CN109374538A - A kind of colorimetric card determining Blueberry maturity grade - Google Patents
A kind of colorimetric card determining Blueberry maturity grade Download PDFInfo
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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
- G01N21/29—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using visual detection
- G01N21/293—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands using visual detection with colour charts, graduated scales or turrets
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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
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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
- G01N21/33—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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- G01N31/16—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using titration
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- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
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- 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
- G01N2021/3129—Determining multicomponents by multiwavelength light
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Abstract
The present invention relates to field of agricultural science and technology, more particularly to a kind of colorimetric card for determining Blueberry maturity grade, the relationship of the color and fruit quality according to Blueberry, by outside the change of fruit internal physiological biochemistry and fruit color and structure feature contact the colour atla for being able to reflect Blueberry difference mature period established.The Blueberry of differing maturity is divided into 6 stages as research sample according to color by the present invention, measure size, quality, hardness, titratable acid, soluble solid, pectin, the total phenol, flavones, anthocyanidin of different times Blueberry, ABA content, analysis Blueberry color and fruit quality contact, the colour atla for being able to reflect Blueberry difference mature period is developed, to select suitable harvesting or time buying to provide theoretical foundation.Reflect the maturity of fruit, concisely convenient for users to selecting the fruit for being suitble to oneself.
Description
Technical field
The present invention relates to field of agricultural science and technology, more particularly to a kind of colorimetric card for determining Blueberry maturity grade.
Background technique
Blueberry nutrient is abundant, is not only rich in conventional nutrients, but also contain flavonoids extremely abundant and polysaccharide
Compound, therefore it is otherwise known as " fruit queen " and " king of berry ".Blueberry pulp is fine and smooth, unique flavor, moderately sour and sweet, and has
There is fragrant pleasant fragrance of feeling well.But Blueberry normal temperature condition is not easy to store, and can only place 5d or so after adopting at room temperature,
Shelf life is shorter, is not easy to transport, therefore sales range surrounds around the place of production more, is unfavorable for distally providing sale.So selection
Suitable plucking time can maximized Shelf-life, while again nutritional ingredient is influenced less, to become and extend blueberry storage
Hide root cause problems of the time other than changing gene.There is no stopping vital movements after fruit is picked from tree, but
Continue complicated physiology and biochemical reaction.With these react progress, the color and nutritional ingredient of fruit also into
The corresponding variation of row.Orchard worker does not have instrument and equipment to measure the maturity of fruit and the change of nutritive value at any time, most direct side
Formula is exactly to judge the mature condition of fruit by observation color change, to extrapolate optimum harvesting according to haulage time
Phase.In addition, consumer can select the blueberry of different colours according to the storage period of oneself, and then help to ensure that consumer is edible
Be all that mouthfeel is best, the highest Blueberry of nutritive value.The relationship between color and fruit maturity is found, can be used as
Judge one of simplest method of fruit maturity.But currently, and having no studies have reported that this kind of equipment or simple and easy method.
Summary of the invention
It is an object of the present invention to overcome the above-mentioned technical problems, and to provide a kind of colorimetrics for determining Blueberry maturity grade
Card, can be fully solved above-mentioned technical problem.
