CN103059095B - 一种黄姜资源综合高效利用的绿色提取技术 - Google Patents
一种黄姜资源综合高效利用的绿色提取技术 Download PDFInfo
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Abstract
一种黄姜资源综合高效利用的绿色提取技术，该技术在提取皂素的同时，综合，高效，同步提取了黄姜中的其它有效成分，如黄姜淀粉，纤维素，冠心宁原料，果胶，农用核酸等。并从水解废液中提取了鼠李糖，整个工艺过程绿色环保，将极大的提升我国皂素的生产水平，提高资源利用效率、节约能源、保护环境，实现经济效益、环境效益和社会效益相统一，推动中国经济和社会的可持续发展。
Description
技术领域
本发明涉及一种黄姜资源综合高效利用的绿色提取技术。
背景技术
黄姜，又名穿地龙，火头根，土名哑边姜，学名盾叶薯蓣，系单子植物薯蓣科植物盾叶薯蓣(DioscoreaZingiberensisC.H.Wright)的根茎，地下根茎是著名淀粉植物，也是滋补食品，又是药用植物。主要分布于鄂、川、陕、辽、吉、黑等地海拔300米以上的山地，丘陵，高寒山区等。我国有50来种，其中17种，1亚种，2变种含皂苷元，占世界含有皂苷元植物的50％以上，含量最高可达16.15％，超过墨西哥小穗花薯蓣15％的记录，是世界上的王牌种类。其块茎有较高的药用和化工价值，经过加工可提炼皂素、双稀等医药、化工用品，皂素的甾体结构加以改造，就可得到各种不同的甾体激素类药物，是激素类药物必不可少的成份，被喻为“激素之母”，并广泛应用于化妆、保健、避孕镇痛、麻醉等类药物，是一种市场需求较为稳定的药材。从黄姜根状茎中提取的最初产品为皂雄酮、醋酸孕酮(单脂)、强的松、可的松等系列产品，以及催产素、甾体避孕药等中间体或药物数千种，我国起步虽较晚，也可以合成近100种产品。以黄姜为原料，可以合成性激素、蛋白同化激素和皮质激素等系列产品，皂素的深加工产品“双烯”、“胱胺酸”、“无水胱胺酸”等多种延伸产品。是目前世界上合成300多种甾体激素和避孕药的原料，因此，医药界称其为“药用黄金”。根状茎在医药、食品、高级化妆品、兽药等行业中也有广泛的用途，如根状茎直接入药，有祛湿、清热解毒之功效，民间用于治疗皮肤急性化脓性感染、软组织损伤、蜂螫、虫咬及各种外科炎症，以及强身壮骨作用。水溶性活性物质可以生产盾叶冠心宁，用于治疗冠心病，效果好，副作用小。薯蓣皂苷还可作为洗涤剂，乳化剂，发泡剂等。除国内需要外，一直俏销世界120多个国家和地区，而我国生产的皂素又占全世界的50％。过去，由于黄姜原料紧缺，很多激素类药物都有采用代用品。一九九二年，联合国卫生组织宣布，禁止使用代用品，只能以黄姜原料为主生产的皂素合成甾体激素类药物。此外，近二十年来，随着人口计划生育政策的推广和激素类药物的广泛使用，世界市场对皂素的需求与日俱增，黄姜原料成为国际市场的紧俏产品，供求矛盾十分突出。黄姜收购价格一路攀升，皂素价格也一涨再涨，开发前景十分可观。世界年用皂素量约2800吨，我国约1000吨。我国目前皂素生产能力约为900吨，年出口量为200吨，国内供求缺口300吨。近几年来，皂素的需求量每年以6～7％的速度递增。八十年代末，皂素价格18万元/吨，九十年代末，皂素吨价达40～45万元。但多年来人们对黄姜野生资源的掠夺性采挖，导致该物种日渐枯竭。在黄姜生产加工中粗放操作经营，导致环境污染严重，生态遭到极大破坏。因而，我国甾体植物即将面临着原料供应不足的局面，这将严重的地威胁到我国甾体药物行业的发展。必须综合高效利用黄姜资源，做到资源节约型，环境友好型。
