CN102879500A - Method for detecting antioxidant ingredient in Lycium ruthenicum Murr fruit juice with DPPH-HPLC method - Google Patents
Method for detecting antioxidant ingredient in Lycium ruthenicum Murr fruit juice with DPPH-HPLC method Download PDFInfo
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- CN102879500A CN102879500A CN2012103818412A CN201210381841A CN102879500A CN 102879500 A CN102879500 A CN 102879500A CN 2012103818412 A CN2012103818412 A CN 2012103818412A CN 201210381841 A CN201210381841 A CN 201210381841A CN 102879500 A CN102879500 A CN 102879500A
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Abstract
The invention relates to a method for detecting an antioxidant ingredient in Lycium ruthenicum Murr fruit juice with a DPPH-HPLC method. The method comprises the steps of reacting an antioxidant 1,1-dibenzyl-2-trinitrophenylhydrazine and the Lycium ruthenicum Murr fruit juice in a dark place to obtained reacted Lycium ruthenicum Murr fruit juice; respectively injecting unreacted Lycium ruthenicum Murr fruit juice and the reacted Lycium ruthenicum Murr fruit juice in a high performance liquid chromatography (HLPC) for detection to obtain a chromatogram of the unreacted Lycium ruthenicum Murr fruit juice and a chromatogram of the reacted Lycium ruthenicum Murr fruit juice; and finally comparing peak areas of corresponding peaks in the chromatogram of the unreacted Lycium ruthenicum Murr fruit juice and the chromatogram of the reacted Lycium ruthenicum Murr fruit juice, and if the peal areas of the reacted Lycium ruthenicum Murr fruit juice are smaller than those of and the unreacted Lycium ruthenicum Murr fruit juice, the result indicates that the Lycium ruthenicum Murr fruit juice contains the antioxidant ingredient. The method has the characteristics of little reagent, simple and convenient operation, high sensitivity, high precision, good stability and the like, and can realize effectiveness evaluation for the antioxidant ingredient in the Lycium ruthenicum Murr fruit juice.
Description
Technical field
The present invention relates to a kind of detection method of fruit juice antioxidant content, relate in particular to the method that a kind of DPPH-HPLC method detects antioxidant content in the black fruit lyceum juice.
Background technology
Black fruit lyceum is with the fruit of Solanaceae Lycium plant black fruit lyceum (Lycium ruthenicum Murr.), is aubergine, and is nontoxic, has stomach invigorating tonifying spleen, nourishing Yin and moistening lung, the effect such as help digestion of invigorating blood circulation.That modern scientific research proof lycium ruthenicum fruit has is anti-oxidant, antifatigue, anti-anoxic, reducing blood lipid, prevent and treat the effect such as artery sclerosis, is widely used in medicine, food and cosmetics.
Mainly contain the antioxidant contents such as anthocyanin class material, total polyphenols in the black fruit lyceum juice.But present anti-oxidant experimental implementation complex steps, precision are poor, can not belong to oxidation resistant composition.The black fruit lyceum juice antioxidant content does not also have complete detection method, is unfavorable for guaranteeing the quality of black fruit lyceum juice.
Summary of the invention
Technical matters to be solved by this invention provide a kind of simply, the DPPH-HPLC method detects the method for antioxidant content in the black fruit lyceum juice fast and accurately.
For addressing the above problem, a kind of DPPH-HPLC method of the present invention detects the method for antioxidant content in the black fruit lyceum juice, it is characterized in that: the method at first is 0.01 ~ 5% antioxidant 1 with mass concentration, 1-diphenyl-2 trinitrophenyl-hydrazine (DPPH) and black fruit lyceum juice are that lucifuge is reacted 15min ~ 30min under 25 ~ 30 ℃ the condition in temperature by the volume ratio of 100:1 ~ 250:1, obtain reacting rear black fruit lyceum juice; Then respectively black fruit lyceum juice after unreacted black fruit lyceum juice and the described reaction is injected high performance liquid chromatography (HPLC) and detect, obtain respectively the chromatogram of unreacted black fruit lyceum juice and the chromatogram of the rear black fruit lyceum juice of reaction; At last, the peak area at corresponding each peak in the chromatogram of black fruit lyceum juice after the chromatogram of more described unreacted black fruit lyceum juice and the described reaction; If the peak that peak area reduces namely illustrates to have antioxidant content in the black fruit lyceum juice.
Described black fruit lyceum juice is the fruit juice behind original fruit juice or the 5000 dalton's organic membrane filters.
