CN110954626A - Method for simultaneously evaluating oxidation resistance and content of gallic acid and ellagic acid in myrtle roots by using ABTS-UPLC method - Google Patents
Method for simultaneously evaluating oxidation resistance and content of gallic acid and ellagic acid in myrtle roots by using ABTS-UPLC method Download PDFInfo
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- CN110954626A CN110954626A CN201911271284.7A CN201911271284A CN110954626A CN 110954626 A CN110954626 A CN 110954626A CN 201911271284 A CN201911271284 A CN 201911271284A CN 110954626 A CN110954626 A CN 110954626A
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- 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
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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
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Abstract
The invention relates to a method for simultaneously evaluating the oxidation resistance and the content of gallic acid and ellagic acid in myrtle roots by using an ABTS-UPLC-PAD method, which comprises the following steps: respectively carrying out detection on the sample solution before and after reaction with ABTS + by using an ultra-high performance liquid chromatograph; calculating the removal rate of gallic acid and ellagic acid according to the areas of the gallic acid and ellagic acid in the sample solution obtained by UPLC determination before and after ABTS + reaction, determining gallic acid and ellagic acid reference substance solutions with different concentrations by UPLC, and calculating the contents of gallic acid and ellagic acid in the myrtle root by respectively taking the concentrations of the gallic acid and ellagic acid reference substances as abscissa and the corresponding areas as ordinate as standard curve. The method can simultaneously detect the oxidation resistance and the contents of the gallic acid and the ellagic acid in the myrtle root, and has the advantages of high analysis speed, high sensitivity, good separation degree, simple and convenient operation and less reagent consumption.
Description
Technical Field
The invention relates to the field of analysis of antioxidant active ingredients of traditional Chinese medicinal materials, in particular to a method for simultaneously evaluating the antioxidant property and the content of gallic acid and ellagic acid which are effective ingredients in myrtle roots by using an ABTS-UPLC-PAD method.
Background
Gallic acid and ellagic acid are polyphenols, and are main effective components of radix Rhodomyrti. Gallic acid and ellagic acid are known to have strong oxidation resistance, so that the evaluation of each antioxidant active ingredient and the content thereof in the myrtle root medicinal material can provide a theoretical basis for screening the antioxidant active ingredients in the myrtle and further developing and applying the myrtle medicinal material.
At present, the method for evaluating the oxidation resistance of the effective ingredients in the traditional Chinese medicinal materials is mainly a UV method, but the UV method can only measure the total oxidation resistance of the traditional Chinese medicinal materials or the oxidation resistance of certain substances, can not screen specific certain antioxidant substances, and is complex, large in reagent consumption, long in time consumption and poor in reproducibility.
Disclosure of Invention
The invention aims to solve the technical problems that the oxidation resistance and the content of gallic acid and ellagic acid in myrtle root can be quickly, simply and conveniently evaluated at the same time based on a UPLC method and by utilizing the principle that after ABTS + free radical ions react with antioxidants, the content of the antioxidants is reduced and the area of a chromatographic peak is reduced.