盾叶薯蓣中化学成分很多，薯蓣皂苷在薯蓣科植物中含量丰富，它有祛痰、脱敏、抗炎、降脂、抗肿瘤等作用。薯蓣皂苷口服后经肠道菌群代谢产生的薯蓣皂苷元是发挥作用的真正有效成分，而且它还是合成甾体激素类药物的重要原料。其药理作用如下：1)溶血作用。薯蓣皂苷属于甾体皂苷，甾体皂苷水溶液与血液接触后会不同程度的破坏红细胞，产生溶血现象，因此皂苷及含有皂苷的生药不能用于静脉注射。2)抗衰老作用。林刚等研究证明适当剂量的盾叶薯蓣皂苷可明显降低老龄小鼠血浆LPO和肝脏LF的含量。试验结果表明，盾叶薯蓣具有一定的抗衰老作用。3)抗肿瘤作用。薯蓣皂苷的前皂苷配基A、薯蓣皂苷和纤细皂苷在体外对肿瘤细胞系K562具有细胞毒作用。能显著抑制Hela细胞的生长。王丽娟等研究证明薯蓣皂营元对三种小鼠移植肿瘤肉瘤-180(S-180)，肝癌腹水型(HepA)，小鼠宫颈癌-14(U14)均有明显的抑制作用。而在离体条件下，薯蓣皂苷元对小鼠肺上皮癌细胞(L129)，人宫颈癌细胞(Hela)人乳腺癌细胞(MCF)3种肿瘤细胞具有明显的抑制作用。霍锐等通过实验发现在离体条件下薯蓣皂苷元诱导人黑素瘤细胞A375-S2凋亡。Moalic等研究了薯蓣皂苷元能抑制人骨肉瘤1547细胞系的生长，使细胞分裂周期停止在G1期，诱导细胞凋亡。4)杀灭钉螺的作用。刘汉成等进行盾叶薯蓣杀湖北钉螺卵的实验研究，糜留西等应用盾叶薯蓣植物根粉进行小规模的现场灭螺试验，崔天义等发现盾叶薯蓣根茎具有显著的灭螺活性，结果表明，对卵龄螺卵具有良好的杀灭作用，且有效的抑制钉螺上爬。5)防治心血管疾病的作用。实验证实薯蓣皂苷元在体外均有明显的抗血小板聚集活性，在降血脂方面，马海英等实验证明，薯蓣皂苷元能明显降低血中胆固醇含量，临床证明，薯蓣皂苷片具有调节脂质代谢，改善血液流变学作用，明显降低血清总胆固醇、甘油三酯、低密度脂蛋白和氧化修饰低密度脂蛋白含量，降低全血粘度以及血浆粘度，因此可以减轻动脉壁脂质浸润及斑块形成，从而防治动脉粥样硬化。地奥心血康胶囊就是从黄山药中提取的甾体皂苷精制而成的纯中药制剂。药效学实验证明，其治疗冠心病的有效部分是8种甾体皂苷，含量在90％以上，其中3种呋甾皂苷含量较高，另外5种为薯蓣皂苷元的衍生苷，地奥心血康对冠心病心绞痛发作疗效显著。盾叶冠心宁为盾叶薯蓣根茎水溶性皂苷制剂，临床治疗冠心病、心绞痛效果较好，现已投入生产。6)其他作用。薯蓣皂苷本身有肝脏保护、治疗骨质疏松、抗糖尿病和消炎等功能。朱广慧等通过试验说明薯蓣皂苷元对油菜生长具有一定的促进作用，可能具有与甾体激素相似的作用。江洪等通过抑菌活性测定试验发现，从盾叶薯蓣中分离出的甾体皂苷对大多数植物病原真菌有广谱高效的抑制作用，而其根茎的粗提物抑菌效果最好，说明该粗提物中含有其他有待进一步分离提纯的活性很高的皂苷成分。