The technical solution for solving above-mentioned technical problem is as follows:
The present invention devises a kind of colorimetric card for determining Blueberry maturity grade, is the color according to Blueberry and fruit
The relationship of real quality, by outside the change of fruit internal physiological biochemistry and fruit color and structure feature contact established energy
Enough reflect the colour atla of Blueberry difference mature period, comprising the following steps:
(1) no disease and pests harm picked the 4-7 month and the Blueberry without machinery wound are chosen, classifies according to maturity, is divided into 6
Class, respectively immature green fruit start mature greenbelt blush fruit, half-mature red tape micro- green fruit, half-mature red
Fruit, most of mature red tape blue fruit and all mature blue fruit；It is to be measured that above-mentioned Blueberry is put into 20 DEG C of insulating boxs；
The blueberry is two kinds of Du Ke and Lan Feng；
(2) it is detected as follows respectively, and then obtains the indices feature of the Blueberry in the different stages of ripeness:
A) fruit growth index, the fruit growth index include: transverse diameter, vertical diameter, quality and hardness；
With the transverse diameter and vertical diameter of the vernier caliper measurement fruit that precision is 0.02mm；Wherein: six kinds of maturations of blue rich kind
The vertical diameter of the Blueberry in stage respectively is: 9.928 ± 0.312a, 10.41 ± 0.536ab, 10.548 ± 0.289ab,
10.588 ± 0.322ab, 10.936 ± 0.788bc, 11.376 ± 0.825c；Transverse diameter respectively is 12.986 ± 0.222a,
14.282 ± 0.811b, 14.337 ± 0.221b, 14.364 ± 0.563b, 14.896 ± 1.022b, 16.636 ± 1.245c；Du
The vertical diameter of the Blueberry in six kinds of stages of ripeness of gram kind respectively is: 9.365 ± 0.386a, 10.036 ± 0.254ab,
10.787 ± 0.525bc, 11.241 ± 0.249c, 11.770 ± 0.683c, 13.092 ± 1.182d；Transverse diameter respectively is:
13.313 ± 1.209a, 13.793 ± 0.050ab, 14.147 ± 0.500abc, 15.107 ± 0.508bc, 15.347 ±
0.480c, 17.227 ± 1.148d；The unit of the transverse diameter and vertical diameter is mm；
Fruit quality is weighed using a ten thousandth balance；It is bent to produce influence of the different stages of ripeness to Blueberry quality
Line；
The hardness of fruit is measured using Texture instrument, probe diameter 8mm, traverse distance 7mm, and finding speed is
0.5mm/s；The different stages of ripeness are produced to the influence curve of Blueberry hardness；
B) nutrition content changes, and the nutrition content includes: that titratable acid content, soluble solid contain
Amount, pectin content, total phenol content of material, Flavonoid Content and anthocyanidin content；
The detection method of the titratable acid content is as follows:
Each 5g of different times fruit is chosen, is placed in mortar and is fully ground, fruit homogenate is transferred completely into 100mL
In volumetric flask, and it is settled to scale with distilled water, shaken up, filtered after standing 30 minutes；
It takes 20.00mL filtrate in triangular flask, two drop phenolphthalein indicators is added, are dripped with calibrated sodium hydroxide solution
It is fixed, data, in triplicate parallel laboratory test are recorded, and do blank control group；
In formula: V --- sample extracting solution total volume, mL；
V2--- taken filtrate volume, mL when titration；
C --- sodium hydroxide titration liquid concentration, mol/L；
V1--- the sodium hydroxide solution volume of titration filtrate consumption, mL；
V0--- the sodium hydroxide solution volume of titration blank control group consumption, mL；
The quality of m --- sample, g；
F --- conversion factor, the most citric acid conversion factor of content is 0.064g/mmoL in blueberry；
The detection method of the soluble solid content is as follows:
It takes 3g Blueberry to be placed in mortar to be fully ground, be detected with 4 layers of filtered through gauze homogenate drop in WYT Refractometer
On mirror, record reading indicates soluble solid content in fruit with mass fraction；Parallel laboratory test in triplicate；
The detection method of the pectin content is as follows:
Pectin substance content is measured using colorimetry of carbazole.The Blueberry of 5g differing maturity, mistake are ground in mortar
Filtrate is transferred in 50mL centrifuge tube after filter, each ethyl alcohol that 25mL95% is added is heated 30min, taken out cold on boiling water bath
But to room temperature；It is put into centrifuge, discards supernatant liquid after 8000r/min centrifugation 15min；The ethyl alcohol of 25mL95% is respectively added again,
