CN113416184A - 一种从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法 - Google Patents
一种从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法 Download PDFInfo
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- CN113416184A CN113416184A CN202110829023.3A CN202110829023A CN113416184A CN 113416184 A CN113416184 A CN 113416184A CN 202110829023 A CN202110829023 A CN 202110829023A CN 113416184 A CN113416184 A CN 113416184A
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- isoschaftoside
- vitexin
- alkaloid
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Abstract
本发明公开了一种从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:以魔芋属植物的各部位或其加工剩余物作为原料,经过预处理或脱毒处理,再经浸提得到异夏佛塔苷、牡荆素和生物碱的提取液,提取液经过分离纯化后得到产品。与产异夏佛塔苷和牡荆素的传统中草药原料相比,魔芋茎叶及提取剩余物是生物量极大,成本极低的丰富资源,作为制备异夏佛塔苷、牡荆素和生物碱的原料,这些魔芋原料和提取剩余物的利用有利于消除因废弃造成的环境污染,提高种植户的经济效益,经提取和加工后可得到高纯度的异夏佛塔苷、牡荆素和生物碱。
Description
技术领域
本发明涉及化学物质提取方法,特别涉及一种从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法。
背景技术
天南星科魔芋属植物为多年生草本。成年的块茎常扁球形,稀萝卜状或长圆柱形,顶部中央下凹。叶柄光滑或粗糙;叶片3全裂,小叶羽状分裂或二次羽状分裂,或二歧分裂后再羽状分裂,最后小裂片多少为长圆形。花序1,通常有长柄,稀为短梗;佛焰苞宽卵形或长圆形,基部漏斗形或钟形,席卷;肉穗花序长或短于佛焰苞,花单性,雌花在下部,附属器粗长;花无花被;雄蕊1~6个;雌花心皮1~4,子房近球形或倒卵形,1~4室,每室胚珠1粒。浆果有1个或少数种子。该属主要分布于东半球。在中国主产于江南各地。(中国植物志,1979,V13,85-86)。
全世界每年生产魔芋粉量约为25000吨,中国和日本都有大面积的魔芋种植(刁英,滕彩珠,吴金平,向发云,顾玉成,胡中立.魔芋的研究进展[J].安徽农学通报,2006(06):137-139+147),中国每年魔芋粉产量约15000吨,位居第一,成为世界上产量最大的魔芋生产国,且形成了涵盖种植、加工和制品为一体的完善的魔芋产业链。日本年产魔芋粉量6500吨,位居第二。中国近年的魔芋种植面积约43万亩,鲜芋产量达20万吨(魔芋产量很高,仅块茎亩产量就达数吨,最大的魔芋块茎达40多公斤一个),工业魔芋精粉(葡甘露聚糖)产量一般仅占魔芋块茎量的20%,因此魔芋精粉加工业产生了大量的茎叶(主要弃在田间地头,日晒雨淋污染水源)和工业提取剩余物急等综合利用,魔芋产品开发目前仅以葡甘露聚糖为代表的多糖(精粉),以食品应用为主,如凝胶食品、增稠剂、乳化剂。
魔芋块茎可加工成豆腐供疏食。有的魔芋品种其干片含多糖较多,达42.05%,吸水膨胀可增大至原体积的80-100倍,粘着力强,可用作浆纱、造纸、瓷器或建筑等的胶粘剂。块茎入药能解毒消肿,健胃,消饱胀。治流火、疔疮、无名肿毒、瘰疬、眼镜蛇咬伤、烫火伤、间日疟、乳痈、腹中痞块、疔癀高烧、疝气等。
Tsukasa Iwashinaa等对巨魔芋叶中黄酮化合物和黄原酮进行了鉴定,发现了16种成份,但未对这16种成份的具体含量进行定量,因此无从判断这16种成份的一种或多种或全部是否达到了商业应用价值。因为魔芋中毒性成份的存在,使人们对魔芋多糖之外的成份利用难以进行或没有进行工业化利用,对其中生物碱的关注也多限于研究层面(Tsukasa Iwashinaa,Destrib,Flavonoids and xanthones from the leaves ofAmorphophallus titanum(Araceae).[J].Biochemical Systematics and Ecology,2020,23(5):21-25)。
