CN104749279B - The HPLC assay methods of the rutinoside of corn flower 3 in blackcurrant - Google Patents
The HPLC assay methods of the rutinoside of corn flower 3 in blackcurrant Download PDFInfo
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- CN104749279B CN104749279B CN201510142303.1A CN201510142303A CN104749279B CN 104749279 B CN104749279 B CN 104749279B CN 201510142303 A CN201510142303 A CN 201510142303A CN 104749279 B CN104749279 B CN 104749279B
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- corn flower
- blackcurrant
- rutinoside
- rutinosides
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Abstract
The HPLC assay methods of the rutinoside of corn flower 3, comprise the following steps in a kind of blackcurrant：Step 1: ripe fresh blackcurrant fruit is chosen, fully squeezing mashing after cleaning up；Step 2: step one gained slurries and blackcurrant fruit residue are fully extracted into the rutinoside of corn flower 3 using ethanol；Step 3: the filtrate containing the rutinoside of corn flower 3 obtained by step 2 is utilized into high performance liquid chromatograph Agilent 2600, using Zorbax SB C18 posts；UV-detector：VWD, G1314F, Detection wavelength are 520nm；Mobile phase A liquid chooses the aqueous formic acid of concentration 1.6%, and B liquid chooses the formic acid acetonitrile solution of concentration 1.6%；Column oven temperature 30；DEG C sample size 10ul；Flow velocity 1ml/min, carries out gradient elution；Step 4: setting up the rutinoside standard curve regression equation of corn flower 3 according to gradient elution data.The inventive method is easy, accurate, available for blackcurrant fruit quality control.Average recovery rate is respectively 96.4%.
Description
Technical field
The present invention relates to Pharmaceutical Analysis extractive technique field, corn flower -3- rutinosides in more particularly to a kind of blackcurrant
HPLC assay methods.
Background technology
Blackcurrant scientific name elements of black currant (Ribes nigrum L.) also known as currant, black bean fruit, purple plum, Saxifragales tea
Fischer scarabaeidae currant belongs to, small shrub, and its ripening fruits is black small berries, interior rich in vitamin C, anthocyanidin, edible.
Fruit juice can also be processed into, the food such as jam.Blackcurrant contains very abundant vitamin C, phosphorus, magnesium, potassium, calcium, anthocyanidin, phenol
Class material.The health-care efficacy for the blackcurrant known at present includes prevention gout, anaemia, oedema, arthritis, rheumatism, mouth
Chamber and pharyngolaryngitis, cough etc..
Blackcurrant is the main berry product of northern China, and yield is higher, and fruits nutrition enriches and has a variety of anthocyanidin
Former times, also with important physiological function and healthcare function, with good market prospects.
The assay method of corn flower -3- rutinosides in existing blackcurrant, complex steps, accuracy rate is low, and is determining
During blackcurrant fruit can be caused compared with lossy, cause product economy inefficiencies.
Corn flower -3- rutinosides are the primary pigments compositions in blackcurrant fruit.The present invention is in prior art basis
Establish a kind of HPLC assay methods of Cyanidin -3-O- rutinoside contents in blackcurrant fruit.
The content of the invention
To solve the problem of above-mentioned prior art is present, it is an object of the invention to provide corn flower in a kind of blackcurrant-
The HPLC assay methods of 3- rutinosides.
To reach above-mentioned purpose, the technical scheme is that：
The HPLC assay methods of corn flower -3- rutinosides, comprise the following steps in a kind of blackcurrant：
Step 1: ripe fresh blackcurrant fruit is chosen, fully squeezing mashing after cleaning up；
Step 2: step one gained slurries and blackcurrant fruit residue are fully extracted into corn flower -3- rues using ethanol
Glucosides；
Step 3: the filtrate containing corn flower -3- rutinosides obtained by step 2 is utilized into high performance liquid chromatograph
Agilent 2600, using Zorbax SB C18 posts；UV-detector：VWD, G1314F, Detection wavelength are 520nm；Mobile phase
A liquid chooses the aqueous formic acid of concentration 1.6%, and B liquid chooses the formic acid acetonitrile solution of concentration 1.6%；Column oven temperature 30；DEG C enter
Sample amount 10ul；Flow velocity 1ml/min, carries out gradient elution；
Step 4: setting up corn flower -3- rutinoside standard curve regression equations according to gradient elution data.
Further, in the step 3, the design parameter of Zorbax SB C18 posts is 250mm × 4.60mm, 5 μm.
Further, in the step 3, the concrete operations of gradient elution are：0-5min, 85%A liquid, 15%B liquid；5-
10min, 80%A liquid, 20%B liquid；10-30min, 72%A liquid, 28%B liquid；30-36min, 100%B liquid.
Further, in the step 4：Corn flower -3- rutinoside standard curve regression equations are：Y=2 ×
107X-33120 (r=0.9998), linear relationship is good in 0.1041~1.041 μ g ranges, and average recovery rate is respectively
96.4%, Cyanidin -3-O- rutinosides content range is 5.263~12.829mg/g.
