CN105572255A - Method for simultaneous determination of naringenin and eriodictyol content in fresh-cut water chestnut etiolation organization - Google Patents
Method for simultaneous determination of naringenin and eriodictyol content in fresh-cut water chestnut etiolation organization Download PDFInfo
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- CN105572255A CN105572255A CN201510982643.5A CN201510982643A CN105572255A CN 105572255 A CN105572255 A CN 105572255A CN 201510982643 A CN201510982643 A CN 201510982643A CN 105572255 A CN105572255 A CN 105572255A
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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
- G01N30/02—Column chromatography
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- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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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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Abstract
The present invention discloses a method for simultaneous determination of naringenin and eriodictyol content in fresh-cut water chestnut etiolation organization, and the method comprises the six steps of preparation of a standard naringenin stock solution, preparation of a standard eriodictyol stock solution, preparation of a mixed naringenin and eriodictyol standard solution, production of a standard curve, preparation of a to-be-tested sample solution, and calculation of naringenin and eriodictyol content in a fresh-cut water chestnut etiolation organization sample. According to the method, the naringenin and the eriodictyol in the fresh-cut water chestnut etiolation organization can be effectively separated, the naringenin and eriodictyol content in the fresh-cut water chestnut etiolation organization can be simultaneously determined, the method is simple, fast, accurate and reliable, and can exclude interference produced by other substance chromatography and other substances contained in the fresh-cut water chestnut etiolation organization.
Description
Technical field
The invention belongs to constituent content determination techniques field, be specifically related to a kind of method of naringenin and eriodictyol content in Simultaneously test fresh-cut water chestnut yellow tissue.
Background technology
Water chestnut (Eleocharistuberosa), another name water chestnut, an ancient name for water chestnut, Di Li, black taro etc. are the underground bulb that per nnial herb Cyperaceae Eleocharis is formed.The water chestnut not only crisp succulence of matter, nutritious, but also can be used as medicine, various diseases is had to the effect of supplemental treatment.Have the good reputation of " underground snow pear " since ancient times, northerner regards it as " south of the River ginseng ".But water chestnut is after fresh-cut process, cutting surfaces can turn yellow in a short time, has had a strong impact on exterior quality and other value.Early-stage Study finds, causes the main cause of yellow to be that in Fresh-cut Chinese Water-chestnut, Cucumber generates caused by Flavonoid substances through phenylpropyl alcohol alkane metabolic pathway, along with the intensification of yellow, and the corresponding increase of the content of general flavone in fresh-cut water chestnut.Carry out separation and purification and Structural Identification further by yellowed matter, show that the yellow of fresh-cut water chestnut cutting surfaces may be main relevant with naringenin two kinds of Flavonoid substances with eriodictyol.Therefore, set up one can in Simultaneously test fresh-cut water chestnut yellow tissue eriodictyol and naringenin content significant to research fresh-cut water chestnut xanthating machine fixture.
In recent years, have both at home and abroad and detect and the independent bibliographical information of assay about naringenin, eriodictyol, mainly high performance liquid chromatography (HPLC), or with additive method coupling, but HPLC method mainly concentrates on naringenin in Chinese herbal medicine, eriodictyol detects and the mensuration of content, and these conditions might not be applicable to naringenin in fresh-cut water chestnut yellow tissue and eriodictyol detection and assay thereof.In addition, also there is not yet the report that HPLC method measures naringenin or eriodictyol in fresh-cut water chestnut yellow tissue at present, more do not had the report of naringenin and eriodictyol in Simultaneously test fresh-cut water chestnut yellow tissue.
Summary of the invention
The object of this invention is to provide a kind of method of naringenin and eriodictyol content in Simultaneously test fresh-cut water chestnut yellow tissue, the method can the content of naringenin and eriodictyol in Simultaneously test fresh-cut water chestnut yellow tissue.