巨魔芋叶中的16种黄酮和黄原酮成份(Tsukasa,2020)
夏佛塔苷和异夏佛塔苷(分子式C26H28O14,分子量564.49,同分异构体)是中草药半夏、天南星、毛鸡骨草、广金钱草等中的重要功效成份,两者均具有保肝护肝、抗炎、清热祛湿的优良功效。虽然在多种植物中均有分布,但含量都很低,而异夏佛塔苷(Isoschaftoside),是比夏佛塔苷(Isoschaftoside)含量更低、更稀少的成份,因此价格高昂。广金钱草是制备异夏佛塔苷的传统原料之一(谢冬养.一种从广金钱草中提取异夏佛塔苷的方法[P].CN2018107674066A)。此外,毛鸡骨草是另一种制备异夏佛塔苷的原料,但异夏佛塔苷在毛鸡骨草中含量仅0.4mg/g(刘东锋,杨成东.一种从毛鸡骨草中制备异夏佛塔苷的方法[P],CN201210115084),广金钱草和毛鸡骨草都是生物量很少的植物,因此无法满足制备大量异夏佛塔苷药物的需求,本发明经大量的实验对多种魔芋的成份进行鉴定和定性、定量分析及分离纯化,发现与魔芋中其他黄酮成份相比,异夏佛塔苷和牡荆素这两种黄酮及生物碱含量都很高,具有良好的工业开发价值,可以替代传统的生物量小、含量低的制备这三种功能成份的中草药资源。
牡荆素具有活血化瘀、理气通脉的作用,主要用于治疗心血管疾病,如冠心病绞痛、高脂血症、心动脉供血不足等。牡荆素具有抗氧化活性,对人类成纤维细胞紫外线照射后产生的自由基有很强的清除能力,有效地防止紫外照射给皮肤造成的不良反应,进而减少皮肤损伤(Kim J H,Lee B C,Kim J H,et al.The isolation and antioxidativeeffects of vitexin from Acer palmatum[J].Archives ofPharmacal Research,2005,28(2):195-202.)。牡荆素还是抗癌抗肿瘤的天然药物成分,研究表明,牡荆素对U937细胞显示出抗肿瘤活性(Lee C Y,Chien Y S,Chiu T H,et al.Apoptosis triggered byvitexin in U937 humanleukemia cells via a mitochondrial signaling pathway[J].Oncology Reports,2012,28(5):1883-1888)。Diaz等报道从黄荆和牡荆中得到的紫花牡荆素在生物活性测定中对人类癌细胞显示了广泛的细胞毒性作用(Díaz Fredyc,ChávezDaniel,Lee Dongho,Mi Qiuwen,Chai Hee-Byung,Tan Ghee T,Kardono Leonardus B S,Riswan Soedarsono,Fairchild Craig R,Wild Robert,Farnsworth Norman R,CordellGeoffrey A,Pezzuto John M,Kinghorn A Douglas.Cytotoxic flavone analogues ofvitexicarpin,a constituent of the leaves of Vitex negundo[J].Journal ofnatural products,2003,66(6):865-867);Massateru等的研究发现,紫花牡荆素在MTT试验中对人肺癌细胞(PC-12)和人结肠癌细胞(HCT116)的生长有抑制作用(百度百科,牡荆素词条)。
牡荆素能够明显降低由蔗糖诱导的糖尿病患者的血糖含量(Choo CY,Sulong NY,Man F.et a1.Vitexin and isovitexin from the Leaves of Ficus deltoidea,within-vivo,α-glucosidase inhibition[J].Journal of Ethnopharmacology,2012,142(3):776),其在治疗糖尿病中有重要的作用。牡荆素不仅能够清除自由基、螯合金属离子,还能够抑制AGE。因此,牡荆素是一种治疗糖尿病及其并发症的良好药物。牡荆素还具有抗病毒活性,能够抑制流感病毒、轮状病毒(Dong L Y,Chen Z W,Guo Y,et al.Mechanisms ofvitexin preconditioning effects on cultured neonatal rat cardiomyocytes withanoxia and reoxygenation[J].American Journal of Chinese Medicine,2008,36(2):385),但是对艾滋病毒的抑制作用较弱,对植物类病毒中的烟草花叶病毒也具有一定抑制作用(E,Rusak G,N,et al.Inhibition of tobacco mosaic virusinfection by quercetin and vitexin[J].Acta Virologica,2008,52(2):119-124)。
