CN107907498A - The detection method of bata-carotene content in a kind of green grass or young crops cumquat fruit powder - Google Patents

The detection method of bata-carotene content in a kind of green grass or young crops cumquat fruit powder Download PDF

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CN107907498A
CN107907498A CN201711040202.9A CN201711040202A CN107907498A CN 107907498 A CN107907498 A CN 107907498A CN 201711040202 A CN201711040202 A CN 201711040202A CN 107907498 A CN107907498 A CN 107907498A
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carotene
fruit powder
beta
content
sample
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刘红
马金爽
张晓青
何云啸
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Hainan Normal University
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Hainan Normal University
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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
    • G01N21/17—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
    • G01N21/35—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light
    • G01N21/3563—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using infrared light for analysing solids; Preparation of samples therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2201/00—Features of devices classified in G01N21/00
    • G01N2201/12—Circuits of general importance; Signal processing
    • G01N2201/129—Using chemometrical methods

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Abstract

The invention belongs to bata-carotene content detection technical field, disclose a kind of detection method that bata-carotene content in blue or green cumquat fruit powder is measured using near infrared spectroscopy, first using the content of bata-carotene in conventional chemical gauging sample, the spectral value of blue or green cumquat fruit powder sample is being measured using near infrared spectrometer.Spectroscopic data and chemical score are combined by Chemical Measurement, modeling, cross validation and forecast analysis, show that RMSE is respectively less than 0.01, effective coefficient of determination R numerical value is between 0.9055~0.966, illustrate that testing result is more accurate, actual value and predicted value correlation are good.

Description

The detection method of content beta-carotene in a kind of green grass or young crops cumquat fruit powder
Technical field
The invention belongs to beta carotene in content beta-carotene detection technique field, more particularly to a kind of blue or green cumquat fruit powder The detection method of content.
Background technology
Blue or green cumquat (lime), originates in China Hainan and Vietnam more, is a kind of evergreen shrubs.Though its volume is small, it is rich in A variety of nutriments such as vitamin C, A, P and carotenoid, there is very powerful medicinal dietotherapy effect, and blue or green cumquat is drying into powder Main edible way, for probe into it is drying after blue or green cumquat powder nutritive value, carotenoid (Carotenoids) is a kind of The general name of natural pigment, belongs to terpene compound and is prevalent in animal, plant, fungi, algae and bacterium, be common in food In carotenoids be known as many as 50~60 kinds, such as beta carotene, lutein, zeaxanthin, kryptoxanthin, astacin and tomato red Element etc..Since carotenoid in Medicines and Health Product, Fine Chemical and feedstuff industry has all multipurposes, it is faced with Substantial amounts of raw materials requirement, so as to trigger the fine market prospects of carotenoid.Carotenoid annual volume is within 2004 8.87 hundred million dollars, it is contemplated that annual average rate of increase is up to 2.9%, will be more than 1,000,000,000 dollars to market scale in 2009 ---《State Inside and outside carotenoid market and Research progress》.Carotenoid is by the how unsaturated of its own so very unstable, chance Light, acid, oxygen and high temperature etc. are degradable, therefore to control these influence factors as far as possible during extraction.Carotenoid has lipophilicity, Water is practically insoluble in, is soluble in organic solvent, therefore generally use organic solvent extraction carotenoid.Required by this method Equipment is simple, and operation is easy, but time-consuming.Secondly ultrasonic extraction, its extraction pigment used time is short, mild condition, but extracts Take process unclear to the extent of the destruction of pigment.In addition, supercritical fluid extraction is also a kind of high selectivity and automation behaviour Make the higher sample preparation methods of degree, and extract organic solvent-free remains, but quantity of sample handling is small, and equipment cost is high, Therefore also there is application in the extraction of carotenoid.Near-infrared spectral analysis technology has developed most since being the 1990s Fast and most noticeable spectral analysis technique, has quick, efficient, pollution-free, unique points such as sampling is simple and lossless Analyse advantage.The fruit powder fast quantitative analysis model for the spectrometry that diffuses based on near-infrared is established using radar stealthy materials, is realized to fruit powder On-line checking.The content of vitamin is directly measured using the near-infrared spectral technology that diffuses, benefit be sample without pretreatment, and Method is easy, reliable results.
In conclusion problem existing in the prior art is：The side of content beta-carotene in the existing blue or green cumquat fruit powder of detection Method, complicated, time-consuming, of high cost, destroys sample etc..And near-infrared is the detection of quick nondestructive developed in recent years Method, and it is easy to operate, the time is short, can realize on-line checking.
