印楝素的提取方法技术领域 TECHNICAL FIELD extracting azadirachtin

本发明涉及一种植物性农药的提取方法，尤其涉及一种印楝素的提取方法。 The present invention relates to a method for extracting botanical pesticides, and more particularly to a method for extracting azadirachtin.

背景技术 Background

印楝种仁中含有大量印楝油，所以印楝种仁直接粉碎十分困难，常常易发粘结块导致粉碎机无法正常运转，并且容易造成种子发热，导致印楝素部分分解； 文献报道中还有采用先压榨去油然后粉碎，这样虽然可解决粉碎结块的问题，但是在压榨过程中部分印楝素溶解在压榨油中，并且在压榨过程中易造成升温导致印楝素部分分解。 Neem seeds contain a lot of neem oil, neem seeds directly crushed so very difficult, and often prone to cause bond block grinder can not function properly, and likely to cause the seed heat, resulting in partial decomposition of azadirachtin; reported in the literature Also using the first press to the oil and then pulverized, so although caked solve the problem, but part of the press during the press azadirachtin dissolved in oil, and in the press process is easy to cause warming leads to partial decomposition of azadirachtin.

发明内容 DISCLOSURE

本发明提供一种工艺简单、提取率高、成本低、产品质量高的粉碎提取工艺。 The present invention provides a simple process, extraction rate, low cost, high quality grinding extraction process. 本发明采用如下技术方案-一种印楝素的提取方法，包括以下歩骤， The present invention adopts the following technical solution - a method for extracting azadirachtin, comprising the step ho,

第一步，用3-5倍去皮印楝种仁质量的极性有机溶剂浸泡去皮印楝种仁6-12小 The first step, with a polar organic solvent, peeled 3-5 times neem seeds quality neem seeds soaked peeled 6-12 small

时， When,

第二步，在上述极性有机溶剂中，采用高速刀片切削搅拌粉碎提取印楝素， 再过滤，得到提取液和滤饼， The second step in the polar organic solvent, a high-speed stirring and crushing blade cutting extract azadirachtin, filtered, and the filter cake obtained extract,

第三步，将提取液在10~40°(:下真空浓縮，除去极性有机溶剂得到含印楝素的浓縮物。 The third step, the extract was 10 ~ 40 ° (: concentrated under vacuum to remove the polar organic solvent to give a concentrate containing azadirachtin.

本发明中还可在第二步得到滤饼后，在滤饼中加入用3-5倍去皮印楝种仁质量的极性有机溶剂继续搅拌提取3-5小时，过滤，得到提取液和滤饼，再将滤饼按照相同的方法重复提取2-5次，再将提取液合并。 The present invention may also be in the second step to obtain a cake, adding the filter cake with a polar organic solvent, peeled 3-5 times neem seeds extract quality continue stirring for 3-5 hours, filtered to obtain an extract and The filter cake, then the cake according to the same extraction procedure was repeated 2-5 times, then the combined extracts.

本发明中，根据权利要求l所述的印楝素的提取方法，其特征在于将权利要求l 得到的含印楝素的浓縮物提纯，步骤为：用去皮印楝种仁质量的3-5倍的正己烷或石油醚，有白色絮状物沉淀产生，过滤，用与浸泡去皮印楝种仁相同的溶剂洗涤沉淀、干燥后，得到含印楝素的白色干粉。 The present invention as claimed azadirachtin extraction method according to claim l, wherein the resulting concentrate contains l will claim azadirachtin purification steps are: neem seeds with peeled quality 3 -5 times as hexane or petroleum ether, white floc precipitate, filtration, sedimentation and soak peeled neem seeds washed with the same solvent and dried to obtain a white powder containing azadirachtin.

本发明在第二步中，在极性有机溶剂介质中采用高速刀片切削搅拌粉碎印楝种子时，为避免高速搅拌粉碎时产生过热可采用夹套冷却或盘管冷却外部冷却。 In the second step of the present invention, the use of high-speed blade in a polar organic solvent medium cutting stirring and crushing neem seeds can be used to generate superheated jacket cooling or external cooling when cooling coil in order to avoid the high-speed stirring and crushing.

本发明获得如下技术效果： The present invention achieves the following technical effects:

1. 本发明采用了在极性有机溶剂介质中利用高速刀片切削搅拌粉碎提取技术， 克服了种仁直接粉碎易发粘结块造成粉碎困难，以及压榨去油造成印楝素丢失的缺点。 1. The present invention uses a high-speed cutting blade in a polar organic solvent medium, stirring and crushing and extraction technology to overcome the seeds directly bonded grinding prone to cause crushing block difficulties, as well as to the oil squeeze caused by the loss of azadirachtin shortcomings.

