CN112321663A - 一种从桑黄中同时纯化提取类固醇和总三萜的方法 - Google Patents

一种从桑黄中同时纯化提取类固醇和总三萜的方法 Download PDF

Info

Publication number
CN112321663A
CN112321663A CN202011130975.8A CN202011130975A CN112321663A CN 112321663 A CN112321663 A CN 112321663A CN 202011130975 A CN202011130975 A CN 202011130975A CN 112321663 A CN112321663 A CN 112321663A
Authority
CN
China
Prior art keywords
steroid
phellinus igniarius
cof
total triterpene
extracting
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
CN202011130975.8A
Other languages
English (en)
Inventor
不公告发明人
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jinan Qiyuan Information Technology Co ltd
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to CN202011130975.8A priority Critical patent/CN112321663A/zh
Publication of CN112321663A publication Critical patent/CN112321663A/zh
Withdrawn legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07J—STEROIDS
    • C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
    • B01J20/26—Synthetic macromolecular compounds
    • B01J20/262—Synthetic macromolecular compounds obtained otherwise than by reactions only involving carbon to carbon unsaturated bonds, e.g. obtained by polycondensation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
    • B01J20/26—Synthetic macromolecular compounds
    • B01J20/264—Synthetic macromolecular compounds derived from different types of monomers, e.g. linear or branched copolymers, block copolymers, graft copolymers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28002—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their physical properties
    • B01J20/28004—Sorbent size or size distribution, e.g. particle size
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
    • B01J20/28042—Shaped bodies; Monolithic structures
    • B01J20/28045—Honeycomb or cellular structures; Solid foams or sponges
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
    • C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
    • C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
    • C07C29/76—Separation; Purification; Use of additives, e.g. for stabilisation by physical treatment
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C29/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom not belonging to a six-membered aromatic ring
    • C07C29/74—Separation; Purification; Use of additives, e.g. for stabilisation
    • C07C29/88—Separation; Purification; Use of additives, e.g. for stabilisation by treatment giving rise to a chemical modification of at least one compound
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C07C51/42—Separation; Purification; Stabilisation; Use of additives
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C07C51/42—Separation; Purification; Stabilisation; Use of additives
    • C07C51/47—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
    • C07C51/42—Separation; Purification; Stabilisation; Use of additives
    • C07C51/487—Separation; Purification; Stabilisation; Use of additives by treatment giving rise to chemical modification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D301/00—Preparation of oxiranes
    • C07D301/32—Separation; Purification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D303/00—Compounds containing three-membered rings having one oxygen atom as the only ring hetero atom
    • C07D303/02—Compounds containing oxirane rings
    • C07D303/38—Compounds containing oxirane rings with hydrocarbon radicals, substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D303/40—Compounds containing oxirane rings with hydrocarbon radicals, substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals by ester radicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2220/00—Aspects relating to sorbent materials
    • B01J2220/40—Aspects relating to the composition of sorbent or filter aid materials
    • B01J2220/48—Sorbents characterised by the starting material used for their preparation
    • B01J2220/4812—Sorbents characterised by the starting material used for their preparation the starting material being of organic character
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07—Optical isomers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/02—Systems containing only non-condensed rings with a three-membered ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/14—The ring being saturated
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/16—Systems containing only non-condensed rings with a six-membered ring the ring being unsaturated

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Analytical Chemistry (AREA)
  • Diabetes (AREA)
  • Immunology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Mycology (AREA)
  • Obesity (AREA)
  • Hematology (AREA)
  • Biotechnology (AREA)
  • Endocrinology (AREA)
  • Epidemiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medical Informatics (AREA)
  • Botany (AREA)
  • Emergency Medicine (AREA)
  • Microbiology (AREA)
  • Rheumatology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Cardiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Pulmonology (AREA)
  • Pain & Pain Management (AREA)
  • Steroid Compounds (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

本发明公开了一种从桑黄中同时纯化提取类固醇和总三萜的方法，该方法采用复合酶酶解、多孔共价有机框架材料COF‑701富集、多频超声辅助提取的工艺，通过优化上样液、洗脱液和超声频率等参数，实现了对桑黄中类固醇和总三萜的同时高效纯化富集。该方法具有稳定性好、选择性高、吸附量大、操作简便可控等优点，可用于各种复杂样品中类固醇和总三萜的纯化富集。