The technical scheme adopted by the invention for solving the technical problems is as follows: a method for simultaneously evaluating oxidation resistance and contents of gallic acid and ellagic acid as effective components in radix Rhodomyrti by ABTS-UPLC-PAD method comprises the following steps:
(1) identifying a sample myrtle root medicinal material;
(2) preparing a standard solution: accurately weighing a gallic acid reference substance and an ellagic acid reference substance respectively, placing the gallic acid reference substance and the ellagic acid reference substance into a brown volumetric flask with the volume of 25-100 mL, adding methanol to dissolve the gallic acid and the ellagic acid reference substance, and diluting the gallic acid and the ellagic acid reference substance to scales to prepare a mixed reference substance solution containing 50-500 mu g/mL of gallic acid and 50-500 mu g/mL of ellagic acid; when in use, the standard solutions are respectively diluted by methanol in sequence to prepare standard working solutions with different concentrations;
(3) preparing ABTS + working solution:
ABTS stock solution: accurately weighing 5-100 mg of ABTS in a 10-100 mL brown volumetric flask, adding ultrapure water for dissolving, and carrying out constant volume to prepare an ABTS stock solution with the concentration of 7.4-37.0 mmol/L;
K2S2O8stock solution: accurately weighing K2S2O85-50 mg of K is dissolved in 10-100 mL of brown volumetric flask by adding ultrapure water and then prepared into K with the concentration of 2.6-13.0 mmol/L by constant volume2S2O8A stock solution;
ABTS stock solution and K2S2O8Mixing the stock solutions in an equal volume ratio, reacting at room temperature in a dark place for 12-16 h, and diluting according to experiment requirements for use;
(4) myrtle root sample solution: pulverizing dried radix Rhodomyrti, sieving with pharmacopeia No. 4 sieve to obtain radix Rhodomyrti medicinal material sample powder, and storing in a dryer; accurately weighing 0.1-5.0 g of myrtle root powder, placing the myrtle root powder in a 50-250 mL conical flask with a plug, precisely adding 10-200 mL of methanol, weighing, extracting for 10-120 min by ultrasonic (50-1500W, 25-100 kHz, 25-60 ℃), cooling, filtering, volatilizing, dissolving residues by methanol, and fixing the volume to a 25mL volumetric flask. Precisely absorbing 1-5 mL of the sample solution into a centrifuge tube, respectively adding ultrapure water and ABTS + working solution which have the same volume with the sample, shaking up, reacting for 5-60 min at room temperature in a dark place to obtain sample solutions before ABTS + reaction and after ABTS + reaction, and measuring after all the sample solutions pass through a 0.2 mu m filter membrane.
(5) And (3) detecting by using ultra-high performance liquid chromatography, wherein the chromatographic conditions are as follows:
the instrument comprises the following steps: UPLC-PDA (Waters, usa);
a chromatographic column: waters BEH C18 chromatography column (2.1 mm. times.100 mm, 1.7 μm); mobile phase: acetonitrile-0.1% formic acid; gradient elution conditions: 0-2 min, 6% acetonitrile; 2-3 min, 6-16% acetonitrile; 3-8 min, 16-20% acetonitrile; 8-10 min, 6% acetonitrile; detection wavelength: 210-400 nm; flow rate: 0.1-0.5 mL/min; column temperature: 25-45 ℃; the injection volume is 0.1-10 mu L.
(6) Component content and antioxidant effect evaluation: recording areas of gallic acid and ellagic acid peaks of the sample solution obtained by UPLC determination before and after ABTS + reaction, and calculating clearance rate according to the following formula: clearance rate is 1-A/A0(A0The peak area of the compound before ABTS + reaction, and the peak area of the compound after ABTS + reaction); and respectively taking the concentrations of the gallic acid and the ellagic acid as abscissa and the corresponding peak areas as ordinate, drawing a standard curve, and calculating the contents of the gallic acid and the ellagic acid in the myrtle root.
Compared with the prior art, the method for simultaneously evaluating the oxidation resistance and the content of the gallic acid and the ellagic acid which are effective components in the myrtle root by using the ABTS-UPLC-PAD method has the following advantages and positive effects:
(1) the method can simultaneously determine the oxidation resistance and the content of the gallic acid and the ellagic acid in the myrtle root medicinal material;
(2) the invention has the advantages of high analysis speed, high sensitivity, good separation degree, simple and convenient operation and less reagent consumption.
(3) The method can be used for evaluating the oxidation resistance and the content of the myrtle root medicinal material, and can provide a theoretical basis for screening and further developing the antioxidant active ingredients in the myrtle root.
Drawings
Fig. 1 is UPLC chromatograms of an ABTS + pre-reaction sample solution and an ABTS + post-reaction sample solution in the present invention, wherein a is an ABTS + pre-reaction sample, B is an ABTS + post-reaction sample, peak 1 is gallic acid, and peak 2 is ellagic acid.
Detailed Description
The technical solution of the present invention is further described below with reference to specific embodiments.
