CN110507577A - 一种从圣女果中制备糖类、有机酸和类黄酮的方法 - Google Patents
一种从圣女果中制备糖类、有机酸和类黄酮的方法 Download PDFInfo
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Classifications
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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- Medicines Containing Plant Substances (AREA)
- Molecular Biology (AREA)
- Alternative & Traditional Medicine (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
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- Birds (AREA)
Abstract
本发明公开了一种从圣女果中制备糖类、有机酸和类黄酮的方法,包括下述步骤:以圣女果为原料,粉碎打浆,酶解,醇提后,离心,上清液通过超滤后,从超滤截留液中浓缩得到类黄酮,超滤透过液浓缩后,通过树脂分离获得糖类与有机酸。相对于传统生产方法,本发明方法具有原料利用率高,生产效率高,节能环保等优点。本发明方法推动了圣女果资源的综合利用,为圣女果的提质增效开发开辟新的途径,具有积极经济价值。
Description
技术领域
本发明属于农产品综合加工与天然产物提取技术领域,具体涉及一种从圣女果中制备糖类、有机酸和类黄酮的方法。
背景技术
圣女果,也称樱桃番茄,是茄科番茄属中的一年生草本植物的果实,其色泽鲜艳、酸甜适口、营养丰富,是一种水果型蔬菜。由于国内消费需求旺盛,我国圣女果产量迅速增加,带动整个圣女果产业发展。国内圣女果加工起步较晚,加工水平处于初级加工阶段,大量圣女果经清理、筛选、分级后出售,消费形式以鲜食为主。这种加工水平限制了圣女果资源的充分利用;同时由于加工链条短,造成产品附加值低,且产品种类少,受市场波动影响大,从而影响圣女果产业的可持续发展。
研究表明,圣女果中含有丰富糖类、有机酸和类黄酮,其中糖类占圣女果干重的60%,由葡萄糖、果糖和少量蔗糖组成;有机酸占圣女果干重的3-4%,由乙酸、富马酸、柠檬酸组成;圣女果中的类黄酮物质包括花青素、芦丁、山奈酚芸香苷等组成。圣女果中的糖类、有机酸不但可以作为甜味剂、酸味剂应用于食品工业中,类黄酮物质具有抗氧化、清除自由基、降低血脂、抑制癌细胞生长等有益功能,因而可以应用于医药、保健、化妆品等领域,因此,开发以上产品具有广阔的市场前景。
目前,国内从圣女果中提取功能成分仍处于初级阶段,提取方法往往只针对单一目标物,原料利用率低、生产成本高、经济效益差;且提取方法为传统有机溶剂提取,存在污染环境、产品中溶剂残留的问题。加之圣女果生产基地成熟时间较为集中,不能及时运输,会迅速腐烂变质,而储存过程中会有损耗。
发明内容
本发明的目的在于提供一种从圣女果中制备糖类、有机酸和类黄酮的方法,实现圣女果资源的充分高效开发,该方法具有生产效率高、原料利用率高、节能环保、条件温和可控等优点。
为了实现上述目的,本发明通过以下技术方案来实现:一种从圣女果中制备糖类、有机酸和类黄酮的方法,包括下述步骤:
1)圣女果预处理:
将圣女果除杂清洗后,破碎打浆后,加入酶水解,随后按照料液比加入0.5-4倍乙醇溶液转移到反应罐中,超声提取,醇提温度为20-60℃,时间为0.5-4h,低温避光备用;
其中,圣女果是成熟未霉变的果实。调节pH所用的酸可以是磷酸、柠檬酸、乙酸,所用的碱可以是氢氧化钠、碳酸氢钠。低温是指0-10℃。装载圣女果浆液的容器材质可以是玻璃、不锈钢、塑料。
2)类黄酮物质制备
将步骤1)中的醇提物采用配置目数为100-400目滤布的平板离心机以2000-5000rpm离心,收集上清液,上清液通过微滤后,采用超滤处理,分别收集超滤截留液和透过液,超滤截留液真空干燥获得类黄酮;
其中,微滤膜的材质可以是陶瓷、石英、有机复合膜。
3)有机酸、糖类制备