It heats on boiling water bath, is so repeated 3 times, then precipitating is transferred in former test tube, 20 milliliters of distilled water are respectively added, in 50 DEG C of water-baths
Middle heat preservation 30min；Taking-up is put into centrifuge after being cooled to room temperature, 8000r/min is centrifuged 15min, and supernatant is transferred to 100mL and is held
In measuring bottle, with a small amount of distillation water washing precipitating, supernatant is gone in volumetric flask together after centrifugation, to equipped with through distilling water washing
25mL0.5mol/L sulfuric acid solution is respectively added in the centrifuge tube of sediment afterwards, 1h is heated in boiling water bath, taking-up is cooled to room temperature
Afterwards, it is put into centrifuge 8000r/min centrifugation 15min, in the 100mL volumetric flask before supernatant is transferred to, with distilled water constant volume
And it shakes up；1mL extracting solution is drawn from each volumetric flask, is added separately in 25mL scale test tube, it is each that the dense sulphur of 6.0mL is added
Acid heats 20min in boiling water bath, each that 0.2mL1.5g/L carbazole-ethanol solution is added after being cooled to room temperature, and shakes up；Dark
After 30min is placed at place, measuring solution in wavelength with ultraviolet specrophotometer is the light absorption value at 530nm；
The detection method of the total phenol substance, flavonoids and anthocyanidin content is as follows:
2g blueberry is weighed, the hydrochloric acid-methanol solution that the volume fraction by pre-cooling is 1% a little is added in mortar,
It is all transferred in 25mL scale test tube after grinding homogenate under condition of ice bath, is settled to scale, mixed, be protected from light extraction in 4 DEG C
During which 20min shakes for several times, after filtering, collection filtrate is to be measured, makees the zeroing of blank reference with 1% hydrochloric acid-methanol solution, takes filter
Liquid measures the absorbance value of solution at wavelength 280nm, 325nm, 600nm and 530nm, records data, as total phenol object
The content of matter, flavonoids and anthocyanidin, in triplicate parallel laboratory test.Standard curve is made with various concentration gallic acid, is calculated
Total phenol content of material indicates flavonoid substances content in wavelength with every gram of Blueberry for the absorbance value at 325nm, i.e.,
ODxs/g；Indicate anthocyanidin content (U) in the difference that wavelength is the absorbance value at 530nm and 600nm, i.e. U=(ODso-ODn)/
g；
C) plant abscisic acid (ABA) changes of contents
The detection of the plant ABA content is as follows:
The Blueberry for weighing 3g differing maturity respectively is placed in mortar, and the PBS of 0.01mol/L is used to grind as homogenate
Fruit is ground, and the homogenate after grinding is transferred completely into 50mL centrifuge tube with the PBS of 0.01mol/L, every group of use
The total amount of 0.01mol/LPBS is 27mL；Homogenate is centrifuged 30min in the case where 4000r/min, takes supernatant to be measured；According to
The application method of plant abscisic acid (ABA) enzyme-linked immunoassay kit is operated, according to obtained standard curve and sample
OD value, calculate every gram of fruit contained by ABA amount；
(3) it according to the curve of the detection data of step (2) and production, is counted using Excel2010 and SPSS22 software
According to statistics with analysis, mapping comparison is carried out with origin8.1 software, carries out color levels figure with Photoshop2018 software
It draws；As a result it is indicated with average value ± standard deviation (x ± s)；
(4) content further according to step (3) develops the colour atla of Blueberry respectively.
The beneficial effects of the present invention are:
The Blueberry of differing maturity is divided into 6 stages as research sample according to color by the present invention, and measurement is different
The size of period Blueberry, quality, hardness, titratable acid, soluble solid, pectin, total phenol, flavones, anthocyanidin, ABA
Contacting for content, analysis Blueberry color and fruit quality, develops the color for being able to reflect Blueberry difference mature period
Card, to select suitable harvesting or time buying to provide theoretical foundation.The maturity for concisely reflecting fruit, conveniently makes
User selects the fruit for being suitble to oneself.
Detailed description of the invention
The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.
