CN108120779B - 一种快速鉴定广西莪术中化学成分的分析方法 - Google Patents
一种快速鉴定广西莪术中化学成分的分析方法 Download PDFInfo
- Publication number
- CN108120779B CN108120779B CN201711398435.6A CN201711398435A CN108120779B CN 108120779 B CN108120779 B CN 108120779B CN 201711398435 A CN201711398435 A CN 201711398435A CN 108120779 B CN108120779 B CN 108120779B
- Authority
- CN
- China
- Prior art keywords
- curcuma
- database
- kwangsiensis
- automatically
- built
- 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.)
- Active
Links
- 239000000126 substance Substances 0.000 title claims abstract description 44
- 241000963421 Curcuma kwangsiensis Species 0.000 title claims abstract description 40
- 235000003397 Curcuma kwangsiensis Nutrition 0.000 title claims abstract description 40
- 238000004458 analytical method Methods 0.000 title claims abstract description 15
- 239000000284 extract Substances 0.000 claims abstract description 19
- 239000012530 fluid Substances 0.000 claims abstract description 15
- 235000014375 Curcuma Nutrition 0.000 claims abstract description 13
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 claims abstract description 13
- 150000001875 compounds Chemical class 0.000 claims abstract description 11
- 239000000203 mixture Substances 0.000 claims abstract description 11
- 229930014626 natural product Natural products 0.000 claims abstract description 10
- 239000012634 fragment Substances 0.000 claims abstract description 9
- 240000009138 Curcuma zedoaria Species 0.000 claims abstract description 8
- 238000012545 processing Methods 0.000 claims abstract description 5
- 241000407170 Curcuma Species 0.000 claims abstract 4
- 238000000605 extraction Methods 0.000 claims description 24
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 18
- 244000164480 Curcuma aromatica Species 0.000 claims description 13
- 150000002500 ions Chemical class 0.000 claims description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- 239000001569 carbon dioxide Substances 0.000 claims description 9
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 9
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 8
- 235000003405 Curcuma zedoaria Nutrition 0.000 claims description 7
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 6
- 244000008991 Curcuma longa Species 0.000 claims description 5
- 238000000194 supercritical-fluid extraction Methods 0.000 claims description 5
- 235000003373 curcuma longa Nutrition 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 230000003068 static effect Effects 0.000 claims description 4
- 239000003643 water by type Substances 0.000 claims description 4
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 3