Described high performance liquid chromatography (HPLC) detection method refers to that mobile phase A is methyl alcohol, and Mobile phase B is pure water; Flow velocity is 0.8 ~ 1.5mL/min, and detecting wavelength is 210 nm ~ 390 nm; The mobile phase condition is: 0 min ~ 15min, 5%A ~ 5%A, 95%B ~ 95%B; 15 min ~ 25 min, 5 %A ~ 15 %A, 95%B ~ 85%B; 25 min ~ 40 min, 15% A ~ 25% A, 85%B ~ 75%B.
The present invention compared with prior art has the following advantages:
1, because the present invention adopts the DPPH-HPLC method to measure antioxidant content in the black fruit lyceum juice, and this method has characteristics such as the agents useful for same amount is few, simple to operation, highly sensitive, the precision high stability is good, therefore, the suitable black fruit lyceum juice antioxidant content of measuring, thus the efficiency evaluation of health care is carried out in realization to black fruit lyceum juice.
2, because the present invention adopts the DPPH-HPLC method to measure antioxidant content in the black fruit lyceum juice, and this method only needs the peak area at more corresponding peak can judge antioxidant content.Be the chromatogram of black fruit lyceum juice such as Fig. 1, namely do not add the black fruit lyceum juice chromatogram of DPPH solution.Fig. 2 is the chromatogram of the black fruit lyceum juice behind the adding DPPH.Each chromatographic peak in the comparison colours spectrogram, wherein the peak area of chromatographic peak A, B, C, D, E, F reduces in the chromatogram, illustrates that compd A, B, C, D, E, F are antioxidant content.
3, the present invention is simple, quick, is easy to promote.
Description of drawings
Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail.
Fig. 1 is not for adding antioxidant DPPH chromatogram.
Fig. 2 is for adding antioxidant DPPH chromatogram.
Embodiment
Embodiment 1A kind of DPPH-HPLC method detects the method for antioxidant content in the black fruit lyceum juice, the method at first is 0.01% antioxidant 1 with mass concentration, 1-diphenyl-2 trinitrophenyl-hydrazine (DPPH) and black fruit lyceum original fruit juice are that lucifuge is reacted 15min under 30 ℃ the condition in temperature by the volume ratio (mL/mL) of 100:1, obtain reacting rear black fruit lyceum juice; Then respectively the rear black fruit lyceum juice of unreacted black fruit lyceum juice and reaction is injected high performance liquid chromatography (HPLC) and detect, obtain respectively the chromatogram of unreacted black fruit lyceum juice and the chromatogram of the rear black fruit lyceum juice of reaction; At last, the peak area at corresponding each peak in the chromatogram of black fruit lyceum juice after the chromatogram of unreacted black fruit lyceum juice and the reaction relatively; If the peak that peak area reduces namely illustrates to have antioxidant content in the black fruit lyceum juice.
Wherein: high performance liquid chromatography (HPLC) detection method refers to that mobile phase A is methyl alcohol, and Mobile phase B is pure water; Flow velocity is 0.8mL/min, and detecting wavelength is 210 nm nm; The mobile phase condition is: 0 min ~ 15min, 5%A ~ 5%A, 95%B ~ 95%B; 15 min ~ 25 min, 5 %A ~ 15 %A, 95%B ~ 85%B; 25 min ~ 40 min, 15% A ~ 25% A, 85%B ~ 75%B.
Embodiment 2A kind of DPPH-HPLC method detects the method for antioxidant content in the black fruit lyceum juice, the method at first is 5% antioxidant 1 with mass concentration, black fruit lyceum juice behind 1-diphenyl-2 trinitrophenyl-hydrazine (DPPH) and the 5000 dalton's organic membrane filters is that lucifuge is reacted 30min under 25 ℃ the condition in temperature by the volume ratio (mL/mL) of 250:1, obtains reacting rear black fruit lyceum juice; Then respectively the rear black fruit lyceum juice of unreacted black fruit lyceum juice and reaction is injected high performance liquid chromatography (HPLC) and detect, obtain respectively the chromatogram of unreacted black fruit lyceum juice and the chromatogram of the rear black fruit lyceum juice of reaction; At last, the peak area at corresponding each peak in the chromatogram of black fruit lyceum juice after the chromatogram of unreacted black fruit lyceum juice and the reaction relatively; If the peak that peak area reduces namely illustrates to have antioxidant content in the black fruit lyceum juice.
Wherein: high performance liquid chromatography (HPLC) detection method refers to that mobile phase A is methyl alcohol, and Mobile phase B is pure water; Flow velocity is 0.8mL/min, and detecting wavelength is 360 nm; The mobile phase condition is: 0 min ~ 15min, 5%A ~ 5%A, 95%B ~ 95%B; 15 min ~ 25 min, 5 %A ~ 15 %A, 95%B ~ 85%B; 25 min ~ 40 min, 15% A ~ 25% A, 85%B ~ 75%B.
