CN110609006A - Method for determining carotenoid in capsanthin - Google Patents

Method for determining carotenoid in capsanthin Download PDF

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Publication number
CN110609006A
CN110609006A CN201910824904.9A CN201910824904A CN110609006A CN 110609006 A CN110609006 A CN 110609006A CN 201910824904 A CN201910824904 A CN 201910824904A CN 110609006 A CN110609006 A CN 110609006A
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China
Prior art keywords
solution
extracting solution
distilled water
volume
volumetric flask
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Withdrawn
Application number
CN201910824904.9A
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Chinese (zh)
Inventor
谈振永
张波
赵可建
崔波
王晶玲
胡桂川
胡乾辉
柏莹莹
于滨
卢艳敏
徐丙涛
马玲
李锐
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Yongren Yinong Food Co Ltd
Qilu University of Technology
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Yongren Yinong Food Co Ltd
Qilu University of Technology
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Application filed by Yongren Yinong Food Co Ltd, Qilu University of Technology filed Critical Yongren Yinong Food Co Ltd
Priority to CN201910824904.9A priority Critical patent/CN110609006A/en
Publication of CN110609006A publication Critical patent/CN110609006A/en
Withdrawn legal-status Critical Current

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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01Arrangements or apparatus for facilitating the optical investigation
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/25Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
    • G01N21/31Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
    • G01N21/33Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light

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  • Physics & Mathematics (AREA)
  • Spectroscopy & Molecular Physics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)

Abstract

The method for measuring carotenoid in capsanthin comprises the steps of extracting a reagent by using a mixed reagent of n-hexane, acetone, ethanol, diethyl ether and dichloromethane; the saponified solution is prepared by mixing potassium hydroxide, distilled water and methanol; the diluted solution comprises a sodium sulfate solution and a sodium chloride solution; the mobile phase is formed by mixing normal hexane and acetone, a sample is dissolved by extracting solution and then stands for layering, supernatant liquid is removed and added with the extracting solution, distilled water and saponification liquid, the mixture is placed in a water bath for saponification reaction, the saponification liquid is centrifugally separated in a centrifugal machine, finally the extracting solution is used as blank reference at the wavelength of 474nm of an ultraviolet visible spectrophotometer for comparison and zeroing, and the solution is poured into an absorption cell to read the light absorption value for calculation at the wavelength of 474 nm. The invention uses less instruments and equipment, only uses a photometer and a centrifuge, the instruments and equipment are simple to operate, the requirement on the operating skill of the method is looser, the relative error of the data detected by the method is smaller, the detected data is stable, and the numerical value can not float greatly.