- 241000196324 Embryophyta Species 0.000 claims description 3
- 238000001514 detection method Methods 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 3
- 235000019253 formic acid Nutrition 0.000 claims description 3
- 235000003398 Curcuma aromatica Nutrition 0.000 claims description 2
- 241000963390 Curcuma wenyujin Species 0.000 claims description 2
- 235000003394 Curcuma wenyujin Nutrition 0.000 claims description 2
- 239000003607 modifier Substances 0.000 claims description 2
- 239000012085 test solution Substances 0.000 claims 2
- 238000004949 mass spectrometry Methods 0.000 claims 1
- 239000000243 solution Substances 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 11
- 239000004615 ingredient Substances 0.000 abstract description 5
- 230000004907 flux Effects 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 238000003815 supercritical carbon dioxide extraction Methods 0.000 abstract 1
- 239000012071 phase Substances 0.000 description 13
- 239000007789 gas Substances 0.000 description 7
- 244000163122 Curcuma domestica Species 0.000 description 5
- 235000008216 herbs Nutrition 0.000 description 5
- 238000001819 mass spectrum Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- 238000004808 supercritical fluid chromatography Methods 0.000 description 5
- 239000001812 curcuma zedoaria berg. rosc. Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 235000019509 white turmeric Nutrition 0.000 description 4
- 150000002009 diols Chemical class 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000004811 liquid chromatography Methods 0.000 description 3
- 208000002193 Pain Diseases 0.000 description 2
- 238000012356 Product development Methods 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229930004069 diterpene Natural products 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 229930003658 monoterpene Natural products 0.000 description 2
- 150000002773 monoterpene derivatives Chemical class 0.000 description 2
- 235000002577 monoterpenes Nutrition 0.000 description 2
- SYSQUGFVNFXIIT-UHFFFAOYSA-N n-[4-(1,3-benzoxazol-2-yl)phenyl]-4-nitrobenzenesulfonamide Chemical class C1=CC([N+](=O)[O-])=CC=C1S(=O)(=O)NC1=CC=C(C=2OC3=CC=CC=C3N=2)C=C1 SYSQUGFVNFXIIT-UHFFFAOYSA-N 0.000 description 2
- 150000007965 phenolic acids Chemical class 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 229930004725 sesquiterpene Natural products 0.000 description 2
- -1 sesquiterpene compounds Chemical class 0.000 description 2
- 239000000341 volatile oil Substances 0.000 description 2
- QXDWRXCXHXYLNC-UHFFFAOYSA-N 4-phenylheptan-4-ylbenzene Chemical class C=1C=CC=CC=1C(CCC)(CCC)C1=CC=CC=C1 QXDWRXCXHXYLNC-UHFFFAOYSA-N 0.000 description 1
