CN105717241A - Determination method of chlorogenic acid in wild chrysanthemum flower - Google Patents
Determination method of chlorogenic acid in wild chrysanthemum flower Download PDFInfo
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- CWVRJTMFETXNAD-FWCWNIRPSA-N 3-O-Caffeoylquinic acid Natural products O[C@H]1[C@@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-FWCWNIRPSA-N 0.000 title claims abstract description 17
- PZIRUHCJZBGLDY-UHFFFAOYSA-N Caffeoylquinic acid Natural products CC(CCC(=O)C(C)C1C(=O)CC2C3CC(O)C4CC(O)CCC4(C)C3CCC12C)C(=O)O PZIRUHCJZBGLDY-UHFFFAOYSA-N 0.000 title claims abstract description 17
- CWVRJTMFETXNAD-KLZCAUPSSA-N Neochlorogenin-saeure Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O CWVRJTMFETXNAD-KLZCAUPSSA-N 0.000 title claims abstract description 17
- CWVRJTMFETXNAD-JUHZACGLSA-N chlorogenic acid Chemical compound O[C@@H]1[C@H](O)C[C@@](O)(C(O)=O)C[C@H]1OC(=O)\C=C\C1=CC=C(O)C(O)=C1 CWVRJTMFETXNAD-JUHZACGLSA-N 0.000 title claims abstract description 17
- 229940074393 chlorogenic acid Drugs 0.000 title claims abstract description 17
- FFQSDFBBSXGVKF-KHSQJDLVSA-N chlorogenic acid Natural products O[C@@H]1C[C@](O)(C[C@@H](CC(=O)C=Cc2ccc(O)c(O)c2)[C@@H]1O)C(=O)O FFQSDFBBSXGVKF-KHSQJDLVSA-N 0.000 title claims abstract description 17
- 235000001368 chlorogenic acid Nutrition 0.000 title claims abstract description 17
- BMRSEYFENKXDIS-KLZCAUPSSA-N cis-3-O-p-coumaroylquinic acid Natural products O[C@H]1C[C@@](O)(C[C@@H](OC(=O)C=Cc2ccc(O)cc2)[C@@H]1O)C(=O)O BMRSEYFENKXDIS-KLZCAUPSSA-N 0.000 title claims abstract description 17
- 235000008495 Chrysanthemum leucanthemum Nutrition 0.000 title abstract description 20
- 244000035851 Chrysanthemum leucanthemum Species 0.000 title abstract description 20
- 238000000034 method Methods 0.000 title abstract description 13
- 238000000605 extraction Methods 0.000 claims abstract description 30
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 230000003068 static effect Effects 0.000 claims abstract description 11
- 238000004811 liquid chromatography Methods 0.000 claims abstract description 10
- 239000000284 extract Substances 0.000 claims abstract description 9
- 239000000203 mixture Substances 0.000 claims abstract description 9
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 claims abstract description 7
- 238000001514 detection method Methods 0.000 claims abstract description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000007788 liquid Substances 0.000 claims abstract description 4
- 239000012528 membrane Substances 0.000 claims abstract description 4
- 238000010790 dilution Methods 0.000 claims abstract 3
- 239000012895 dilution Substances 0.000 claims abstract 3
- 238000000227 grinding Methods 0.000 claims abstract 2
- 238000007873 sieving Methods 0.000 claims abstract 2
- 238000003556 assay Methods 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 6
- 238000004458 analytical method Methods 0.000 claims description 5
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 claims description 3
- 239000007791 liquid phase Substances 0.000 claims description 3
- 239000012071 phase Substances 0.000 claims description 3
- 238000000638 solvent extraction Methods 0.000 claims description 3
- 238000010926 purge Methods 0.000 claims description 2
- 241000628997 Flos Species 0.000 claims 6
- 229930003944 flavone Natural products 0.000 claims 3
- 150000002213 flavones Chemical class 0.000 claims 3
- 235000011949 flavones Nutrition 0.000 claims 3
- 238000004587 chromatography analysis Methods 0.000 claims 1
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 5
- 238000010992 reflux Methods 0.000 abstract description 4
- 238000011010 flushing procedure Methods 0.000 abstract description 2
- 239000002904 solvent Substances 0.000 abstract description 2
- 238000002156 mixing Methods 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 6
- 239000005909 Kieselgur Substances 0.000 description 3
- 238000007865 diluting Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 239000002253 acid Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- GPWDPLKISXZVIE-UHFFFAOYSA-N cyclo[18]carbon Chemical compound C1#CC#CC#CC#CC#CC#CC#CC#CC#C1 GPWDPLKISXZVIE-UHFFFAOYSA-N 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
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- G—PHYSICS
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- 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
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- G—PHYSICS
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- 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
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- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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- 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
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Abstract
Description
技术领域technical field
本发明涉及实验室检测领域，特别是一种野菊花中的绿原酸的测定方法。The invention relates to the field of laboratory testing, in particular to a method for measuring chlorogenic acid in wild chrysanthemums.