The content of the invention
In view of the problems of the existing technology, the present invention provides a kind of inspection of content beta-carotene in blue or green cumquat fruit powder Survey method.
The present invention is achieved in that content beta-carotene in a kind of blue or green cumquat fruit powder using near infrared spectroscopy measure Detection method, first with the content of beta carotene in conventional chemical method detection sample, utilizing near-infrared determination sample Spectral value, is afterwards being combined spectral value and chemical score by Chemical Measurement, and beta carotene is near in the blue or green cumquat fruit powder of foundation Infrared spectrum analysis model, and pls verifications are carried out to model, draw the prediction model of blue or green cumquat fruit powder beta carotene, realization pair The quick detection of content beta-carotene in blue or green cumquat；Wherein, RMSE is less than 1%, and effective coefficient of determination R numerical value is between 0.9055 Between~0.966, illustrate that testing result is more accurate, model is better.
Further, the detection method of content beta-carotene weighs the beta carotene mark of 0.005g in the blue or green cumquat fruit powder Quasi- sample, makes it be dissolved in 60 DEG C -90 DEG C of petroleum ether, and for constant volume in the volumetric flask of 10mL browns, it is 0.50mg/ to obtain concentration The standard solution of mL, step by step dilution are configured to the standard solution of various concentrations；
The standard solution of above 0.050mg/mL is taken, is to be scanned in 350~600nm in wave-length coverage, measures β-carrot The abosrption spectrogram Detection wavelength selection 450nm of plain standard sample.
Further, the drafting of the standard curve includes：0.04,0.08,0.12,0.16,0.20 and is measured respectively The beta carotene standard items of the 0.05mg/mL of 0.24mL in 25mL brown volumetric flasks, are added chloroform and are settled to scale using liquid Line, using water as blank, carries out the measure of absorbance in the spectrophotometer that wavelength is 450nm respectively；The recurrence of standard curve Equation is A=0.3089 ρ -0.0088；r2=0.9995.
Further, the detection method of content beta-carotene comprises the following steps in the blue or green cumquat fruit powder：
Step 1, takes blue or green cumquat fruit powder in 50mL beakers respectively, be respectively placed in 25 DEG C, 30 DEG C, 40 DEG C, 50 DEG C, 60 In DEG C environment, sample is taken out every 0.5h and is measured；
Step 2, measures 5.0g green grass or young crops cumquat fruit powders and is placed in conical flasks of the 100mL with glass stopper, add the acetone prepared Petroleum ether 3:7 mixed liquors, cover static 5min after glass stopper shaking 1min, extracting solution are transferred to equipped with 5% anhydrous sulphur of 100mL In the separatory funnel of acid sodium solution, with 10mL acetone-petroleum ethers 3:7 mixed liquors continue extraction 2-3 time, up to extracting solution is to colourless Untill, it is static, discard lower aqueous solution after solution layering, then washed for several times with 5% anhydrous sodium sulfate of 15mL, to no longer muddy Untill turbid；
Step 3, extracting solution is transferred in the 500mL aryballos supporting with rotating additional issue device, rotary spherical bottle is connect, 55 Depressurized in DEG C water-bath and be distilled to recover petroleum ether, until removing aryballos not when there is drop to drip in condenser pipe, adding immediately 2mL petroleum ethers carry out constant volume；
Step 4, a baseline is done at away from filter paper lower end 4cm, and equidistant 4 points of A, B, C, D is taken on baseline；With Digitalized electric pipettor takes the sample concentration liquid of 0.4mL in AB point-to-point transmissions and CD point-to-point transmissions band application back and forth rapidly, until point It is complete；After sampling liquid on filter paper evaporates into do completely, filter paper is rolled into tubular, is inserted at baseline positioned at low side and fills petroleum ether It is unfolded in the chromatography cylinder that saturation is crossed；After beta carotene and other pigments are completely isolated, filter paper is taken out, treats petroleum ether Evaporate into completely after doing, cut the carrotene chromatographic band positioned at solvent the top with scissors, chromatographic band is cut into fragment immediately It is put into the tool match test tube for filling 5mL petroleum ethers, firmly shakes, until carrotene is completely dissolved in petroleum ether；
Sample eluent, is carried out the measure of absorbance, in standard by step 5 in the case where wavelength is the spectrophotometer of 450nm The content of corresponding beta carotene is found out on curve；
Step 6, the relevance model of near infrared spectrum and sample actual value is established using micro- with the method for Chemical Measurement The scope of type near infrared spectrum scanning wavelength is 900~1700nm, and Unscrambler analysis near infrared spectrums and spectrophotometric are real The relation of data is tested, is modeled by PLS, cross validation and forecast analysis；Effective coefficient of determination R2Close to 1, model is better；
Step 7, scans actual blue or green cumquat fruit powder, its actual carotene carotene content is calculated according to model.
