阅读说明：本技术 一种蒜氨酸提取工艺优化方法 (Alliin extraction process optimization method ) 是由 于桂琴 常彦龙 刘相 黄国生 董翔 牛雁宁 万元 蒋伟 李静 于 2019-10-15 设计创作，主要内容包括：本发明提供一种蒜氨酸提取工艺优化方法,涉及化合物提取技术领域,包括如下步骤：S1.大蒜的预处理；S2.提取分离：将储罐中的溶剂通过离心泵导入提取罐中,密封储罐,调节减压阀,打开氮气瓶的第一阀门通过向提取罐中充入氮气达到设定压力后进行保压提取,打开第二阀门排出提取液；S3.浓缩结晶：将提取液倒入减压蒸馏浓缩罐中,将浓缩液使用超滤膜过滤除去胶状物,将过滤液通过离子交换柱进行纯化；S4.溶剂的回收：减压蒸馏浓缩罐中的溶剂注入精馏塔中,精馏后得到95％乙醇并收集；S5.洗脱液的处理：经过减压蒸馏浓缩罐减压蒸馏后的洗脱液进行中和反应后,进行收集或排入下水道。本发明化学污染小,操作方便,成本低,效率高,提取率高,再现性好。(The invention provides an alliin extraction process optimization method, relates to the technical field of compound extraction, and comprises the following steps: s1, preprocessing garlic; s2, extraction and separation: introducing the solvent in the storage tank into an extraction tank through a centrifugal pump, sealing the storage tank, adjusting a pressure reducing valve, opening a first valve of a nitrogen bottle, filling nitrogen into the extraction tank to reach a set pressure, performing pressure-maintaining extraction, and opening a second valve to discharge an extracting solution; s3, concentrating and crystallizing: pouring the extract into a reduced pressure distillation concentration tank, filtering the concentrated solution by using an ultrafiltration membrane to remove jelly, and purifying the filtrate by using an ion exchange column; s4, solvent recovery: injecting the solvent in the reduced pressure distillation concentration tank into a rectifying tower, rectifying to obtain 95% ethanol, and collecting; s5, treating the eluent: and neutralizing the eluent after the vacuum distillation in the vacuum distillation concentration tank, and collecting or discharging the eluent into a sewer. The method has the advantages of small chemical pollution, convenient operation, low cost, high efficiency, high extraction rate and good reproducibility.)

1. An alliin extraction process optimization method is characterized by comprising the following steps:

s1, pretreatment of garlic: peeling off the garlic clove, cutting the garlic clove into two halves along the longitudinal direction by a blade, and immediately washing the effluent liquid at the garlic cutting opening by using purified water;

s2, extraction and separation: placing garlic in a dry and clean place for 15min, then pouring the garlic into an extraction tank, introducing a solvent in the storage tank into the extraction tank through a centrifugal pump, sealing the storage tank, adjusting a pressure reducing valve, opening a first valve of a nitrogen bottle, replacing air in the extraction tank by filling nitrogen into the extraction tank for 10min, closing the first valve, continuously filling nitrogen into the extraction tank to pressurize the interior of the extraction tank, performing pressure-maintaining extraction after the pressure reaches a set pressure, opening the first valve to ensure that the pressure inside and outside the extraction tank is the same, and then opening a second valve to discharge an extracting solution;

s3, concentrating and crystallizing: pouring the extracting solution into a reduced pressure distillation concentration tank, carrying out reduced pressure distillation at the temperature of 50 ℃, obtaining concentrated solution when the residual amount of the extracting solution is one third of the amount before evaporation, filtering the concentrated solution by using an ultrafiltration membrane to remove jelly, purifying the filtrate by using an ion exchange column, adsorbing alliin in the filtrate onto cation resin, eluting and collecting the alliin by using eluent, carrying out reduced pressure distillation on the eluent containing the alliin by using the reduced pressure distillation concentration tank to obtain a crude alliin, and recrystallizing the crude alliin to obtain a pure alliin;

s4, solvent recovery: distilling the solvent in the concentration tank under reduced pressure with ethanol and water as main components, introducing the solvent into a rectifying tower, rectifying to obtain 95% ethanol, and collecting;

s5, treating the eluent: the main component of the eluent after the vacuum distillation of the vacuum distillation concentration tank is water, and the eluent is collected or discharged into a sewer after being subjected to neutralization reaction by using acid solution or alkali solution.