目前，提取皂素方法主要有以下几种：1)直接酸水解法。传统的提取薯蓣皂苷元的方法是采用Rothrock法，即直接将盾叶薯蓣根茎用硫酸水解成苷元，然后用有机溶剂提取薯蓣皂苷元。用盐酸作为水解酸比硫酸好，工艺简单，效果差异不大，加压法比常压法水解苷元收率高，缺点是盐酸对不锈钢设备有严重腐蚀。直接酸水解法仅能提取1/4的皂苷元，且此法费时很多，薯蓣皂素收率较低，由于使用溶剂汽油，易发生危险；另外，盾叶薯蓣中的淀粉和纤维素等其它成分也在酸水解过程中被破坏而不能利用，因此该工艺不够理想。2)预发酵法。一般认为预发酵法可提高薯蓣皂苷元的收率。预发酵法有自然发酵法、酶解法、微生物发酵法。预发酵法既提高了皂苷收率，反应条件也比较温和，保持了有效成分的本来理化性质，但自然发酵法影响因素较多，产品质量不稳定。3)CO2超临界萃取法。葛发欢等报道用CO2超临界萃取法从盾叶薯蓣中提取薯蓣皂苷元的研究，和传统的汽油法比较，收率提高了1.5倍，生产周期大大缩短，避免了使用汽油引起易燃易爆的危险，且成本相差不大。但存在设备一次投资过大的缺点。4)其他方法。利用超声波破碎细胞壁也可提高薯蓣皂苷元的收率。用盐酸一丙酮/乙醇混合液直接加热提取薯蓣皂苷元的工艺虽可节约物料，缩短工艺过程，但对资源的综合利用不足，在实际生产中应用该工艺的还未见报道。
盾叶薯蓣的根状茎含含薯蓣皂甙元。薯蓣皂甙元的含量分1.1％～16.15％，45％左右的淀粉、40％的纤维素以及一些水溶性苷类、生物碱类、黄酮苷类、强心苷类、生物碱、单宁、色素等化学成分。黄姜除提取皂素外，还含有45～50％的淀粉，可用于酿造工业生产酒精、酵母粉、肌苷粉、葡萄糖等。所含40～50％的纤维素，可生产羧甲基纤维素。提取皂素的废液，可提取农用核酸，是优质肥料。根状茎在医药、食品、高级化妆品、兽药等行业中也有广泛的用途，如根状茎直接入药，有祛湿、清热解毒之功效，民间用于治疗皮肤急性化脓性感染、软组织损伤、蜂螫、虫咬及各种外科炎症，以及强身壮骨作用。水溶性活性物质可以生产盾叶冠心宁，用于治疗冠心病，效果好，副作用小。成都生物所研制的地奥心血康，具有调节新陈代谢机能，治疗动脉硬化以及对心血管系统的疾病都有较好的效疗，畅销国内外市场。另外，中科院武汉植物研究所研究发现，黄姜活性物质是杀灭钉螺，防血吸虫的理想药物，不仅灭螺效果好，而且又不污染环境，保持生态平衡。
本发明采用预处理的方法先分理处淀粉，粗纤维等，然后再断开薯蓣皂苷元与糖连接的苷键，使薯蓣皂苷元游离出来，利用它的亲脂性，用石油醚或高标号的汽油或6#溶剂油把它萃取出来。通过预处理，分离了粗纤维，去除了淀粉，克服了直接水解造成黄姜资源中其他成分破坏，水解不彻底，收率低，从黄姜植物资源中综合提取了淀粉，纤维素，鼠李糖，皂素，冠心宁原料，果胶，农用核酸等。具有高效，节能，得率高，无污染的特点。
发明内容
第一打浆备料：将黄姜水浸泡，打浆，过筛，去渣，筛下物沉降，去清液备用，下层浆状物为提取原料。
第二水液处理：打浆沉降的上清液减压浓缩后，以正丁醇萃取，得到有机相和水相。水相减压浓缩，得到果胶混合物，用于制作酵母等。有机相浓缩，干燥，得到皂苷类混合物，用于生产冠心宁。
第三打浆过筛渣处理：滤渣以氢氧化钠煮沸，过滤，水洗滤饼，得到粗纤维。
