CN108185429A - 一种利用超高压设备提取鲜人参的方法及其在健康食品中的用途 - Google Patents
一种利用超高压设备提取鲜人参的方法及其在健康食品中的用途 Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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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
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/36—Caryophyllaceae (Pink family), e.g. babysbreath or soapwort
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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
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K2236/30—Extraction of the material
- A61K2236/37—Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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Abstract
本发明提供了一种利用超高压技术提取鲜人参的方法,简言之,选优质4‑5年生具有完整外表皮的新鲜人参,加0.5‑1倍水,人参与水置入塑料袋中然后密封,将塑料袋放入超高压舱中,400兆帕压力,提取5分钟,取提取液过滤,得鲜人参提取液。提取过程中样品材料不需粉碎,保持整支鲜人参原型,外表皮完好,从而可以利用人参外皮这一天然滤膜的性质,只有相对较小分子量物质通过外皮,阻止部分大分子化合物的通过,保证了鲜人参原液的澄明。利用鲜人参提取液可制备鲜人参口服液或者饮品,具有明显降低糖尿病小鼠空腹血糖,明显改善2型糖尿病小鼠口服糖耐量,同时并不升高血胰岛素水平。可作为制备防治2型糖尿病的药品、食品或保健品。
Description
技术领域
本发明涉及超高压提取技术制备鲜人参及鲜人参饮料制备方法。
背景技术
人参是我国传统名贵药材,具有大补元气,复脉固脱,补脾益肺,生津止渴,安神益智的功效。经过国内外学者不断地研究与探索发现人参中的主要活性是人参皂苷、人参多糖,此外还有蛋白质、多肽、氨基酸、微量元素等化学成分。这些物质为人参发挥其提高机体免疫力、增强记忆力、抗焦虑、抗癌、降血糖等药理活性提供了基础[1,2]。人参自古就有用于治疗“消渴” 的记载,虽然消渴不完全等同于糖尿病,但现代药理学实验已经证明人参中的人参皂苷、人参多糖、多肽等物质都具有对抗糖尿病及其并发症的作用[3-5]。
胰岛素抵抗(Insulin Resistance,IR)是近些年来学术界争相探讨的热点之一,随着研究的不断升温,有关胰岛素抵抗的研究领域逐渐从内分泌学科扩展到心血管病学科、肾病学科和神经病学科等多个交叉学科之中。据报道,目前对于治疗胰岛素抵抗比较有效的药物是烷二酮类。如吡格列酮,它可以通过改善2型糖尿病患者的肌肉、脂肪、肝脏的胰岛素抵抗,从而达到保护胰岛β细胞的功能[6]。但这些西药对于胰岛素抵抗的治疗都具有一定的副作用[7]。因此,寻找一种天然,低副作用的药物具有重要的意义。Sheth等[8]指出脱落酸(ABA)对胰岛素抵抗大鼠有调节改善的作用。最近研究表明[9] 口服0.5-1 μg / kg的ABA显降低大鼠和人体血糖和胰岛素血症。因此,体内低剂量ABA的降血糖作用并不取决于胰岛素释放增加,低剂量ABA摄入量可作为有助于改善糖尿病患者胰岛素抵抗或改善糖耐量。在体外分别用ABA刺激人β胰岛细胞和大鼠成肌细胞来增加胰岛素释放和糖的摄取,最低的ABA起效浓度分别是1.0和0.1nM,这可能显示成肌细胞对ABA具有更高的敏感性。