The near infrared spectrometer technology of the present invention with its is lossless, conveniently, immediately, stablize, accuracy rate is high the features such as, become 20 One of most rapid detection technique is developed since the nineties in century, is widely used in including agriculture and animal husbandry, food, chemical industry, pharmacy, cigarette Many fields including grass etc..The a collection of sample (i.e. modeling collection) for being used to establish model is collected first, and sample must have certain Representativeness；Secondly, sample is measured with the near infrared spectrum of near infrared spectroscopy instrument collecting sample and with chemical analysis method Actual value；Then, the relevance model of near infrared spectrum and sample actual value is established with the method for Chemical Measurement；Finally, The mathematical model prediction unknown sample established with oneself.Using spectrophotometry, beta carotene is carried out under 450 nanometers of wavelength The measure of content；The scope of miniature near infrared spectrum scanning wavelength is 900~1700nm, and Unscrambler analyzes near infrared light Spectrum and the relation of spectrophotometric experimental data, by modeling, cross validation and forecast analysis, show that RMSE is average 1%, effectively Coefficient of determination R2 numerical value illustrates that testing result is more accurate between 0.8 to 0.95, and actual value and predicted value correlation are good It is good.
Brief description of the drawings
Fig. 1 is the detection method flow chart of content beta-carotene in blue or green cumquat fruit powder provided in an embodiment of the present invention.
Fig. 2 is the near infrared spectrum schematic diagram of beta carotene provided in an embodiment of the present invention.
Fig. 3 is the standard curve schematic diagram of beta carotene provided in an embodiment of the present invention.
Fig. 4 is the PLS model schematics of content beta-carotene provided in an embodiment of the present invention.
Fig. 5 is the near infrared spectrum schematic diagram of blue or green cumquat fruit powder provided in an embodiment of the present invention.
Fig. 6 is the PLS verification model schematics of content beta-carotene provided in an embodiment of the present invention.
Fig. 7 is the PLS prediction model schematic diagrames of content beta-carotene provided in an embodiment of the present invention.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to Limit the present invention.
The application principle of the present invention is explained in detail below in conjunction with the accompanying drawings.
As shown in Figure 1, in green grass or young crops cumquat fruit powder provided in an embodiment of the present invention the detection method of content beta-carotene include with Lower step：
S101：Blue or green cumquat fruit powder is taken to be respectively placed in 25 DEG C, 30 DEG C, 40 DEG C, 50 DEG C, 60 DEG C in 50mL beakers respectively In environment, sample is taken out every 0.5h and is measured；
S102：Measure 5.0g green grass or young crops cumquat fruit powders to be placed in conical flasks of the 100mL with glass stopper, add the acetone stone prepared Oily ether (3:7) mixed liquor, covers static 5min after glass stopper shaking 1min, extracting solution is transferred to equipped with 5% anhydrous sulphur of 100mL In the separatory funnel of acid sodium solution, with 10mL acetone-petroleum ethers (3:7) mixed liquor continues extraction 2-3 times, up to extracting solution to nothing It is static untill color, lower aqueous solution is discarded after solution layering, then washed for several times with 5% anhydrous sodium sulfates of 15mL, to no longer Untill muddiness；
S103：Extracting solution is transferred in the 500mL aryballos supporting with rotating additional issue device, rotary spherical bottle is connect, at 55 DEG C Depressurized in water-bath and be distilled to recover petroleum ether, until removing aryballos not when there is drop to drip in condenser pipe, adding immediately 2mL petroleum ethers carry out constant volume；
S104：A baseline is done at away from filter paper lower end 4cm, equidistant 4 points of A, B, C, D is taken on baseline；With number Word electric pipettor takes the sample concentration liquid of 0.4mL in AB point-to-point transmissions and CD point-to-point transmissions band application back and forth rapidly, until having put； After sampling liquid on filter paper evaporate into completely it is dry after, filter paper is rolled into tubular, is inserted at baseline positioned at low side and is filled petroleum ether and satisfy It is unfolded with chromatography cylinder excessively；After beta carotene and other pigments are completely isolated, filter paper is taken out, treats that petroleum ether is complete Evaporate into entirely after doing, cut the carrotene chromatographic band positioned at solvent the top with scissors, chromatographic band is cut into fragment puts immediately Enter to the tool for filling 5mL petroleum ethers and match in test tube, firmly shake, until carrotene is completely dissolved in petroleum ether；
S105：Sample eluent is carried out to the measure of absorbance in the case where wavelength is the spectrophotometer of 450nm, it is bent in standard The content of corresponding beta carotene is found out on line；
S106：The relevance model of near infrared spectrum and sample actual value is established using miniature with the method for Chemical Measurement The scope of near infrared spectrum scanning wavelength is 900~1700nm, and Unscrambler analysis near infrared spectrums are tested with spectrophotometric The relation of data, is modeled, cross validation and forecast analysis by PLS；Effective coefficient of determination R2Close to 1, model is better；
S107：Actual blue or green cumquat fruit powder is scanned, its actual carotene carotene content is calculated according to model.