将步骤2)中获得超滤透过液采用反渗透进行浓缩后,进行树脂吸附,首先采用去离子水以10-100mL/min流速洗脱树脂,去离子水的使用量为柱体积1-3倍,将去离子水溶液采用反渗透浓缩后,干燥获得糖类;随后加入无水乙醇以10-100mL/min流速对树脂进行洗脱,乙醇的使用量为柱体积的2-5倍,将乙醇溶液采用反渗透浓缩后,干燥获得有机酸。
本发明一具体实施例中,步骤1)中酶为纤维素酶、半纤维素酶、果胶酶的任意两种或两种以上的混合酶;酶解条件为加酶量是原料质量的0.1%-5%,酶解温度为30-50℃,pH为3-5,酶解时间为0.5-3.0h。
本发明一具体实施例中,步骤1)中乙醇溶液的浓度为50%-100%;超声醇提采用的频率为20kHZ、40kHZ、59kHZ或其组合,超声强度为20-200W/kg,超声方式为连续或间歇。
本发明一具体实施例中,步骤2)中微滤孔径在0.45-5μm;超滤使用的超滤膜截留分子量在1000-5000道尔顿,超滤方式为死端或错流过滤,操作压力为0.5-3.0MPa,浓缩至原体积的3%-10%;真空干燥的条件是30-65℃干燥2-8h。
本发明一具体实施例中,步骤3)中反渗透采用死端或错流方式,采用的反渗透膜对氯化钠截留率大于99.00%,操作压力为1.0-6.0MPa,浓缩至原体积的1-5%;树脂选择非极性大孔树脂或聚酰胺树脂,树脂粒度为60-150目。
本发明一具体实施例中,步骤3)中糖类、有机酸的干燥可采用鼓风干燥、喷雾干燥、真空干燥中的任一种。
进一步的,鼓风干燥和真空干燥的温度在40-70℃,喷雾干燥时进风温度为150-190℃,出风温度为80-120℃。
其中所制备糖类、有机酸和类黄酮检测方法如下:
①类黄酮检测采用三氯化铝分光光度法。
取0.05g类黄酮粗品加入100mL容量瓶,采用50%甲醇定容,从中取2.5mL待测溶液置于试管中,加入5%NaNO2溶液0.5mL混匀放置5min加入10%Al(NO3)3溶液0.5mL混匀放置5min再加入2ml 20%NaOH溶液,加4.5mL 50%甲醇至混匀静置15min,506nm测定吸光值。以芦丁为标品,做标准曲线,计算样品中类黄酮质量。
②有机酸测定采用滴定法。
称取10g有机酸粗品粉末于100mL烧杯中,加入60mL蒸馏水,60℃加热搅拌30min,将溶液转移入250mL离心杯中,用60mL蒸馏水分三次冲洗烧杯,并将洗液倒入离心杯中,5000rpm离心15min,将离心上清液转移入250mL容量瓶中,定容。取试液3份,每份50mL,分别置于250mL三角瓶中,滴入酚酞指示剂2-3滴。用0.02mol/L标准NaOH(邻苯二甲酸氢钾标定)滴定。采用如下公式计算:
有机酸(g)=(V*C*N*F)/V1
式中,V为样品稀释总体积(mL),V1为滴定时取样品体积(50mL),C为消耗NaOH毫升数(mL),N为NaOH摩尔浓度(0.02mol/L),F为转换系数,取苹果酸0.067g/mmol。
③糖类测定采用硫酸苯酚法。
取0.5g粗糖粉加入到50mL烧杯中,加30mL蒸馏水,50℃加热搅拌30min,过滤,期间用70mL蒸馏水分三次冲洗烧杯,将洗液过滤、合并,转移到250mL容量瓶中,蒸馏水定容。从中取2mL样品液加入试管,随后加入0.15mL 80%苯酚,5mL浓硫酸,混匀,水浴冷却至室温,静置30min,于490nm测定吸光度。以葡萄糖为标品,做标准曲线,计算粗品中糖类质量。
上述制备方法集成目前先进的超声波辅助提取、膜分离、生物酶解等技术,在提高生产效率的同时,也降低生产成本,减少环境污染与能源消耗,并提高产品品质与安全性。该方法为实现圣女果资源的提质增效加工提供可靠的途径,延长圣女果产品的加工链,促进圣女果产业的良性发展。
与现有技术相比,本发明具有如下有益效果:
1)由于采用生物酶解技术,增加原料中目标物的溶出率,从而提高分离效率的同时,避免使用化学试剂,从而减少环境污染。
2)通过采用膜分离技术,实现对低含量物质的经济回收与浓缩,提高了经济效益,并且由于膜分离易于放大,便于后续生产规模增加。
3)制备的类黄酮产品,安全性好,不但可以进一步纯化生产保健药品,也可以作为原料应用于食品、化妆品等行业。
4)本发明对圣女果深加工可以增加产品附加值,减少资源浪费。
具体实施方式
下面详细说明本发明的具体实施方式,其作为本说明书的一部分,通过实施例来说明本发明的原理,本发明的其他方面、特征及其优点通过该详细说明将会变得一目了然。