The technical solution adopted in the present invention is, the method for naringenin and eriodictyol content in Simultaneously test fresh-cut water chestnut yellow tissue, specifically implements according to following steps:
Step 1, precision takes standard items naringenin 0.70 ± 0.01mg and is placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and makes standard items naringenin storing solution;
Precision takes standard items eriodictyol 0.40 ± 0.01mg and is placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and makes standard items eriodictyol storing solution;
Accurate measuring standard items naringenin storing solution and each 1mL of standard items eriodictyol storing solution are placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and obtaining concentration is naringenin 28 ± 0.4 μ gmL
-1with eriodictyol 16 ± 0.4 μ gmL
-1hybrid standard product solution, 4 DEG C keep in Dark Place, for subsequent use;
Step 2, get naringenin and the eriodictyol standard items mixed solution of preparation in step 1, being mixed with concentration is respectively 5.60 μ gmL
-1, 11.20 μ gmL
-1, 16.80 μ gmL
-1, 22.40 μ gmL
-1, 28.00 μ gmL
-1naringenin, concentration be 3.20 μ gmL
-1, 6.40 μ gmL
-1, 9.60 μ gmL
-1, 12.80 μ gmL
-1, 16.00 μ gmL
-1eriodictyol series of standards product mixed solution, adopts high effective liquid chromatography for measuring chromatographic peak area; Respectively with naringenin and eriodictyol concentration X for horizontal ordinate, chromatographic peak area Y is that ordinate carries out linear regression and obtains typical curve;
Step 3, get fresh-cut water chestnut yellow tissue surface and prepare testing sample solution, then high effective liquid chromatography for measuring chromatographic peak area A, then by checking in chromatographic peak area A corresponding naringenin concentration X1 and eriodictyol concentration X2 from the typical curve of step 2, naringenin and eriodictyol content in fresh-cut water chestnut yellow tissue sample is calculated.
Feature of the present invention is also,
In step 2 and 4, the chromatographic condition of high performance liquid chromatography (HPLC) is as follows:
Chromatographic column: AgilentEclipseXDB-C
18(150mm × 4.6mm, 5 μm); Column temperature: 40 DEG C; Mobile phase: 0.5% glacial acetic acid solution (A)-acetonitrile (B), gradient elution (0 ~ 10min, 10% → 20%B; 10 ~ 15min, 20% → 30%B; 15 ~ 25min, 30% → 40%B; 25 ~ 30min, 40% → 45%B; 30 ~ 35min, 45% → 50%B), mobile phase needs through 0.22 μm of miillpore filter suction filtration and ultrasonic degas before using; Flow velocity: 1mLmin
-1; Sample size: 10 μ L; Determined wavelength: 280nm.With this understanding, calculate should be not less than 5000 by the theoretical cam curve of naringenin and eriodictyol, peak degree of separation should be greater than 1.5.
In step 3, the preparation process of testing sample solution is as follows: fresh water chestnut peeling, cutting, preserve under being placed in 16 ± 1 DEG C of temperature conditions, treat the surface yellow of fresh-cut water chestnut, accurately take yellow surface 29.60 ~ 30.40g, be placed in triangular flask, be that 1:5 adds absolute ethyl alcohol by solid-liquid ratio, ultrasonic extraction 30 ~ 35min at 30 ~ 40 DEG C of temperature, 4000 ~ 5000r/min centrifuging, 15 ~ 20min, get supernatant, residue adopts same method to extract, merge supernatant, rotary evaporation is concentrated into dry, concentrating part methyl alcohol is dissolved and is settled to 5mL, shake up, 0.22 μm of filtering with microporous membrane, filtrate is testing sample solution.
The invention has the beneficial effects as follows establish can in Simultaneously test fresh-cut water chestnut yellow tissue naringenin and eriodictyol content assay method---high performance liquid chromatography (HPLC).Current HPLC method is mainly used in naringenin in Chinese herbal medicine, eriodictyol and detects respectively and the mensuration of content, not about the case of naringenin, eriodictyol content in its Simultaneously test fresh-cut water chestnut yellow tissue, and its condition is not suitable for the assay measuring naringenin and eriodictyol in fresh-cut water chestnut yellow tissue yet.Method in the present invention just can be separated with eriodictyol the naringenin in fresh-cut water chestnut yellow tissue effectively, go back its naringenin of Simultaneously test and eriodictyol content in addition, and the method is simple and efficient, accurately and reliably, can get rid of the interference that in fresh-cut water chestnut yellow tissue, other material chromatograms contained and other materials produce.Thus, the present invention also can be naringenin and eriodictyol content Simultaneously test in other fresh-cut fruit and vegetables and provides certain foundation.