脱落酸是一种植物激素,它普遍存在于哺乳动物和植物体内,各种人和动物组织和细胞在生理或刺激条件下释放ABA,而免疫细胞,心血管组织和细胞,胰腺细胞和干细胞具有重要的调控作用,被认为是生物世界 广泛的一般信号因子,可以治疗许多疾病,例如,抗炎,癌症,糖尿病等。在大多植物中含量甚微,本专利揭示鲜人参中存在丰富的脱落酸。揭示本专利技术获得的鲜人参提取物主要活性因子为脱落酸,具有降低2型糖尿病血糖、改善糖耐量,同时降低血中胰岛素水平。一种利用超高压设备提取植物中有效成分的方法(ZL201510231951.4)专利指出,将粉状植物原材料装入容器中,加入水或5%-60% 酒精浸泡,料液比为1:1-1:3,充分搅拌,浸泡4-8小时,使植物细胞充分吸水膨胀,借鉴此专利基础上,作者将鲜人参保持原型状态不切碎,保持整支人参外表皮完好,利用人参外皮这一天然膜的性质,使许多大分子物质,例如蛋白质、多糖等较少通过,这一性质保证了提取液澄明更加有利于口服饮料的制作,从而节省了大量的加热、浓缩、过滤、沉淀等工序,达到节能减少污染,属于环境友好型的加工工艺;选择料液比小于1,可以最大限度增加提取液中活性因子脱落酸的浓度,并且由于超高压提取过程是在室温或者可以保持在更低温度下进行,从而最大限度保持鲜人参所特有各种活性物质。
发明内容
本发明公开了一种鲜人参提取物的制备方法。
方法如下:一种鲜人参提取物的制备方法,方法如下:挑选优质新鲜4-5年生新鲜人参,每支重量在20-40g,要求无病疤、无锈病、无污染、无农药残留,保持整支人参外表皮完好,洗净,加水0.5-1倍,将人参与水置入塑料袋中,密封,将塑料袋放入超高压舱中,加压400 兆帕,保持压力时间5 分钟,取出,排出袋中溶液,整个提取过程是在室温或者更低的温度下进行,再加入0.5-1倍水,重复上述过程,合并2次提取液,过滤,得鲜人参提取液。
本发明公开了以鲜人参提取物主要含有脱落酸,还有人参皂苷、游离氨基酸等活性物质。
鲜人参中含有脱落酸,水提取鲜人参每公斤最高可达15mg脱落酸。
本发明公开了鲜人参提取物以脱落酸为基本活性成分在降低血糖、改善葡萄糖耐量方面的应用。该鲜人参提取物含有脱落酸,具有防止胰岛素抵抗、抗糖尿病作用,可以应用于糖尿病预防及治疗药品的开发,也可应用于相关领域的食品及保健品的开发。
本发明公开了含脱落酸的鲜人参提取物能降血糖同时不升高血胰岛素水平,可降低糖尿病患者的血糖及提高葡萄糖耐受量,且不会导致高血胰岛素症,可以成为预防糖尿病和辅助治疗糖尿病的天然制品。
本发明药物组合物中还可以加入一种或几种目前常见的具有保健意义的功能性食品,从而可以起到维持或者增强本发明的食品组合物的作用。
附图说明
附图1:脱落酸对照品的HPLC-MS谱图
附图2:鲜人参水提物HPLC-MS谱图
附图3: 脱落酸对照品的高效液相色谱图
附图4:鲜人参水提物脱落酸的高效液相色谱图
附图5:人参提取物对2型糖尿病模型小鼠葡萄糖耐受量(OGTT)的影响
注:空白对照组(A),模型组(B),二甲双胍组(C),人参提取物低、中、高剂量组(D、E、F)与空白组,#P<0.05,##P<0.01;与模型组,*P<0.05,**P<0.01,附图3和附图4中1为脱落酸。
参考文献
[1] 黎阳,张铁军,刘素香,等. 人参化学成分和药理研究进展[J]. 中草药, 2009,40(1):164-166.
[2] 曹智,张燕娣,许永华,等. 人参有效成分及其药理作用研究进展[J]. 人参研究,2012, 2:39-43.
[3]Banz WJ, Iqbal MJ, Bollaert M, et al. Ginseng modifies the diabeticphenotype and genes associated with diabetes in the male ZDF rat[J].Phytomedicine,2007,14(10):681-689.
[4]Liu Z, Li W, Li X, et al. Antidiabetic effects of malonyl ginsenosidesfrom Panax ginseng on type 2 diabetic rats induced by high-fat diet andstreptozotocin[J]. J Ethnopharmacol, 2013,145(1):233-240.
[5]马萍,胡荣,王小英,等. 中药人参治疗糖尿病的研究进展[J]. 湖北中医药大学学报,2013,15(1):63-65.
[6]陈莉明. 噻唑烷二酮类药物治疗2型糖尿病的临床认识[J]. 中国实用内科杂志,2014, 34(10): 947-951.
[7]张士勇. 噻唑烷二酮类药物致不良反应文献分析[J]. 中国药物评价, 2014, 31(4): 223-225.