2. The optimization method of claim 1, wherein in step S2, the solvent in the storage tank is 70% ethanol, the first valve is closed, and nitrogen is continuously filled to pressurize the interior of the extraction tank, and the pressure is maintained for extraction for 120min after the set pressure of 0.6MPa is reached.

3. The method for optimizing the process of extracting alliin according to claim 2, wherein the bottom of the extracting tank is provided with a liquid outlet pipe, the liquid outlet pipe is internally provided with a filter screen, and the second valve is arranged above the filter screen.

4. The optimization method of the alliin extraction process according to claim 3, wherein the garlic after extraction treatment is poured into a pulping machine filled with 70% ethanol for full pulping, and after pulping, the garlic is extracted under pressure for 120min at room temperature and under the nitrogen pressure of 0.6MPa, and then purified by using cation resin.

5. The optimization method of the alliin extraction process according to claim 4, wherein in step S3, absolute ethyl alcohol is added into the crude alliin, the mixture is placed for 30min for recrystallization and then filtered, the obtained solid matter is dissolved by using water, then the absolute ethyl alcohol is added for 30min, and the filtration is carried out to obtain a recrystallized product, namely the pure alliin.

Technical Field

The invention belongs to the technical field of compound extraction, and relates to an alliin extraction process optimization method.

Background

Alliin, a non-protein sulfur-containing amino acid, is one of the major bioactive substances in garlic, and approximately 0.6% -2% of the dry weight of garlic is the precursor substance of allicin. Is very easy to dissolve in water, is not dissolved in pure absolute ethyl alcohol, acetone, chloroform, ether and benzene, and can obtain white needle cluster crystals by crystallizing with dilute acetone and ethyl alcohol. Modern medical research proves that sulfur-containing amino acids represented by alliin have unique pharmacological activity. Has obvious effects of reducing blood fat, improving body immunity, sterilizing, inhibiting bacteria, resisting cold, resisting aging, promoting blood circulation, preventing cancer and resisting cancer and the like after being taken for a long time. It has the functions of inhibiting and killing various pathogenic bacteria harmful to human body, is natural phytoncide, and has therapeutic and preventive values for bacterial, fungal and protozoal infections. Alliin has a molecular formula of C₆H₁₁NO₃S, molecular weight: 177.2214, isoelectric point: PI ═ 4.86, melting point: 163-165 ℃, color: dissolving with dilute acetone or ethanol to obtain white needleCluster crystals.

Alliin has special pharmacological activity and has obvious effects on resisting cancer, reducing blood fat, resisting thrombus and the like. However, the extraction of pure alliin is difficult due to harsh conditions, so that high-purity alliin products and high-purity medicinal alliin have high commercial value. The existing alliin synthesis and extraction methods have various defects, and the chemical synthesis method has the disadvantages of chemical pollution and higher product cost and is greatly different from natural alliin. Cell tissue culture methods are costly and are only suitable for laboratory studies. According to the traditional alliin extraction report, high temperature or microwave enzyme deactivation is carried out, then solvent extraction, column chromatography and recrystallization are carried out for purification to obtain an alliin pure product, however, the method is high in cost and low in extraction rate, and solvent residues are difficult to generally apply to medical intermediates due to the use of organic solvents.

Therefore, an alliin extraction process optimization method with small chemical pollution, convenient operation, low cost, high efficiency, high extraction rate and good reproducibility is urgently needed.

Disclosure of Invention

In order to solve the problem of optimization of the existing alliin extraction process, the invention provides an alliin extraction process optimization method.