Accompanying drawing explanation
Fig. 1 is eriodictyol and naringenin standard items HPLC chromatogram;
Fig. 2 is eriodictyol and naringenin HPLC chromatogram in fresh-cut water chestnut yellow tissue sample.
Embodiment
Below in conjunction with the drawings and specific embodiments, the present invention is described in detail.
The method of naringenin and eriodictyol content in Simultaneously test fresh-cut water chestnut yellow tissue of the present invention, specifically implement according to following steps:
Step 1, precision takes standard items naringenin 0.70 ± 0.01mg and is placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and makes standard items naringenin storing solution;
Precision takes standard items eriodictyol 0.40 ± 0.01mg and is placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and makes standard items eriodictyol storing solution;
Accurate measuring standard items naringenin storing solution and each 1mL of standard items eriodictyol storing solution are placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and obtaining concentration is naringenin 28 ± 0.4 μ gmL
-1with eriodictyol 16 ± 0.4 μ gmL
-1hybrid standard product solution, 4 DEG C keep in Dark Place, for subsequent use;
Step 2, get naringenin and the eriodictyol standard items mixed solution of preparation in step 1, being mixed with concentration is respectively 5.60 μ gmL
-1, 11.20 μ gmL
-1, 16.80 μ gmL
-1, 22.40 μ gmL
-1, 28.00 μ gmL
-1naringenin, concentration be 3.20 μ gmL
-1, 6.40 μ gmL
-1, 9.60 μ gmL
-1, 12.80 μ gmL
-1, 16.00 μ gmL
-1eriodictyol series of standards product mixed solution, adopts high effective liquid chromatography for measuring chromatographic peak area; Respectively with naringenin and eriodictyol concentration X for horizontal ordinate, chromatographic peak area Y is that ordinate carries out linear regression and obtains typical curve;
The chromatographic condition of high performance liquid chromatography (HPLC) is as follows:
Chromatographic column: AgilentEclipseXDB-C
18(150mm × 4.6mm, 5 μm); Column temperature: 40 DEG C; Mobile phase: 0.5% glacial acetic acid solution (A)-acetonitrile (B), gradient elution (0 ~ 10min, 10% → 20%B; 10 ~ 15min, 20% → 30%B; 15 ~ 25min, 30% → 40%B; 25 ~ 30min, 40% → 45%B; 30 ~ 35min, 45% → 50%B), mobile phase needs through 0.22 μm of miillpore filter suction filtration and ultrasonic degas before using; Flow velocity: 1mLmin
-1; Sample size: 10 μ L; Determined wavelength: 280nm.With this understanding, calculate should be not less than 5000 by the theoretical cam curve of naringenin and eriodictyol, peak degree of separation should be greater than 1.5.Condition of gradient elution is in table 1.
Table 1 condition of gradient elution
Step 3, fresh water chestnut peeling, cutting, preserve under being placed in 16 ± 1 DEG C of temperature conditions, treat the surface yellow of fresh-cut water chestnut, accurately take yellow surface 29.60 ~ 30.40g, be placed in triangular flask, be that 1:5 adds absolute ethyl alcohol by solid-liquid ratio, ultrasonic extraction 30 ~ 35min at 30 ~ 40 DEG C of temperature, 4000 ~ 5000r/min centrifuging, 15 ~ 20min, get supernatant, residue adopts same method to extract, merge supernatant, rotary evaporation is concentrated into dry, concentrating part methyl alcohol is dissolved and is settled to 5mL, shake up, 0.22 μm of filtering with microporous membrane, filtrate is testing sample solution,
Step 4, get the testing sample solution that step 3 obtains, then high effective liquid chromatography for measuring chromatographic peak area A, chromatographic condition is identical with step 2, then by checking in chromatographic peak area A corresponding naringenin concentration X1 and eriodictyol concentration X2 from the typical curve of step 2, naringenin and eriodictyol content in fresh-cut water chestnut yellow tissue sample is calculated.