Description

一种从桑黄中同时纯化提取类固醇和总三萜的方法
技术领域
本发明涉及一种同时纯化提取桑黄中类固醇和总三萜的方法，属于天然有机化学领域。
背景技术
桑黄是我国一类传统珍稀药用真菌的俗称，具有多种药理作用，如抗肿瘤、抗氧化、提升免疫力等。桑黄的天然子实体内含有丰富的高分子类多糖体，三萜类、蛋白质、矿物质、氨基酸和固醇类等化合物。三萜类化合物具有调节免疫力、抑制癌细胞，调整血压、保护肝脏，降低血脂肪、抗过敏与发炎等多种作用，其药理作用与临床效果深受肯定。固醇类化合物是由胆固醇衍生而来，具有抗发炎，调节体内的钠、钾比例，使其保持恒定，调节血糖含量，避免过高或不足以及调节性功能，同样促进第二性征成熟的作用。目前，对于桑黄多糖的提取、分离纯化研究较多，而类固醇和三萜类化合物的研究较少。
发明内容
本发明的技术任务是为了弥补现有技术的不足，提供一种从桑黄中同时纯化提取类固醇和总三萜的方法，该方法耗时短、成本低、操作简单，具有很好的工业前景。
为实现上述目的，本发明采用的技术方案如下：
(1)将除杂后的桑黄在60℃下烘干后粉碎，过40目筛网，得到桑黄干粉；
(2)向桑黄干粉中加入磷酸氢二钠缓冲液，混合均匀，再按桑黄干粉质量的0.2～1.0％加入复合酶，在45～55℃下酶解30～90分钟，酶解液经离心分离得到上清液和沉淀物；
(3)在步骤(2)中上清液中加入共价有机框架材料COF-701，其添加量为桑黄干粉质量的1％，进行多频超声萃取，经离心分离得到COF-701富集材料；
(4)COF-701富集材料置于离心管中，加入洗脱液，超声振荡1分钟，经离心分离得到上清液和COF-701；上清液经旋转蒸发仪减压浓缩后得到类固醇和总三萜产品。
所述磷酸氢二钠缓冲液,成分为磷酸氢二钠和柠檬酸，pH为4.0～6.0，其添加量为桑黄干粉质量的5～10倍。
所述复合酶，由纤维素酶和葡萄糖氧化酶组成，其中，纤维素酶和葡萄糖氧化酶质量比为3：1～1：3。
所述共价有机框架材料COF-701，由2,4,6-三甲基-1,3,5-三嗪和4,4′-联苯二甲醛在三氟乙酸催化下于150℃聚合得到,比表面积是1715m2g-1，孔径为1.1nm，在强酸性和碱性条件下可以保持其组成和结晶度，其结构式如下：
Figure RE-GDA0002863579120000021
所述多频超声萃取，超声频率为20-80kHz，两两组合或三种同时组合，超声处理功率为100-1000W，萃取时间为30-300s。
所述离心分离，在700～2000rpm下离心10～30min。
所述洗脱液，由乙醇、甘油、碳酸二甲酯、柠檬烯、乙酸乙酯中的一种或二种及以上混合；洗脱液添加量为COF-701富集材料质量的2～5倍。
所述超声振荡,超声频率为25kHz，超声处理功率为600W。
所述类固醇和总三萜产品为含有Phellilane L(图1所示1)、Phellidene E(图1所示 2)、Phellinulin D(图1所示3)、Phellinulin I(图1所示4)、Phellinulin K(图1 所示5)、啤酒甾醇(图1所示6)、β-谷甾醇(图1所示7)和齿孔醇(图1所示8)的混合物，提取率为60％-80％。
相对于现有技术，本发明所述的从桑黄中同时纯化提取类固醇和总三萜的方法具有以下优势：
(1)本发明在桑黄的类固醇和总三萜提取中加入了多频超声协同提取方法，显著缩短提取时间，减少溶剂用量，提高提取率；且得到的类固醇和总三萜提取物纯度高，无需后续分离处理，提取物具有广阔的药用和食用的应用前景。
(2)COF-701是由轻质元素C和N连接而成的高度有序多孔晶态材料，具有低的密度和高的比表面积(1715m2g-1)，在强酸性和碱性条件下也可以稳定存在，其作为富集材料可反复循环利用；孔径为1.1nm，孔道内含有π电子共轭体系，对类固醇和总三萜化合物具有高的选择性吸附性能；与其他多孔性吸附材料相比，具有无法比拟的优势。
附图说明
图1是提取的类固醇和总三萜产品种类，图1所示1为Phellilane L、图1所示2 为Phellidene E、图1所示3为Phellinulin D、图1所示4为Phellinulin I、图1 所示5为Phellinulin K、图1所示6为啤酒甾醇、图1所示7为β-谷甾醇、图1所示 8为齿孔醇。
具体实施方式
除有定义外，以下实施例中所用的技术术语具有与本发明所属领域技术人员普遍理解的相同含义，所述实验方法，如无特殊说明，均为常规方法。
下面结合实施例来详细说明本发明。
实施例1一种从桑黄中同时纯化提取类固醇和总三萜的方法
(1)将除杂后的桑黄在60℃下烘干后粉碎，过40目筛网，得到桑黄干粉；
(2)向桑黄干粉中加入磷酸氢二钠缓冲液，混合均匀，再按桑黄干粉质量的0.2％加入复合酶，在55℃下酶解90分钟，酶解液经离心分离得到上清液和沉淀物；
(3)在步骤(2)中上清液中加入共价有机框架材料COF-701，其添加量为桑黄干粉质量的1％，进行多频超声萃取，经离心分离得到COF-701富集材料；
