CN103558164A - Method for simultaneously determining proanthocyanidins and resveratrol in wine - Google Patents
Method for simultaneously determining proanthocyanidins and resveratrol in wine Download PDFInfo
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- CN103558164A CN103558164A CN201310387469.0A CN201310387469A CN103558164A CN 103558164 A CN103558164 A CN 103558164A CN 201310387469 A CN201310387469 A CN 201310387469A CN 103558164 A CN103558164 A CN 103558164A
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Abstract
The invention relates to the food examination and analysis fields, and concretely relates to a method for simultaneously determining proanthocyanidins and resveratrol in a wine. The invention aims to directly determine proanthocyanidins and resveratrol in the wine by adopting a high performance liquid chromatography tandem diode-array detector and a fluorescence detector. The fluorescence detector is characterized in that the excitation wavelength and the emission wavelength can be adjusted to reduce the interferences of impurities, so the fluorescence detector has a high selectivity, has a far higher sensitivity than an ultraviolet detector, can accurately and reliably quantitatively analyze substances, and can accurately and reliably detect and analyze the two efficacy components comprising proanthocyanidins and resveratrol in the wine.
Description
Technical field
The present invention relates to Food Inspection and analysis field, be specially a kind of method of measuring procyanidin and resveratrol in grape wine.
Background technology
Procyanidin (procyanidins, referred to as PC) is the general name of flavan-3-alcohol derivant, and the monomer, dimer, tripolymer to ten aggressiveness that mainly contain catechin combine.Its feature is efficient, low toxicity, high bioavailability, and half lethal dose is LD50=3g/kg, is the continuous a kind of extremely strong activity in vivo function factor of research and development in recent years.Much research shows, PC removes the very strong antioxidant of free radical, and it is anti-oxidant, and the ability of removing free radical is 50 times of VE, and 20 times of Vc have caused people's very big concern.It is not only distributed widely in plant, as ginkgo, Japanese Thuja, North America arborvitae, Turkey arbor-vitae, flower chess pine, boorishly life thorn of white birch certain herbaceous plants with big flowers, Japanese thick grass, ear leaf cassia angustifolia, maritime pine, rheum officinale, larch, lotus pod etc., also be present in some beverage, vegetables, fruit and grain, as grape, grape wine, apple, cider, beer, tealeaves, barley, Fructus Hordei Germinatus, jowar, legume, manaca, weeds poison, cherry etc.In the numerous natural product relating to, the PC rich content in grape and goods thereof, therefore study more.The method for quantitatively determining of this material comprises titrimetry, optical spectroscopy, electrochemical process, chemoluminescence method, Capillary Electrophoresis and the HPLC method on molysite acidified degradation basis at present.Resveratrol (resveratrol) also claim resvertrol, is a Plant Hormone, it should be noted that it can be in brewing grape process, due to the catalytic reaction generation of biochemical enzyme.The content of high-quality wine resveratrol is higher, and blended liquor (grape juice adds alcohol) Resveratrol content is zero.At present, research shows that it has antitumor, anti-cardiovascular disease, treatment sudden change, anti-oxidant, antimicrobial antiphlogistic, protects the liver, the bio-pharmacology such as cell death inducing and estrogen adjusting is active.The assay method of current this material has high performance liquid chromatography, fluorescence method, chemoluminescence method, capillary electrophoresis, electrochemical process.
Application number is that 201210184125.5 Chinese patent had once been announced a kind of method that HPLC of utilization measures 17 kinds of aldehydes matters in grape and citrusfruit, adopt high performance liquid chromatography to the gallic acid in grape and citrusfruit, chlorogenic acid, caffeic acid, syringic acid, coumaric acid, rutin, forulic acid, aurantiamarin, aurantiin, salicylic acid, Quercetin, Nobiletin, hesperetin, catechin, epicatechin, benzoic acid, resveratrol is measured, by UV-detector qualitative and quantitative analysis, but employing UV-detector, need carry out to sample the extraction and purification program of series of complex, impurity disturbs more, and owing to having adopted loaded down with trivial details extraction pretreatment technology, sample is lost, and measuring accuracy is not high.