2. 縮短了工艺，粉碎提取合二为一，减少了植物材料的浪费，提高了印楝素的提取率，降低了成本。 2. shorten the process, crushed to extract combined, reducing the waste plant material to improve the extraction rate of azadirachtin, reducing costs.

3. 本发明提供了以对印楝素溶解性较好的极性有机溶剂浸泡软化去皮印楝种仁，然后在极性有机溶剂介质中采用高速刀片切削搅拌粉碎，—粉碎提取合二为一，为避免高速搅拌粉碎时产生过热可采用夹套冷却或盘管冷却加以解决。 3. The present invention provides a better solubility of azadirachtin polar organic solvent peeled neem seeds soaked to soften, then use high-speed blade cutting stirring and crushing in a polar organic solvent medium, - smash together two of the extraction First, overheating or cooling jacket can be resolved when the cooling coil in order to avoid the high-speed stirring and crushing. 该工艺的特征是印楝种子粉碎容易、粉碎后的物料具有较好的流动性和较细的颗粒有利于进一步的提取，克服了已有粉碎工艺的缺点，缩短了工艺过程， 提高了印楝素提取率。 The process is characterized by easy neem seed crushing, the crushed material has good fluidity and finer particles help to further extraction, to overcome the shortcomings of the existing grinding process and shorten the process and improve the neem Prime extraction rate.

本发明的原理如下：去皮种仁直接粉碎由于其含油较多而发粘结块，若在粉碎时加入适当溶剂使之分散，降低其粘度即可达到粉碎的目的。 Principles of the present invention is as follows: peeled crushed seeds directly due to its large oil and fat binding blocks, if the addition of a suitable solvent at the time of grinding to disperse, reducing its viscosity to achieve the purpose of crushing.

附图说明 Brief Description

图1为印楝素A萃取品的高压液相色谱(HPLC)图图2为印楝素A标准品的高压液相色谱(HPLC)图 Figure 1 is a product of the extraction of azadirachtin A high pressure liquid chromatography (HPLC) Figure 2 is azadirachtin A standard high pressure liquid chromatography (HPLC) FIG.

具体实施方式 DETAILED DESCRIPTION

实施例l Example l

一种印楝素的提取方法，包括以下步骤， Azadirachtin extraction method comprises the following steps,

第--步，用3-5倍去皮印楝种仁质量的极性有机溶剂浸泡去皮印楝种仁6-12小时， 第二步，在上述极性有机溶剂中，采用高速刀片切削搅拌粉碎提取印楝素，再过滤，得到提取液和滤饼， The first - step, with a polar organic solvent 3-5 times neem seeds peeled peeled quality neem seeds soaked 6-12 hours, the second step in the polar organic solvent, using high-speed cutting blade stirring and crushing and extracting azadirachtin, and then filtered to obtain extract and cake

第三步，将提取液在10〜4(A:下，真空浓缩除去极性有机溶剂得到含印楝素的浓縮物。 The third step, the extract was 10~4 (A: under, and concentrated in vacuo to remove the organic polar solvent to give a concentrate containing azadirachtin.

上述极性有机溶剂为为醇、酮、酯、醚、腈中的一种或它们的混合物，混合物的配比为任意配比，如甲醇、乙醇、丙酮、丁酮、乙酸乙酯、乙酸甲酯、甲基叔丁基醚、乙腈等或其混合物。 The polar organic solvent is an alcohol, ketone, ester, ether, nitrile or one of their mixtures, the mixture ratio of an arbitrary ratio, such as methanol, ethanol, acetone, methyl ethyl ketone, ethyl acetate, methyl ester, methyl tert-butyl ether, acetonitrile and the like or mixtures thereof.