Fig. 1 is influence curve of the blueberry different stages of ripeness to Blueberry quality；
Fig. 2 is influence curve of the blueberry different stages of ripeness to Blueberry hardness；
Fig. 3 is influence curve of the blueberry different stages of ripeness to Blueberry titratable acid content；
Fig. 4 is influence curve of the blueberry different stages of ripeness to Blueberry soluble solid content；
Fig. 5 is influence curve of the Du Ke kind blueberry different stages of ripeness to Blueberry pectin content；
Fig. 6 is the influence curve of the different stage of ripeness Blueberry total phenol content of material of blueberry；
Fig. 7 is the influence curve of the different stage of ripeness Blueberry Flavonoid Contents of blueberry；
Fig. 8 is the influence curve of the different stage of ripeness Blueberry anthocyanidin contents of blueberry；
Fig. 9 is the influence curve of the different stage of ripeness Blueberry ABA contents of blue rich kind blueberry；
Figure 10 is the colour atla of Du Ke kind blueberry；
Figure 11 is the colour atla of blue rich kind blueberry；
Figure 12 is blueberry colour atla；
Specific embodiment
Embodiment 1:
A kind of colorimetric card of judgement Blueberry maturity grade as shown in figs. 1-12, be according to the color of Blueberry with
The relationship of fruit quality, by outside the change of fruit internal physiological biochemistry and fruit color and structure feature contact and established
It is able to reflect the colour atla of Blueberry difference mature period, comprising the following steps:
(1) no disease and pests harm picked the 4-7 month and the Blueberry without machinery wound are chosen, classifies according to maturity, is divided into 6
Class, respectively immature green fruit start mature greenbelt blush fruit, half-mature red tape micro- green fruit, half-mature red
Fruit, most of mature red tape blue fruit and all mature blue fruit；It is to be measured that above-mentioned Blueberry is put into 20 DEG C of insulating boxs；
The blueberry is Du Ke kind；
(2) it is detected as follows respectively, and then obtains the indices feature of the Blueberry in the different stages of ripeness:
A) fruit growth index, the fruit growth index include: transverse diameter, vertical diameter, quality and hardness；
With the transverse diameter and vertical diameter of the vernier caliper measurement fruit that precision is 0.02mm；Six kinds of stages of ripeness of Du Ke kind
The vertical diameter of Blueberry respectively is: 9.365 ± 0.386a, 10.036 ± 0.254ab, 10.787 ± 0.525bc, and 11.241
± 0.249c, 11.770 ± 0.683c, 13.092 ± 1.182d；Transverse diameter respectively is: 13.313 ± 1.209a, 13.793 ±
0.050ab, 14.147 ± 0.500abc, 15.107 ± 0.508bc, 15.347 ± 0.480c, 17.227 ± 1.148d；Transverse diameter and
The unit of vertical diameter is mm；
Fruit quality is weighed using a ten thousandth balance；It is bent to produce influence of the different stages of ripeness to Blueberry quality
Line；As shown in Figure 1.As shown in Figure 1, ascendant trend (p < 0.05) is presented with the increase of maturity in the quality of Blueberry.?
Fruit mezzanine level, fruit mass are about 0.85~0.887g.In growth after the 3rd stage, the rich quality of Du Ke BELAND
Increase fast, final stage of ripeness Du Ke fruit mass is close to 2.5g.
The hardness of fruit is measured using Texture instrument, probe diameter 8mm, traverse distance 7mm, and finding speed is
0.5mm/s；The different stages of ripeness are produced to the influence curve of Blueberry hardness；As shown in Figure 2.As shown in Figure 2, for the first time
It is measured as fruit belt leather hardness, the i.e. hardness of fruit.It is measured as fruit not belt leather hardness, i.e. flesh firmness for the second time.With fruit
Continuous maturation, Blueberry hardness and flesh firmness be presented decreasing trend (p < 0.05), immature green fruit hardness
Very big, when measurement when probe penetrates fruit, the clear and melodious rupture sound of audible pericarp, the hardness of fruit is up to 4.34N, about at
More than twice of the ripe hardness of fruit (1.25N)；The immature fruit hardness that belt leather does not measure is 1.81N, is ripening fruits not belt leather
More than 8 times of hardness (0.22N).