发明内容
发明目的:本发明目的是提供高纯度的异夏佛塔苷、牡荆素和生物碱的提取方法。
技术方案:本发明提供的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,以魔芋属植物的各部位或其加工物作为原料,经过预处理或脱毒处理,再经浸提得到异夏佛塔苷、牡荆素和生物碱的提取液,提取液经过分离纯化后得到产品。
进一步地,所述魔芋属植物的各部位或其加工物为块茎、茎、叶、提取多糖之后的剩余物或者生产魔芋精粉过程中产生的飞粉。
进一步地,所述预处理或脱毒处理方法:微生物发酵、酶处理。
进一步地,所述微生物为真菌、细菌或者放线菌。
进一步地,所述酶为纯酶、酶曲或混合酶。
进一步地,所述魔芋属植物为A.aberrans、A.abyssinicus、A.adamsensis、A.albispathus、A.albus、A.amygdaloides、A.andranogidroensis、A.angolensis、A.angulatus、A.angustispathus、A.ankarana、A.annulifer、A.antsingyensis、A.aphyllus、A.arcuspadix、A.asper、A.asterostigmatus、A.atrorubens、A.atroviridis、A.bangkokensis、A.bankokensis、A.bannanensis、A.barbatus、A.barthlottii、A.baumannii、A.beccarii、A.bequaertii、A.bhandarensis.A.bognerianus、A.bolikhamxayensis、A.bonaccordensis、A.borneensis、A.boyceanus、A.brachyphyllus、A.brevipetiolatus、A.brevispathus、A.bubenensis、A.bufo、A.bulbifer、A.calabaricus、A.canaliculatus、A.carneus、A.chlorospathus、A.cicatricirer、A.cirriler、A.claudelii、A.coaetaneus、A.commutatus、A.consimilis、A.corrugatus、A.costatus、A.coudercii、A.crispifolius、A.croatii、A.cruddasianus、A.curvistylis、A.dactylifer、A.declinatus、A.decus-silvae、A.discophorus、A.dracontioides、A.dunnii、A.dzui、A.eburneus、A.echinatus、A.eichleri、A.elatus、A.elegans、A.elliottii、A.erythrororrhachis、A.excentricus、A.fallax、A.ferruginosus、A.flotoi、A.forbesii、A.fuscus、A.galbra、A.gallaensis、A.gallowayi、A.gigantiflorus、A.gigasTeijsm、A.glaucophyllus、A.gliruroides、A.glossophyllus、A.goetzei、A.gomboczianus、A.gracilior、A.gracilis、A.haematospadix、A.harmandii、A.hayiHett、A.hemicryptus、A.henryi、A.hetterscheidii、A.hewittii、A.hildebrandtii、A.hirsutus、A.hirtus、A.hohenackeri、A.hottae、A.impressus、A.incurvatus、A.infundibuliformis、A.interruptus、A.johnsonii、A.josefbogneri、A.julaihii、A.juliae、A.kachinensis、A.khammouanensis、A.kienluongensis、A.kiusianus、A.konjac、A.konkanensis、A.koratensis、A.krausei、A.kuznetso训i、A.lacou行i、A.lambii、A.lanceolatus、A.lanuginosus、A.laoticus、A.lewallei、A.linearis、A.linguiformis、A.longicomus、A.longiconnectivus、A.longispathaceus、A.longistylus、A.longituberosus、A.lunatus、A.luzoniensis、A.lyratus、A.macrophyllus、A.macrorhizus、A.mairei、A.mangelsdorffii、A.manta、A.margaritifer、A.matrgretae、A.maximus、A.maxwellii、A.mekongensis、A.mellii、A.merrillii、A.microappendiculatus、A.mildbraedii、A.minor、A.mossambicensis、A.muelleriBlume、A.mullendersii、A.myosuroides、A.mysorensis、A.napalensis、A.napiger、A.natolii、A.niahensis、A.nicolaii、A.nicolsonianus、A.niimurai、A.obovoideus、A.obscurus、A.ochroleucus、A.oncophyllus、A.ongsakulii、A.operculatus、A.opertus、A.paeoniifolius、A.palawanensis、A.paucisectus、A.pendulus、A.perakensis、A.perrieri、A.pilosus、A.plicatus、A.polyanthus、A.prainii、A.preussii、A.prolificus、A.pulchcllus、A.purpurascens、A.pusillus、A.putii、A.pygmaeus、A.ranchanensis、A.reflexus、A.rhizomatosus、A.richardsiae、A.rivierei、A.rostratus、A.rugosus、A.sagittarius、A.salmoneus、A.saraburensis、A.saururus、A.scaber、A.schmidtiae、A.scutatus、A.serrulatus、A.shyamsalilianus、A.sinensis、A.sinuatus、A.sizemoreae、A.smithsonianus、A.sparsiflorus、A.spectabilis、A.staudtii、A.stipitatus、A.stuhlmannii、A.subcymbiformis、A.sumawongii、A.suwidjianus、A.sylvaticus、A.symonianus、A.synandrifer、A.taurostigma、A.tenuispadix、A.tenuistylis、A.terrestris、A.teuszii、A.thaiensis、A.tinekeae、A.titanum、A.tonkinensis、A.tuberculatus、A.variabilis、A.venustus、A.verticillatus、A.virosus、A.vogelianus、A.xiei、A.yuloensis、A.yunnanensis、A.zengianus、A.zenkeri。
进一步地,所述魔芋属品种为A.albus、A.atroviridis、A.bankokensis、A.bannanensis、A.corrugatus、A.bubenensis、A.hirtus、A.niimurai、A.virosus、A.mekongensis、A.yunnanensis、A.dunnii、A.stipitatus、A.variabilis、A.henryi、A.oncophyllus、A.bulbifer、A.kiusianus、A.tonkinensis、A.kachinensis、A.coaetaneus、A.hayi、A.krausei、A.konjac、A.paeoniifolius、A.titanum、A.xiei、A.yuloensis、A.zengianus。
进一步地,所述分离方法为:水或有机溶剂提取、大孔树脂吸附、聚酰胺层析、硅胶层析、液相色谱或高效逆流色谱。