背景技术Background technique
HeraldofmedicineVOL28No8Augest2009公开了高效液相色谱法测定野菊花中绿原酸的含量，其采用回流提取的方法提取得到绿原酸，然后通过高效液相色谱法进行测量，其测量时间较长且提取测量精度有待提高。HeraldofmedicineVOL28No8August2009 disclosed the content of chlorogenic acid in the wild chrysanthemum by high-performance liquid chromatography, which adopts the method of reflux extraction to extract chlorogenic acid, and then measures it by high-performance liquid chromatography. improve.
随着快速检测技术的发展，通过超声萃取法、快速溶剂萃取法可以得到中药中的多种有效成分，难点在于，如何有效调整各参数达到对各组分准确测试的目的。With the development of rapid detection technology, a variety of active ingredients in traditional Chinese medicine can be obtained through ultrasonic extraction and rapid solvent extraction. The difficulty lies in how to effectively adjust each parameter to achieve the purpose of accurate testing of each component.
发明内容Contents of the invention
本发明的目的是提供一种野菊花中的绿原酸的测定方法，该方法相比于回流提取法提取并进行高效液相色谱法检测得到结果更加精确。The object of the present invention is to provide a method for assaying chlorogenic acid in wild chrysanthemum, which is more accurate than the method extracted by reflux extraction and detected by high performance liquid chromatography.
本发明提供的技术方案为：一种野菊花中的绿原酸的测定方法，包括以下步骤：The technical scheme provided by the invention is: a method for assaying chlorogenic acid in wild chrysanthemums, comprising the following steps:
步骤1：将野菊花粉碎后过筛，取0.25重量份的野菊花粉与0.3g硅藻土混合，加入到预先铺设有滤膜的萃取池中进行静态萃取获得萃取液，萃取条件为：萃取溶剂为重量比为0.1:9.9的乙二胺和乙醇的混合物，萃取温度100℃，静态萃取时间为5min，冲洗体积100％，静态循环次数为1次，萃取压力1500psi；Step 1: crush the wild chrysanthemum flower and sieve it, mix 0.25 parts by weight of wild chrysanthemum pollen with 0.3 g of diatomaceous earth, add it to an extraction pool with a filter membrane in advance, and perform static extraction to obtain an extract. The extraction conditions are: extraction solvent It is a mixture of ethylenediamine and ethanol with a weight ratio of 0.1:9.9, the extraction temperature is 100°C, the static extraction time is 5min, the flushing volume is 100%, the number of static cycles is 1, and the extraction pressure is 1500psi;
步骤2：将萃取液稀释离心后，得到上清液；Step 2: After diluting and centrifuging the extract, obtain the supernatant;
步骤3：将上清液采用液相色谱法进行分析；Step 3: analyzing the supernatant by liquid chromatography;
其中，液相色谱法的分析条件为：Wherein, the analysis condition of liquid chromatography is:
a)仪器：液相色谱仪a) Instrument: liquid chromatograph
b)色谱柱C183μm3×100mmb) Column C 18 3μm3×100mm
c)柱温：40℃c) Column temperature: 40°C
d)流速：0.5mL/mind) Flow rate: 0.5mL/min
e)流动相：乙腈-水，体积比为4:6e) Mobile phase: acetonitrile-water, the volume ratio is 4:6
f)检测波长：326nm。f) Detection wavelength: 326nm.
在上述的野菊花中的绿原酸的测定方法中，所述的萃取池的体积为5ml，所述的静态萃取的吹扫时间为60s。In the above method for measuring chlorogenic acid in wild chrysanthemum, the volume of the extraction pool is 5ml, and the purging time of the static extraction is 60s.
在上述的野菊花中的绿原酸的测定方法中，步骤2具体为：将萃取液用重量比为0.1:9.9的乙二胺和乙醇的混合物稀释后在15000r/min下离心5min，取上清液。In the method for measuring chlorogenic acid in the above-mentioned wild chrysanthemum, step 2 is specifically: after diluting the extract with a mixture of ethylenediamine and ethanol with a weight ratio of 0.1:9.9, centrifuge at 15000r/min for 5min, and take the upper Serum.
在上述的野菊花中的绿原酸的测定方法中，液相色谱分析所用的仪器为：ThermoU-3000双三元液相色谱仪，色谱柱为ThermoSyncronisC18色谱柱。In the assay method of chlorogenic acid in the above-mentioned wild chrysanthemum, the instrument used for liquid chromatography analysis is: Thermo U-3000 dual-ternary liquid chromatography, and the chromatographic column is ThermoSyncronis C18 chromatographic column.