B) nutrition content changes, and the nutrition content includes: that titratable acid content, soluble solid contain
Amount, pectin content, total phenol content of material, Flavonoid Content and anthocyanidin content；
The detection method of the titratable acid content is as follows:
Each 5g of different times fruit is chosen, is placed in mortar and is fully ground, fruit homogenate is transferred completely into 100mL
In volumetric flask, and it is settled to scale with distilled water, shaken up, filtered after standing 30 minutes；
It takes 20.00mL filtrate in triangular flask, two drop phenolphthalein indicators is added, are dripped with calibrated sodium hydroxide solution
It is fixed, data, in triplicate parallel laboratory test are recorded, and do blank control group；
In formula: V --- sample extracting solution total volume, mL；
V2--- taken filtrate volume, mL when titration；
C --- sodium hydroxide titration liquid concentration, mol/L；
V1--- the sodium hydroxide solution volume of titration filtrate consumption, mL；
V0--- the sodium hydroxide solution volume of titration blank control group consumption, mL；
The quality of m --- sample, g；
F --- conversion factor, the most citric acid conversion factor of content is 0.064g/mmoL in blueberry；
Testing result is as shown in Figure 3；
The detection method of the soluble solid content is as follows:
It takes 3g Blueberry to be placed in mortar to be fully ground, be detected with 4 layers of filtered through gauze homogenate drop in WYT Refractometer
On mirror, record reading indicates soluble solid content in fruit with mass fraction；Parallel laboratory test in triplicate；As a result such as Fig. 4
It is shown:
The detection method of the pectin content is as follows:
Pectin substance content is measured using colorimetry of carbazole.The Blueberry of 5g differing maturity, mistake are ground in mortar
Filtrate is transferred in 50mL centrifuge tube after filter, each ethyl alcohol that 25mL95% is added is heated 30min, taken out cold on boiling water bath
But to room temperature；It is put into centrifuge, discards supernatant liquid after 8000r/min centrifugation 15min；The ethyl alcohol of 25mL95% is respectively added again,
It heats on boiling water bath, is so repeated 3 times, then precipitating is transferred in former test tube, 20 milliliters of distilled water are respectively added, in 50 DEG C of water-baths
Middle heat preservation 30min；Taking-up is put into centrifuge after being cooled to room temperature, 8000r/min is centrifuged 15min, and supernatant is transferred to 100mL and is held
In measuring bottle, with a small amount of distillation water washing precipitating, supernatant is gone in volumetric flask together after centrifugation, to equipped with through distilling water washing
25mL0.5mol/L sulfuric acid solution is respectively added in the centrifuge tube of sediment afterwards, 1h is heated in boiling water bath, taking-up is cooled to room temperature
Afterwards, it is put into centrifuge 8000r/min centrifugation 15min, in the 100mL volumetric flask before supernatant is transferred to, with distilled water constant volume
And it shakes up；1mL extracting solution is drawn from each volumetric flask, is added separately in 25mL scale test tube, it is each that the dense sulphur of 6.0mL is added
Acid heats 20min in boiling water bath, each that 0.2mL1.5g/L carbazole-ethanol solution is added after being cooled to room temperature, and shakes up；Dark
After 30min is placed at place, measuring solution in wavelength with ultraviolet specrophotometer is the light absorption value at 530nm；As a result as shown in Figure 5；
Pectin can be conducive to the growth of beneficial bacteria in enteron aisle, promote the excretion of fat and steroids, while can adsorb food
Heavy metal in object, without being digested liquid digestion, so internal noxious material can be excluded.As shown in Figure 5, Du Ke blueberry is preceding
3 stage pectin contents are all very low, there was only 0.00463mg in the 1st every gram of fruit of stage, only have in the 2nd every gram of fruit of stage
0.00934mg, only 0.02688mg in the 3rd every gram of fruit of stage.3 stage fruits start large area and turn red turn of indigo plant, pectin afterwards
Content rises rapidly.Pectin content is up to 0.48456mg in 4th every gram of fruit of stage, pectin content in the 5th every gram of fruit of stage
Up to 1.90978mg, and pectin content is up to 3.70434mg in the 6th every gram of fruit of stage.