实施例2 Example 2

一种印楝素的提取方法，用3-5倍去皮印楝种仁质量的极性有机溶剂常温(例如可以为20摄氏度或25摄氏度)浸泡软化去皮印楝种仁6-12小时，其中极性有机溶剂的质量可以是去皮印楝种仁质量的3.2倍，4倍，4.2倍，4.8倍，浸泡时间可以为7小时，8.5小时，9.5小时，11小时，11.5小时，然后在上述极性有机溶剂中采用高速刀片切削搅拌粉碎(可采用豆浆机，转速可以为10000-22000 转/分钟）提取印楝素，过滤后得到提取液和滤饼，在滤饼中加入2-5倍去皮印楝种仁质量的极性有机溶剂继续常温机械搅拌(在豆浆机中进行，搅拌提取的速度为2000-8000转/分)提取3-5小时，其中极性有机溶剂的重量可以是去皮印楝种仁质量的2.5倍，3倍，3.2倍，4倍，4.2倍，4.8倍，时间可以为3.3小时， 4.2小时，4.8小时，如此重复2-5次，例如可以为3或4次，再将提取液合并， 在1(TC〜40。C下真空浓縮，除去极性有机溶剂，得到含印楝素的浓缩物，其中真空浓縮温度可以选取为15°C， 22°C， 28°C， 32'C, 38°C，压力可以为0.07-0. 095MPa。 A method of extracting azadirachtin, neem with 3-5 seeds peeled quality polar organic solvent at room temperature (for example, 20 degrees or 25 degrees Celsius) to soften peeled neem seeds soak for 6-12 hours, wherein the quality polar organic solvent may be 3.2 times neem seeds peeled quality, 4x, 4.2 times, 4.8 times, soaking time for 7 hours, 8.5 hours, 9.5 hours, 11 hours, 11.5 hours, then the polar organic solvent used in the high-speed stirring and crushing blade cutting (Soymilk can be used, the rotational speed of 10000-22000 rev / min) to extract azadirachtin, the extract was filtered and the cake, add 2-5 filter cake polar organic solvents times neem seeds peeled quality mechanical stirring continued at room temperature (at Soymilk be stirred extraction speed 2000 to 8000 rev / min) for 3-5 hours to extract the weight of the polar organic solvent which can Neem seeds are peeled mass 2.5 times, 3 times, 3.2 times, four times, 4.2 times, 4.8 times, time for 3.3 hours, 4.2 hours, 4.8 hours, repeat 2-5 times, for example, 3 or 4 times, then the extracts were combined, concentrated in a vacuum (under TC~40.C, removing the polar organic solvent, to give a concentrate containing azadirachtin wherein the vacuum concentration temperature can be chosen to be 15 ° C, 22 ° C, 28 ° C, 32'C, 38 ° C, pressure may be 0.07-0. 095MPa.

本实施例中，将权利要求l得到的含印楝素的浓縮物提纯，步骤为：浓縮物加入4-8倍种仁质量石油醚（沸程为60-卯Gc)中，有白色絮状物产生，过滤，用非极性有机溶剂如石油醚（沸程为60-9()OC)洗涤滤饼、干燥得含印楝素的白色千粉。 Concentrate containing azadirachtin present embodiment, l get the claims purification steps are: concentrate was added quality seeds 4-8 times with petroleum ether (boiling range 60 to d Gc) and a white floc produced, filtered, washed with a non-polar organic solvent such as petroleum ether (boiling range 60-9 () OC) filter cake was washed and dried to give a white one thousand azadirachtin-containing powder.

本实施例中在极性有机溶剂介质中采用高速刀片切削搅拌粉碎印楝种子时， 为避免高速搅拌粉碎时产生过热可采用夹套冷却或盘管冷却。 The implementation of high-speed stirring and crushing blade cutting neem seed, or overheating jacket cooling coil can be cooled in order to avoid the use of high speed stirring and crushing example in a polar organic solvent medium.

上述极性有机溶剂为醇、酮、酯、醚、腈中的一种或它们的混合物'如甲醇、 乙醇、丙酮、丁酮、乙酸乙酯、乙酸甲酯、甲基叔丁基醚、乙腈等或其混合物，混合物的配比可以是任意配比。 The polar organic solvent is an alcohol, a ketone, an ester, an ether, a nitrile of one or a mixture of 'such as methanol, ethanol, acetone, methyl ethyl ketone, ethyl acetate, methyl acetate, methyl tert-butyl ether, acetonitrile, and the like or mixtures thereof, the ratio of the mixture may be any ratio. 实施例3 Example 3