The producing areas of the myrtle root medicinal materials adopted in the 2 examples are Guangxi Yulin and Guangxi Pingle respectively, and the myrtle root medicinal materials are identified as myrtle roots by Liuwei high-level veterinarians of Guangxi veterinary institute.
Example 1
(1) Preparation of standard solution: accurately weighing appropriate amount of gallic acid reference substance and ellagic acid reference substance respectively, placing into 100mL brown volumetric flask, adding methanol to dissolve and dilute to scale, and making into mixed reference substance solution containing gallic acid 80.12 μ g/mL and ellagic acid 80.44 μ g/mL; when in use, the standard solutions are respectively diluted by methanol in sequence to prepare standard working solutions with different concentrations;
(2) preparing ABTS + working solution:
ABTS stock solution: accurately weighing ABTS 40.60mg in a 10mL brown volumetric flask, adding ultrapure water for dissolving, and fixing the volume to prepare ABTS stock solution with the concentration of 7.4 mmol/L;
K2S2O8stock solution: accurately weighing K2S2O86.99 mg of the K is dissolved in a 10mL brown volumetric flask by adding ultrapure water and prepared into the K with the concentration of 2.6mmol/L by constant volume2S2O8A stock solution;
ABTS stock solution and K2S2O8Mixing the stock solutions in an equal volume ratio, reacting at room temperature in a dark place for 12 hours, and diluting by 10 times for use;
(3) myrtle root sample solution: pulverizing dried myrtle root which is produced in Yulin of Guangxi, sieving by a pharmacopoeia No. 4 sieve to obtain myrtle root medicinal material sample powder, accurately weighing 1g of fine powder, placing the fine powder into a 100mL conical flask with a plug, precisely adding 50mL of methanol, weighing, ultrasonically (350W, 53kHz, 35 ℃) for 30min, cooling, filtering, volatilizing, dissolving residues by the methanol, and fixing the volume to a 25mL volumetric flask. Precisely absorbing 1mL of the sample solution into a centrifuge tube, respectively adding ultrapure water and ABTS + working solution which have the same volume with the sample, shaking up, reacting for 5min at room temperature in a dark place to obtain sample solutions before and after ABTS + reaction, and injecting all the sample solutions into a UPLC instrument for determination after passing through a 0.2 mu m filter membrane.
(4) Chromatographic conditions are as follows: the instrument comprises the following steps: UPLC-PDA (Waters, usa); a chromatographic column: waters BEH C18 chromatography column (2.1 mm. times.100 mm, 1.7 μm); mobile phase: acetonitrile-0.1% formic acid; gradient elution conditions: 0-2 min, 6% acetonitrile; 2-3 min, 6-16% acetonitrile; 3-8 min, 16-20% acetonitrile; 8-10 min, 6% acetonitrile; detection wavelength: 260 nm; flow rate: 0.25 mL/min; column temperature: 35 ℃; injection volume 1. mu.L.
(5) Component content and antioxidant effect evaluation: recording areas of gallic acid and ellagic acid peaks of the sample solution obtained by UPLC determination before and after ABTS + reaction, and calculating clearance rate according to the following formula: clearance rate is 1-A/A0(A0ABTS + peak area of pre-reacted compound, A is ABTS + peak area of post-reacted compound). The calibration curves of the gallic acid and the ellagic acid are respectively 4516.5x +608.2(R2 is 0.9991) and 6032.9x +369.99(R2 is 0.9989), and the corresponding areas of the gallic acid and the ellagic acid peak before ABTS + reaction are respectively substituted into the calibration curves to calculate the content. The content of gallic acid and ellagic acid in the root of Myrtus communis of Yulin of Guangxi and the clearance rate thereof are shown in Table 1. The result shows that the gallic acid and the ellagic acid in the myrtle root have stronger oxidation resistance, wherein the oxidation resistance of the ellagic acid is higher than that of the gallic acid.
TABLE 1 Gallic acid and ellagic acid content and clearance (n ═ 3)
Example 2
(1) The standard solution was prepared as in example 1.
(2) ABTS + working solution was prepared as in example 1.
