CN110609006A - Determination method of carotenoids in capsanthin - Google Patents

Abstract

The determination method of carotenoids in capsanthin, the extraction reagent is a mixed reagent of n-hexane, acetone, ethanol, ether and dichloromethane; the saponification solution is made by mixing potassium hydroxide, distilled water and methanol; the dilution solution includes sodium sulfate solution and sodium chloride solution; the mobile phase is a mixture of n-hexane and acetone, the sample is dissolved in the extract and allowed to stand for stratification, the supernatant is taken out and the extract, distilled water and saponification solution are added, and placed in a water bath for saponification reaction , the saponification solution is centrifuged in a centrifuge, and finally the extract is used as a blank reference comparison at a wavelength of 474nm in a UV-visible spectrophotometer to set zero, and the solution is poured into an absorbance cell to read the absorbance value at a wavelength of 474nm for calculation. The present invention uses less instruments and equipment, only photometers and centrifuges are used, the instruments and equipment are easy to operate, the method requires relatively loose operating skills, the relative error of the data detected by this method is small, the detection data is stable, and the value will not appear large of float.

Description

Determination method of carotenoids in capsanthin

technical field

The invention relates to the technical field of food detection, in particular to a method for determining carotenoids in capsanthin.

Background technique

The chromogenic substance of pepper is mainly capsanthin. Capsanthin is a carotenoid pigment present in peppers, accounting for 0.2%-0.5% of pepper peels. Carotenoids are very important to human health, mainly to prevent the oxidation of "bad" cholesterol, protect blood vessels, and avoid plaque and vascular lesions. At present, the detection method of carotenoids is mainly based on liquid chromatography, which can quantitatively detect carotenoids with different structures, but it has the following defects: liquid chromatography is a precision analytical instrument, and the working environment and operation The technical requirements are very strict, and the deviation of the detection value is very easy to appear; the chromatographic column used will have a certain deviation compared with the previous chromatographic column after each replacement; the cost of the carotenoid standard used is high; and the whole process is complicated and cumbersome. Any small detail may lead to large errors.

Contents of the invention

Aiming at the shortcomings of the prior art, the invention provides a method for determining carotenoids in capsanthin.

The technical solution adopted by the present invention to solve its technical problems is:

A kind of assay method of carotenoid in capsanthin, extraction reagent adopts the mixed reagent of n-hexane, acetone, ethanol, ether and dichloromethane, n-hexane: acetone: ethanol: ether: dichloromethane=30:21:18 :21:10 (volume ratio); saponification solution is made by mixing potassium hydroxide, distilled water and methanol, among which potassium hydroxide: distilled water: methanol = 40:20:40 (mass: volume: volume); diluting solution includes sodium sulfate solution and sodium chloride solution, the sodium sulfate solution is made of 30g anhydrous sodium sulfate by distilled water to 1000ml, and the sodium chloride solution is made by 30g sodium chloride by distilled water to 1000ml; the mobile phase is made of n-hexane and acetone Mixed, the mixing mass ratio of n-hexane and acetone is 81:19, and the specific process steps are as follows:

Weigh 0.1-0.4g sample into a 100ml brown volumetric flask, add 80ml of extract to dissolve, then use the extract to make up to 100ml, let it stand for stratification; pipette 15ml of supernatant into a 100ml volumetric flask, add 15ml of extract, 1ml Distilled water, 2ml saponification solution with a concentration of 40%, ultrasonication for 5 minutes, then saponification reaction in a water bath at 55°C-57°C for 20 minutes, cooling; first take 5ml extract solution and place it in a 250ml separatory funnel, then put the above saponification solution Transfer to a separatory funnel, then pipette 25ml of extract to clean the saponification volumetric flask and combine them into the separatory funnel, add 40ml of 3% sodium sulfate solution, shake and stand for stratification; reduce the lower layer of water after stratification Into another 100ml separating funnel, add 30ml of extracting solution, after shaking and standing for stratification, discard the lower layer, separate the upper layer and wash the separating funnel with 5ml of extracting solution, then centrifuge at 2500 rpm in a centrifuge for 1 Minutes, combine the upper layer liquid for 2 times into the separatory funnel; add 50ml of 3% sodium sulfate solution to the above supernatant liquid to wash and shake, then let it stand for stratification, discard the lower layer liquid; then use distilled water to wash the upper layer liquid repeatedly 3 times , put the upper liquid in a 100ml volumetric flask, and use 5-10ml of the extract to clean the separatory funnel and put the cleaning solution into the volumetric flask, use the extract to make up to 100ml, seal it with a rubber stopper and shake it evenly; then take 2ml of the above liquid to 50ml In the volumetric flask, use the extract to make up the volume; use the extract to make a blank reference comparison at a wavelength of 474nm in the UV-Vis spectrophotometer and set it to zero, then pour the diluted solution into an absorbance cell and read the absorbance at a wavelength of 474nm for calculation.

Calculation formula:

The present invention uses less instruments and equipment, only uses one photometer and one centrifuge, the instruments and equipment are easy to operate, and the method requires relatively loose operating skills, the relative error of the data detected by the method is small, the detection data is stable, and the numerical value There will be no large fluctuations.