Embodiment 3A kind of DPPH-HPLC method detects the method for antioxidant content in the black fruit lyceum juice, the method at first is 0.1% antioxidant 1 with mass concentration, 1-diphenyl-2 trinitrophenyl-hydrazine (DPPH) and black fruit lyceum original fruit juice are that lucifuge is reacted 20min under 25 ℃ the condition in temperature by the volume ratio (mL/mL) of 150:1, obtain reacting rear black fruit lyceum juice; Then respectively the rear black fruit lyceum juice of unreacted black fruit lyceum juice and reaction is injected high performance liquid chromatography (HPLC) and detect, obtain respectively the chromatogram of unreacted black fruit lyceum juice and the chromatogram of the rear black fruit lyceum juice of reaction; At last, the peak area at corresponding each peak in the chromatogram of black fruit lyceum juice after the chromatogram of unreacted black fruit lyceum juice and the reaction relatively; If the peak that peak area reduces namely illustrates to have antioxidant content in the black fruit lyceum juice.
Wherein: high performance liquid chromatography (HPLC) detection method refers to that mobile phase A is methyl alcohol, and Mobile phase B is pure water; Flow velocity is 1.2mL/min, and detecting wavelength is 254 nm; The mobile phase condition is: 0 min ~ 15min, 5%A ~ 5%A, 95%B ~ 95%B; 15 min ~ 25 min, 5 %A ~ 15 %A, 95%B ~ 85%B; 25 min ~ 40 min, 15% A ~ 25% A, 85%B ~ 75%B.
Embodiment 4A kind of DPPH-HPLC method detects the method for antioxidant content in the black fruit lyceum juice, the method at first is 1% antioxidant 1 with mass concentration, black fruit lyceum juice behind 1-diphenyl-2 trinitrophenyl-hydrazine (DPPH) and the 5000 dalton's organic membrane filters is that lucifuge is reacted 25min under 32 ℃ the condition in temperature by the volume ratio (mL/mL) of 180:1, obtains reacting rear black fruit lyceum juice; Then respectively the rear black fruit lyceum juice of unreacted black fruit lyceum juice and reaction is injected high performance liquid chromatography (HPLC) and detect, obtain respectively the chromatogram of unreacted black fruit lyceum juice and the chromatogram of the rear black fruit lyceum juice of reaction; At last, the peak area at corresponding each peak in the chromatogram of black fruit lyceum juice after the chromatogram of unreacted black fruit lyceum juice and the reaction relatively; If the peak that peak area reduces namely illustrates to have antioxidant content in the black fruit lyceum juice.
Wherein: high performance liquid chromatography (HPLC) detection method refers to that mobile phase A is methyl alcohol, and Mobile phase B is pure water; Flow velocity is 1mL/min, and detecting wavelength is 390 nm; The mobile phase condition is: 0 min ~ 15min, 5%A ~ 5%A, 95%B ~ 95%B; 15 min ~ 25 min, 5 %A ~ 15 %A, 95%B ~ 85%B; 25 min ~ 40 min, 15% A ~ 25% A, 85%B ~ 75%B.
Embodiment 5A kind of DPPH-HPLC method detects the method for antioxidant content in the black fruit lyceum juice, the method at first is 2% antioxidant 1 with mass concentration, black fruit lyceum juice behind 1-diphenyl-2 trinitrophenyl-hydrazine (DPPH) and the 5000 dalton's organic membrane filters is that lucifuge is reacted 24min under 35 ℃ the condition in temperature by the volume ratio (mL/mL) of 200:1, obtains reacting rear black fruit lyceum juice; Then respectively the rear black fruit lyceum juice of unreacted black fruit lyceum juice and reaction is injected high performance liquid chromatography (HPLC) and detect, obtain respectively the chromatogram of unreacted black fruit lyceum juice and the chromatogram of the rear black fruit lyceum juice of reaction; At last, the peak area at corresponding each peak in the chromatogram of black fruit lyceum juice after the chromatogram of unreacted black fruit lyceum juice and the reaction relatively; If the peak that peak area reduces namely illustrates to have antioxidant content in the black fruit lyceum juice.
Wherein: high performance liquid chromatography (HPLC) detection method refers to that mobile phase A is methyl alcohol, and Mobile phase B is pure water; Flow velocity is 1.5mL/min, and detecting wavelength is 290 nm; The mobile phase condition is: 0 min ~ 15min, 5%A ~ 5%A, 95%B ~ 95%B; 15 min ~ 25 min, 5 %A ~ 15 %A, 95%B ~ 85%B; 25 min ~ 40 min, 15% A ~ 25% A, 85%B ~ 75%B.