- 206010000077 Abdominal mass Diseases 0.000 description 1
- 201000000736 Amenorrhea Diseases 0.000 description 1
- 206010001928 Amenorrhoea Diseases 0.000 description 1
- 206010002383 Angina Pectoris Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 201000004569 Blindness Diseases 0.000 description 1
- 235000003392 Curcuma domestica Nutrition 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010061876 Obstruction Diseases 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 241000234299 Zingiberaceae Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 231100000540 amenorrhea Toxicity 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000006184 cosolvent Substances 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 150000004141 diterpene derivatives Chemical class 0.000 description 1
- 125000000567 diterpene group Chemical group 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000004089 microcirculation Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 235000009048 phenolic acids Nutrition 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 150000004354 sesquiterpene derivatives Chemical class 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000000115 thoracic cavity Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000010415 tropism Effects 0.000 description 1
- 235000013976 turmeric Nutrition 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
本发明提供了一种快速鉴定广西莪术中化学成分的分析方法，所述广西莪术提取物通过超临界二氧化碳萃取得到。将提取物通过超高效超临界流体色谱‑四极杆飞行时间质谱联用仪检测，UNIFI软件可自动进行数据处理，自动在UNIFI内置的天然产物数据库和自建姜黄属化学成分数据库内检索，鉴定结果自动显示，并且结构是否合理也自动通过二级碎片结构匹配进行确证。无法通过内置的天然产物数据库和自建姜黄属化学成分数据库鉴定的成分，可自动搜索在线数据库，然后自动通过二级碎片结构匹配来确证化合物。该方法快速、准确地实现了对广西莪术中化学成分的检测与鉴定，具有准确、操作简单、高通量、高效率和绿色环保等优点。
Description
技术领域
本发明涉及植物提取物的化学成分鉴定分析方法，具体涉及一种快速鉴定广西莪术中化学成分的分析方法。
背景技术
广西莪术(Curcuma kwangsiensis S.G.Lee&C.F.Liang)为姜科姜黄属植物，多年生草本，产我国广西、云南，栽培或野生于山坡草地及灌木丛中。本种的根茎即为中药莪术的一种。2015年版《中国药典》记载其性味与归经为辛、苦，温。归肝、脾经。功能与主治为行气破血，消积止痛。用于癥瘕痞块，血瘀经闭，胸痹心痛，食积胀痛。现代药理研究发现，中药莪术具有抗癌、抗菌、抗病毒、抗氧化、抗溃疡、抗炎及促进微循环等作用。广西莪术中一般含有两大类活性成分，二苯庚烷类化合物和挥发油类化合物。除此之外，还含有二萜类、多糖类、酚酸类和甾醇类等化学成分。广西莪术富含挥发油，是其有效成分群，多为单萜和倍半萜类化合物。
中药的功效已为几千年的历史和现代临床实验所肯定，但中药作用的物质基础和作用机理的研究还面临诸多挑战。中药作用物质基础的研究首先是要了解中药的化学成分。所有天然产物的研究都要源于化学成分分析，成分分析的研究一直以来也面临着许多挑战。传统的化学成分分析方法复杂，耗时且低效。目前，对广西莪术化学成分研究有以下3种方法：1.购买相关对照品，与样品中的组分进行对比，这种方法的缺点是成本高，且绝大多数化学成分无对照品。2.采用各种分离制备手段对组分进行分离纯化，这种方法的缺点是盲目性大，耗时太长。3.目前最常见的液质联用解决方法是对每个色谱峰逐一鉴定，通过检索数据库寻找可能的结构信息，然后再查文献进一步匹配二级碎片的数据与裂解途径，最终确定化合物的结构。这些方法和局限性是耗时长、效率低，同时对检测人员的中药及化学背景要求很高。因此，如何对广西莪术及其产品的化学成分的快速分析对广西莪术的研究、产品开发以及产品质量控制具有重要的意义。
超高效超临界流体色谱是以超临界二氧化碳为流动相，配以亚2微米颗粒色谱柱的色谱。超临界二氧化碳为流动相，它比液相所使用的液体流动相以及气相所使用的载气有更多突出的优点。CO2单独使用或与少量助溶剂共同使用作为流动相，流体粘度小，比液相色谱中所使用的液体流动相扩散率更高、更有利于传质。与气相色谱相比，CO2作流动相可在更低的温度下实现分离。超高效超临界流体色谱能够更加精确地改变流动相的强度、系统压力和温度，从而调整出系统的分离度和选择性。当分析结构类似物、异构体、对映体和非对映异构体混合物时，能够更好地控制分析物的保留，从而更好地分离，而这些化合物以任何其他方法分离通常都是很困难的。