Relative to prior art, beneficial effects of the present invention are：
The inventive method is easy, accurate, available for blackcurrant fruit quality control.As a result show and utilize present invention arrow car
Chrysanthemum -3- rutinoside standard curve regression equations are：Y=2 × 107X-33120 (r=0.9998), in 0.1041~1.041 μ
Linear relationship is good in g range, and average recovery rate is respectively 96.4%, and Cyanidin -3-O- rutinoside content ranges are
5.263~12.829mg/g.
Embodiment
Technical solution of the present invention is described in further detail with reference to embodiment：
Test example：
Instrument：Agilent 2600；
Detection wavelength：515nm, detector：VWD(G1314F)；
Column oven temperature：30 DEG C, TCC (G1316A)；
Sample size：10ul
Flow velocity：1ml/min
Mobile phase：A liquid：1.6% aqueous formic acid；B liquid：1.6% formic acid methanol solution
Operation program：
| Time | A liquid (%) | B liquid (%) |
| 0 | 85 | 15 |
| 5 | 80 | 20 |
| 10 | 72 | 28 |
| 30 | 40 | 60 |
| 36 | 100 |
Corn flower -3- rutinosides retention time is 13.5min
The content of Cyanidin -3-O- rutinosides in blackcurrant fruit is determined using HPLC methods, INSTRUMENT MODEL is
Agilent 2600；Using Zorbax SB C18 posts (250mm × 4.60mm, 5 μm)；UV-detector (VWD, G1314F), inspection
Survey wavelength is 520nm；Mobile phase A liquid is 1.6% aqueous formic acid, and B liquid is 1.6% formic acid acetonitrile solution；Column oven temperature 30；
DEG C sample size 10ul；Flow velocity 1ml/min.Carry out gradient elution：0-5min, 85%A liquid, 15%B liquid；5-10min, 80%A liquid,
20%B liquid；10-30min, 72%A liquid, 28%B liquid；30-36min, 100%B liquid.
As a result：Corn flower -3- rutinoside standard curve regression equations are：Y=2 × 107X-33120 (r=
0.9998), linear relationship is good in 0.1041~1.041 μ g ranges, and average recovery rate is respectively 96.4%, and Cyanidin-
3-O- rutinosides content range is 5.263~12.829mg/g.
The foregoing is only a specific embodiment of the invention, but protection scope of the present invention is not limited thereto, any
The change or replacement expected without creative work, should all be included within the scope of the present invention.Therefore, it is of the invention
Protection domain should be determined by the scope of protection defined in the claims.
Claims (1)
1. the HPLC assay methods of corn flower -3- rutinosides in a kind of blackcurrant, it is characterised in that comprise the following steps：
Step 1: ripe fresh blackcurrant fruit is chosen, fully squeezing mashing after cleaning up；
Step 2: step one gained slurries and blackcurrant fruit residue are fully extracted into corn flower -3- rutinosides using ethanol；
Step 3: the filtrate containing corn flower -3- rutinosides obtained by step 2 is utilized into high performance liquid chromatograph Agilent
2600, using Zorbax SB C18 posts；UV-detector：VWD, G1314F, Detection wavelength are 520nm；Mobile phase A liquid is chosen
The aqueous formic acid of concentration 1.6%, B liquid chooses the formic acid acetonitrile solution of concentration 1.6%；30 DEG C of column oven temperature；Sample size
10ul；Flow velocity 1ml/min, carries out gradient elution；The concrete operations of gradient elution are：0-5min, 85%A liquid, 15%B liquid；5-
10min, 80%A liquid, 20%B liquid；10-30min, 72%A liquid, 28%B liquid；30-36min, 100%B liquid；
Step 4: setting up corn flower -3- rutinoside standard curve regression equations, corn flower -3- rues according to gradient elution data
Fragrant glucosides standard curve regression equation is：Y=2 × 107X-33120, r=0.9998, in 0.1041~1.041 μ g ranges
Linear relationship is good, and average recovery rate is 96.4%, and corn flower -3- rutinosides content range is 5.263~12.829mg/g；
In the step 3, the design parameter of Zorbax SB C18 posts is 250mm × 4.60mm, 5 μm.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510142303.1A CN104749279B (en) | 2015-03-27 | 2015-03-27 | The HPLC assay methods of the rutinoside of corn flower 3 in blackcurrant |
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| CN201510142303.1A CN104749279B (en) | 2015-03-27 | 2015-03-27 | The HPLC assay methods of the rutinoside of corn flower 3 in blackcurrant |
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| CN104749279B true CN104749279B (en) | 2017-09-29 |
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| CN106290607B (en) * | 2016-07-25 | 2018-08-21 | 西南大学 | A method of quickly detecting Application of Mulberry Anthocyanins and flavones using UPLC-TUV |
| CN107525861A (en) * | 2017-07-28 | 2017-12-29 | 中国农业科学院郑州果树研究所 | The Sparklet testing method of anthocyanin and its application in a kind of apricot pericarp |
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Granted publication date: 20170929 Termination date: 20180327 |