The detection method of the total phenol substance, flavonoids and anthocyanidin content is as follows:
2g blueberry is weighed, the hydrochloric acid-methanol solution that the volume fraction by pre-cooling is 1% a little is added in mortar,
It is all transferred in 25mL scale test tube after grinding homogenate under condition of ice bath, is settled to scale, mixed, be protected from light extraction in 4 DEG C
During which 20min shakes for several times, after filtering, collection filtrate is to be measured, makees the zeroing of blank reference with 1% hydrochloric acid-methanol solution, takes filter
Liquid measures the absorbance value of solution at wavelength 280nm, 325nm, 600nm and 530nm, records data, as total phenol object
The content of matter, flavonoids and anthocyanidin, in triplicate parallel laboratory test.Standard curve is made with various concentration gallic acid, is calculated
Total phenol content of material indicates flavonoid substances content in wavelength with every gram of Blueberry for the absorbance value at 325nm, i.e.,
ODxs/g；Indicate anthocyanidin content (U) in the difference that wavelength is the absorbance value at 530nm and 600nm, i.e. U=(ODso-ODn)/
g；As shown in Figure 6；Polyphenol in Blueberry has inoxidizability, can be antitumor.According to Fig. 6 as can be seen that Blueberry
Total phenol content is changed significantly (p < 0.05) in maturation.Du Ke fruit total phenol content rises steadily.In immature green
Stage, fruit contain less total phenol, and content is 185.99mg/g or so.After red occurs in fruit, total phenol substance increases therewith
Long, content is in 198.76mg/g or so.3rd stage slightly slightly slowed down to the 5th stage total phenol content growth trend, when fruit is in the
When 6 stage, total phenol content of material increases comparatively fast, and content is up to 228.74mg/g or so.
As shown in Figure 7, Du Ke flavonoids in maturation content totally present ascendant trend, but ascensional range compared with
Small, only final stage of ripeness content goes up by more significant (p < 0.05).In prematurity period in the 1st stage, the content of flavonoids
It just can reach 2.22mg/g, be 2.31mg/g in the content of final stage of ripeness flavonoids.And in final stage of ripeness fruit
Flavonoid Content is relatively stable, standard deviation very little.
As shown in Figure 8, in fruit anthocyanidin content with fruit maturation present ascendant trend (p < 0.05).It is in fruit color
The mezzanine level of green, the content of Du Ke anthocyanidin are 0.22mg/g, and the Du Ke anthocyanidin content of second stage increases less
For 0.24mg/g, the 2nd stage to the 4th stage, fruit color is red by green turn, then turns blue, and anthocyanidin content significantly rises, the 6th rank
Duan Guoshi full maturity.The light absorption value of anthocyanidin is green fruit anthocyanidin light absorption value when Du Ke fruit final full maturity
13.5 times.
C) plant abscisic acid (ABA) changes of contents
The detection of the plant ABA content is as follows:
The Blueberry for weighing 3g differing maturity respectively is placed in mortar, and the PBS of 0.01mol/L is used to grind as homogenate
Fruit is ground, and the homogenate after grinding is transferred completely into 50mL centrifuge tube with the PBS of 0.01mol/L, every group of use
The total amount of 0.01mol/LPBS is 27mL；Homogenate is centrifuged 30min in the case where 4000r/min, takes supernatant to be measured；According to
The application method of plant abscisic acid (ABA) enzyme-linked immunoassay kit is operated, according to obtained standard curve and sample
OD value, calculate every gram of fruit contained by ABA amount；ABA is one of the important hormone for accelerating fruit abscission, it can inhibit fruit
Growth increases resistance, stomata can be caused to close in fruit water shortage, reduce the transpiration of fruit.
(3) it according to the curve of the detection data of step (2) and production, is counted using Excel2010 and SPSS22 software
According to statistics with analysis, mapping comparison is carried out with origin8.1 software, carries out color levels figure with Photoshop2018 software
It draws；As a result it is indicated with average value ± standard deviation (x ± s)；
(4) content further according to step (3) develops the colour atla of Blueberry respectively.