有益效果:与产异夏佛塔苷和牡荆素的传统中草药原料相比,魔芋茎叶及提取剩余物是生物量极大,成本极低,作为制备异夏佛塔苷、牡荆素和生物碱的原料既有利于环保又充分利用了资源,极大地提高魔芋种植业和加工业的经济效益。经加工可得到异夏佛塔苷、牡荆素和生物碱这些极有价值的药用功能性成分,本发明对充分利用魔芋资源、开拓制备异夏佛塔苷、牡荆素和生物碱的新原料来源及提升中药现代化效益都有着显著的应用价值
附图说明
图1夏佛塔苷标品UV扫描;
图2白魔芋提取物的UV扫描;
图3疣柄魔芋提取物UV扫描;
图4疣柄魔芋叶提取液HPLC分析;
图5花魔芋叶提取液HPLC-MS二级质谱(异夏佛塔苷);
图6东亚魔芋叶提取液HPLC-MS二级质谱(牡荆素);
图7未加夏佛塔苷标品的魔芋提取液HPLC分析;
图8加有夏佛塔苷标品的魔芋提取液HPLC分析;
图9异夏佛塔苷氢谱。
具体实施方式
实施例1
A.virosus块茎原料100kg,按固液比1∶50(w/w)加入体积浓度60%的乙醇,超声辅助提取30min,经过固液分离脱乙醇并浓缩后得到浸膏3.8kg,经过D101树脂分离和硅胶层析,获得纯度95%的异夏佛塔苷2.81Kg,82%的牡荆素0.29Kg,在经过石油醚-乙酸乙酯洗脱得到66%的神经酰胺0.05kg。
实施例2
A.paeoniifolius块茎原料100kg,加入用柠檬酸调PH至6.0的乙醇,按固液比1∶40(w/w),微波辅助提取30min,经过分离得到含异夏佛塔苷\牡荆素\生物碱的提取液,回收溶剂得到浸膏,经过D201树脂分离和聚酰胺分离,得到纯度95%的异夏佛塔苷1.86Kg,纯度94%牡荆素0.23kg,提取完后的提取液再经过分离后得到纯度72%木脂素0.32kg。
实施例3
A.albus叶100kg,按固液比1∶60(w/w)加入体积浓度60%的乙醇溶液,加热回流提取30min,经过固液分离得到异夏佛塔苷的提取液,回收溶剂得到浸膏,经过D101树脂和聚酰胺分离,得到纯度90%异夏佛塔苷1.87kg,纯度94%牡荆素0.194kg,提取后的提取液经过高效逆流色谱分离出纯度65%的氰苷0.25kg。
实施例4
A.yunnanensis叶原料100kg,按固液比1∶30(w/w)加入70%乙醇,,加热回流提取60min,经过分离得到提取液,回收溶剂得到浸膏,经过聚酰胺柱,分离得到纯度87%的牡荆素0.42kg,纯度93%异夏佛塔苷2.13kg。
实施例5
A.hirtus叶柄原料100kg,经黑曲霉发酵4天,按照固液比1∶50(w/w)加入90%乙醇溶液,微波辅助提取90min,经分离得到夏佛塔苷的提取液,回收溶剂得到浸膏5.0Kg,经过ADS-17分离得到纯度91%的异夏佛塔苷1.66Kg,纯度85%的牡荆素0.47kg,纯度62%木脂素0.31Kg,提取剩余物经脉孢菌发酵成为饲料产品。
实施例6
A.kachinensis块茎原料100kg,经过纤维素酶处理2小时,按固液比1:50(w/w)加入体积浓度70%的甲醇溶液,超声辅助提取60min,经固液分离后得到浸膏,经过硅胶柱层析得到纯度95%的异夏佛塔苷1.67kg和81%的牡荆素0.21kg。
实施例7
A.yuloensis叶原料100kg,经炮制处理,按固液比1∶30(w/w)加入Ph5.0的体积浓度50%的甲醇溶液,在超声辅助提取120min,经过分离得到异夏佛塔苷的提取液,提取液经过高效逆流色谱分离纯化,分离得到纯度为96%的异夏佛塔苷1.43kg,90%的牡荆素0.29kg。
实施例8
A.konjac叶原料100kg,经过果胶酶生物发酵,发酵后按照固液比1∶30(w/w)加入Ph6.0的体积浓度60%的甲醇溶液,,经过分离得到异夏佛塔苷的提取液,提取液经过高效逆流色谱分离纯化,分离得到纯度为92%的异夏佛塔苷1.59kg。
实施例9
A.corrugatus块茎原料100kg,经过晒4h,按照固液比1∶60(w/w)加入体积浓度70%的甲醇溶液,在超声辅助提取180min,经过固液分离后得到浸膏经过硅胶柱层析分离获得纯度95%的异夏佛塔苷1.89kg和93%的牡荆素0.35kg。
实施例10
A.corrugatus块茎原料100kg,经过久煎2h,按照固液比1∶60(w/w)加入体积浓度70%的甲醇溶液,在超声辅助提取下提取60min,经过固液分离后得到浸膏经过硅胶柱层析分离获得纯度95%的异夏佛塔苷1.19kg。提取剩余物经脉孢菌发酵成为饲料产品。
实施例11
与实施例1相同,原料选择A.titanum,前处理选择用纤维素酶发酵,提取液经过D301大孔树脂经分离纯化后,分别得到纯度91%的异夏佛塔苷1.34kg和纯度93%的牡荆素0.35kg,再经过石油醚-乙酸乙酯洗脱得到纯度55%的神经酰胺0.07Kg。
实施例12