实施例1:
一种从圣女果中制备糖类、有机酸和类黄酮的方法,包括下述步骤:
1)圣女果预处理:
收集圣女果,剔除发霉与未成熟果实,用自来水清洗除去泥沙,沥干后,称重10kg,破碎打浆后,加入100g纤维素酶与果胶酶的复合酶(两者重量比例为1:1),并用2mol/L磷酸将体系pH调节到4.0,500r/min搅拌下,40℃酶解1.5h后,按照料液比(m/V)为1:2加入20L80%乙醇溶液,转移入超声波提取仪中,采用500r/min搅拌下,20kHZ频率以1500w强度在50℃连续超声60min。超声结束后,将料液转移入不锈钢桶中,4℃备用。
2)类黄酮物质制备:
将步骤1)中的醇提物采用配置目数为300目滤布的平板离心机以5000rpm离心,收集滤过液。滤过液采用孔径1um微滤膜过滤,以除去蛋白与多糖;随后采用错流超滤,超滤膜截留分子量为3000道尔顿,操作压力1.5MPa,超滤至截留液占原液体积的5%时,结束超滤。将截留液转移入真空干燥箱,采用50℃干燥4h,获得2.87g类黄酮产品。
3)有机酸、糖类制备:
将步骤2)中获得超滤透过液采用死端反渗透进行浓缩后,其中反渗透膜对氯化钠截留率大于99.50%,操作压力为5MPa,当截留液至原体积的3%时,终止反渗透。将反渗透截留液上样到120目非极性大孔树脂柱上,采用20mL/min去离子水洗脱树脂柱,去离子水使用量为柱体积的2倍,收集水溶液采用反渗透浓缩至原体积的3%,50℃鼓风干燥,获得300.53g糖类产品。对树脂采用15mL/min无水乙醇洗脱时,无水乙醇使用量为3倍柱体积,收集醇溶液通过反渗透浓缩至原体积的3%,60℃真空干燥,获得20.48g有机酸。
实施例2:
一种从圣女果中制备糖类、有机酸和类黄酮的方法,包括如下步骤:
1)圣女果预处理:
收集圣女果,剔除发霉与未成熟果实,清洗除去泥沙,沥干后,称重20kg,破碎打浆后,加入300g纤维素酶、半纤维素酶、果胶酶的复合酶(三者重量比例为2:1:2),并用3mol/L柠檬酸将体系pH调节到4.5,400r/min搅拌下,45℃酶解1.5h后,按照料液比(m/V)为1:3加入30L无水乙醇,转移入超声波提取仪中,采用600r/min搅拌下,40kHZ频率以3000w强度在45℃间歇超声90min,每次超声工作10s后,停止5s。超声结束后,将料液转移入不透明的塑料桶中,4℃备用。
2)类黄酮物质制备:
将步骤1)中的醇提物采用配置目数为400目滤布的平板离心机以5000rpm离心,收集滤过液。滤过液采用孔径2um微滤膜过滤,以除去蛋白与多糖;随后采用死端超滤,超滤膜截留分子量为1000道尔顿,操作压力3.0MPa,超滤至截留液占原液体积的3%时,结束超滤。将截留液转移入真空干燥箱,采用60℃干燥3h,获得6.37g类黄酮产品。
3)有机酸、糖类制备:
将步骤2)中获得超滤透过液采用错流反渗透进行浓缩后,其中反渗透膜对氯化钠截留率大于99.80%,操作压力为3MPa,当截留液至原体积的5%时,终止反渗透。将反渗透截留液上样到160目聚酰胺树脂柱上,采用10mL/min去离子水洗脱树脂柱,去离子水使用量为柱体积的2倍,收集水溶液采用反渗透浓缩至原体积的5%,55℃鼓风干燥,获得599.67g糖类产品。对树脂采用15mL/min无水乙醇洗脱时,无水乙醇使用量为3倍柱体积,收集醇溶液通过反渗透浓缩至原体积的5%,65℃真空干燥,获得42.67g有机酸。
实施例3:
一种从圣女果中制备糖类、有机酸和类黄酮的方法,包括下述步骤:
1)圣女果预处理:
收集圣女果,剔除发霉与未成熟果实,清洗除去泥沙、枯叶、树枝,沥干后,称重50kg,破碎打浆后,加入1000g纤维素酶、果胶酶的复合酶(两者重量比例为1:2),并用3mol/L乙酸将体系pH调节到4.8,600r/min搅拌下,50℃酶解2.0h后,加入125L 75%乙醇溶液,转移入超声波提取机中,采用800r/min搅拌下,59kHZ频率以3000w强度在50℃连续超声75min。超声结束后,将料液转移入不透明的塑料桶中,2℃备用。
2)类黄酮物质制备:
将步骤1)中的醇提物采用配置目数为200目滤布的平板离心机以3000rpm离心,收集滤过液。滤过液采用孔径0.5um微滤膜过滤,以除去蛋白与多糖;随后采用错流超滤,超滤膜截留分子量为1000道尔顿,操作压力1.0MPa,超滤至截留液占原液体积的6%时,结束超滤。将截留液转移入真空干燥箱,采用45℃干燥6h,获得13.98g类黄酮产品。