中药中有大量的结构类似物和同分异构体。应用液相色谱法、气相色谱法、气质联用法和液质联用法对中药中的结构类似物和同分异构体的分离鉴定通常都是很困难的。广西莪术中化合物种类众多，极性差别大、含量大小不一，并且有大量的同分异构体，对其进行分离鉴定尤其困难。现行的方法不能解决复杂基质广西莪术的物质基质研究。本发明利用超高效超临界流体色谱-四极杆飞行时间质谱联用仪结合自建姜黄属化学成分数据库可快速鉴定广西莪术中的化学成分。
发明内容
本发明的目的是在于提供一种快速、准确，能同时分离鉴定多种化学成分的分析方法，该方法具有准确、操作简单、快速、高通量和绿色环保等优点，可应用于广西莪术及其产品的物质基础研究、质量控制、产品开发等各种与广西莪术相关的分析领域。
为实现上述目的，本发明采用如下技术方案：
一种广西莪术的提取物，所述广西莪术提取物按如下步骤制备得到：
超临界流体萃取：将干燥的广西莪术根茎粉碎，然后放入到萃取罐中，应用超临界流体萃取仪中进行萃取。萃取背压15MPa，萃取剂为超临界二氧化碳，改性剂为乙醇，其比例为0-30％，在温度45-55℃和压力15-25MPa下保持超临界流体充满萃取罐5-10min进行静态萃取；再通过流路切换阀切换，萃取罐中的萃取物在5-10min内由流经萃取罐的超临界流体洗脱，完成动态萃取。萃取次数(静态萃取+动态萃取)1-3次。完成动态萃取后，收集广西莪术萃取液，得到广西莪术提取物，可供超高效超临界流体色谱-四极杆飞行时间质谱联用仪分析。
一种所述的快速鉴定广西莪术中化学成分的分析方法，所述的快速鉴定分析方法为：
取广西莪术提取物，进样至超高效超临界流体色谱-四极杆飞行时间质谱联用仪，所述超高效超临界流体色谱-四极杆飞行时间质谱联用仪的实验条件为：
色谱条件：色谱柱为Torus Diol 130
(100*3mm，1.7μm)；流动相A为CO2，流动相B：甲醇/异丙醇(3∶1)+0.1％甲酸。梯度洗脱：0-2min，98％A；2-4min，92％A；4-9min，60％A；9-9.5min，98％A；9.5-11min，98％A；9.5-11min，98％A。补偿溶剂为甲醇。进样体积为1μL，流速0.4mL/min，柱温：45℃。
质谱条件：离子化模式：ESI+/ESI-，毛细管电压：2.5kV(ESI+)、2.0kV(ESI-)，锥孔电压：40V，离子源温度：120℃，雾化气温度：500℃，雾化气流速：800L/h，锥孔气流速：50L/h。
姜黄属化学成分数据库的建立：查阅国内外报道姜黄属化学成分的相关文献，姜黄属包括广西莪术、莪术(Curcuma phaeocaulis Valeton)、温郁金(Curcuma wenyujinY.H.Chen&C.Ling)、姜黄(Curcuma longa Linnaeus)、郁金(Curcuma aromaticaSalisbury)等药用植物，把这些化学成分的英文名称、分子式和精确分子量录入到Excel表，并画出每种化学成分的结构式，再把Excel表与结构式导入Waters的UNIFI软件中，建立姜黄属化学成分数据库。姜黄属化学成分数据库共收集了211种化合物，包括二苯庚烷类、单萜、倍半萜、二萜、多糖和酚酸类等化合物。
化合物鉴定：把自建姜黄属化学成分数据库导入UNIFI软件后，UNIFI软件可自动进行数据处理，自动在UNIFI内置的天然产物数据库和自建姜黄属化学成分数据库内检索，鉴定结果自动显示，并且结构是否合理也可通过二级碎片结构自动确证。无法通过内置的天然产物数据库和自建姜黄属化学成分数据库鉴定的成分，可自动搜索在线数据库，然后自动通过二级碎片结构来确证化合物。
本发明的有益效果在于：
(1)本发明采用超临界二氧化碳萃取广西莪术，有效地提取和采集广西莪术中的有效成分，成本低，效率高，绿色环保，易于工业化生产和应用。
(2)广西莪术萃取物中成分众多，极性差别大，同分异构体多，通过超高效超临界流体色谱和优化条件获得更好地分离广西莪术提取物中各个组分的方法，为广西莪术中的化学成分鉴定奠定了基础。
(3)通过超高效超临界流体色谱-四极杆飞行时间质谱联用仪对广西莪术提取物的检测，结合自建的姜黄属化学成分数据库、仪器内置天然产物数据库和在线数据库，应用于广西莪术化学成分的检测分析，能实现对广西莪术中化学成分快速准确的分离鉴定，具有操作简单、效率高和高通量等优点。可广泛应用于广西莪术及其产品的分析。该方法也可以拓展到姜黄属的其它药用植物提取物的化学成分快速分析。
(4)超临界萃取应用超临界二氧化碳作为萃取剂，超临界流体色谱应用超临界二氧化碳作为流动相，两者都应用了成本低且无毒超临界二氧化碳，把有毒溶剂的使用和废液处理降到最低水平，极大地节省了成本，同时保护了环境和实验人员健康。
附图说明
图1为广西莪术提取物正负的总离子流色谱图(TIC)：(a)为正离子扫描TIC，(b)为负离子扫描TIC。
图2为233。1538的精确提取离子色谱图(EIC)。
图3为8-Hydroxyarturmerone的质谱图：(a)一级质谱图，(b)二级质谱图。
具体实施方式
以下通过具体实施例对本例发明进行进一步说明，但本发明的保护范围并不仅限于此。
1.仪器、试剂与材料
仪器：超临界流体萃取仪(岛津公司，SFE-30A)；超高效超临界流体色谱-四极杆飞行时间质谱联用仪，超高效超临界流体色谱仪(Waters，ACQUITY UPC2)，四极杆飞行时间质谱仪(Waters，Xevo G2-XS QTof)，色谱柱为Torus Diol 130
(100*3mm，1.7μm)。
气体：CO2。
试剂：乙醇、甲醇和异丙醇均为色谱纯。
材料：广西莪术根茎的干燥粉末。
2.广西莪术提取物
将2.0g广西莪术根茎干燥粉末放进5mL萃取罐中，用超临界流体萃取仪中进行萃取。萃取背压15MPa，萃取压力20MPa，改进剂为乙醇，其比例为15％，萃取温度50℃，静态萃取时间10min，动态萃取时间10min，萃取次数1次。
3.色谱条件：色谱柱为Torus Diol 130
