CN110609006A - Method for determining carotenoid in capsanthin - Google Patents
Method for determining carotenoid in capsanthin Download PDFInfo
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- 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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- distilled water
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- 238000000034 method Methods 0.000 title claims abstract description 16
- 235000021466 carotenoid Nutrition 0.000 title claims abstract description 13
- 150000001747 carotenoids Chemical class 0.000 title claims abstract description 13
- VYIRVAXUEZSDNC-TXDLOWMYSA-N (3R,3'S,5'R)-3,3'-dihydroxy-beta-kappa-caroten-6'-one Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC(=O)[C@]1(C)C[C@@H](O)CC1(C)C VYIRVAXUEZSDNC-TXDLOWMYSA-N 0.000 title claims abstract description 10
- VYIRVAXUEZSDNC-LOFNIBRQSA-N Capsanthyn Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC(=O)C2(C)CC(O)CC2(C)C VYIRVAXUEZSDNC-LOFNIBRQSA-N 0.000 title claims abstract description 10
- WRANYHFEXGNSND-LOFNIBRQSA-N capsanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC(=O)C2(C)CCC(O)C2(C)C WRANYHFEXGNSND-LOFNIBRQSA-N 0.000 title claims abstract description 10
- 235000018889 capsanthin Nutrition 0.000 title claims abstract description 10
- 235000012658 paprika extract Nutrition 0.000 title claims abstract description 10
- 239000001688 paprika extract Substances 0.000 title claims abstract description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 239000007788 liquid Substances 0.000 claims abstract description 24
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims abstract description 21
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 21
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims abstract description 21
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims abstract description 21
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims abstract description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 20
- 239000012153 distilled water Substances 0.000 claims abstract description 20
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims abstract description 16
- 229910052938 sodium sulfate Inorganic materials 0.000 claims abstract description 13
- 235000011152 sodium sulphate Nutrition 0.000 claims abstract description 13
- 239000006228 supernatant Substances 0.000 claims abstract description 13
- 238000007127 saponification reaction Methods 0.000 claims abstract description 12
- 239000011780 sodium chloride Substances 0.000 claims abstract description 10
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 8
- 238000010521 absorption reaction Methods 0.000 claims abstract description 4
- 230000031700 light absorption Effects 0.000 claims abstract description 4
- 239000000203 mixture Substances 0.000 claims abstract description 4
- 238000000870 ultraviolet spectroscopy Methods 0.000 claims abstract description 4
- 238000005406 washing Methods 0.000 claims description 15
- 238000004140 cleaning Methods 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 238000009210 therapy by ultrasound Methods 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 238000000605 extraction Methods 0.000 claims 1
- 238000001514 detection method Methods 0.000 description 4
- 235000002566 Capsicum Nutrition 0.000 description 3
- 239000006002 Pepper Substances 0.000 description 3
- 241000722363 Piper Species 0.000 description 3
- 235000016761 Piper aduncum Nutrition 0.000 description 3
- 235000017804 Piper guineense Nutrition 0.000 description 3
- 235000008184 Piper nigrum Nutrition 0.000 description 3
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000216 vascular lesion Toxicity 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating 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
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:。
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Cited By (2)
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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2019
- 2019-09-02 CN CN201910824904.9A patent/CN110609006A/en not_active Withdrawn
Cited By (2)
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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