取去皮未发霉印楝种仁100g ,用200ml乙酸乙酯浸泡软化12小时，后在乙酸乙酯介质中用九阳家用豆浆机搅拌粉碎提取，搅拌提取四次；滤液合并，真空浓縮除去乙酸乙酯,在浓缩物中加入4-8倍种仁质量石油醚(沸程为60-90。C), 有白色絮状物产生，过滤，用石油醚洗涤滤饼、干燥得含印楝素23%的白色干粉， 印楝素提取率达0.29%。 Take peeled not moldy neem seeds 100g, soaked to soften with 200ml of ethyl acetate for 12 hours after the media with ethyl acetate stirring and crushing nine-yang Soymilk household extract, stirring extracted four times; the filtrate were combined, concentrated under vacuum to remove Ethyl acetate was added 4-8 times the seeds quality petroleum ether in concentrate (boiling range 60-90.C), white floc generation, filtration, the filter cake washed with petroleum ether, and dried to yield containing neem - 23% of the white powder, azadirachtin extraction rate of 0.29%.

而用压榨脱脂粉碎种仁为原料按同样方法提取，即用乙酸乙酯浸泡12小时， 后搅拌提取四次，滤液合并，真空浓縮除去乙酸乙酯，在浓縮物中加入4-8倍种仁质量石油醚（沸程为60-90QC),有白色絮状物产生，过滤，用石油醚（沸程为60-9()OC)洗涤滤饼、干燥得产品，其印楝素提取率只有0.17%。 The material extracted by the same method, i.e., 12 hours with ethyl acetate soaked defatted by pressing the crushed seeds, after stirring extracted four times, the filtrates were combined and concentrated in vacuo to remove ethyl acetate, was added in 4-8-fold concentrate Seeds quality petroleum ether (boiling range 60-90QC), white floc produced, filtered, washed with petroleum ether (boiling range 60-9 () OC) cake was washed and dried to obtain the product, the azadirachtin extract rate of only 0.17%. 实施例4 Example 4

取去皮未发霉印楝种仁100g ,用200ml乙醇浸泡软化12小时，后在乙醇介质中用九阳家用豆浆机切削搅拌粉碎，搅拌提取四次，滤液合并，真空浓缩除去乙醇，加入4-8倍种仁质量石油醚（沸程为60-9()Gc)，，有白色絮状物产生， 过滤，用石油醚（沸程为60-90QC)洗涤滤饼、干燥得含印楝素19%的白色粉末， 印楝素提取率达0.46%。 Take peeled not moldy neem seeds 100g, softened with 200ml ethanol for 12 hours, after using ethanol medium household Soymilk Joyoung cutting stirring and crushing, stirring extracted four times, and the filtrate were combined, concentrated under vacuum to remove the ethanol, 4 8x seeds quality petroleum ether (boiling range 60-9 () Gc) ,, white floc generation, filtered, washed with petroleum ether (boiling range 60-90QC) cake was washed and dried to give containing azadirachtin 19 percent of white powder, azadirachtin extraction rate of 0.46%.

而用压榨脱脂粉碎种仁为原料按同样方法提取，其印楝素提取率只有0. 28%。 Extracting the same manner with the press defatted crushed seeds as raw material, its azadirachtin extraction rate of only 0.28%. 实施例5 Example 5

取去皮未发霉印楝种仁100g ，用200ml甲醇浸泡软化，后在甲醇介质中用九阳家用豆浆机切削搅拌粉碎，搅拌提取四次，滤液合并，真空一浓縮除去甲醇， 在浓缩液中加入适当水和适当乙酸乙酯进行液相-液相分配，水相继续用乙酸乙酯萃取，乙酸乙酯相合并，用脱水剂干燥，真空除去乙酸乙酯，加入4-8倍种仁质量石油醚（沸程为60-9(fC)，混合液中有白色絮状物产生，过滤，用石油醚洗涤滤饼、干燥得含印棟素20%的白色干粉，印楝素提取率达0.54%。 Take peeled not moldy neem seeds 100g, soaked to soften with 200ml of methanol, methanol medium after home with nine positive Soymilk cutting stirring and crushing, stirring extracted four times, and the filtrate were combined, concentrated under a vacuum to remove methanol, the concentrate Water was added to the appropriate and adequate ethyl acetate phase - liquid phase distribution, the aqueous phase was extracted with ethyl continue, ethyl acetate layers were combined, dried with a dehydrating agent, ethyl acetate was removed in vacuo, added 4-8 times seeds quality petroleum ether (boiling range 60-9 (fC), a mixture of white floc generation, filtration, the filter cake washed with petroleum ether and dried to give 20% of pigment-containing printing houses of white powder, azadirachtin extraction rate up to 0.54%.