(100*3mm，1.7μm)；流动相A为CO2，流动相B：甲醇/异丙醇(3∶1)+0.1％甲酸，梯度洗脱：0-2min，98％A；2-4min，92％A；4-9min，60％A；9-9.5min，98％A；9.5-11min，98％A；9.5-11min，98％A。补偿溶剂为甲醇。进样体积为1μL，流速0.4mL/min，柱温：45℃
4.质谱条件：离子化模式：ESI+/ESI-，毛细管电压：2.5kV(ESI+)、2.0kV(ESI-)，锥孔电压：40V，离子源温度：120℃，雾化气温度：500℃，雾化气流速：800L/h，锥孔气流速：50L/h。
5.把自建姜黄属化学成分数据库导入UNIFI软件后，UNIFI软件可自动进行数据处理，自动在UNIFI内置的天然产物数据库和自建姜黄属化学成分数据库内检索，鉴定结果自动显示，并且结构是否合理也可自动通过二级碎片结构匹配来确证。无法通过内置的天然产物数据库和自建姜黄属化学成分数据库鉴定的成分，可自动搜索在线数据库，然后自动通过二级碎片结构匹配来确证化合物。共鉴定出48种化合物，详见表1。
表1：鉴定广西莪术提取物的化学成分
Claims (2)
1.一种快速鉴定广西莪术中化学成分的分析方法，其特征在于：它包括如下步骤：
(1)广西莪术提取物的制备：
将干燥的广西莪术根茎粉碎，然后放入到萃取罐中，应用超临界流体萃取仪进行萃取。萃取背压15MPa，萃取剂为超临界二氧化碳，改性剂为乙醇，其比例为0-30％，在温度45-55℃和压力15-25MPa下保持超临界流体充满萃取罐5-10min进行静态萃取；再通过流路切换阀切换，萃取罐中的萃取物在5-10min内由流经萃取罐的超临界流体洗脱，完成动态萃取，萃取次数1-3次，完成动态萃取后，收集广西莪术萃取液，制备得到广西莪术提取物作为供试品溶液；
(2)检测：取供试品溶液，应用超高效超临界流体色谱-四极杆飞行时间质谱联用仪检测；
色谱条件如下：
色谱柱：Torus Diol
规格为100*3mm，1.7μm；流动相：流动相A为CO2，流动相B：甲醇∶异丙醇＝3∶1+0 .1％甲酸；梯度洗脱：0-2min，98％A；2-4min，92％A；4-9min，60％A；9-9.5min，98％A；9.5-11min，98％A；9.5-11min，98％A；补偿溶剂：甲醇；流速：0.4mL/min；柱温：45℃；进样量：1μL；
质谱条件如下：
离子化模式：电喷雾离子源正负离子ESI+/ESI-，毛细管电压：正离子ESI+：2.5kV、负离子ESI-：2.0kV，锥孔电压：40V，离子源温度：120℃，雾化气温度：500℃，雾化气流速：800L/h，锥孔气流速：50L/h；
(3)姜黄属植物化学成分数据库的建立：
把已报道姜黄属的广西莪术、莪术、温郁金、姜黄和郁金5种植物的化学成分的英文名称、精确分子量、分子式和结构式，导入UNIFI软件中，建立姜黄属植物化学成分数据库；
(4)化合物鉴定：
UNIFI软件可自动进行数据处理，自动对UNIFI软件内置的天然产物数据库和自建姜黄属化学成分数据库进行检索，鉴定结果自动显示，并且结构是否合理也可通过二级碎片结构自动匹配来确证；无法通过内置的天然产物数据库和自建姜黄属化学成分数据库所鉴定的成分，可通过搜索在线数据库，然后通过二级碎片结构是否匹配来鉴定化合物。
2.根据权利要求1所述的分析方法，其特征在于：检测采用Waters超高效超临界流体色谱-四极杆飞行时间质谱联用仪。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711398435.6A CN108120779B (zh) | 2017-12-21 | 2017-12-21 | 一种快速鉴定广西莪术中化学成分的分析方法 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711398435.6A CN108120779B (zh) | 2017-12-21 | 2017-12-21 | 一种快速鉴定广西莪术中化学成分的分析方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108120779A CN108120779A (zh) | 2018-06-05 |
| CN108120779B true CN108120779B (zh) | 2022-09-27 |
Family
ID=62231230
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711398435.6A Active CN108120779B (zh) | 2017-12-21 | 2017-12-21 | 一种快速鉴定广西莪术中化学成分的分析方法 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108120779B (zh) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109212119A (zh) * | 2018-09-27 | 2019-01-15 | 成都中医药大学 | 一种基于uplc-q-tof/ms技术快速鉴定黄丝郁金的方法 |
| CN110208401B (zh) * | 2019-05-24 | 2021-11-02 | 岛津企业管理（中国）有限公司 | 固相脱水萃取-超临界流体色谱-质谱在线分析系统及方法 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651366A (zh) * | 2004-11-30 | 2005-08-10 | 河北科技大学 | 超临界co2流体萃取－精馏提取莪术中榄香烯的方法 |
| CN101144802A (zh) * | 2006-08-01 | 2008-03-19 | 烟台大学 | 莪术油的分析方法 |
| CN101400359A (zh) * | 2006-03-17 | 2009-04-01 | 草药科学新加坡私人有限公司 | 姜黄属提取物及方法 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060110468A1 (en) * | 2004-11-24 | 2006-05-25 | Liang Liu | Herbal formulations for arthritis |