(4)COF-701富集材料置于离心管中，加入洗脱液，超声振荡1分钟，经离心分离得到上清液和COF-701；上清液经旋转蒸发仪减压浓缩后得到类固醇和总三萜产品。
所述磷酸氢二钠缓冲液,成分为磷酸氢二钠和柠檬酸，pH为4.0，其添加量为桑黄干粉质量的5倍；
所述复合酶，由纤维素酶和葡萄糖氧化酶组成，其中，纤维素酶和葡萄糖氧化酶质量比为3：1；
所述共价有机框架材料COF-701，由2,4,6-三甲基-1,3,5-三嗪和4,4′-联苯二甲醛在三氟乙酸催化下于150℃聚合得到,比表面积是1715m2g-1，孔径为1.1nm，在强酸性和碱性条件下可以保持其组成和结晶度，其结构式如下：
Figure RE-GDA0002863579120000031
所述多频超声萃取，超声频率为20kHz和80kHz，超声处理功率为1000W，萃取时间为300s；
所述离心分离，在2000rpm下离心10min；
所述洗脱液，由乙醇和柠檬烯混合，洗脱液添加量为COF-701富集材料质量的2倍；所述超声振荡,超声频率为25kHz，超声处理功率为600W。
实施例2一种从桑黄中同时纯化提取类固醇和总三萜的方法
(1)将除杂后的桑黄在60℃下烘干后粉碎，过40目筛网，得到桑黄干粉；
(2)向桑黄干粉中加入磷酸氢二钠缓冲液，混合均匀，再按桑黄干粉质量的0.6％加入复合酶，在50℃下酶解60分钟，酶解液经离心分离得到上清液和沉淀物；
(3)在步骤(2)中上清液中加入共价有机框架材料COF-701，其添加量为桑黄干粉质量的1％，进行多频超声萃取，经离心分离得到COF-701富集材料；
(4)COF-701富集材料置于离心管中，加入洗脱液，超声振荡1分钟，经离心分离得到上清液和COF-701；上清液经旋转蒸发仪减压浓缩后得到类固醇和总三萜产品。
所述磷酸氢二钠缓冲液,成分为磷酸氢二钠和柠檬酸，pH为5.0，其添加量为桑黄干粉质量的7.5倍；
所述复合酶，由纤维素酶和葡萄糖氧化酶组成，其中，纤维素酶和葡萄糖氧化酶质量比为1：1；
所述共价有机框架材料COF-701，由2,4,6-三甲基-1,3,5-三嗪和4,4′-联苯二甲醛在三氟乙酸催化下于150℃聚合得到,比表面积是1715m2g-1，孔径为1.1nm，在强酸性和碱性条件下可以保持其组成和结晶度，其结构式如下：
Figure RE-GDA0002863579120000041
所述多频超声萃取，超声频率为30kHz和70kHz，超声处理功率为550W，萃取时间为135s；
所述离心分离，在1400rpm下离心20min；
所述洗脱液，由乙醇、甘油和乙酸乙酯混合，洗脱液添加量为COF-701富集材料质量的3.5倍；
所述超声振荡,超声频率为25kHz，超声处理功率为600W。
实施例3一种从桑黄中同时纯化提取类固醇和总三萜的方法
(1)将除杂后的桑黄在60℃下烘干后粉碎，过40目筛网，得到桑黄干粉；
(2)向桑黄干粉中加入磷酸氢二钠缓冲液，混合均匀，再按桑黄干粉质量的1.0％加入复合酶，在45℃下酶解30分钟，酶解液经离心分离得到上清液和沉淀物；
(3)在步骤(2)中上清液中加入共价有机框架材料COF-701，其添加量为桑黄干粉质量的1％，进行多频超声萃取，经离心分离得到COF-701富集材料；
(4)COF-701富集材料置于离心管中，加入洗脱液，超声振荡1分钟，经离心分离得到上清液和COF-701；上清液经旋转蒸发仪减压浓缩后得到类固醇和总三萜产品。
所述磷酸氢二钠缓冲液,成分为磷酸氢二钠和柠檬酸，pH为6.0，其添加量为桑黄干粉质量的10倍；
所述复合酶，由纤维素酶和葡萄糖氧化酶组成，其中，纤维素酶和葡萄糖氧化酶质量比为1：3；
所述共价有机框架材料COF-701，由2,4,6-三甲基-1,3,5-三嗪和4,4′-联苯二甲醛在三氟乙酸催化下于150℃聚合得到,比表面积是1715m2g-1，孔径为1.1nm，在强酸性和碱性条件下可以保持其组成和结晶度，其结构式如下：
Figure RE-GDA0002863579120000051
所述多频超声萃取，超声频率为20kHz,50kHz和80kHz，超声处理功率为100W，萃取时间为30s；
所述离心分离，在700rpm下离心30min；
所述洗脱液，由乙醇、甘油、碳酸二甲酯混合；洗脱液添加量为COF-701富集材料质量的5倍；
所述超声振荡,超声频率为25kHz，超声处理功率为600W。
实施例4共价有机框架材料COF-701的制备方法
(1)0.06g 2,4,6-三甲基-1,3,5-三嗪和0.154g 4,4'-联苯二醛溶解在4.5mL三甲基苯，4.5mL1,4-二恶烷，2mL三氟乙酸和0.25mL乙腈中；
(2)混合溶液置于高压釜中在150℃下进行72小时，得到红色固体；
(3)红色固体用0.05mol/L氨水溶液中和，并用乙醇清洗3次；
(4)在120℃真空条件下干燥48h，得到黄色粉末COF-701，产率为85％；
循环利用的共价有机框架材料COF-701需要在120℃真空条件下活化24h。