Summary of the invention
For addressing the above problem, the present invention is intended to adopt high performance liquid chromatography series connection secondary array tube detecting device and fluorescence detector, directly measures procyanidin and resveratrol in grape wine.Wherein fluorescence detector can reduce the interference of impurity by the adjustment of excitation wavelength and emission wavelength, thereby has high selectivity, and its sensitivity is far above UV-detector, can be accurately, reliably material is carried out to quantitative test.
Concrete technical scheme is as follows: a kind of method of measuring procyanidin and resveratrol in grape wine, comprises the following steps:
(1) sample preparation
Get the organic filter membrane standby of the appropriate mistake of grape wine and be used as testing sample;
(2) mensuration of high performance liquid chromatograph system
Getting appropriate testing sample adds in high performance liquid chromatograph, utilize mobile phase to carry out gradient elution to it, then utilize diode array detector to carry out qualitative analysis to the procyanidin in sample and resveratrol, the principle of work of diode array detector is to enter injection device after polychromatic light is absorbed by compositional selecting by sample cell again, be radiated at and on diode array device, make the light intensity of each nano wave length be transformed into corresponding electrical signal intensity, obtain the absorption spectrum of component, thereby obtain the structural information of specific components, contribute to the structure of unknown component or complex component to determine, therefore, can be according to the chromatographic retention (R of object
t) with standard substance retention time tentatively more qualitative to compound, utilize the absorption spectrum of object and standard substance more qualitative simultaneously, dual qualitative more accurately and reliably, finally recycle fluorescence detector the procyanidin in sample and resveratrol are carried out to quantitative test, fluorescence detector is to utilize the compound with special construction to receive ultraviolet excitation (E
ex) after can launch the light (E longer than excitation source wavelength
em) fluorescent characteristic detect, many organic compounds have natural fluoresence activity, wherein with the compound of aromatic group, there is fluorescence activity very strong, at certain condition, fluorescence intensity is directly proportional to material concentration, the concentration that can determine material by glimmering light intensity size, it is particularly suitable for condensed-nuclei aromatics, steride etc., and procyanidin and resveratrol all have condensed-nuclei aromatics structure.Fluorescence detector is highly sensitive, selectivity good, than UV-detector, exceeds 2-3 the order of magnitude.
(3) quantitative by typical curve
Take respectively appropriate procyanidin and resveratrol standard items, utilize methyl alcohol to dissolve and constant volume, configure successively the mixed solution of two standard items, wherein, procyanidin concentration is followed successively by 0.0100, 0.0500, 0.100, 0.150, 0.200, 0.250mg/ml, the concentration of resveratrol is followed successively by 1.00, 5.00, 10.0, 15.0, 20.0, 25.0, 30.0 μ g/ml, mixed solution repeating step (2) is measured, the data obtained be take peak area as ordinate, concentration is horizontal ordinate, obtain peak area-concentration standard curve, curve is determined the content of procyanidin and resveratrol in sample thus.
During high performance liquid chromatograph system measurement, liquid phase chromatogram condition is: chromatographic column is ultimate XB-C18 chromatographic column, and its specification is 150mm * 4.6mm, i.d. 5 μ m; Mobile phase A is 0.1% formic acid, and Mobile phase B is acetonitrile, and gradient elution becomes program: 0min-10min, 10%B, 10min-11min, 10%B → 25%B, 11min-23min, 25%B, 23min-24min, 25%B → 10%B, 24min-30min, 10%B; Flow velocity is 1ml/min, and column temperature is 30 ℃, and sample size is 20 μ l; The scanning wavelength of diode array detector is 210nm-600nm; Fluorescence detector condition is: excitation wavelength during 0min-11min and emission wavelength are respectively 280nm, 305nm, and the excitation wavelength of 11min-30min and emission wavelength are respectively 320nm, 370nm.