The invention provides the following technical scheme:

an alliin extraction process optimization method comprises the following steps:

s1, pretreatment of garlic: peeling off the garlic clove, cutting the garlic clove into two halves along the longitudinal direction by a blade, and immediately washing the effluent liquid at the garlic cutting opening by using purified water;

s2, extraction and separation: placing garlic in a dry and clean place for 15min, then pouring the garlic into an extraction tank, introducing a solvent in the storage tank into the extraction tank through a centrifugal pump, sealing the storage tank, adjusting a pressure reducing valve, opening a first valve of a nitrogen bottle, replacing air in the extraction tank by filling nitrogen into the extraction tank for 10min, closing the first valve, continuously filling nitrogen into the extraction tank to pressurize the interior of the extraction tank, performing pressure-maintaining extraction after the pressure reaches a set pressure, opening the first valve to ensure that the pressure inside and outside the extraction tank is the same, and then opening a second valve to discharge an extracting solution;

s3, concentrating and crystallizing: pouring the extracting solution into a reduced pressure distillation concentration tank, carrying out reduced pressure distillation at the temperature of 50 ℃, obtaining concentrated solution when the residual amount of the extracting solution is one third of the amount before evaporation, filtering the concentrated solution by using an ultrafiltration membrane to remove jelly, purifying the filtrate by using an ion exchange column, adsorbing alliin in the filtrate onto cation resin, eluting and collecting the alliin by using eluent, carrying out reduced pressure distillation on the eluent containing the alliin by using the reduced pressure distillation concentration tank to obtain a crude alliin, and recrystallizing the crude alliin to obtain a pure alliin;

s4, solvent recovery: distilling the solvent in the concentration tank under reduced pressure with ethanol and water as main components, introducing the solvent into a rectifying tower, rectifying to obtain 95% ethanol, and collecting;

s5, treating the eluent: the main component of the eluent after the vacuum distillation of the vacuum distillation concentration tank is water, and the eluent is collected or discharged into a sewer after being subjected to neutralization reaction by using acid solution or alkali solution.

Preferably, in step S2, the solvent in the storage tank is 70% ethanol, the first valve is closed, and nitrogen is continuously filled to pressurize the interior of the extraction tank, and the pressure is maintained for extraction for 120min after the set pressure is reached to 0.6 MPa.

Preferably, the bottom of extracting the jar is equipped with the drain pipe, and the inside of drain pipe is equipped with the filter screen, and the top of filter screen is located to the second valve.

Preferably, the garlic after extraction treatment is poured into a beater filled with 70% ethanol for full beating, and after beating, the garlic is extracted under pressure for 120min at room temperature and under the nitrogen pressure of 0.6MPa, and is purified by using cation resin.

Preferably, in step S3, absolute ethanol is added to the crude alliin, and the mixture is left for 30min to recrystallize and then filtered, the obtained solid matter is dissolved by water, then absolute ethanol is added to the obtained solid matter and left for 30min, and the recrystallized product is the pure alliin product after filtering.

The invention has the beneficial effects that: the ethanol solvent has little chemical pollution and little organic solvent residue; the method has the advantages of no need of cell tissue culture and other steps, convenient operation, low cost, high efficiency, high extraction rate and good reproducibility, and is suitable for being widely applied to medical intermediates.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:

FIG. 1 is a graph of alliin production versus ethanol concentration;

FIG. 2 is a graph showing the trend of alliin yield with pressure;

FIG. 3 is a graph showing the trend of the change of the yield of alliin with time;

FIG. 4 is a standard curve of alliin solution;

FIG. 5 is an HPLC chromatogram of an alliin standard sample;

FIG. 6 is a spectrum of the purified solution of alliin extracted;

FIG. 7 is a MS spectrum of an alliin sample;

FIG. 8 is a 1H NMR chart of an alliin standard;

FIG. 9 is a 1H NMR chart of the alliin extract product;

FIG. 10 is a UV absorption spectrum of alliin standard;

FIG. 11 is a UV absorption spectrum of an alliin sample;

FIG. 12 is an infrared spectrum of the final alliin product;

FIG. 13 is a thermogravimetric plot of the final alliin product.

Detailed Description