(3) Myrtle root sample solution: pulverizing dried radix Rhodomyrti in Guangxi Pingle, sieving with pharmacopeia No. 4 sieve to obtain radix Rhodomyrti medicinal material sample powder, accurately weighing 0.5g of fine powder, placing in 100mL conical flask with plug, adding 30mL of methanol precisely, weighing, extracting with ultrasound (350W, 53kHz, 45 deg.C) for 40min, cooling, filtering, volatilizing filtrate, re-dissolving the obtained residue with methanol, and fixing volume to 25mL brown volumetric flask. Precisely absorbing 2mL of the sample solution into a centrifuge tube, adding ultrapure water and ABTS + working solution which have the same volume with the sample, shaking up, reacting for 10min at room temperature in a dark place to obtain sample solutions before and after ABTS + reaction, and injecting all the sample solutions into a UPLC instrument for determination after passing through a 0.2 mu m filter membrane.
(4) The chromatographic conditions were the same as in example 1.
(5) Component content and antioxidant effect evaluation: the clearance was calculated according to the following formula: clearance rate is 1-A/A0(A0ABTS + peak area of pre-reacted compound, A is ABTS + peak area of post-reacted compound). The calibration curves of the gallic acid and the ellagic acid are respectively 4516.5x +608.2(R2 is 0.9991) and 6032.9x +369.99(R2 is 0.9989), and the corresponding areas of the gallic acid and the ellagic acid peak before ABTS + reaction are respectively substituted into the calibration curves to calculate the content. The contents of gallic acid and ellagic acid in Guangxi Guiping Myrtus communis root and their clearance rates are shown in Table 2. The result shows that the gallic acid and the ellagic acid in the myrtle root have stronger oxidation resistance, wherein the oxidation resistance of the ellagic acid is higher than that of the gallic acid.
TABLE 2 Gallic acid and ellagic acid content and clearance (n ═ 3)
Claims (5)
1. A method for simultaneously evaluating the oxidation resistance and the content of gallic acid and ellagic acid which are effective components in myrtle roots by using an ABTS-UPLC-PAD method is characterized by comprising the following steps:
(1) identifying a sample myrtle root medicinal material;
(2) preparing a gallic acid reference substance and an ellagic acid reference substance standard solution;
(3) preparing ABTS + working solution: ABTS solution with the concentration of 7.4-37.0 mmol/L and K with the concentration of 2.6-13.0 mmol/L2S2O8Mixing the solutions according to the equal volume ratio, reacting at room temperature in a dark place for 12-16 h, and diluting to obtain ABTS + working solution;
(4) preparing a myrtle root sample solution: accurately weighing 0.1-5.0 g of myrtle root powder, placing the myrtle root powder into a 50-250 mL conical flask with a plug, precisely adding 10-200 mL of methanol, weighing, carrying out ultrasonic extraction for 10-120 min at the temperature of 25-60 ℃ under the conditions of 50-1500W, 25-100 kHz, cooling, filtering, volatilizing, dissolving residues with methanol, and fixing the volume to a 25mL volumetric flask;
(5) precisely absorbing 1-5 mL of the sample solution obtained in the step (4) into two centrifuge tubes, adding ultrapure water and ABTS + working solution which are equal in volume to the sample solution, shaking up, and reacting for 5-60 min at room temperature in a dark place to obtain sample solutions before and after ABTS + reaction;
(6) respectively carrying out detection on the sample solution before and after the ABTS + reaction by using an ultra-high performance liquid chromatograph under the following chromatographic conditions:
a chromatographic column: 2.1mm × 100mm, 1.7 μm Waters BEH C18 chromatography column; mobile phase: acetonitrile-0.1% formic acid; gradient elution conditions: 0-2 min, 6% acetonitrile; 2-3 min, 6-16% acetonitrile; 3-8 min, 16-20% acetonitrile; 8-10 min, 6% acetonitrile; detection wavelength: 210-400 nm; flow rate: 0.1-0.5 mL/min; column temperature: 25-45 ℃; the sample injection volume is 0.1-10 mu L;
(7) component content and antioxidant effect evaluation: recording areas of gallic acid and ellagic acid peaks of the sample solution obtained by UPLC determination before and after ABTS + reaction, and calculating clearance rate according to the following formula: clearance rate is 1-A/A0Wherein A is0The peak area of the compound before ABTS + reaction, A is the peak area of the compound after ABTS + reaction; and respectively taking the concentrations of the gallic acid and the ellagic acid as abscissa and the corresponding peak areas as ordinate, making a standard curve, and calculating the contents of the gallic acid and the ellagic acid in the myrtle root.