By technique scheme, assay method provided by the present invention has high selectivity, high sensitivity, can accurately and reliably detect procyanidin and these two kinds of functional components of resveratrol of analyzing in grape wine.
Accompanying drawing explanation
Fig. 1 is standard items diode array detector chromatogram;
Fig. 2 is sample diode array detector chromatogram;
Fig. 3 is standard items diode array detector spectrogram;
Fig. 4 is sample diode array detector spectrogram;
Fig. 5 is standard items fluorescence detector chromatogram;
Fig. 6 is fluorescent detecting device chromatogram;
Fig. 7 is procyanidin typical curve;
Fig. 8 is resveratrol typical curve.
Embodiment
One, mensuration institute's employing equipment and medicine are as follows: Waters 2695 type high performance liquid chromatograph systems, Waters 2996 type diode array detector, waters 2475 fluorescence detectors, Empower chromatographic work station and data handling system; Procyanidin (> 95%, the natural research and development of Tianjin spike company limited); Resveratrol (Nat'l Pharmaceutical & Biological Products Control Institute, lot number: 111535); One bottle of saint coeur board grape wine.
Two, determination step is as follows:
(1) sample preparation
After respectively getting commercially available saint coeur board grape wine 1ml and crossing the organic filter membrane of 0.22 μ m, standby organic filter membrane standby is used as testing sample;
(2) mensuration of high performance liquid chromatograph system
Get 1ml testing sample and add in high performance liquid chromatograph, utilize mobile phase to carry out gradient elution to it, liquid phase chromatogram condition is: chromatographic column is ultimate XB-C18 chromatographic column, and its specification is 150mm * 4.6mm, i.d. 5 μ m; Mobile phase A is 0.1% formic acid, and Mobile phase B is acetonitrile, and gradient elution becomes program: 0min-10min, 10%B, 10min-11 min, 10%B → 25%B, 11min-23min, 25%B, 23min-24min, 25%B → 10%B, 24min-30min, 10% B; Flow velocity is 1ml/min, and column temperature is 30 ℃, and sample size is 20 μ l.
Then utilize diode array detector to carry out qualitative analysis to the procyanidin in sample and resveratrol, when this is measured, the scanning wavelength 210nm-600nm of diode array detector; Finally recycle fluorescence detector the procyanidin in sample and resveratrol are carried out to quantitative test.When this is measured, the condition of fluorescence detector is: excitation wavelength during 0min-11min (Eex) and emission wavelength (Eem) are respectively 280nm, 305nm, and the excitation wavelength of 11min-30min (Eex) and emission wavelength (Eem) are respectively 320nm, 370nm.
(3) quantitative by typical curve
Precision takes procyanidin 0.0300g in 10ml volumetric flask, with methyl alcohol, dissolves and constant volume.Obtain 3.00mg/ml procyanidin standard solution.Precision takes 0.0150g resveratrol standard items in 10ml volumetric flask, with methyl alcohol, dissolves and constant volume, joins to obtain 1.50 mg/ml resveratrol standard reserving solutions, then obtains standard solution 0.300mg/ ml with 5 times of methyl alcohol dilutions.With methyl alcohol dilution, configure successively the mixed solution of two standard items.Wherein, procyanidin concentration is 0.0100,0.0500,0.100,0.150,0.200, the standard operation liquid of 0.250mg/ml, the concentration of resveratrol is 1.00,5.00,10.0,15.0,20.0,25.0,30.0 μ g/ml, mixed solution repeating step (2) is measured, the data obtained be take peak area as ordinate, concentration is horizontal ordinate, obtains peak area-concentration standard curve.
(4) data analysis
As Fig. 1 and Fig. 2 contrast known, on diode array detector chromatogram, the retention time of standard items procyanidin and resveratrol is respectively 8.489 minutes and 20.108 minutes, and the retention time of procyanidin and resveratrol is respectively 8.504 minutes and 20.504 minutes in sample, this and standard items approach; As shown in Figure 3 and Figure 4, on diode array detector spectrogram, the maximum absorption wavelength of standard items procyanidin and resveratrol is respectively 280nm and 306nm, and in sample, the maximum absorption wavelength of procyanidin and resveratrol is respectively 279nm and 306nm, this and standard items approach, therefore, by spectrum and chromatogram, carried out these two kinds of compositions of qualitative analysis sample.