The application principle of the present invention is further described with reference to experiment.
1 experimental section
1.1 laboratory apparatus, material and reagent
1.1.1 laboratory apparatus
1.1.2 experiment material
Blue or green cumquat original powder South of Hainan sends Industrial Co., Ltd.
1.1.3 experiment reagent
The content of beta carotene in the blue or green cumquat fruit powder of 2 chemical methodes detection
2.1 survey its light absorption value using spectrophotometer to the standard beta carotene of various concentrations, make concentration C and extinction The standard curve of angle value A.Thin layer is used from blue or green cumquat powder by the feature that beta carotene polarity is minimum, development rate is most fast It is extracted constant volume with other pigment separateds and surveys its absorbance by chromatography, its mass concentration is found out by standard curve.
The accurate beta carotene standard sample for weighing 0.005g, makes it be dissolved in 60~90 DEG C of petroleum ether, constant volume in In the volumetric flask of 10mL browns, the standard solution that concentration is 0.50mg/mL is obtained, dilution step by step is configured to the standard of various concentrations Solution.
The standard solution of above 0.050mg/mL is taken, is to be scanned in 350~600nm in wave-length coverage, measures β-carrot The abosrption spectrogram (Fig. 2) of plain standard sample, as seen from Figure 2 beta carotene standard sample have at 450nm and 475nm A absworption peak two-by-two, wherein have maximum absorbance value at 450nm, therefore Detection wavelength selection 450nm.
The drafting of 2.2 standard curves
It is accurate respectively measure 0.04,0.08,0.12,0.16,0.20, the beta carotene standard of the 0.05mg/mL of 0.24mL Product in 25mL brown volumetric flasks, are added chloroform and are settled to graduation mark, be respectively 450nm in wavelength using water as blank using liquid Spectrophotometer in carry out absorbance measure.Such as table 1..Gained standard curve such as Fig. 3, the regression equation of standard curve is A =0.3089 ρ -0.0088；r2=0.9995。
2.3 experimental procedure
2.3.1 processing early period of sample
Appropriate blue or green cumquat fruit powder is taken to be respectively placed in 25 DEG C, 30 DEG C, 40 DEG C, 50 DEG C, 60 DEG C of rings in 50mL beakers respectively In border, sample is taken out every 0.5h and is measured.
2.3.2 the preparation of sample
The accurate 5.0g green grass or young crops cumquat fruit powders that measure are placed in conical flasks of the 100mL with glass stopper, add the acetone stone prepared Oily ether (3:7) mixed liquor, covers static 5min after glass stopper shaking 1min, extracting solution is transferred to equipped with 5% anhydrous sulphur of 100mL In the separatory funnel of acid sodium solution, with 10mL acetone-petroleum ethers (3:7) mixed liquor continues extraction 2-3 times, up to extracting solution to nothing It is static untill color, discard lower aqueous solution after solution layering, then washed for several times with 15mL5% anhydrous sodium sulfates, to no longer muddy Untill turbid.
2.3.3 constant volume
Extracting solution is transferred in the 500mL aryballos supporting with rotating additional issue device, rotary evaporator is connect, in 55 DEG C of water-baths Depressurize and be distilled to recover petroleum ether, until removing evaporator not when there is drop to drip in condenser pipe, add 2mL oil immediately Ether carries out constant volume.