| JP2011517686A (ja) * | 2008-04-11 | 2011-06-16 | ボード オブ スーパーバイザーズ オブ ルイジアナ ステイト ユニバーシティー アンド アグリカルチュラル アンド メカニカル カレッジ | 天然の可溶化剤としてのジテルペングリコシド |
-
2017
- 2017-12-21 CN CN201711398435.6A patent/CN108120779B/zh active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1651366A (zh) * | 2004-11-30 | 2005-08-10 | 河北科技大学 | 超临界co2流体萃取－精馏提取莪术中榄香烯的方法 |
| CN101400359A (zh) * | 2006-03-17 | 2009-04-01 | 草药科学新加坡私人有限公司 | 姜黄属提取物及方法 |
| CN101144802A (zh) * | 2006-08-01 | 2008-03-19 | 烟台大学 | 莪术油的分析方法 |
Non-Patent Citations (7)
| Title |
|---|
| Application of Supercritical Fluid Extraction and Chromatography to the Analysis of Turmeric;M. Marsin Sanagi等;《Journal of Chromatographic Science》;19930131;第31卷;20-25 * |
| Efficient separation of curcumin, demethoxycurcumin, and bisdemethoxycurcumin from turmeric using supercritical fluid chromatography: From analytical to preparative scale;Wei Song等;《J. Sep. Sci.》;20151231;第38卷;3305-3486 * |
| 一年蓬超临界提取物的GC-MS测定;赵昱玮等;《中国医药指南》;20101210(第34期);221-223 * |
| 不同提取方法对莪术挥发油成分的影响;佟磊等;《湖北中医学院学报》;20101231(第06期);35-37 * |
| 提取技术对温莪术挥发油化学成分及其体外抗肿瘤活性的影响;聂小华等;《药物生物技术》;20030830(第03期);152-154 * |
| 温郁金超临界萃取物的化学成分研究;钱华丽等;《天然产物研究与开发》;20071015(第05期);814-818 * |
| 超临界流体萃取分析蓬莪术挥发性成分;陈淑莲等;《中草药》;20001225(第12期);902-904 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108120779A (zh) | 2018-06-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gao et al. | Overview of the quality standard research of traditional Chinese medicine | |
| Pang et al. | The applications and features of liquid chromatography-mass spectrometry in the analysis of traditional Chinese medicine | |
| Liang et al. | Localization of ginsenosides in the rhizome and root of Panax ginseng by laser microdissection and liquid chromatography–quadrupole/time of flight-mass spectrometry | |
| Xu et al. | Simultaneous determination of eight anthraquinones in Semen Cassiae by HPLC‐DAD | |
| Zhu et al. | Recent development in mass spectrometry and its hyphenated techniques for the analysis of medicinal plants | |
| Shan et al. | Metabolomic study of raw and processed Atractylodes macrocephala Koidz by LC–MS | |
| Wei et al. | Analysis of tilianin and acacetin in Agastache rugosa by high-performance liquid chromatography with ionic liquids-ultrasound based extraction | |
| CN103789094A (zh) | 一种从沉香木屑中提取沉香精油的方法 | |
| Peng-Peng et al. | Metabolomics analysis and rapid identification of changes in chemical ingredients in crude and processed Astragali Radix by UPLC-QTOF-MS combined with novel informatics UNIFI platform | |
| Jiang et al. | Authentication of Schisandra chinensis and Schisandra sphenanthera in Chinese patent medicines | |
