CN105911173A - Determination method for chlorogenic acid in guiling jelly - Google Patents
Determination method for chlorogenic acid in guiling jelly Download PDFInfo
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- CN105911173A CN105911173A CN201610232065.8A CN201610232065A CN105911173A CN 105911173 A CN105911173 A CN 105911173A CN 201610232065 A CN201610232065 A CN 201610232065A CN 105911173 A CN105911173 A CN 105911173A
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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
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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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- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a determination method for chlorogenic acid in guiling jelly. The method sequentially comprises the steps of: 1) preparation of a reference solution; 2) drawing of a calibration curve; and 3) preparation of a sample solution: taking a paste and stirring the paste into a slurry, accurately weighing 20g of the slurry, adding methanol and ethanol in a volume ratio of 1:1 to a constant volume in a 100mL volumetric flask, performing extraction at 100DEG C for 2h, conducting cold discharge, supplementing methanol and ethanol in a volume ratio of 1:1 to a scale, shaking the mixture evenly, performing standing, and conducting filtering with a 0.45 micrometer filter membrane so as to obtain the sample solution, injecting the sample solution into a high performance liquid chromatograph, determining the peak area, and calculating the chlorogenic acid content of the tested solution by means of the calibration curve.
Description
Technical field
The present invention relates to the assay method of a kind of chlorogenic acid, specifically, be that a kind of mensuration GUILING GAO medium green is former
The method of acid, belongs to technical field of chemical detection.
Background technology
GUILING GAO is the Traditional health care food in the south of the Five Ridges, with heat-clearing and toxic substances removing, skin maintenance and famous, dry over 100 years
Pole is liked by Guangdong and Guangxi Provinces, Taiwan, Hong Kong and Macau and the Southeast Asia people.Sweep the country the most especially.Wuzhou, Guangxi is Testudinis
The authentic place of production of Siberian cocklebur cream, to national applications geography symbol product.Chlorogenic acid is the main Chinese medicinal materials in GUILING GAO
One of the characteristic component of Flos Lonicerae.Chlorogenic acid chemical name: 3 one caffeoyl guinic acids.Chlorogenic acid have antibacterial,
Antiinflammatory, removing toxic substances, function of gallbladder promoting, blood pressure lowering and leukocyte increasing and dramatically increase gastrointestinal peristalsis and promote gastric secretion etc.
Pharmacological action, has stronger inhibitory action to escherichia coli, gold-coloured staphylococci, streptococcus pneumoniae and virus；
Acute laryngopharyngitis and dermatosis there is obvious curative effects.The primary bioactivity of chlorogenic acid has (1) to hyaluronic acid
Enzyme and the inhibitory action of glucose 1 phosphatase；(2) removing and the lipoid peroxidization resistant to free radical；
(3) antimutagenesis；(4) hepatic cholagogic effect；(5) effect such as antibacterial, antiviral and spasmolytic.
Therefore, measure the content of GUILING GAO Content of Chlorogenic Acid, the product quality to monitoring GUILING GAO, there is reality
Meaning.
Summary of the invention
For the problems referred to above, it is simple that the purpose that the present invention provides is to provide a kind of detection method, Detection accuracy
Height, the method for the mensuration GUILING GAO Content of Chlorogenic Acid that test result is reproducible.
For solving above-mentioned technical problem, the technical scheme that the present invention provides is such that
A kind of method measuring GUILING GAO Content of Chlorogenic Acid, comprises the steps: successively
1) preparation of reference substance solution
Accurately weigh chlorogenic acid reference substance, be placed in brown measuring bottle, add Methanamide and make solution, to obtain final product；
2) drafting of calibration curve
Take chlorogenic acid formamide solution that concentration is 20ug/mL 5.0,10.0,15.0,20.0,25.0uL, note
Entering high performance liquid chromatograph, measure its peak area, with peak area as vertical coordinate, concentration is that abscissa draws school
Directrix curve, obtains regression equation；
3) preparation of sample solution
Take mastic stirring pulping, accurately weigh 20g, add the methanol of volume ratio 1:1 and ethanol constant volume in 100mL
In volumetric flask, 100 degrees Celsius are extracted 2 hours, let cool, and mend the methanol of volume ratio 1:1 and ethanol to scale,
Shaking up, place, through 0.45um membrane filtration, obtain sample solution, note sample enters high performance liquid chromatograph, measures
Its peak area, is calculated detected solution Content of Chlorogenic Acid by calibration curve.
Further, the method for above-mentioned mensuration GUILING GAO Content of Chlorogenic Acid, described high performance liquid chromatography
Parameter is as follows: described chromatographic condition is:
Liquid phase instrument: Agilent 1290；
Chromatographic column: Thermo syncronls aQ Dim.mm50 × 2.1；
Flowing phase: acetonitrile-1% phosphoric acid solution, volume ratio: 9.9:90.1；
Flow velocity: 0.3ml/min；
Wavelength: 327nm；
Column temperature: 40 DEG C；
Retention time: 13.5min.
Compared with prior art, the detection method that the present invention provides is simple, and Detection accuracy is high, test result
Reproducible, under conditions of sharp separation, meet the theoretical cam curve needed for standard.
Accompanying drawing explanation
Fig. 1 is the ortho acid reference colour spectrogram that the present invention provides.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the claim of the present invention is described in further detail, but not
Constitute any limitation of the invention, any limited number of time done in the claims in the present invention protection domain
Amendment, still within the claims of the present invention.
