogenic acid within 24 h,good repeatability and ideal recovery rate.Finally,it is determined that HPLC is completely feasible to determine chlorogenic acid content.(2)Study on the optimal extraction process parameters of chlorogenic acidBy single factor experiment and orthogonal experiment,the optimum technological parameters of extracting chlorogenic acid from flowers,stems and leaves of "Nanyin No.1" new strain of Lonicerae japonica were obtained,and the content of chlorogenic acid was extracted and measured respectively.The optimal extraction parameters of chloric acid from Lonicerae japonicae were as follows: the particle size of Lonicerae japonicae was 60 mesh,the extraction solvent was 75% methanol solution,the solvent p H value was 4,the extraction time was 1.5h,the extraction times were 2 times,the ratio of solid to liquid was 1:20,and the extraction temperature was 70℃.The optimal extraction parameters of chlorogenic acid from stems were as follows:the stem particle size was 80 mesh,the extraction solvent was 75% methanol solution,the solvent p H value was 4,the extraction time was 1.5h,the extraction times were 2times,the ratio of solid to liquid was 1:20,and the extraction temperature was 70℃.The optimal extraction parameters of chlorogenic acid from leaves were as follows:leaf size 40 mesh,extraction solvent 75% methanol solution,solvent p H 4,extraction time 75 min,extraction times 2 times,solid-liquid ratio 1:20,extraction temperature70℃,alcohol reflux extraction method.The contents of chlorogenic acid in the flowers,stems and leaves of the new strain "Nanyin No.1" were 1.07/10 g,0.187/10 g and 0.679/10 g,respectively.The sum of chlorogenic acid content in stem and leaf is equivalent to 86% chlorogenic acid content in flower bud of honeysuckle.(3)Study on the optimum decolorization process parameters of chlorogenic acidThe extract of Lonicerae japonicae obtained in(1)was decolorized by activated carbon,and the optimal decolorization process parameters were obtained by single factor test.The optimal decolorization process parameters of chlorogenic acid in flowers,stems and leaves of "Nanyin No.1" new strain of Lonicerae japonica were as follows:activated carbon content 2.4%,decolorization temperature 60℃,decolorization time60 min,and decolorization p H =3.(4)Study on the optimal separation and purification process parameters of chlorogenic acidThe content of chlorogenic acid in the crude extract of honeysuckle is low and contains many impurities,so it needs to be separated and purified.The traditional methods have low separation degree and complicated operation.The separation of chlorogenic acid by polyamide chromatography and coagulant chromatography is costly and the effect is not ideal.In this paper,macroporous resin was used to separate and extract chlorogenic acid from liquid.Single factor method and orthogonal experiment method were used to study the factors affecting the separation effect,and the optimal separation and purification process parameters were obtained:D-101 macroporous resin was selected as the best macroporous resin for chlorogenic acid adsorption and desorption.The optimal adsorption conditions for chlorogenic acid by D-101 macroporous resin were as follows: concentration 665μg/m L,injection flow rate 3BV/h,adsorption p H =4.The optimum elution conditions of D-101 macroporous resin for chlorogenic acid were as follows: 30% ethanol,dosage of4 BV,flow rate of 4BV/h,p H =7. |
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