The development of Du Ke colour atla:
As shown in Figure 10, Figure 10 is the blueberry colour atla according to the variation production of Du Ke fruit color.1st stage fruit has been presented
Full green, except the hardness of fruit, flesh firmness and titratable acid content it is higher in addition to, volume, quality, soluble solid, pectin,
Total phenol substance, flavonoids and anthocyanidin are lower content.2nd stage was just to start the period for red color occur, fruit transverse diameter
And quality growth is more significant, vertical diameter growth is smaller, and the hardness of fruit, flesh firmness and titratable acid content have significant decrease,
Soluble solid content increases, and pectin content, flavonoids and anthocyanidin content increase more gentle, total phenol content of material growth
It is larger, increase by 27.3% compared to green fruit total phenol content of material.3rd stage fruit red area increases, and reaches 90% or so
Red change, volume of fruits and quality have more significant growth, and the hardness of fruit slightly reduces, flesh firmness and titratable acid content
There is more significant reduction, soluble solid content significantly rises, and pectin and total phenol content of material growth rate are smaller, and flavonoids contains
Amount has small size rise, and anthocyanidin content has conspicuousness raising.4th stage fruit becomes red completely and gradually heightens the color, body
Long-pending to have conspicuousness increase with quality, the hardness of fruit and flesh firmness significantly reduce titratable acid content and soluble solid content
Variation is smaller, and especially titratable acid content is almost horizontal with holding on last stage, and pectin content then has conspicuousness to increase increasing
Long, content is about 17 times of previous stage, and total phenol substance and Flavonoid Content have small range growth, and variation is not very significant, but
Anthocyanidin content is compared conspicuousness increase on last stage.5th stage fruit is substantially blue, slightly partially red at carpopodium, fruit
Volume and quality increment are less, and the hardness of fruit and flesh firmness reduce, and titratable acid content has more significant reduction, solvable
Property solid content increases compared to the increment in the 4th stage, and pectin and total phenol content of material have significant growth, flavonoids
Content only has small range growth, and anthocyanidin content continues to keep steadily significant and increases, and growth rate is up to 69.93%.6th rank
Duan Guoshi becomes blue completely, and volume significantly increases, and quality significantly improves, and the hardness of fruit has a more significant reduction, and flesh firmness
Variation is smaller, and titratable acid content reduces extremely significant, and soluble solid content and pectin content have conspicuousness raising, always
Phenol substance increasing degree is smaller, and Flavonoid Content possesses the highest increment of entire maturation, is the 5th stage growth rate
4.3 times, anthocyanidin content growth rate reduces in fruit, but still has conspicuousness growth.
The analysis instrument service condition is as follows:
FA-004 electronic analytical balance, Science and Technology Ltd. of upper Nereid section；CT14RD high speed freezing centrifuge, Shanghai day beauty
Biochemical Co., Ltd；Table model high speed centrifuge, Changsha Xiang Yi Instrument Ltd.；DKS26 thermostat water bath, the upper macro experiment of Nereid
Instrument company；Italian FT-327 type Texture instrument；WYT Refractometer；UV-1810 ultraviolet-uisible spectrophotometer, Beijing are general
Analyse general scientific & technical corporation；IMark microplate reader, Bio-Rad company of the U.S.；Gloves；Mask；Glass sample bottle (band rubber stopper)；It grinds
Alms bowl；Volumetric flask；Pipette；Plug test tube (25mL)；Rubber head dropper；Liquid-transfering gun；Base buret；Triangular flask；Funnel；Electricity
Magnetic furnace；Iron stand；Glass bar；Water-bath；Timer；Vernier caliper；Plant abscisic acid (ABA) enzyme-linked immunoassay kit
Deng.
Phenolphthalein indicator；Sodium hydroxide；Potassium Hydrogen Phthalate；1% hydrochloric acid-methanol solution；1.5%g/L carbazole-second
Alcoholic solution；100 μ g/mL galacturonic acid standard solution；The concentrated sulfuric acid；0.5mol/L sulfuric acid solution；6mol/L sulfuric acid solution；95%
Ethyl alcohol；0.01mol/L PBS；Gallic acid etc..