Claims (8)

1.一种从桑黄中同时纯化提取类固醇和总三萜的方法，其特征在于，步骤如下：
(1)将除杂后的桑黄在60℃下烘干后粉碎，过40目筛网，得到桑黄干粉；
(2)向桑黄干粉中加入磷酸氢二钠缓冲液，混合均匀，再按桑黄干粉质量的0.2～1.0％加入复合酶，在45～55℃下酶解30～90分钟，酶解液经离心分离得到上清液和沉淀物；
(3)在步骤(2)中上清液中加入共价有机框架材料COF-701，其添加量为桑黄干粉质量的1％，进行多频超声萃取，经离心分离得到COF-701富集材料；
(4)COF-701富集材料置于离心管中，加入洗脱液，超声振荡1分钟，经离心分离得到上清液和COF-701；上清液经旋转蒸发仪减压浓缩后得到类固醇和总三萜产品。
2.根据权利要求1所述的一种从桑黄中同时纯化提取类固醇和总三萜方法，其特征在于：所述磷酸氢二钠缓冲液,成分为磷酸氢二钠和柠檬酸，pH为4.0～6.0，其添加量为桑黄干粉质量的5～10倍。
3.根据权利要求1所述的一种从桑黄中同时纯化提取类固醇和总三萜方法，其特征在于：所述复合酶，由纤维素酶和葡萄糖氧化酶组成，其中，纤维素酶和葡萄糖氧化酶质量比为3：1～1：3。
4.根据权利要求1所述的一种从桑黄中同时纯化提取类固醇和总三萜方法，其特征在于：所述共价有机框架材料COF-701，由2,4,6-三甲基-1,3,5-三嗪和4,4′-联苯二甲醛在三氟乙酸催化下于150℃聚合得到,比表面积是1715m2g-1，孔径为1.1nm，在强酸性和碱性条件下可以保持其组成和结晶度，其结构式如下：
Figure FDA0002735146030000021
5.根据权利要求1所述的一种从桑黄中同时纯化提取类固醇和总三萜方法，其特征在于：所述多频超声萃取，超声频率为20-80kHz，两两组合或三种同时组合，超声处理功率为100-1000W，萃取时间为30-300s。
6.根据权利要求1所述的一种从桑黄中同时纯化提取类固醇和总三萜方法，其特征在于：所述离心分离，在700～2000rpm下离心10～30min。
7.根据权利要求1所述的一种从桑黄中同时纯化提取类固醇和总三萜方法，其特征在于：所述洗脱液，由乙醇、甘油、碳酸二甲酯、柠檬烯、乙酸乙酯中的一种或二种及以上混合；洗脱液添加量为COF-701富集材料质量的2～5倍。
8.根据权利要求1所述的方法，其特征在于：所述超声振荡,超声频率为25kHz，超声处理功率为600W。
CN202011130975.8A 2020-10-21 2020-10-21 一种从桑黄中同时纯化提取类固醇和总三萜的方法 Withdrawn CN112321663A (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202011130975.8A CN112321663A (zh) 2020-10-21 2020-10-21 一种从桑黄中同时纯化提取类固醇和总三萜的方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202011130975.8A CN112321663A (zh) 2020-10-21 2020-10-21 一种从桑黄中同时纯化提取类固醇和总三萜的方法