Description

Method for determining carotenoid in capsanthin
Technical Field
The invention relates to the technical field of food detection, in particular to a method for determining carotenoid in capsanthin.
Background
The color substance of the pepper is mainly capsanthin. The capsanthin is carotenoid pigment existing in pepper, and accounts for 0.2-0.5% of pepper peel. Carotenoids are very important to human health, and mainly can prevent oxidation of 'bad' cholesterol, protect blood vessels and avoid plaque and vascular lesions. The existing carotenoid detection method mainly adopts a liquid chromatograph to test, and the liquid chromatograph can carry out quantitative detection on carotenoids with different structures, but the method has the following defects: the liquid chromatograph is a precise analytical instrument, the requirements on the working environment and the operating skill are very strict, and the deviation of the detection value is very easy to occur; the chromatographic column used has certain deviation compared with the prior chromatographic column after each replacement; the carotenoid standards used are costly; moreover, the whole process is complicated and tedious, and any small detail can cause a large error.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a method for measuring carotenoid in capsanthin.
The technical scheme adopted by the invention for solving the technical problems is as follows:
a method for determining carotenoid in capsanthin comprises the following steps of extracting a reagent by using a mixed reagent of n-hexane, acetone, ethanol, diethyl ether and dichloromethane, wherein the n-hexane: acetone: ethanol: diethyl ether: dichloromethane = 30:21:18:21:10 (volume ratio); the saponification liquid is prepared by mixing potassium hydroxide, distilled water and methanol, wherein the weight ratio of potassium hydroxide: distilled water: methanol =40:20:40 (mass: volume); the diluted solution comprises a sodium sulfate solution and a sodium chloride solution, wherein the sodium sulfate solution is prepared by using 30g of anhydrous sodium sulfate and making the volume of distilled water to 1000ml, and the sodium chloride solution is prepared by using 30g of sodium chloride and making the volume of distilled water to 1000 ml; the mobile phase is formed by mixing n-hexane and acetone according to the mixing mass ratio of 81:19, and the specific process steps are as follows:
weighing 0.1-0.4 g of sample in a 100ml brown volumetric flask, adding 80ml of extracting solution for dissolving, then fixing the volume to 100ml by using the extracting solution, standing and layering; transferring 15ml of supernatant into a 100ml volumetric flask, adding 15ml of extracting solution, 1ml of distilled water and 2ml of saponification solution with the concentration of 40%, performing ultrasonic treatment for 5 minutes, then placing the mixture in a water bath kettle at the temperature of 55-57 ℃ for saponification reaction for 20 minutes, and cooling; firstly taking 5ml of extracting solution, placing the extracting solution into a 250ml separating funnel, transferring the saponified solution into the separating funnel, then transferring 25ml of extracting solution, cleaning a saponified volumetric flask I, combining the volumetric flask I with the separating funnel, adding 40ml of sodium sulfate solution with the concentration of 3%, shaking, standing and layering; reducing the separated lower-layer water phase to separate into another 100ml separating funnel, adding 30ml of extracting solution, shaking, standing and layering, discarding the lower-layer liquid, separating the upper-layer liquid, washing the separating funnel with 5ml of extracting solution, then centrifuging for 1 minute in a centrifuge, and combining the upper-layer liquid for 2 times into the separating funnel; adding 50ml of 3% sodium sulfate solution into the supernatant, washing, shaking, standing for layering, and removing the lower layer liquid; then repeatedly washing the supernatant with distilled water for 3 times, placing the supernatant in a 100ml volumetric flask, washing a separating funnel with 5-10 ml of extracting solution, putting the washing solution into the volumetric flask, fixing the volume to 100ml with the extracting solution, sealing with a rubber stopper, and uniformly shaking; taking 2ml of the liquid to a 50ml volumetric flask, and fixing the volume by using the extracting solution; using the extracting solution as blank reference at the wavelength of 474nm of an ultraviolet-visible spectrophotometer to perform contrast zeroing, then pouring the diluted solution into an absorption cell to read the light absorption value at the wavelength of 474nm for calculation,
calculating the formula:
the invention uses less instruments and equipment, only uses 1 photometer and 1 centrifuge, the instrument and equipment are simple to operate, the requirement on the operating skill of the method is loose, the relative error of the detected data is small, the detected data is stable, and the numerical value can not float greatly.
Detailed Description
A method for determining carotenoid in capsanthin comprises the following steps of extracting a reagent by using a mixed reagent of n-hexane, acetone, ethanol, diethyl ether and dichloromethane, wherein the n-hexane: acetone: ethanol: diethyl ether: dichloromethane = 30:21:18:21:10 (volume ratio); the saponification liquid is prepared by mixing potassium hydroxide, distilled water and methanol, wherein the weight ratio of potassium hydroxide: distilled water: methanol =40:20:40 (mass: volume); the diluted solution comprises a sodium sulfate solution and a sodium chloride solution, wherein the sodium sulfate solution is prepared by using 30g of anhydrous sodium sulfate and making the volume of distilled water to 1000ml, and the sodium chloride solution is prepared by using 30g of sodium chloride and making the volume of distilled water to 1000 ml; the mobile phase is formed by mixing n-hexane and acetone according to the mixing mass ratio of 81:19, and the specific process steps are as follows:
weighing 0.1-0.4 g of sample in a 100ml brown volumetric flask, adding 80ml of extracting solution for dissolving, then fixing the volume to 100ml by using the extracting solution, standing and layering; transferring 15ml of supernatant into a 100ml volumetric flask, adding 15ml of extracting solution, 1ml of distilled water and 2ml of saponification solution with the concentration of 40%, performing ultrasonic treatment for 5 minutes, then placing the mixture in a water bath kettle at the temperature of 55-57 ℃ for saponification reaction for 20 minutes, and cooling; firstly taking 5ml of extracting solution, placing the extracting solution into a 250ml separating funnel, transferring the saponified solution into the separating funnel, then transferring 25ml of extracting solution, cleaning a saponified volumetric flask I, combining the volumetric flask I with the separating funnel, adding 40ml of sodium sulfate solution with the concentration of 3%, shaking, standing and layering; reducing the separated lower-layer water phase to separate into another 100ml separating funnel, adding 30ml of extracting solution, shaking, standing and layering, discarding the lower-layer liquid, separating the upper-layer liquid, washing the separating funnel with 5ml of extracting solution, then centrifuging for 1 minute in a centrifuge, and combining the upper-layer liquid for 2 times into the separating funnel; adding 50ml of 3% sodium sulfate solution into the supernatant, washing, shaking, standing for layering, and removing the lower layer liquid; then repeatedly washing the supernatant with distilled water for 3 times, placing the supernatant in a 100ml volumetric flask, washing a separating funnel with 5-10 ml of extracting solution, putting the washing solution into the volumetric flask, fixing the volume to 100ml with the extracting solution, sealing with a rubber stopper, and uniformly shaking; taking 2ml of the liquid to a 50ml volumetric flask, and fixing the volume by using the extracting solution; using the extracting solution as blank reference at the wavelength of 474nm of an ultraviolet-visible spectrophotometer to perform contrast zeroing, then pouring the diluted solution into an absorption cell to read the light absorption value at the wavelength of 474nm for calculation,
calculating the formula:

Claims (1)

1. a method for determining carotenoid in capsanthin is characterized in that an extraction reagent adopts a mixed reagent of n-hexane, acetone, ethanol, diethyl ether and dichloromethane, wherein the n-hexane: acetone: ethanol: diethyl ether: dichloromethane = 30:21:18:21:10 (volume ratio); the saponification liquid is prepared by mixing potassium hydroxide, distilled water and methanol, wherein the weight ratio of potassium hydroxide: distilled water: methanol =40:20:40 (mass: volume); the diluted solution comprises a sodium sulfate solution and a sodium chloride solution, wherein the sodium sulfate solution is prepared by using 30g of anhydrous sodium sulfate and making the volume of distilled water to 1000ml, and the sodium chloride solution is prepared by using 30g of sodium chloride and making the volume of distilled water to 1000 ml; the mobile phase is formed by mixing n-hexane and acetone according to the mixing mass ratio of 81:19, and the specific process steps are as follows:
weighing 0.1-0.4 g of sample in a 100ml brown volumetric flask, adding 80ml of extracting solution for dissolving, then fixing the volume to 100ml by using the extracting solution, standing and layering; transferring 15ml of supernatant into a 100ml volumetric flask, adding 15ml of extracting solution, 1ml of distilled water and 2ml of saponification solution with the concentration of 40%, performing ultrasonic treatment for 5 minutes, then placing the mixture in a water bath kettle at the temperature of 55-57 ℃ for saponification reaction for 20 minutes, and cooling; firstly taking 5ml of extracting solution, placing the extracting solution into a 250ml separating funnel, transferring the saponified solution into the separating funnel, then transferring 25ml of extracting solution, cleaning a saponified volumetric flask I, combining the volumetric flask I with the separating funnel, adding 40ml of sodium sulfate solution with the concentration of 3%, shaking, standing and layering; reducing the separated lower-layer water phase to separate into another 100ml separating funnel, adding 30ml of extracting solution, shaking, standing and layering, discarding the lower-layer liquid, separating the upper-layer liquid, washing the separating funnel with 5ml of extracting solution, then centrifuging for 1 minute in a centrifuge, and combining the upper-layer liquid for 2 times into the separating funnel; adding 50ml of 3% sodium sulfate solution into the supernatant, washing, shaking, standing for layering, and removing the lower layer liquid; then repeatedly washing the supernatant with distilled water for 3 times, placing the supernatant in a 100ml volumetric flask, washing a separating funnel with 5-10 ml of extracting solution, putting the washing solution into the volumetric flask, fixing the volume to 100ml with the extracting solution, sealing with a rubber stopper, and uniformly shaking; taking 2ml of the liquid to a 50ml volumetric flask, and fixing the volume by using the extracting solution; using the extracting solution as blank reference at the wavelength of 474nm of an ultraviolet-visible spectrophotometer to perform contrast zeroing, then pouring the diluted solution into an absorption cell to read the light absorption value at the wavelength of 474nm for calculation,
calculating the formula:
CN201910824904.9A 2019-09-02 2019-09-02 Method for determining carotenoid in capsanthin Withdrawn CN110609006A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113588577A (en) * 2021-07-30 2021-11-02 河北东之星生物科技股份有限公司 Method for detecting content of lutein in chrysanthemum granules
CN113624707A (en) * 2021-09-17 2021-11-09 云南博瑞生物科技有限公司 Quantitative detection method for radish red smell

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113588577A (en) * 2021-07-30 2021-11-02 河北东之星生物科技股份有限公司 Method for detecting content of lutein in chrysanthemum granules
CN113624707A (en) * 2021-09-17 2021-11-09 云南博瑞生物科技有限公司 Quantitative detection method for radish red smell

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