Claims (3)
1. a DPPH-HPLC method detects the method for antioxidant content in the black fruit lyceum juice, it is characterized in that: the method at first is 0.01 ~ 5% antioxidant 1 with mass concentration, 1-diphenyl-2 trinitrophenyl-hydrazine and black fruit lyceum juice are lucifuge reaction 15min ~ 30min under 25 ~ 30 ℃ the condition in temperature by the volume ratio of 100:1 ~ 250:1, obtain reacting rear black fruit lyceum juice; Then respectively black fruit lyceum juice after unreacted black fruit lyceum juice and the described reaction is injected high performance liquid chromatography and detect, obtain respectively the chromatogram of unreacted black fruit lyceum juice and the chromatogram of the rear black fruit lyceum juice of reaction; At last, the peak area at corresponding each peak in the chromatogram of black fruit lyceum juice after the chromatogram of more described unreacted black fruit lyceum juice and the described reaction; If the peak that peak area reduces namely illustrates to have antioxidant content in the black fruit lyceum juice.
2. a kind of DPPH-HPLC method as claimed in claim 1 detects the method for antioxidant content in the black fruit lyceum juice, and it is characterized in that: described black fruit lyceum juice is the fruit juice behind original fruit juice or the 5000 dalton's organic membrane filters.
3. a kind of DPPH-HPLC method as claimed in claim 1 detects the method for antioxidant content in the black fruit lyceum juice, and it is characterized in that: described high-efficiency liquid chromatography method for detecting refers to that mobile phase A is methyl alcohol, and Mobile phase B is pure water; Flow velocity is 0.8 ~ 1.5mL/min, and detecting wavelength is 210 nm ~ 390 nm; The mobile phase condition is: 0 min ~ 15min, 5%A ~ 5%A, 95%B ~ 95%B; 15 min ~ 25 min, 5 %A ~ 15 %A, 95%B ~ 85%B; 25 min ~ 40 min, 15% A ~ 25% A, 85%B ~ 75%B.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105758977A (en) * | 2016-03-09 | 2016-07-13 | 西华大学 | Method for detecting quality of Lycium ruthenicum Murr fruit or Lycium ruthenicum Murr fruit extracts |
| CN108303485A (en) * | 2018-01-30 | 2018-07-20 | 中国中医科学院中药研究所 | A kind of method of winter pear tea quality conformance assessment |
| CN109444281A (en) * | 2018-11-28 | 2019-03-08 | 南京林业大学 | A kind of method of antioxidant content in detection Qingqian Willow leaf |
| CN110108808A (en) * | 2019-04-12 | 2019-08-09 | 广西壮族自治区兽医研究所 | Method that is a kind of while evaluating oxidation-resistant active ingredient and content in downy rosemyrtle root |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101858859A (en) * | 2010-05-20 | 2010-10-13 | 中国科学院西北高原生物研究所 | Quality control method of lycium ruthenicum pigment |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101858859A (en) * | 2010-05-20 | 2010-10-13 | 中国科学院西北高原生物研究所 | Quality control method of lycium ruthenicum pigment |
Non-Patent Citations (4)
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| JIE ZHENG等: "Anthocyanins composition and antioxidant activity of wild Lycium ruthenicum Murr. from Qinghai-Tibet Plateau", 《FOOD CHEMISTRY》 * |
| 张强: "植物抗氧化活性成分检测新方法及杜仲抗氧化活性研究", 《中国博士学位论文全文数据库农业科技辑》 * |
| 白红进等: "黑果枸杞色素的提取及其清除DPPH自由基作用的研究", 《西北农业学报》 * |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105758977A (en) * | 2016-03-09 | 2016-07-13 | 西华大学 | Method for detecting quality of Lycium ruthenicum Murr fruit or Lycium ruthenicum Murr fruit extracts |
| CN108303485A (en) * | 2018-01-30 | 2018-07-20 | 中国中医科学院中药研究所 | A kind of method of winter pear tea quality conformance assessment |
| CN109444281A (en) * | 2018-11-28 | 2019-03-08 | 南京林业大学 | A kind of method of antioxidant content in detection Qingqian Willow leaf |
| CN109444281B (en) * | 2018-11-28 | 2022-04-26 | 南京林业大学 | Method for detecting antioxidant components in cyclocarya paliurus leaves |
| CN110108808A (en) * | 2019-04-12 | 2019-08-09 | 广西壮族自治区兽医研究所 | Method that is a kind of while evaluating oxidation-resistant active ingredient and content in downy rosemyrtle root |
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Application publication date: 20130116 |