2. The method according to claim 1, wherein in the step (2), the specific preparation method of the standard solutions of the gallic acid reference substance and the ellagic acid reference substance is as follows: accurately weighing a gallic acid reference substance and an ellagic acid reference substance respectively, placing the gallic acid reference substance and the ellagic acid reference substance into a brown volumetric flask with the volume of 25-100 mL, adding methanol to dissolve the gallic acid and the ellagic acid reference substance, and diluting the gallic acid and the ellagic acid reference substance to scales to prepare a mixed reference substance solution containing 50-500 mu g/mL of gallic acid and 50-500 mu g/mL of ellagic acid; when the standard working solution is used, the standard working solutions with different concentrations are prepared by sequentially diluting the standard solutions with methanol respectively.
3. According to claim 1The method is characterized in that in the step (3), the preparation method of the ABTS solution is as follows: accurately weighing 5-100 mg of ABTS in a 10-100 mL brown volumetric flask, adding ultrapure water for dissolving, and carrying out constant volume to prepare an ABTS stock solution with the concentration of 7.4-37.0 mmol/L; k2S2O8The preparation method of the solution comprises the following steps: accurately weighing K2S2O85-50 mg of K is dissolved in 10-100 mL of brown volumetric flask by adding ultrapure water and then prepared into K with the concentration of 2.6-13.0 mmol/L by constant volume2S2O8And (4) stock solution.
4. The method according to claim 1, wherein in the step (3), the dilution is 10-fold.
5. The method of claim 1, wherein in step (5), after obtaining the sample solutions before and after the ABTS + reaction, all sample solutions are filtered through a 0.2 μm filter and subjected to UPLC assay.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100215783A1 (en) * | 2009-02-23 | 2010-08-26 | Nutragenesis, Llc | Antioxidant Compositions, Beverage Formulations Thereof, and Methods of Increasing Antioxidant Capability |
| CN104407087A (en) * | 2014-12-04 | 2015-03-11 | 广西壮族自治区兽医研究所 | Method for simultaneously detecting gallic acid and ellagic acid in radix rhodomyrti |
| 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 |
| CN110196290A (en) * | 2019-04-02 | 2019-09-03 | 贵州医科大学 | Ellagic acid content assaying method in a kind of Rosa roxburghii Tratt |
-
2019
- 2019-12-12 CN CN201911271284.7A patent/CN110954626A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100215783A1 (en) * | 2009-02-23 | 2010-08-26 | Nutragenesis, Llc | Antioxidant Compositions, Beverage Formulations Thereof, and Methods of Increasing Antioxidant Capability |
| CN104407087A (en) * | 2014-12-04 | 2015-03-11 | 广西壮族自治区兽医研究所 | Method for simultaneously detecting gallic acid and ellagic acid in radix rhodomyrti |
| CN110196290A (en) * | 2019-04-02 | 2019-09-03 | 贵州医科大学 | Ellagic acid content assaying method in a kind of Rosa roxburghii Tratt |
| 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 |
Non-Patent Citations (3)
| Title |
|---|
| 罗爽妍 等: "野生桃金娘主要抗氧化成分及其抗氧化能力", 《食品科学》 * |
| 袁媛 等: "桃金娘花青素和黄酮的提取方法比较及其抗氧化能力研究", 《中国食品学报》 * |
| 郑晓燕 等: "桃金娘果实多酚抗氧化活性及乙酰胆碱酯酶抑制活性研究", 《中国农学通报》 * |
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