3)有机酸、糖类制备:
将步骤2)中获得超滤透过液采用错流反渗透进行浓缩后,其中反渗透膜对氯化钠截留率大于99.00%,操作压力为3MPa,当截留液至原体积的5%时,终止反渗透。将反渗透截留液上样到100目聚酰胺树脂柱上,采用25mL/min去离子水洗脱树脂柱,去离子水使用量为柱体积的2倍,收集水溶液采用反渗透浓缩至原体积的5%,45℃鼓风干燥,获得1531.49g糖类产品。对树脂采用20mL/min无水乙醇洗脱时,无水乙醇使用量为3倍柱体积,收集醇溶液通过反渗透浓缩至原体积的5%,喷雾干燥,进风温度为180℃,出风温度为100℃,获得103.38g有机酸。
需要说明的是,圣女果的水分含量在90%左右,因而本发明的滤渣很少,制备过程中产生的滤渣可作为饲料用。
综上可看出:相较于传统单一提取工艺,本发明方法具有原料利用率高、生产成本低、处理量大、操作简便、节能环保等优点。本发明方法为推动圣女果的综合加工,延长了圣女果加工链条,丰富提供给消费者的圣女果相关产品,也为加工企业与种植农户带来收益。本发明对圣女果深加工可以增加产品附加值,减少资源浪费。
以上所述是本发明的优选实施方式而已,当然不能以此来限定本发明之权利范围,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和变动,这些改进和变动也视为本发明的保护范围。
Claims (7)
1.一种从圣女果中制备糖类、有机酸和类黄酮的方法,其特征在于,包括下述步骤:
1)圣女果预处理:
将圣女果除杂清洗后,破碎打浆后,加入酶水解,随后按照料液比加入0.5-4倍乙醇溶液转移到反应罐中,超声提取,醇提温度为20-60℃,时间为0.5-4h,低温备用;
2)类黄酮物质制备
将步骤1)中的醇提物采用配置目数为100-400目滤布的平板离心机以2000-5000rpm离心,收集上清液,上清液通过微滤后,采用超滤处理,分别收集超滤截留液和透过液,超滤截留液真空干燥获得类黄酮;
3)有机酸、糖类制备
将步骤2)中获得超滤透过液采用反渗透进行浓缩后,进行树脂吸附,
首先采用去离子水以10-100mL/min流速洗脱树脂,去离子水的使用量为柱体积1-3倍,将去离子水溶液采用反渗透浓缩后,干燥获得糖类;
随后加入无水乙醇以10-100mL/min流速对树脂进行洗脱,乙醇的使用量为柱体积的2-5倍,将乙醇溶液采用反渗透浓缩后,干燥获得有机酸。
2.如权利要求1所述从圣女果中制备糖类、有机酸和类黄酮的方法,其特征在于,
步骤1)中酶为纤维素酶、半纤维素酶、果胶酶的任意两种或两种以上的混合酶;
酶解条件为加酶量是原料质量的0.1%-5%,酶解温度为30-50℃,pH为3-5,酶解时间为0.5-3.0h。
3.如权利要求1所述从圣女果中制备糖类、有机酸和类黄酮的方法,其特征在于,
步骤1)中乙醇溶液的浓度为50%-100%;超声醇提采用的频率为20kHZ、40kHZ、59kHZ或其组合,超声强度为20-200W/kg,超声方式为连续或间歇。
4.如权利要求1所述从圣女果中制备糖类、有机酸和类黄酮的方法,其特征在于,
步骤2)中微滤孔径在0.45-5μm;
超滤使用的超滤膜截留分子量在1000-5000道尔顿,超滤方式为死端或错流过滤,操作压力为0.5-3.0MPa,浓缩至原体积的3%-10%;
真空干燥的条件是30-65℃干燥2-8h。
5.如权利要求1所述从圣女果中制备糖类、有机酸和类黄酮的方法,其特征在于,
步骤3)中反渗透采用死端或错流方式,采用的反渗透膜对氯化钠截留率大于99.00%,操作压力为1.0-6.0MPa,浓缩至原体积的1-5%;
树脂选择非极性大孔树脂或聚酰胺树脂,树脂粒度为60-150目。
6.如权利要求1所述从圣女果中制备糖类、有机酸和类黄酮的方法,其特征在于,
步骤3)中糖类、有机酸的干燥可采用鼓风干燥、喷雾干燥、真空干燥中的任一种。
7.如权利要求6所述从圣女果中制备糖类、有机酸和类黄酮的方法,其特征在于,
鼓风干燥和真空干燥的温度在40-70℃,喷雾干燥时进风温度为150-190℃,出风温度为80-120℃。
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