Embodiment
Step 1, precision takes standard items naringenin 0.70 ± 0.01mg respectively, standard items eriodictyol 0.40 ± 0.01mg is placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and makes standard items naringenin storing solution and standard items eriodictyol storing solution;
Precision criterion product naringenin storing solution and each 1mL of standard items eriodictyol storing solution are placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and obtaining concentration is naringenin 28 ± 0.4 μ gmL
-1, eriodictyol 16 ± 0.4 μ gmL
-1hybrid standard product solution, 4 DEG C keep in Dark Place, for subsequent use.
Step 2, gets naringenin and the eriodictyol standard items mixed solution of preparation in step 1, is mixed with naringenin (5.60 μ gmL
-1, 11.20 μ gmL
-1, 16.80 μ gmL
-1, 22.40 μ gmL
-1, 28.00 μ gmL
-1), eriodictyol (3.20 μ gmL
-1, 6.40 μ gmL
-1, 9.60 μ gmL
-1, 12.80 μ gmL
-1, 16.00 μ gmL
-1) the standard items mixed solution of a series of concentration, measure chromatographic peak area.Respectively with naringenin and eriodictyol concentration X (μ gmL
-1) be horizontal ordinate, chromatographic peak area Y is that ordinate carries out linear regression and obtains regression equation: naringenin Y=2767.3X+6.81 (r=1.0000), the range of linearity 5.60 ~ 28 μ gmL
-1; Eriodictyol Y=2925.7X+0.9 (r=0.9999), the range of linearity 3.20 ~ 16 μ gmL
-1.
Step 3, get with a collection of fresh-cut water chestnut yellow surface sample 3 parts (every part of 30.00g), preparation yellow sample solution, then sample introduction analysis under the chromatographic condition determined after optimization, by the qualitative eriodictyol of standard items HPLC chromatogram and naringenin, and the typical curve drawn by step 2 calculates naringenin and eriodictyol content in fresh-cut water chestnut yellow tissue sample, the results are shown in Table 2.Standard items HPLC chromatogram is shown in Fig. 1, and sample HPLC chromatogram is shown in Fig. 2.
Naringenin and eriodictyol content in table 2 fresh-cut water chestnut yellow surface
As can be seen from table 2 and Fig. 2, although containing other much different components in fresh-cut water chestnut yellow surface, wherein really there is naringenin and eriodictyol, and content is higher.
Claims (3)
1. the method for naringenin and eriodictyol content in Simultaneously test fresh-cut water chestnut yellow tissue, is characterized in that, specifically implement according to following steps:
Step 1, precision takes standard items naringenin 0.70 ± 0.01mg and is placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and makes standard items naringenin storing solution;
Precision takes standard items eriodictyol 0.40 ± 0.01mg and is placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and makes standard items eriodictyol storing solution;
Accurate measuring standard items naringenin storing solution and each 1mL of standard items eriodictyol storing solution are placed in the brown volumetric flask of 5mL, and methyl alcohol dissolves and is diluted to scale, shakes up, and obtaining concentration is naringenin 28 ± 0.4 μ gmL
-1with eriodictyol 16 ± 0.4 μ gmL
-1hybrid standard product solution, 4 DEG C keep in Dark Place, for subsequent use;
Step 2, get naringenin and the eriodictyol standard items mixed solution of preparation in step 1, being mixed with concentration is respectively 5.60 μ gmL
-1, 11.20 μ gmL
-1, 16.80 μ gmL
-1, 22.40 μ gmL
-1, 28.00 μ gmL
-1naringenin, concentration be 3.20 μ gmL
-1, 6.40 μ gmL
-1, 9.60 μ gmL
-1, 12.80 μ gmL
-1, 16.00 μ gmL
-1eriodictyol series of standards product mixed solution, adopts high effective liquid chromatography for measuring chromatographic peak area; Respectively with naringenin and eriodictyol concentration X for horizontal ordinate, chromatographic peak area Y is that ordinate carries out linear regression and obtains typical curve;
Step 3, get fresh-cut water chestnut yellow tissue surface and prepare testing sample solution, then high effective liquid chromatography for measuring chromatographic peak area A, then by checking in chromatographic peak area A corresponding naringenin X1 and eriodictyol concentration X2 from the typical curve of step 2, naringenin and eriodictyol content in fresh-cut water chestnut yellow tissue sample is calculated.