[8]Sheth D B, Tirgar P R, Chavda J R. et al. Effect of Abscisic acid infructose induced insulin resistant rats[J]. IJPI’s Journal of Pharmacologyand Toxicology, 2012, 2(8): 9-15.
[9]Magnone M, Ameri P, Salis A, et al. Microgram amounts of abscisic acidin fruit extracts improve glucose tolerance and reduce insulinemia in ratsand in humans[J]. Faseb Journal Official Publication of the Federation ofAmerican Societies for Experimental Biology, 2015, 29(12)。
具体实施方案
实施例1
鲜人参提取物的制备:挑选优质新鲜5年生鲜人参,100g,要求无病疤、无锈病、无污染、无农药残留,保持鲜人参外表皮完整,洗净,加水90ml,将人参与水置入塑料袋中,密封,将塑料袋放入超高压舱中,加压400 兆帕,保持压力时间5 分钟,卸压,取出,排出袋中溶液,再加入90ml水,将人参与水置入塑料袋中,密封,将塑料袋放入超高压舱中,加压400 兆帕,保持压力时间5 分钟,卸压,取出,排出袋中溶液,然后取出人参再用90 ml水浸泡5h,合并3次提取液,过滤,得滤液约260ml。经检测100g鲜参提出1.5mg脱落酸, 15μg/g,按提出液体积计算:260ml溶液中含有脱落酸1.5mg ,5.8μg/ml;经检测100g鲜参提出人参总皂苷80mg,0.8mg/g,按提出液体积计算:260ml溶液中含有人参总皂苷80mg,0.3mg/ml;经检测100g鲜参提出游离氨基酸2002 mg,20.02mg/g, 按提出液体积计算:260ml溶液中含有游离氨基酸2002mg,7.7mg/ml。
实施例2
鲜人参提取物的制备:挑选优质新鲜5年生鲜人参,100g,要求无病疤、无锈病、无污染、无农药残留,洗净,尤其注意不要破坏人参外皮,加水70ml,将人参与水置入塑料袋中,密封,将塑料袋放入超高压舱中,加压400 兆帕,保持压力时间5 分钟,卸压,取出,排出袋中溶液,再加入70ml水,将人参与水置入塑料袋中,密封,将塑料袋放入超高压舱中,加压400兆帕,保持压力时间5 分钟,卸压,取出,排出袋中溶液,然后取出人参再用70 ml水浸泡5h,合并3次提取液,过滤,过滤,得滤液200ml。经检测100g鲜参提出1.35mg脱落酸,13.5μg/g,按提出液体积计算:200ml溶液中含有脱落酸1.35mg,6.75μg/ml;经检测100g鲜参提出人参总皂苷60mg,0.6mg/g,按提出液体积计算:200ml溶液中含有人参总皂苷60mg,0.3mg/ml;经检测100g鲜参提出游离氨基酸1820mg,18.2mg/g, 按提出液体积计算:200ml溶液中同时还含有游离氨基酸1820mg,9.1mg/ml。
实施例3
鲜人参提取物的制备:挑选优质新鲜5年生鲜人参,100g,要求无病疤、无锈病、无污染、无农药残留,洗净,尤其注意不要破坏人参外皮,加水90ml,将人参与水置入塑料袋中,密封,将塑料袋放入超高压舱中,加压400 兆帕,保持压力时间5 分钟,卸压,取出袋中溶液,过滤,得滤液约90ml,加水使成100ml。经检测100g鲜参提出0.625mg脱落酸,6.25μg/g,按定容液体积计算:100ml溶液中含有脱落酸0.625mg,6.25μg/ml,经检测100g鲜参提出人参总皂苷41mg,0.41mg/g,按定容液体积计算:100ml溶液中含有人参总皂苷41mg,0.41mg/ml;经检测100g鲜参提出游离氨基酸1200mg,15.00mg/g, 按定容液体积计算:100ml溶液中含有游离氨基酸1200mg,12mg/ml;经检测精氨酸糖苷(AF)每ml溶液含有0.3mg。