Publications (1)

Publication Number Publication Date
CN112321663A true CN112321663A (zh) 2021-02-05

Family

ID=74311849

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202011130975.8A Withdrawn CN112321663A (zh) 2020-10-21 2020-10-21 一种从桑黄中同时纯化提取类固醇和总三萜的方法

Country Status (1)

Country Link
CN (1) CN112321663A (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114716502A (zh) * 2022-04-15 2022-07-08 菏泽学院 一种桑黄三萜类化合物的提取方法

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114716502A (zh) * 2022-04-15 2022-07-08 菏泽学院 一种桑黄三萜类化合物的提取方法

Similar Documents

Publication Publication Date Title
US12109509B2 (en) Ultrasonic composite acidic water extraction method for cordyceps polysaccharide and cordycepin in Cordyceps militaris
CN101768614B (zh) 一种从姬松茸中同时高效提取多糖和多酚的方法
US9447198B2 (en) Method for extracting polysaccharides from higher plants and fungi through microwave chemical treatment
CN102948758B (zh) 一种从荞麦麸皮中提取荞麦黄酮的方法
CN104211827B (zh) 一种从猴头菇菌渣中提取多糖的方法
CN103613674A (zh) 一种黄芪多糖的提取方法
CN101613418A (zh) 一种提取双孢蘑菇多糖的方法
CN101705275A (zh) 薯蓣皂素生产工艺及提取薯蓣皂素后盾叶薯蓣的处理方法
CN114774493A (zh) 一种桑黄活性物质多糖的提取及纯化方法
CN103627510A (zh) 一种从香菇中综合获取有效成分的方法
CN107970270A (zh) 一种从杜仲叶中分步提取总黄酮和多糖的方法
CN114949915A (zh) 一种猴头菇复合提取物及其制备方法
CN106432529A (zh) 一种高纯度米糠多糖的制备方法
CN113750135B (zh) 一种牛大力中总皂苷的超声-微波协同提取方法
CN102002072A (zh) 一种从枣核中提取黄酮的工艺方法
CN118388677B (zh) 一种猴头菇多糖的提取方法及其应用
CN116535536B (zh) 一种桑黄多糖的提取方法
CN112321662A (zh) 一种从灵芝中纯化提取总三萜的方法
CN109762866B (zh) 利用醋酸杆菌全细胞催化提高人参总皂苷中稀有皂苷含量的方法与应用
CN114716502B (zh) 一种桑黄三萜类化合物的提取方法
CN114560959B (zh) 一种菌菇类提取物的制备方法
CN102718831B (zh) 一种用于制造蛋白丝的蚕蛹蛋白的制备方法
CN111388512A (zh) 一种桑黄活性成分的提取方法及桑黄中药饮片
CN105838770A (zh) 一种氧化微杆菌发酵二醇组人参皂苷制备稀有人参皂苷ck的方法
CN110075134A (zh) 一种灵芝三萜酸的提取方法

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
TA01 Transfer of patent application right

Effective date of registration: 20210414

Address after: Room 1-2512, Zhonghai International Mansion, Shizhong District, Jinan City, Shandong Province

Applicant after: Jinan Qiyuan Information Technology Co.,Ltd.

Address before: No. 28, sangyuan Road, Licheng District, Jinan City, Shandong Province

Applicant before: Song Jinghua

TA01 Transfer of patent application right
WW01 Invention patent application withdrawn after publication

Application publication date: 20210205

WW01 Invention patent application withdrawn after publication