与实施例1相同,原料选择A.stipitatus,将提取溶剂换为甲基乙基酮溶液,用微波辅助提取60min,分离纯化同实施例1,得到纯度88%的异夏佛塔苷1.87kg、纯度90%的牡荆素0.27Kg和纯度52%的神经酰胺0.07kg。
实施例13
与实施例1相同,原料选择A.zengianus,将提取溶剂换为异丙醇溶液,用微波辅助提取30min,提取三次,分离纯化同实施例1,得到纯度87%的异夏佛塔苷1.67kg、纯度88%的牡荆素0.31kg和纯度51%的神经酰胺0.06kg。
实施例14
与实施例1相同,原料选择A.atroviridis,前处理选择用纤维素酶发酵,再经过蜜灸处理,提取液经过AB-8树脂经分离纯化后,分离方法同实施例1相同,分别得到纯度92%的异夏佛塔苷2.11kg、纯度82%的牡荆素0.21kg。
实施例15
与实施例1相同,原料选择A.bankokensis,将提取溶剂换为甲醇溶液,分离纯化同实施例1,得到纯度93%的异夏佛塔苷1.89kg和纯度91%的牡荆素0.34kg。
实施例16
与实施例1相同,原料选择A.bubenensis,将提取溶剂换为正丁醇溶液,分离纯化同实施例1,得到纯度90%的异夏佛塔苷2.05kg、纯度87%的牡荆素0.37kg和61%纯度的神经酰胺0.06kg。
实施例17
与实施例1相同,原料选择A.niimurai,将提取溶剂换为水,分离纯化同实施例1,得到纯度92%的异夏佛塔苷2.27kg、纯度91%的牡荆素0.35kg和65%纯度的神经酰胺0.05kg。。
实施例18
与实施例2相同,原料选择A.mekongensis,前处理选择用冠突散囊菌菌发酵,提取液经过HPD-722大孔树脂经分离纯化后,分别得到纯度94%的异夏佛塔苷2.46kg、纯度90%的牡荆素0.32kg和纯度69%的木脂素0.07kg。
实施例19
与实施例2相同,原料选择A.dunniii,将提取溶剂换为丙酮溶液,分离纯化同实施例2,分别得到纯度93%的异夏佛塔苷1.97kg、纯度90%的牡荆素0.37kg和纯度64%的木脂素0.07kg。
实施例20
与实施例2相同,原料选择A.variabilis,将提取溶剂换为乙酸乙酯溶液,分离纯化同实施例2,分别得到纯度91%的异夏佛塔苷2.68kg、纯度89%的牡荆素0.29kg和纯度64%的木脂素0.05kg。
实施例21
与实施例2相同,原料选择A.oncophyllus,分离纯化同实施例2,分别得到纯度92%的异夏佛塔苷2.03kg、纯度91%的牡荆素0.31kg和纯度62%的木脂素0.05kg。
实施例22
与实施例2相同,原料选择A.henri,分离纯化同实施例2,分别得到纯度93%的异夏佛塔苷2.03kg、纯度87%的牡荆素0.37kg和纯度62%的木脂素0.07kg。
实施例23
与实施例2相同,原料选择A.bulbifer,分离纯化同实施例2,分别得到纯度89%的异夏佛塔苷2.29kg、纯度89%的牡荆素0.34kg和纯度58%的木脂素0.08kg。
实施例24
与实施例2相同,原料选择A.kiusianus,分离纯化同实施例2,分别得到纯度92%的异夏佛塔苷1.74kg、纯度87%的牡荆素0.29kg和纯度61%的木脂素0.04kg。
实施例25
与实施例2相同,原料选择A.tonkinensis,分离纯化同实施例2,分别得到纯度93%的异夏佛塔苷1.21kg、纯度88%的牡荆素0.23kg和纯度55%的木脂素0.06kg。
实施例26
与实施例2相同,原料选择A.coaetaneus,分离纯化同实施例2,分别得到纯度92%的异夏佛塔苷1.84kg、纯度91%的牡荆素0.27kg和纯度64%的木脂素0.05kg。
实施例27
与实施例3相同,原料选择A.hayi,分离纯化同实施例3,得到纯度89%异夏佛塔苷1.96kg,纯度92%牡荆素0.19kg,提取后的提取液经过高效逆流色谱分离出纯度62%的氰苷0.18kg。
实施例28
与实施例3相同,原料选择A.krausei,分离纯化同实施例3,得到纯度93%异夏佛塔苷1.52kg,纯度90%牡荆素0.21kg,提取后的提取液经过高效逆流色谱分离出纯度61%的氰苷0.15kg。
实施例29
与实施例3相同,原料选择A.xiei,分离纯化同实施例3,得到纯度90%异夏佛塔苷1.87kg,纯度94%牡荆素0.24kg,提取后的提取液经过高效逆流色谱分离出纯度60%的氰苷0.17kg。
实施例30
与实施例7相同,煎制时间为4小时,固液比为1∶100(w/w)
实施例31
与实施例3相同,原料前处理用蛋白酶处理,再用50%的甲醇提取经加热回流提取60min,分离纯化同实施例3。
实施例32
与实施例8相同,固液比为1∶60,pH为5.0.