第四淀粉的去除：浆料加入活性酶，发酵，酶解，过滤，滤液另用，滤饼，滤液用于生产酒精，滤饼提取。
第五水解：滤饼中加入盐酸液加压水解，得到水解液，用石灰乳调中性。
第六萃取：上述水解液以弱极性溶剂萃取，得到萃取液和水液。
第七皂素的制备：上述萃取液加入活性炭脱色，过滤，浓缩，结晶，干燥，得到皂素。
第八水液的处理：上述萃取后的水溶液加入酒曲发酵，过滤，滤液减压浓缩，低温结晶，得到粗品鼠李糖。结晶母液可制备酒精。粗品鼠李糖以无水甲醇溶解后，加入活性炭脱色，过滤，去除大量无机盐和色素，浓缩，结晶，干燥，得到精品鼠李糖。
第九农用核酸提取：农用核苷酸由核糖核酸的降解产生(如核苷酸、核苷、碱基)组成，可以促进作物生长、发育和提高产量。利用处理鼠李糖的母液来培养白地霉以提取农用核苷酸。
附图说明
图1黄姜综合提取工艺图
具体实施方式
取陕南产干黄姜500Kg用水2500Kg浸泡24h，以打浆机打浆，用30～40目筛网过滤，去除粗纤维渣，得到浆状物，将浆状物再以粗滤布过滤，得到乳白色过滤液和沉淀。将过滤液沉淀12～24h，压滤，得到黄姜淀粉，水洗，干燥，得到210Kg淀粉。压滤的水液减压浓缩至比重1.10～1.15后，以95％乙醇沉，直至醇度达75～80％，压滤，得到果胶。滤液可生产酒精。将粗滤布过滤的沉淀(2)以500LpH1～2的盐酸液水解2～4h，降温至20～30℃，以等体积的6#溶剂油(或石油醚，汽油)萃取3次，萃取液以纯化水洗涤3次，加入1～3Kg活性炭在50～60℃脱色30min，过滤，浓缩，结晶，得到皂素69.5Kg，含量99.2％。萃取后的水液以石灰乳调pH中性后，沉降，过滤，过滤液加入3～5％(w/v)的酒曲在35～45℃发酵96～120h，加入3～5％(w/v)硅藻土充分搅拌后过滤，滤液加入1～3Kg活性炭加热至60～70℃，保温搅拌20min，停止搅拌，静置30min，放出脱色液过滤，流出的脱色液基本清亮，很好过滤，直至脱色釜底部留下大量活性炭时停止。然后将脱色釜四周吸附的活性炭以水冲洗干净。滤液减压浓缩至比重1.10～1.15，在5～10℃下结晶120～140h，离心，得到鼠李糖粗品，。将此粗品以5～8倍(w/w)的99％甲醇回流溶解后过滤，除去无机盐，滤液浓缩至酒精度20～30％后，冷却至5～10℃，结晶24～48h后。离心，干燥，得到精品鼠李糖9.5Kg，含量99.8％。结晶后的母液加0.5％白糖、0.1％硫酸铵、0.05％磷酸二氢钾，用浓硫酸调pH2.5～3.0，煮沸15min，再调pH5～6，按种白地霉，培养14h。把白地霉培养液用200目/cm2过滤，将滤渣即湿白地霉放入容器中，加入菌体5倍的水，再加入5％菌体重的pH10的硼砂氢化钠缓冲液，75℃搅拌1h，菌体自溶。再用200目/cm2过滤，滤液煮沸消毒，即得农用核酸。
Claims (9)
1.一种黄姜资源综合高效利用的绿色提取方法，其特征在于，以盾叶薯蓣(黄姜)为原料，该方法依次经以下步骤：
(1)打浆备料：将黄姜水浸泡，打浆，过筛，去渣，得到黄姜粗纤维和浆状物；
(2)浆状物的处理：将浆状物以粗滤布过滤，得到乳白色混浊滤液和滤渣I，将混浊滤液沉降，过滤，得到黄姜淀粉，滤渣用于提取皂素，滤液下步处理；