2.3.4 paper chromatography
A baseline is done at away from filter paper lower end 4cm, equidistant 4 points of A, B, C, D is taken on baseline.With digital electricity Dynamic pipettor takes the sample concentration liquid of 0.4mL in AB point-to-point transmissions and CD point-to-point transmissions band application back and forth rapidly, until having put.It is to be filtered On paper sampling liquid evaporate into completely it is dry after, filter paper is rolled into tubular, is inserted at baseline positioned at low side and fills petroleum ether saturation mistake Chromatography cylinder in be unfolded.After beta carotene and other pigments are completely isolated, filter paper is taken out, treats that petroleum ether is waved completely It is sent to after doing, cuts the carrotene chromatographic band positioned at solvent the top with scissors, chromatographic band is cut into fragment is immediately placed in In the tool match test tube for filling 5mL petroleum ethers, firmly shake, until carrotene is completely dissolved in petroleum ether.
The measure of 2.4 samples
Sample eluent is carried out to the measure of absorbance in the case where wavelength is the spectrophotometer of 450nm, on standard curve Find out the content of corresponding beta carotene.
2.5 interpretation of result
Following table is the content (chemical score) of beta carotene at different conditions in blue or green cumquat powder
Under 1 different condition of table in blue or green cumquat fruit powder beta carotene content
3rd, near infrared spectroscopy measures the content of beta carotene in blue or green cumquat fruit powder
The processing of 3.1 samples
Appropriate amount of sample is taken to carry out near infrared spectrum scanning under conditions of being placed in different temperatures different time, setting wavelength is 900~1700nm, resolution ratio 10.53, number of scan points are 400 points, are scanned 5 times.
3.2 samples establish regression model and cross validation
Extract 20 reagent samples from 63 laboratory samples at random to be modeled, remaining 43 test specimens are used for handing over Fork verification.Blue or green cumquat fruit powder atlas of near infrared spectra under different temperatures is as shown in the figure, beta carotene therein establishes model such as Shown in figure, cross validation model is as Figure 4-Figure 7.
The most fast and most noticeable spectrum analysis that near-infrared spectral analysis technology has developed since being the 1990s Technology.The technical Analysis sample have easily and fast, efficiently, it is accurately relatively low with cost, do not destroy sample, do not consume chemical examination Agent, it is free from environmental pollution the advantages that.Contain substantial amounts of carotenoid in blue or green cumquat, since carotenoid is in Medicines and Health Product, essence Thin chemical industry and feedstuff industry have all multipurposes, are faced with substantial amounts of raw materials requirement, so as to trigger carotenoid beautiful Good market prospects.The traditional detection method of use -- spectrophotometer method measures the content of beta carotene, the drawback is that time-consuming Long, existence time is poor between sample detection and data feedback, samples poor repeatability, and human factor can also influence result.Near infrared light Spectral technology the sixth of the twelve Earthly Branches becomes the hot spot of food service industry concern, which has been used for former material, auxiliary material, intermediate quality testing, powder production Process on-line checking and finished product original position are without destructive analysis.And the near infrared spectroscopy that this experiment uses, by experiment From the point of view of the modeling analysis of data, cross validation and prediction result, RMSE below 1%, effective coefficient of determination R numerical value between Between 0.9055~0.966, illustrate that testing result is more accurate, actual value and predicted value correlation are good.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention All any modification, equivalent and improvement made within refreshing and principle etc., should all be included in the protection scope of the present invention.

Claims (4)

  1. A kind of 1. detection method of content beta-carotene in green grass or young crops cumquat fruit powder, it is characterised in that β-Hu in the green grass or young crops cumquat fruit powder The detection method of radish cellulose content measures the detection of content beta-carotene in blue or green cumquat fruit powder using near infrared spectroscopy, first with normal The content of beta carotene, in the spectral value using near-infrared determination sample, is passing through afterwards in the chemical method detection sample of rule Chemical Measurement combines spectral value and chemical score, establishes the NIR Spectroscopy Analysis Model of beta carotene in blue or green cumquat fruit powder, And model is verified, draw the prediction model of blue or green cumquat fruit powder beta carotene, realize and beta carotene in blue or green cumquat is contained The quick detection of amount；Wherein, RMSE is lower, R2Value illustrates that testing result is more accurate, model is better closer to 1.