Embodiment 1
1) preparation of reference substance solution
Accurately weigh chlorogenic acid reference substance, be placed in brown measuring bottle, add Methanamide and make solution, to obtain final product；
2) drafting of calibration curve
Take chlorogenic acid formamide solution that concentration is 20ug/mL 5.0,10.0,15.0,20.0,25.0uL, note
Entering high performance liquid chromatograph, measure its peak area, with peak area as vertical coordinate, concentration is that abscissa draws school
Directrix curve, obtains regression equation, obtains regression equation y=O.0378x+0.000, linearly dependent coefficient 0.9999.
3) preparation of sample solution
Take mastic stirring pulping, accurately weigh 20g, add the methanol of volume ratio 1:1 and ethanol constant volume in 100mL
In volumetric flask, 100 degrees Celsius are extracted 2 hours, let cool, and mend the methanol of volume ratio 1:1 and ethanol to scale,
Shaking up, place, through 0.45um membrane filtration, obtain sample solution, note sample enters high performance liquid chromatograph, measures
Its peak area, is calculated detected solution Content of Chlorogenic Acid by calibration curve.
The parameter of described high performance liquid chromatography is as follows: liquid phase instrument: Agilent 1290；
Chromatographic column: Thermo syncronls aQ Dim.mm50 × 2.1；
Flowing phase: acetonitrile-1% phosphoric acid solution, volume ratio: 9.7:90.3；
Flow velocity: 0.3ml/min；
Wavelength: 327nm；
Column temperature: 40 DEG C；
Randomly draw 6 parts of GUILING GAO sample, make style solution, take 20u l and inject high performance liquid chromatograph,
Measure its peak area, calibration curve calculate detected solution Content of Chlorogenic Acid.
| GUILING GAO sample | Sample 1 | Sample 2 | Sample 3 | Sample 4 | Sample 5 | Sample 6 |
| Testing result mg. (100g-1) | 0.56 | 0.72 | 1.08 | 0.86 | 0.78 | 0.98 |
According to calibrating curve determining, the chlorogenic acid content of this experimental technique has well between 0.05-0.34ug
Linear relationship, detection line is 0.1mg.kg-1, average RSD% is 0.35%.
Claims (2)
1. the method measuring GUILING GAO Content of Chlorogenic Acid, it is characterised in that comprise the steps: successively
1) preparation of reference substance solution
Accurately weigh chlorogenic acid reference substance, be placed in brown measuring bottle, add Methanamide and make solution, to obtain final product；
2) drafting of calibration curve
Take chlorogenic acid formamide solution that concentration is 20ug/mL 5.0,10.0,15.0,20.0,25.0uL, note
Entering high performance liquid chromatograph, measure its peak area, with peak area as vertical coordinate, concentration is that abscissa draws school
Directrix curve, obtains regression equation；
3) preparation of sample solution
Take mastic stirring pulping, accurately weigh 20g, add the methanol of volume ratio 1:1 and ethanol constant volume in 100mL
In volumetric flask, 100 degrees Celsius are extracted 2 hours, let cool, and mend the methanol of volume ratio 1:1 and ethanol to scale,
Shaking up, place, through 0.45um membrane filtration, obtain sample solution, note sample enters high performance liquid chromatograph, measures
Its peak area, is calculated detected solution Content of Chlorogenic Acid by calibration curve.
The method of mensuration GUILING GAO Content of Chlorogenic Acid the most according to claim 1, it is characterised in that described
The parameter of high performance liquid chromatography as follows: described chromatographic condition is:
Liquid phase instrument: Agilent 1290；
Chromatographic column: Thermo syncronls aQ Dim.mm50 × 2.1；
Flowing phase: acetonitrile-1% phosphoric acid solution, volume ratio: 9.7:90.3；
Flow velocity: 0.3ml/min；
Wavelength: 327nm；
Column temperature: 40 DEG C；
Retention time: 13.5min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201610232065.8A CN105911173A (en) | 2016-04-14 | 2016-04-14 | Determination method for chlorogenic acid in guiling jelly |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610232065.8A CN105911173A (en) | 2016-04-14 | 2016-04-14 | Determination method for chlorogenic acid in guiling jelly |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102706980A (en) * | 2012-06-06 | 2012-10-03 | 福建省农业科学院农业工程技术研究所 | Method utilizing HPLC to test seventeen phenol substances contained in grapes and oranges |
| WO2015168964A1 (en) * | 2014-05-09 | 2015-11-12 | 四川九章生物化工科技发展有限公司 | Chlorogenic acid raw material or active pharmaceutical ingredient and preparation method therefor, and quality testing method |
-
2016
- 2016-04-14 CN CN201610232065.8A patent/CN105911173A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102706980A (en) * | 2012-06-06 | 2012-10-03 | 福建省农业科学院农业工程技术研究所 | Method utilizing HPLC to test seventeen phenol substances contained in grapes and oranges |
| WO2015168964A1 (en) * | 2014-05-09 | 2015-11-12 | 四川九章生物化工科技发展有限公司 | Chlorogenic acid raw material or active pharmaceutical ingredient and preparation method therefor, and quality testing method |
Non-Patent Citations (3)
| Title |
|---|
| 吴婕: "高效液相色谱法测定龟苓膏中的绿原酸", 《光谱实验室》 * |
| 彭翠芳等: "龟苓膏中绿原酸的测定方法、稳定性的研究", 《中国新技术新产品》 * |
| 李亚等: "龟苓膏中绿原酸含量测定及衰减跟踪监测", 《现代食品科技》 * |
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Application publication date: 20160831 |