(3)沉降滤液的处理：滤液减压浓缩后，以正丁醇萃取，得到有机相和水相，水相减压浓缩，得到果胶等，有机相浓缩，干燥，得到皂苷类混合物；
(4)打浆过筛渣处理：滤渣以碱水煮，过滤，水洗，烘干，得到粗纤维；
(5)滤渣I的处理：滤渣中加入盐酸液加压水解，得到水解液，用石灰乳调中性；
(6)萃取：上述水解液以弱极性溶剂萃取，得到萃取液和水液，萃取液以纯化水反萃取，所述的弱极性溶剂为6#溶剂油或60～90°石油醚；
(7)皂素的制备：上述萃取液加入活性炭脱色，过滤，浓缩，结晶，干燥，得到皂素；
(8)水液的处理：上述萃取后的水溶液加入酒曲发酵，过滤，滤液减压浓缩，低温结晶，得到粗品鼠李糖，结晶母液可制备酒精，粗品鼠李糖以无水甲醇溶解后，加入活性炭脱色，过滤，去除大量无机盐和色素，浓缩，结晶，干燥，得到精品鼠李糖；
(9)结晶母液的处理：母液提取农用核酸。
2.根据权利要求1所述的一种黄姜资源综合高效利用的绿色提取方法，其特征在于，步骤(1)中干黄姜用4～5倍(w/w)水室温浸泡，防止淀粉糊化，不利于下步提取，打浆后以30～40目的筛网过滤。
3.根据权利要求1所述的一种黄姜资源综合高效利用的绿色提取方法，其特征在于，步骤(2)中浆状物中加入一倍量的水充分搅拌后用80目的粗滤布过滤。
4.根据权利要求1所述的一种黄姜资源综合高效利用的绿色提取方法，其特征在于，步骤(3)中滤液浓缩至比重1.05～1.08，以等体积正丁醇萃取3～5次。
5.根据权利要求1所述的一种黄姜资源综合高效利用的绿色提取方法，其特征在于，步骤(4)中用pH10～12氢氧化钠溶液煮沸，过滤，得到的滤饼用水洗至中性，烘干。
6.根据权利要求1所述的一种黄姜资源综合高效利用的绿色提取方法，其特征在于，步骤(5)中以pH1～2的盐酸液加压至0.05MPa水解2～4h。
7.根据权利要求1所述的一种黄姜资源综合高效利用的绿色提取方法，其特征在于，步骤(7)中活性炭的用量为黄姜量的0.5％，加热至50～60℃脱色30min，浓缩至比重1.05～1.08，5～10℃结晶。
8.根据权利要求1所述的一种黄姜资源综合高效利用的绿色提取方法，其特征在于，步骤(8)中以酒曲发酵去除葡萄糖，酒曲用量为水液的3～5％，发酵温度35～45℃，发酵时间96～120h，加入3～5％(w/v)硅藻土起助滤作用，滤液加入1～3Kg活性炭加热至60～70℃，保温搅拌20min，停止搅拌，静置30min再过滤，此时活性炭吸附在脱色釜四周，过滤完成后将吸附在脱色釜四周的活性炭用水冲洗掉。
9.根据权利要求1所述的一种黄姜资源综合高效利用的绿色提取方法，其特征在于，步骤(9)中结晶后的母液加0.5％白糖、0.1％硫酸铵、0.05％磷酸二氢钾，用浓硫酸调pH2.5～3.0，煮沸15min，再调pH5～6，接种白地霉，培养14h；把白地霉培养液用200目/cm2过滤，将滤渣即湿白地霉放入容器中，加入菌体5倍的水，再加入5％菌体重的pH10的硼砂氢化钠缓冲液，75℃搅拌1h，菌体自溶；再用200目/cm2过滤，滤液煮沸消毒，即得农用核酸。
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