实施例33
与实施例8相同,提取溶剂为异丙醇。
实施例34
与实施例6相同,煎制时间为5小时。
实施例35
与实施例3相同,树脂换成XAD-2。
实施例36
与实施例1相同,魔芋品种为珠芽魔芋(Amorphophallus bulbifer(Roxb.)Blume),分离纯化同实施例2,分别得到纯度81%的异夏佛塔苷2.26kg、纯度86%的牡荆素0.52kg和纯度67%的木脂素0.012kg。提取剩余物经产朊假丝酵母发酵后成为富含蛋白质和维生素的饲料。
实施例37
与实施例1相同,魔芋品种为花魔芋(Amorphophallus konjac K.Koch)与白魔芋(Amorphophallus albus)的杂交品种,分离纯化同实施例2,分别得到纯度921%的异夏佛塔苷2.59kg、纯度87%的牡荆素0.58kg和纯度87%的神经酰胺0.26kg,提取剩余物经乳酸菌发酵后成为富含乳酸和乳酸菌的饲料。
实验例1
将夏佛塔苷标准品配制为浓度为50μg/ml的溶液,利用高效液相色谱对其进行分析,将实施例1的魔芋叶提取异夏佛塔苷和牡荆素,经过旋转蒸发浓缩提取液,再次利用高效液相色谱进行测定,得到图7,提取液中加入配制好的夏佛塔苷标准品,进行高效液相色谱分析,得到图8,其中出现了夏佛塔苷和异夏佛塔苷的两个峰,根据LC-MS分析其分子量,确定实验所用魔芋叶中含量最高的物质是异夏佛塔苷。
实验例2
将实施例2中的A.paeoniifolius的提取液,稀释10倍后,过0.22μm滤膜,利用高效液相色谱对其进行分析,与牡荆素标品进行比较,LC-MS分析其分子量,比较保留时间,提取液中出峰之一的一级质谱[M-H]-的m/z是431.30,二级质谱产生的离子碎片m/z分别为413.10、341.20、311.20、283.30,这些与牡荆素产生的二级质谱的离子碎片相同。后续经分离纯化后再经UV\IR\NMR鉴定,确定其为牡荆素。
Claims (10)
1.一种从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:以魔芋属植物的各部位或其加工剩余物为原料,经过预处理或脱毒处理,再经浸提或分离得到异夏佛塔苷、牡荆素和生物碱的提取液,分离液或提取液经过进一步的分离纯化得到异夏佛塔苷、牡荆素和生物碱产品。
2.根据权利要求1所述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:所述魔芋属植物的各部位或其加工物为块茎、茎、叶、提取多糖之后的剩余物、提取生物碱或者生产魔芋精粉过程中产生的飞粉。
3.根据权利要求1所述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:所述预处理或脱毒处理方法:粉碎、磨浆、烘烤、蒸煮、炮制、微生物发酵、酶处理。
4.根据权利要求3述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:所述微生物为真菌、细菌或放线菌。
5.根据权利要求3述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:所述酶为纯酶、酶曲或混合酶。
6.权利要求1所述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:所述魔芋属植物为A.aberrans、A.abyssinicus、A.adamsensis、A.albispathus、A.albus、A.amygdaloides、A.andranogidroensis、A.angolensis、A.angulatus、A.angustispathus、A.ankarana、A.annulifer、A.antsingyensis、A.aphyllus、A.arcuspadix、A.asper、A.asterostigmatus、A.atrorubens、A.atroviridis、A.bangkokensis、A.bankokensis、A.bannanensis、A.barbatus、A.barthlottii、A.baumannii、A.beccarii、A.bequaertii、A.bhandarensis.A.bognerianus、A.bolikhamxayensis、A.bonaccordensis、A.borneensis、A.boyceanus、A.brachyphyllus、A.brevipetiolatus、A.brevispathus、A.bubenensis、A.bufo、A.bulbifer、A.calabaricus、A.canaliculatus、A.carneus、A.chlorospathus、A.cicatricifer、A.cirrifer、A.claudelii、A.coaetaneus、A.commutatus、A.consimilis、A.corrugatus、A.costatus、A.coudercii、A.crispifolius、A.croatii、A.cruddasianus、A.curvistylis、A.dactylijer、A.declinatus、A.decus-silvae、A.discophorus、A.dracontioides、A.dunnii、A.dzui、A.eburneus、A.echinatus、A.eichleri、A.elatus、A.elegans、A.elliottii、A.erythrororrhachis、A.excentricus、A.fallax、A.ferruginosus、A.flotoi、A.forbesii、A.fuscus、A.galbra、A.gallaensis、A.gallowayi、A.gigantiflorus、A.gigasTeijsm、A.glaucophyllus、A.gliruroides、A.glossophyllus、A.goetzei、A.gomboczianus、A.gracilior、A.gracilis、A.haematospadix、A.harmandii、A.hayiHett、A.hemicryptus、A.henryi、A.hetterscheidii、A.hewittii、A.hildebrandtii、A.hirsutus、A.hirtus、A.hohenackeri、A.hottae、A.impressus、A.incurvatus、A.infundibuliformis、A.interruptus、A.johnsonii、A.josefbogneri、A.julaihii、A.juliae、A.kachinensis、A.khammouanensis、A.kienluongensis、A.kiusianus、A.konjac、A.konkanensis、A.koratensis、A.krausei、A.kuznetsovii、A.lacourii、A.lambii、A.lanceolatus、A.lanuginosus、A.laoticus、A.lewallei、A.linearis、A.linguiformis、A.longicomus、A.longiconnectivus、A.longispathaceus、A.longistylus、A.longituberosus、A.lunatus、A.luzoniensis、A.lyratus、A.macrophyllus、A.macrorhizus、A.mairei、A.mangelsdorffii、A.manta、A.margaritifer、A.margretae、A.maximus、A.maxwellii、A.mekongensis、A.mellii、A.merrillii、A.microappendiculatus、A.mildbraedii、A.minor、A.mossambicensis、A.muelleriBlume、A.mullendersii、A.myosuroides、A.mysorensis、A.napalensis、A.napiger、A.natolii、A.niahensis、A.nicolaii、A.nicolsonianus、A.niimurai、A.obovoideus、A.obscurus、A.ochroleucus、A.oncophyllus、A.ongsakulii、A.operculatus、A.opertus、A.paeoniifolius、A.palawanensis、A.paucisectus、A.pendulus、A.perakensis、A.perrieri、A.pilosus、A.plicatus、A.polyanthus、A.prainii、A.preussii、A.prolificus、A.pulchellus、A.purpurascens、A.pusillus、A.putii、A.pygmaeus、A.ranchanensis、A.reflexus、A.rhizomatosus、A.richardsiae、A.rivierei、A.rostratus、A.rugosus、A.sagittarius、A.salmoneus、A.saraburensis、A.saururus、A.scaber、A.schmidtiae、A.scutatus、A.serrulatus、A.shyamsalilianus、A.sinensis、A.sinuatus、A.sizemoreae、A.smithsonianus、A.sparsiflorus、A.spectabilis、A.staudtii、A.stipitatus、A.stuhlmannii、A.subcymbiformis、A.sumawongii、A.suwidjianus、A.sylvaticus、A.symonianus、A.synandrifer、A.taurostigma、A.tenuispadix、A.tenuistylis、A.terrestris、A.teuszii、A.thaiensis、A.tinekeae、A.titanum、A.tonkinensis、A.tuberculatus、A.variabilis、A.venustus、A.verticillatus、A.virosus、A.vogelianus、A.xiei、A.yuloensis、A.yunnanensis、A.zengianus、A.zenkeri。
7.根据权利要求1所述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:所述魔芋属品种为:A.albus、A.atroviridis、A.bankokensis、A.bannanensis、A.corrugatus、A.bubenensis、A.hirtus、A.niimurai、A.virosus、A.mekongensis、A.yunnanensis、A.dunnii、A.stipitatus、A.variabilis、A.henryi、A.oncophyllus、A.bulbifer、A.kiusianus、A.tonkinensis、A.kachinensis、A.coaetaneus、A.hayi、A.krausei、A.konjac、A.paeoniifolius、A.titanum、A.xiei、A.yuloensis、A.zengianus。
8.根据权利要求1所述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:所述分离方法为:水或有机溶剂提取、大孔树脂吸附、聚酰胺层析、硅胶层析、制备色谱、高效逆流色谱。
9.根据权利要求1所述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:所述预处理或脱毒处理方法为中药炮制的传统方法,如蒸、煮、晒、酒制、蜜灸、加硝。
10.根据权利要求1所述的从魔芋中提取异夏佛塔苷、牡荆素和生物碱的方法,其特征在于:在提取异夏佛塔苷、牡荆素和生物碱可以在提取魔芋多糖之前或之后或同步,提取多糖和提取异夏佛塔苷、牡荆素和生物碱之后的剩余物可以直接或经发酵制成饲料或肥料。
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