实施例4
鲜人参提取物的制备:挑选优质新鲜5年生鲜人参,100g,要求无病疤、无锈病、无污染、无农药残留,洗净,尤其注意不要破坏人参外皮,加水60ml,将人参与水置入塑料袋中,密封,将塑料袋放入超高压舱中,加压400 兆帕,保持压力时间5 分钟,卸压,取出袋中溶液,过滤,得滤液60ml。经检测100g鲜参提出0.525mg脱落酸,5.25μg/g,按提出液体积计算:60ml溶液中含有脱落酸0.525mg,8.41μg/ml;经检测100g鲜参提出人参总皂苷30mg,0.3mg/g,按提出液体积计算:60ml溶液中含有人参总皂苷30mg,0.5mg/ml;经检测100g鲜参提出游离氨基酸900mg,9.0mg/g;按提出液体积计算:60ml溶液中含有游离氨基酸900mg,15.0mg/ml。
实施例5 鲜人参植物饮料
取实施例1 所得鲜人参提取液200ml,加入800ml纯净水使成1000ml,添加适量植物香精和适量防腐剂,经过滤,灌封,灭菌,每支50ml,每支含有58μg脱落酸,室温条件下存放2年,未发现有沉淀产生,饮品仍然为澄清透明室温液体。
实施例6 鲜人参氨基酸营养液
取实施例3 所得鲜人参提取液100ml,加入400ml纯净水使成500ml,添加0.2g甜叶菊苷,适量香精,每100ml营养液中,含有氨基酸300mg,人参皂苷20mg,脱落酸125μg,精氨酸单糖苷(AF),6mg。过滤,灌封,灭菌,封盖。每支10ml,室温条件下存放2年,未发现有沉淀产生,饮品仍然为澄清透明室温液体。
实施例7 HPLC-MS确定样品中脱落酸
标准脱落酸溶液的配制 标准品溶液 精密称取脱落酸标准品10.00mg,用80%甲醇溶解定容于100ml容量瓶中,0.22μm微孔滤膜过滤,即可制得质量浓度为0.10mg/mL的脱落酸对照品溶液。样品溶液配制 取实施例中不同方法获得的鲜人参提取物5ml放入规格相同的250mL锥形瓶中,加入100mL 80%甲醇,于4℃冰箱浸提,每次四个小时,提取三次,合并三次提取液,将提取液进行减压旋蒸后,调其PH=2.8~3.0,用乙酸乙酯进行萃取,萃取三次,合并三次提取液,再次进行减压旋蒸,蒸干后用80%甲醇定容25mL,用0.22μm微孔滤膜过滤,即得供试品溶液。
质谱条件 ESI离子源,负离子测定模式;保护柱:Phenomenex(4.0×3.0mm,5μm);色谱柱: ODS-MG C18液相色谱柱(4.6×150mm,5μm);流动相:甲醇-水(55:45), 流速:0.8mL/min;柱温:45°;离子源温度:500℃;雾化气:50 psi;辅助气:50 psi;气帘气:30 psi;分流比:1(进质谱):3(分流)。;进样量:5μL。
通过HPLC-MS确定样品中脱落酸分子量为263.1 与标准品一致(附图1和附图2)。
实施例8鲜人参提取物中脱落酸含量测定方法
1 溶液的配制 标准品溶液 精密称取脱落酸标准10.00mg,用80%甲醇溶解定容于100ml容量瓶中,0.22μm微孔滤膜过滤,即可制得质量浓度为0.10mg/mL的脱落酸对照品溶液。
供试品溶液 配制取实施例中不同方法获得的鲜人参提取物1g各自放入规格相同的250mL锥形瓶中,加入100mL 80%甲醇,于4℃冰箱浸提,每次四个小时,提取三次,合并三次提取液,将提取液进行减压旋蒸后,调其PH=2.8~3.0,用乙酸乙酯进行萃取,萃取三次,合并三次提取液,再次进行减压旋蒸,蒸干后用80%甲醇定容25mL,用0.22μm微孔滤膜过滤,即得供试品溶液。
色谱条件 色谱柱:Wakopak C18 (250mm×4.6mm , 5μm);柱温:45℃;流动相:甲醇-2‰磷酸水(V(甲醇):V(水)=55:45);流速:0.8mL/min;检验波长:260nm;进样量:20μL。色谱图见附图3和附图4。
实施例9 人参提取物对糖尿病小鼠血糖、糖耐量、血清胰岛素的影响
药物准备 阳性药溶液:取250mg二甲双胍溶于1‰羧甲基纤维素钠溶液中,配制成25mg·mL-1溶液。