  2. 2. the detection method of content beta-carotene in green grass or young crops cumquat fruit powder as claimed in claim 1, it is characterised in that the green grass or young crops The detection method of content beta-carotene weighs the beta carotene standard sample of 0.005g in cumquat fruit powder, it is dissolved in 60~90 DEG C petroleum ether in, constant volume in the volumetric flask of 10mL browns, obtain concentration be 0.50mg/mL standard solution, dilute step by step It is configured to the standard solution of various concentrations；
    The standard solution of above 0.050mg/mL is taken, is to be scanned in 350~600nm in wave-length coverage, measures beta carotene mark The abosrption spectrogram Detection wavelength selection 450nm of quasi- sample.
  3. 3. the detection method of content beta-carotene in green grass or young crops cumquat fruit powder as claimed in claim 1, it is characterised in that the mark The drafting of directrix curve includes：Measure 0.04 respectively, 0.08,0.12,0.16,0.20, β-carrot of the 0.05mg/mL of 0.24mL Plain standard items in 25mL brown volumetric flasks, are added chloroform and are settled to graduation mark, using water as blank, be in wavelength respectively using liquid The measure of absorbance is carried out in the spectrophotometer of 450nm；The regression equation of standard curve is A=0.3089 ρ -0.0088；r2 =0.9995.
  4. 4. the detection method of content beta-carotene in green grass or young crops cumquat fruit powder as claimed in claim 1, it is characterised in that the green grass or young crops The detection method of content beta-carotene comprises the following steps in cumquat fruit powder：
    Step 1, takes blue or green cumquat fruit powder in 50mL beakers respectively, is respectively placed in 25 DEG C, 30 DEG C, 40 DEG C, 50 DEG C, 60 DEG C of rings In border, sample is taken out every 0.5h and is measured；
    Step 2, measures 5.0g green grass or young crops cumquat fruit powders and is placed in conical flasks of the 100mL with glass stopper, add the acetone oil prepared Ether 3:7 mixed liquors, cover static 5min after glass stopper shaking 1min, extracting solution are transferred to equipped with 5% anhydrous sodium sulfates of 100mL In the separatory funnel of solution, with 10mL acetone-petroleum ethers 3:7 mixed liquors continue extraction 2-3 times, until extracting solution is to colourless Only, it is static, lower aqueous solution is discarded after solution layering, then washed for several times with 5% anhydrous sodium sulfates of 15mL, to no longer muddiness Untill；
    Step 3, extracting solution is transferred in the 500mL aryballos supporting with rotating additional issue device, rotary spherical bottle is connect, in 55 DEG C of water Depressurized in bath and be distilled to recover petroleum ether, until removing aryballos not when there is drop to drip in condenser pipe, adding 2mL immediately Petroleum ether carries out constant volume；
    Step 4, a baseline is done at away from filter paper lower end 4cm, and equidistant 4 points of A, B, C, D is taken on baseline；With numeral Electric pipettor takes the sample concentration liquid of 0.4mL in AB point-to-point transmissions and CD point-to-point transmissions band application back and forth rapidly, until having put；Treat On filter paper sampling liquid evaporate into completely it is dry after, filter paper is rolled into tubular, is inserted at baseline positioned at low side and fills petroleum ether saturation It is unfolded in the chromatography cylinder crossed；After beta carotene and other pigments are completely isolated, filter paper is taken out, treats that petroleum ether is complete Evaporate into after doing, cut the carrotene chromatographic band positioned at solvent the top with scissors, chromatographic band is cut into fragment is immediately placed in Match in test tube to the tool for filling 5mL petroleum ethers, firmly shake, until carrotene is completely dissolved in petroleum ether；
    Sample eluent, is carried out the measure of absorbance, in standard curve by step 5 in the case where wavelength is the spectrophotometer of 450nm On find out the content of corresponding beta carotene；
    Step 6, the relevance model of near infrared spectrum and sample actual value is established using miniature near with the method for Chemical Measurement The scope of IR spectrum scanning wavelength is 900~1700nm, Unscrambler analysis near infrared spectrums and spectrophotometric experiment number According to relation, modeled by PLS, cross validation and forecast analysis；Effective coefficient of determination R2Close to 1, model is better；
    Step 7, scans actual blue or green cumquat fruit powder, its actual carotene carotene content is calculated according to model.
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CN108918456A (en) * 2018-07-04 2018-11-30 安徽科技学院 A kind of Astaxanthin In Haematococcus Pluvialis synthesis process research method based on infrared spectroscopy micro-imaging technique
CN109297918A (en) * 2018-08-21 2019-02-01 广西科技大学 A method of detection Acid red 26
CN114047155A (en) * 2021-09-26 2022-02-15 嘉必优生物技术（武汉）股份有限公司 Method for rapidly detecting carotenoid

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