Detailed ways

A kind of assay method of carotenoid in capsanthin, extraction reagent adopts the mixed reagent of n-hexane, acetone, ethanol, ether and dichloromethane, n-hexane: acetone: ethanol: ether: dichloromethane=30:21:18 :21:10 (volume ratio); saponification solution is made by mixing potassium hydroxide, distilled water and methanol, among which potassium hydroxide: distilled water: methanol = 40:20:40 (mass: volume: volume); diluting solution includes sodium sulfate solution and sodium chloride solution, the sodium sulfate solution is made of 30g anhydrous sodium sulfate by distilled water to 1000ml, and the sodium chloride solution is made by 30g sodium chloride by distilled water to 1000ml; the mobile phase is made of n-hexane and acetone Mixed, the mixing mass ratio of n-hexane and acetone is 81:19, and the specific process steps are as follows:

Weigh 0.1-0.4g sample into a 100ml brown volumetric flask, add 80ml of extract to dissolve, then use the extract to make up to 100ml, let it stand for stratification; pipette 15ml of supernatant into a 100ml volumetric flask, add 15ml of extract, 1ml Distilled water, 2ml saponification solution with a concentration of 40%, ultrasonication for 5 minutes, then saponification reaction in a water bath at 55°C-57°C for 20 minutes, cooling; first take 5ml extract solution and place it in a 250ml separatory funnel, then put the above saponification solution Transfer to a separatory funnel, then pipette 25ml of extract to clean the saponification volumetric flask and combine them into the separatory funnel, add 40ml of 3% sodium sulfate solution, shake and stand for stratification; reduce the lower layer of water after stratification Into another 100ml separating funnel, add 30ml of extracting solution, after shaking and standing for stratification, discard the lower layer, separate the upper layer and wash the separating funnel with 5ml of extracting solution, then centrifuge at 2500 rpm in a centrifuge for 1 Minutes, combine the upper layer liquid for 2 times into the separatory funnel; add 50ml of 3% sodium sulfate solution to the above supernatant liquid to wash and shake, then let it stand for stratification, discard the lower layer liquid; then use distilled water to wash the upper layer liquid repeatedly 3 times , put the upper liquid in a 100ml volumetric flask, and use 5-10ml of extract to clean the separatory funnel and put the cleaning solution into the volumetric flask, use the extract to make up to 100ml, seal the rubber stopper and shake evenly; then take 2ml of the above liquid to 50ml In the volumetric flask, use the extract to make up the volume; use the extract at 474nm wavelength of the UV-Vis spectrophotometer as a blank reference comparison and set to zero, then pour the diluted solution into the light absorption cell and read the absorbance value at 474nm wavelength to calculate,

Calculation formula: .

Claims (1)

1. a kind of assay method of carotenoid in capsanthin, it is characterized in that extraction reagent adopts the mixed reagent of n-hexane, acetone, ethanol, ether and methylene dichloride, n-hexane: acetone: ethanol: ether: methylene dichloride= 30:21:18:21:10 (volume ratio); saponified solution is made by mixing potassium hydroxide, distilled water and methanol, where potassium hydroxide: distilled water: methanol = 40:20:40 (mass: volume: volume); Dilute solution comprises sodium sulfate solution and sodium chloride solution, and sodium sulfate solution adopts 30g anhydrous sodium sulfate to be made by distilled water to 1000ml, and sodium chloride solution adopts 30g sodium chloride to be made by distilled water to 1000ml; mobile phase It is made by mixing n-hexane and acetone. The mixing mass ratio of n-hexane and acetone is 81:19. The specific process steps are as follows: Weigh 0.1-0.4g sample into a 100ml brown volumetric flask, add 80ml of extract to dissolve, then use the extract to make up to 100ml, let it stand for stratification; pipette 15ml of supernatant into a 100ml volumetric flask, add 15ml of extract, 1ml Distilled water, 2ml saponification solution with a concentration of 40%, ultrasonication for 5 minutes, then saponification reaction in a water bath at 55°C-57°C for 20 minutes, cooling; first take 5ml extract solution and place it in a 250ml separatory funnel, then put the above saponification solution Transfer to a separatory funnel, then pipette 25ml of extract to clean the saponification volumetric flask and combine them into the separatory funnel, add 40ml of 3% sodium sulfate solution, shake and stand for stratification; reduce the lower layer of water after stratification Into another 100ml separating funnel, add 30ml of extracting solution, after shaking and standing for stratification, discard the lower layer, separate the upper layer and wash the separating funnel with 5ml of extracting solution, then centrifuge at 2500 rpm in a centrifuge for 1 Minutes, combine the upper layer liquid for 2 times into the separatory funnel; add 50ml of 3% sodium sulfate solution to the above supernatant liquid to wash and shake, then let it stand for stratification, discard the lower layer liquid; then use distilled water to wash the upper layer liquid repeatedly 3 times , put the upper liquid in a 100ml volumetric flask, and use 5-10ml of extract to clean the separatory funnel and put the cleaning solution into the volumetric flask, use the extract to make up to 100ml, seal the rubber stopper and shake evenly; then take 2ml of the above liquid to 50ml In the volumetric flask, use the extract to make up the volume; use the extract at 474nm wavelength of the UV-Vis spectrophotometer as a blank reference comparison and set to zero, then pour the diluted solution into the light absorption cell and read the absorbance value at 474nm wavelength to calculate, Calculation formula: .