| Liu et al. | Differentiation of essential oils in Atractylodes lancea and Atractylodes koreana by gas chromatography with mass spectrometry | |
| Zhang et al. | Quantitative determination of 15 bioactive triterpenoid saponins in different parts of Acanthopanax henryi by HPLC with charged aerosol detection and confirmation by LC–ESI‐TOF‐MS | |
| Wen et al. | Simultaneous determination of twenty-two components in Asari Radix et Rhizoma by ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry | |
| Cheng et al. | Quality assessment of traditional Chinese medicine herb couple by high‐performance liquid chromatography and mass spectrometry combined with chemometrics | |
| CN108120779B (zh) | 一种快速鉴定广西莪术中化学成分的分析方法 | |
| Liang et al. | Structural characterisation and discrimination of the aerial parts of Paris polyphylla var. yunnanensis and Paris polyphylla var. chinensis by UHPLC‐QTOF‐MS coupled with multivariate data analysis | |
| Lv et al. | GC–MS based metabolomics study of stems and roots of Ephedra sinica | |
| CN109917066B (zh) | 一种基于uplc-qtof-ms检测伸筋草生物碱成分的方法 | |
| Li et al. | Rapid quantitative analysis of 12 chemical constituents in wild-simulated and cultivated Astragali Radix based on UHPLC-MS | |
| Jiang et al. | Screening the Q‐markers of TCMs from RA rat plasma using UHPLC‐QTOF/MS technique for the comprehensive evaluation of Wu‐Wei‐Wen‐Tong Capsule | |
| Chen et al. | Characterization of effective phytochemicals in traditional Chinese medicine by mass spectrometry | |
| Yi et al. | Histochemical evaluation of alkaloids in rhizome of Coptis chinensis using laser microdissection and liquid chromatography/mass spectrometry | |
| Duan et al. | High‐throughput LC–MS method for the rapid characterisation and comparative analysis of multiple ingredients of four hawthorn leaf extracts | |
| Xun et al. | Identification and comparison of compounds in commercial Tripterygium wilfordii genus preparations with HPLC-QTOF/MS based on molecular networking and multivariate statistical analysis | |
| Zhao et al. | Gas chromatography–mass spectrometry for quantitative and qualitative analysis of essential oil from Curcuma wenyujin rhizomes |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20180605 Assignee: Guangxi hengderun capsaicin Co.,Ltd. Assignor: GUANGXI BOTANICAL GARDEN OF MEDICINAL PLANTS Contract record no.: X2023980045867 Denomination of invention: A Rapid Analysis Method for Identifying Chemical Constituents in Curcuma zedoaria in Guangxi Granted publication date: 20220927 License type: Common License Record date: 20231107 |
|
| EE01 | Entry into force of recordation of patent licensing contract |