而用压榨脱脂粉碎种仁为原料按同样方法提取，其印楝素提取率只有0. 30%。 Extracting the same manner with the press defatted crushed seeds as raw material, its azadirachtin extraction rate of only 0.30%.

实施例6 Example 6

取去皮未发霉印楝种仁50g ，用100ml乙酸乙酯浸泡软化12小时，后在乙酸乙酯介质中用九阳豆浆机切削搅拌粉碎，过滤滤液，滤饼以100ml乙酸乙酯分散，在一定功率条件下，设置超声方式（超声一分钟，间歇一分钟），进行超声提取，达到设定时间后过滤，滤饼加入新的溶剂重复超声提取，如此重复四次。 Take peeled not moldy neem seeds 50g, soaked to soften with 100ml of ethyl acetate for 12 hours after the media with ethyl acetate 9 Yang Soymilk cutting stirring and crushing, filtered and the filtrate cake was dispersed in 100ml of ethyl acetate, in power under certain conditions, setting mode ultrasound (ultrasound one minute, one minute interval), ultrasonic extraction, after a set time and the filter cake to add new ultrasonic extraction solvent repeat, repeat four times. 后处理的方法同实例一，印楝素提取率为0. 31%。 Post-processing method as in Example A, azadirachtin extraction rate of 0.31%. 实施例7 Example 7

取去皮未发霉印楝种仁50g ，用100ml乙醇浸泡软化，后在乙醇介质中用九阳家用豆浆机切削搅拌粉碎，过滤滤液，滤饼以100ml乙醇分散，在一定功率条件下，设置超声方式（超声一分钟，间歇一分钟），进行超声提取，达到设计时间后过滤，滤饼加入新的溶剂重复超声提取，如此重复四次。 Take peeled not moldy neem seeds 50g, soaked to soften with 100ml of ethanol, with ethanol medium after nine positive Soymilk home cutting stirring and crushing, filtered and the filtrate cake was dispersed in 100ml of ethanol, under certain power conditions, set ultrasound mode (ultrasound one minute, one minute interval), ultrasonic extraction, after reaching design time and the filter cake to add new ultrasonic extraction solvent repeat, repeat four times. 后处理的方法同实例三，印楝素提取率为0. 49%。 Post-processing method as in Example III, azadirachtin extraction rate of 0.49%. 实施例8 Example 8

取去皮未发霉印楝种仁50g ，用100ml甲醇浸泡软化，后在甲醇介质中用九阳家用豆浆机切削搅拌粉碎，过滤滤液，滤饼以100ml甲醇分散，在一定功率条件下，设置超声方式（超声一分钟，间歇一分钟），进行超声提取，达到设计时间后过滤，滤饼加入新的溶剂重复超声提取，如此重复四次。 Take peeled not moldy neem seeds 50g, soaked to soften with 100ml of methanol, methanol medium after home with nine positive Soymilk cutting stirring and crushing, filtered and the filtrate cake was dispersed in 100ml of methanol, under certain power conditions, set ultrasound mode (ultrasound one minute, one minute interval), ultrasonic extraction, after reaching design time and the filter cake to add new ultrasonic extraction solvent repeat, repeat four times. 后处理的方法同实例二，印楝素提取率为0. 56%。 Post-processing method as in Example II, azadirachtin extraction rate of 0.56%.

定型鉴定材料 Shape identification material

选用Kromasil-C18色谱分析柱(4.0 mm x 250 mm, 10 pm),以甲醇：水(V:V)-65 : 35为流动相，选用紫外检测器、检测波长为217nm，流速为lm:7min，常温条件下，对印楝素A的标准对照品及提取样品进行色谱分析，提取样品中的大部分峰均得到有效分离且重现性好，印楝素A的保留时间在5.8min左右。 Selection Kromasil-C18 chromatography column (4.0 mm x 250 mm, 10 pm), with methanol: water (V: V) -65: 35 as the mobile phase, the choice of UV detector, detection wavelength of 217nm, the flow rate was lm: 7min under normal conditions, the reference standards and take samples of azadirachtin A chromatographic analysis, samples were extracted most of the peaks have been effectively isolated and reproducible retention time of azadirachtin A is about 5.8min. 印楝素A的萃取样品及标准对照品的高压液相色谱图如下（见图1，图2)。 High pressure liquid chromatogram of the sample extract azadirachtin A and the reference standards are as follows (see Figure 1, Figure 2).