2. the method for naringenin and eriodictyol content in Simultaneously test fresh-cut water chestnut yellow tissue according to claim 1, it is characterized in that, in step 2 and 4, the chromatographic condition of high performance liquid chromatography (HPLC) is as follows:
Chromatographic column: AgilentEclipseXDB-C
18(150mm × 4.6mm, 5 μm); Column temperature: 40 DEG C; Mobile phase: 0.5% glacial acetic acid solution (A)-acetonitrile (B), condition of gradient elution: 0 ~ 10min, 10% → 20%B; 10 ~ 15min, 20% → 30%B; 15 ~ 25min, 30% → 40%B; 25 ~ 30min, 40% → 45%B; 30 ~ 35min, 45% → 50%B, mobile phase needs through 0.22 μm of miillpore filter suction filtration and ultrasonic degas before using; Flow velocity: 1mLmin
-1; Sample size: 10 μ L; Determined wavelength: 280nm.With this understanding, calculate should be not less than 5000 by the theoretical cam curve of naringenin and eriodictyol, peak degree of separation should be greater than 1.5.
3. the method for naringenin and eriodictyol content in Simultaneously test fresh-cut water chestnut yellow tissue according to claim 1, it is characterized in that, in step 3, the preparation process of testing sample solution is as follows: fresh water chestnut peeling, cutting, preserve under being placed in 16 ± 1 DEG C of temperature conditions, treat the surface yellow of fresh-cut water chestnut, accurately take yellow surface 29.60 ~ 30.40g, be placed in triangular flask, be that 1:5 adds absolute ethyl alcohol by solid-liquid ratio, ultrasonic extraction 30 ~ 35min at 30 ~ 40 DEG C of temperature, 4000 ~ 5000r/min centrifuging, 15 ~ 20min, get supernatant, residue adopts same method to extract, merge supernatant, rotary evaporation is concentrated into dry, concentrating part methyl alcohol is dissolved and is settled to 5mL, shake up, 0.22 μm of filtering with microporous membrane, filtrate is testing sample solution.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064256A (en) * | 2017-05-25 | 2017-08-18 | 济南大学 | A kind of preparation method of naringenin molecular imprinting electrochemical sensor |
| CN109580604A (en) * | 2018-11-28 | 2019-04-05 | 江南大学 | A kind of method of high-throughput detection naringenin |
| CN112268973A (en) * | 2020-11-05 | 2021-01-26 | 上海黄海制药有限责任公司 | Method for measuring naringenin HPLC content in pine pollen |
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| CN104529983A (en) * | 2014-12-29 | 2015-04-22 | 贺州学院 | Method for extracting eriodictyol from water chestnut peel |
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| CN104529983A (en) * | 2014-12-29 | 2015-04-22 | 贺州学院 | Method for extracting eriodictyol from water chestnut peel |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107064256A (en) * | 2017-05-25 | 2017-08-18 | 济南大学 | A kind of preparation method of naringenin molecular imprinting electrochemical sensor |
| CN107064256B (en) * | 2017-05-25 | 2019-03-01 | 济南大学 | A kind of preparation method of naringenin molecular imprinting electrochemical sensor |
| CN109580604A (en) * | 2018-11-28 | 2019-04-05 | 江南大学 | A kind of method of high-throughput detection naringenin |
| CN112268973A (en) * | 2020-11-05 | 2021-01-26 | 上海黄海制药有限责任公司 | Method for measuring naringenin HPLC content in pine pollen |
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