As shown in Figure 5 and Figure 6, the retention time of standard items procyanidin and resveratrol is respectively 8.489 minutes and 20.108 minutes, and in sample, the retention time of procyanidin and resveratrol is respectively 8.517 minutes and 20.237 minutes, as shown in Figure 7 and Figure 8, gained peak area-concentration standard curve is respectively:
Procyanidin: Y=6.00*10
8x+3.54*10
5, coefficient R=0.9989;
Resveratrol: Y=3.00*10
9x+5.60*10
6, coefficient R=0.9991;
Recording thus the content of procyanidin and resveratrol in 1 ml saint coeur board grape wine is 1.326mg/ml and 8.585 ug/ml.
The method that this is measured to procyanidin and resveratrol is carried out methodological study, and acquired results is as following table:
As seen from the above table, it is highly sensitive with method of resveratrol that this measures procyanidin, and separating power is strong, the qualitative analysis is accurate, detectability is low, can detect rapidly the content of procyanidin and resveratrol in grape wine the while.
Claims (4)
1. a method of simultaneously measuring procyanidin and resveratrol in grape wine, is characterized in that, comprises the following steps:
(1) sample preparation
Get the organic filter membrane standby of the appropriate mistake of grape wine and be used as testing sample;
(2) mensuration of high performance liquid chromatograph system
Getting appropriate testing sample adds in high performance liquid chromatograph, utilize mobile phase to carry out gradient elution to it, then utilize diode array detector to carry out qualitative analysis to the procyanidin in sample and resveratrol, finally recycle fluorescence detector the procyanidin in sample and resveratrol are carried out to quantitative test;
(3) quantitative by typical curve
Take respectively appropriate procyanidin and resveratrol standard items, utilize methyl alcohol to dissolve and constant volume, configure successively the mixed solution of two standard items, wherein, procyanidin concentration is followed successively by 0.0100, 0.0500, 0.100, 0.150, 0.200, 0.250mg/ml, the concentration of resveratrol is followed successively by 1.00, 5.00, 10.0, 15.0, 20.0, 25.0, 30.0 μ g/ml, mixed solution repeating step (2) is measured, the data obtained be take peak area as ordinate, concentration is horizontal ordinate, obtain peak area-concentration standard curve, curve is determined the content of procyanidin and resveratrol in sample thus.
2. a kind of method of measuring procyanidin and resveratrol in grape wine as claimed in claim 1, it is characterized in that, during high performance liquid chromatograph system measurement, liquid phase chromatogram condition is: chromatographic column is ultimate XB-C18 chromatographic column, its specification is 150 mm * 4.6 mm, i.d. 5 μ m; Mobile phase A is 0.1% formic acid, and Mobile phase B is acetonitrile, and gradient elution becomes program: 0 min-10 min, 10% B, 10 min-11 min, 10% B → 25% B, 11 min-23 min, 25% B, 23 min-24 min, 25% B → 10% B, 24 min-30min, 10% B; Flow velocity is 1 ml/min, and column temperature is 30 ℃, and sample size is 20 μ l.
3. a kind of method of measuring procyanidin and resveratrol in grape wine as claimed in claim 1, is characterized in that, the scanning wavelength of diode array detector is 210 nm-600 nm.