样品溶液 分别取实施例3人参提取液1ml、2 ml、4 ml分别定容到10ml容量瓶中,即得0.625、1.25、2.50μg·mL-1人参提取液(每ml分别含有0.625、1.25、2.50μg脱落酸)。
实验动物模型建立与分组
ICR雄性小鼠120只,适应性饲养7天后,随机选取10只为空白对照组,喂正常饲料,其他110只,喂高脂饲料30天,于第30天晚间禁食不禁水,第31天随机抽取30只眼眶取血,离心取血清测TC、TG,高于正常值,如表1。腹腔注射100mg/kgSTZ柠檬酸缓冲液(0.1M PH=4.4),继续喂高脂饲料10天,第10天晚间禁食不禁水,于第11天尾部采血测定血糖值,测空腹血糖≥11.1mmol/L为造模成功,选取成模中80只小鼠,随机分为8组,分别为模型组、二甲双胍组(二甲双胍250 mg·kg-1)、人参提取物高、中、低剂量组(25.00、12.50、6.25μg·kg-1脱落酸),每组10只,空白对照组和模型组每天灌胃生理盐水,其他给药组分别灌胃相应药物,0.1mL/10g剂量给药,共给药40天,每天记录小鼠生物利用率和体重。
统计学方法
采用SPSS17.0统计学软件,实验数据用均值±标准差组间比较采用方差分析,采用LSD法进行两两比较。P<0.05表示差异具有统计学意义。
结果
对2型糖尿病模型小鼠空腹血糖及血清胰岛素的影响
造模成功后小鼠空腹血糖值明显升高,与空白对照组有显著性差异(P<0.01),给药治疗后,实验组小鼠空腹血糖值均呈下降趋势。持续给药四周后,与模型组相比,二甲双胍组、人参提取物高剂量组(25.00μg·kg-1脱落酸)和人参提取物中剂量组(12.50μg·kg-1)脱落酸,空腹血糖值显著降低(P<0.01),其他给药组小鼠空腹血糖值也有所降低,但无明显差异,无统计学意义。结果见表1。表1为小鼠血清中INS的测定结果,各给药组的INS与模型组小鼠相比,均有极显著差异(P<0.01),见表1。
表. 1人参提取物对糖尿病小鼠血糖、血清胰岛素的影响
组别 | 2周 | 4周 | INS(mIU/L) |
空白组 | 4.57±0.61 | 5.4±0.86 | 16.79±1.01 |
模型组 | 12.44±8.23## | 16.73±3.88## | 25.23±4.17## |
二甲双胍组 | 9.41±5.08 | 11.87±7.41 | 19.14±1.52** |
人参提取物(高) | 8.77±5.08 | 10.64±8.13* | 20.81±3.26** |
人参提取物(中) | 7.53±4.27* | 11.33±5.63* | 20.49±2.48** |
人参提取物(低) | 8.52±2.05 | 10.53±6.62* | 18.83±3.21** |
与空白组比较## P<0.01;与模型组比较* P<0.05,** P<0.01
对2型糖尿病小鼠葡萄糖耐受量的影响
如附图5所示,人参提取物高剂量组与中剂量组血糖曲线下面积AUC明显低于模型组,且具有极显著性差异(P<0.05)。
Claims (5)
1.一种鲜人参提取物的制备方法,方法如下:挑选优质新鲜4-5年生新鲜人参,重量在20-40g,要求无病疤、无锈病、无污染、无农药残留,保持整支人参外表皮完好,洗净,加水0.5-1倍,将人参与水置入塑料袋中,密封,将塑料袋放入超高压舱中,加压400 兆帕,保持压力时间5 分钟,取出,排出袋中溶液,整个提取过程是在室温或者更低的温度下进行,再加入0.5-1倍水,重复上述过程,合并2次提取液,过滤,得鲜人参提取液。
2.根据权利要求1所述制备方法,其特征在于,鲜人参还可以是鲜西洋参、鲜三七、鲜太子参等根茎类植物。
3.根据权利要求2所述的鲜人参、鲜西洋参,其特征在于为1-6年整支外表皮完好鲜参。
4.根据权利要求1所述的鲜人参提取液,其特征在于提取液中含有脱落酸、游离氨基酸、多肽、精氨酸衍生物、人参皂苷、维生素等小分子物质。
5.根据权利要求4所述的鲜人参提取液,其特征在于可应用于相关领域的食品及保健品的开发。
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