在上述的野菊花中的绿原酸的测定方法中，所述的步骤1中萃取所用到的仪器为ASE350快速溶剂萃取仪In the assay method of the chlorogenic acid in the above-mentioned wild chrysanthemum, the instrument used for extraction in the described step 1 is ASE350 fast solvent extraction instrument
本发明在采用上述技术方案后，其具有的有益效果为：After the present invention adopts above-mentioned technical scheme, the beneficial effect that it has is:
本发明通过ASE静态快速溶液萃取方法对野菊花中的绿原酸行萃取，并调整液相色谱法参数，达到对野菊花中的绿原酸更加快速简便且高精度的测定野菊花中绿原酸的目的。The present invention extracts the chlorogenic acid in the wild chrysanthemum through the ASE static fast solution extraction method, and adjusts the parameters of the liquid chromatography to achieve a more rapid, simple and high-precision determination of the chlorogenic acid in the wild chrysanthemum acid purpose.
具体实施方式detailed description
下面结合具体实施方式，对本发明的技术方案作进一步的详细说明，但不构成对本发明的任何限制。The technical solutions of the present invention will be further described in detail below in conjunction with specific embodiments, but this does not constitute any limitation to the present invention.
实施例1：Example 1:
一种野菊花中的绿原酸的测定方法：将野菊花经粉碎机粉碎，过三号筛，约0.25g，精密称定，与0.5g硅藻土混合均匀，待用，移入到预先放好过滤膜的5ml萃取池中再加入适量硅藻土，轻轻振摇使之与池口在同一水平线上，拧紧萃取池上盖。萃取结束后，把萃取液转移于25ml容量瓶中，用重量比为0.1:9.9的乙二胺和乙醇的混合物稀释至刻度，在15000r/min下离心5min，取上清液，进入LC测定。A method for the determination of chlorogenic acid in wild chrysanthemums: crush wild chrysanthemums through a pulverizer, pass through a No. 3 sieve, weigh about 0.25g, accurately weigh, mix with 0.5g diatomaceous earth, set aside, and transfer to a pre-set good filter membrane Add an appropriate amount of diatomaceous earth to the 5ml extraction pool, shake gently to make it on the same level as the pool mouth, and tighten the upper cover of the extraction pool. After the extraction, transfer the extract to a 25ml volumetric flask, dilute to the mark with a mixture of ethylenediamine and ethanol with a weight ratio of 0.1:9.9, centrifuge at 15000r/min for 5min, take the supernatant, and enter it into LC for determination.
萃取条件(见表1)为：The extraction conditions (see table 1) are:
表1Table 1
分析方法为LC液相色谱法，液相色谱分析条件：Analysis method is LC liquid chromatography, liquid chromatography analysis conditions:
a)仪器：Thermo双三元液相(U-3000)a) Instrument: Thermo double ternary liquid phase (U-3000)
b)色谱柱ThermoSyncronisC183μm3×100mm，即碳18色谱柱，直径3mm，长度100mm，粒径3μm)b) Chromatographic column ThermoSyncronisC183μm3×100mm, that is, a carbon-18 chromatographic column with a diameter of 3mm, a length of 100mm, and a particle size of 3μm)
c)柱温：40℃c) Column temperature: 40°C
d)流速：0.5mL/mind) Flow rate: 0.5mL/min
e)流动相：乙腈-水，20℃条件下的体积比为4:6e) Mobile phase: acetonitrile-water, the volume ratio at 20°C is 4:6
f)检测波长：326nmf) Detection wavelength: 326nm
取3个批号的野菊花进行测试，批号分别为1510020、151001、20150701，采用实施例1的萃取和检测方法进行测试，同时对相同批号的产品按照背景技术记载的回流提取和高效液相色谱法测试。野菊花中的绿原酸的测试结果如下表1：The wild chrysanthemums of 3 batch numbers are taken for testing, the batch numbers are respectively 1510020, 151001, and 20150701, and the extraction and detection methods of Example 1 are used for testing, and the products of the same batch number are extracted according to the reflux extraction and high-performance liquid chromatography recorded in the background technology test. The test results of chlorogenic acid in wild chrysanthemum are shown in Table 1:
表1Table 1
以上所述的仅为本发明的较佳实施例，凡在本发明的精神和原则范围内所作的任何修改、等同替换和改进等，均应包含在本发明的保护范围之内。The above are only preferred embodiments of the present invention, and any modifications, equivalent replacements and improvements made within the spirit and scope of the present invention shall be included within the protection scope of the present invention.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106841478A (en) * | 2017-04-20 | 2017-06-13 | 广西壮族自治区梧州食品药品检验所 | It is a kind of at the same determine chrysanthemum indicum in seven kinds of methods of the content of composition |
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| CN102928523A (en) * | 2011-08-11 | 2013-02-13 | 上海雷允上科技发展有限公司 | Wild chrysanthemum flower fingerprint determination method, its application, and wild chrysanthemum flower quality detection method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106841478A (en) * | 2017-04-20 | 2017-06-13 | 广西壮族自治区梧州食品药品检验所 | It is a kind of at the same determine chrysanthemum indicum in seven kinds of methods of the content of composition |
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