4. a kind of method of measuring procyanidin and resveratrol in grape wine as claimed in claim 1, it is characterized in that, fluorescence detector condition is: excitation wavelength during 0 min-11min and emission wavelength are respectively 280nm, 305nm, and the excitation wavelength of 11min-30 min and emission wavelength are respectively 320 nm, 370 nm.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109682669A (en) * | 2018-12-24 | 2019-04-26 | 久为利华（天津）文化发展有限公司 | The detection method of anthocyanidin content in grape wine |
| CN111579507A (en) * | 2020-05-22 | 2020-08-25 | 国珍健康科技（北京）有限公司 | Sulfuric acid-vanillin method for detecting content of procyanidine |
| CN112033941A (en) * | 2020-06-30 | 2020-12-04 | 中南民族大学 | Method for rapidly detecting resveratrol in food |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS56133649A (en) * | 1980-03-24 | 1981-10-19 | Japan Spectroscopic Co | Method and device for measurig catechol amines using natural fluorescent method |
| WO2008031004A2 (en) * | 2006-09-07 | 2008-03-13 | The Government Of The United States Of America, As Represented By The Secretary Of The Navy | Binding of proanthocyanidins with bacteria and bacterial components |
| CN101750459A (en) * | 2010-01-19 | 2010-06-23 | 中南大学湘雅二医院 | Method for simultaneously determining tryptophan and tyrosine in blood serum by high performance liquid chromatography and fluorescence method |
| CN102211980A (en) * | 2011-04-15 | 2011-10-12 | 湖南三为生物科技有限公司 | Method for preparing resveratrol extract by using waste residue from grape winery as raw material |
| CN102680446A (en) * | 2012-05-25 | 2012-09-19 | 中国人民解放军第三军医大学 | Method for detecting location of resveratrol in target cells |
-
2013
- 2013-08-30 CN CN201310387469.0A patent/CN103558164B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS56133649A (en) * | 1980-03-24 | 1981-10-19 | Japan Spectroscopic Co | Method and device for measurig catechol amines using natural fluorescent method |
| WO2008031004A2 (en) * | 2006-09-07 | 2008-03-13 | The Government Of The United States Of America, As Represented By The Secretary Of The Navy | Binding of proanthocyanidins with bacteria and bacterial components |
| CN101750459A (en) * | 2010-01-19 | 2010-06-23 | 中南大学湘雅二医院 | Method for simultaneously determining tryptophan and tyrosine in blood serum by high performance liquid chromatography and fluorescence method |
| CN102211980A (en) * | 2011-04-15 | 2011-10-12 | 湖南三为生物科技有限公司 | Method for preparing resveratrol extract by using waste residue from grape winery as raw material |
| CN102680446A (en) * | 2012-05-25 | 2012-09-19 | 中国人民解放军第三军医大学 | Method for detecting location of resveratrol in target cells |
Non-Patent Citations (4)
| Title |
|---|
| 张亮亮等: "高效液相色谱-二极管阵列检测法分析原花青素降解产物及副产物", 《分析化学》, vol. 36, no. 9, 30 September 2008 (2008-09-30), pages 1281 - 1284 * |
| 张敏等: "同步荧光检测虎杖提取物中白藜芦醇含量的新方法", 《林产化学与工业》, vol. 25, no. 2, 30 June 2005 (2005-06-30), pages 63 - 66 * |
| 薛洁: "山葡萄酒中白藜芦醇含量的测定", 《酿酒科技》, no. 5, 31 December 2004 (2004-12-31), pages 103 - 104 * |
| 黄声岚等: "HPLC-PDA法测定虎杖中的白藜芦醇", 《海峡药学》, vol. 20, no. 10, 31 December 2008 (2008-12-31), pages 70 - 71 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109682669A (en) * | 2018-12-24 | 2019-04-26 | 久为利华（天津）文化发展有限公司 | The detection method of anthocyanidin content in grape wine |
| CN111579507A (en) * | 2020-05-22 | 2020-08-25 | 国珍健康科技（北京）有限公司 | Sulfuric acid-vanillin method for detecting content of procyanidine |
| CN112033941A (en) * | 2020-06-30 | 2020-12-04 | 中南民族大学 | Method for rapidly detecting resveratrol in food |
| CN112033941B (en) * | 2020-06-30 | 2021-03-30 | 中南民族大学 | Method for rapidly detecting resveratrol in food |
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