Embodiment 2:
Unlike the first embodiment:
The blueberry is blue rich kind；
(2) it is detected as follows respectively, and then obtains the indices feature of the Blueberry in the different stages of ripeness:
A) fruit growth index, the fruit growth index include: transverse diameter, vertical diameter, quality and hardness；
With the transverse diameter and vertical diameter of the vernier caliper measurement fruit that precision is 0.02mm；Six kinds of stages of ripeness of blue rich kind
The vertical diameter of Blueberry respectively is: 9.928 ± 0.312a, 10.41 ± 0.536ab, 10.548 ± 0.289ab, and 10.588
± 0.322ab, 10.936 ± 0.788bc, 11.376 ± 0.825c；Transverse diameter respectively is 12.986 ± 0.222a, and 14.282
± 0.811b, 14.337 ± 0.221b, 14.364 ± 0.563b, 14.896 ± 1.022b, 16.636 ± 1.245c；Transverse diameter and vertical
The unit of diameter is mm；
Fruit quality is weighed using a ten thousandth balance；It is bent to produce influence of the different stages of ripeness to Blueberry quality
Line；As shown in Figure 1；As shown in Figure 1, ascendant trend (p < 0.05) is presented with the increase of maturity in the quality of Blueberry.?
Fruit mezzanine level, fruit mass are about 0.85~0.887g.Then blue rich fruit mass reaches 1.2g or so.3rd stage
Slow down with the 4th stage blue rich fruit quality growth rate, fruit mass is about 1.27g.The final stage of ripeness blue rich fruit mass
For 1.95g.
The hardness of fruit is measured using Texture instrument, probe diameter 8mm, traverse distance 7mm, and finding speed is
0.5mm/s；The different stages of ripeness are produced to the influence curve of Blueberry hardness；As shown in Fig. 2, being measured as fruit for the first time
Belt leather hardness, the i.e. hardness of fruit.It is measured as fruit not belt leather hardness, i.e. flesh firmness for the second time.With the continuous maturation of fruit,
Blueberry hardness and flesh firmness are presented decreasing trend (p < 0.05), and immature green fruit hardness is very big, measurement
When probe penetrates fruit, the clear and melodious rupture sound of audible pericarp, the hardness of fruit is up to 4.34N, about ripening fruits hardness
More than twice of (1.25N)；The immature fruit hardness that belt leather does not measure is 1.81N, is ripening fruits not belt leather hardness (0.22N)
More than 8 times.
From the figure 3, it may be seen that blue rich Blueberry titratable acid content is reduced with fruit maturation degree.Lan Feng with fruit at
The percentage of the raising of ripe degree, titratable acid is gradually reduced to 0.5438% from initial 3.3217%.
Soluble solid refers mainly to soluble carbohydrate material.As shown in Figure 4, as fruit is constantly mature, solubility is solid
Shape object content is gradually increasing.For blueberry in the case where initial green state completely, blue rich soluble solid content is on 5.2% left side
It is right.Lan Feng is 6.7% in the 3rd stage soluble solid content, and the 4th stage amount of increase is smaller, and the 5th stage was 7.5% or so, the
6 stage amounts of increase are larger, and soluble solid content is 9.2% or so.
Polyphenol in Blueberry has inoxidizability, can be antitumor.According to Fig. 6 as can be seen that Blueberry total phenol
Content is changed significantly (p < 0.05) in maturation.Blue Feng Guoshi in entire maturation total phenol content of material fluctuation compared with
Greatly.Total phenol content of material maximum value appears in the 1st stage of fruit maturation, and the 2nd stage content is very fast to be reduced to 176.76mg/g,
With fruit maturation, total phenol content of material gradually goes up again in upper and lower fluctuation, and the 5th stage is up to 209.89mg/g.
As shown in Figure 7, there is maximum value, about 2.45mg/g, the 2nd stage in the 1st stage in blue Feng Guoshi Flavonoid Content
Minimum 2.17mg/g of content or so, the 3rd stage rapid Flavonoid Content rise rapidly, and under the 4th stage and the 5th stage
Drop, final stage of ripeness content are 2.28mg/g.
As shown in Figure 8, in fruit anthocyanidin content with fruit maturation present ascendant trend (p < 0.05).It is in fruit color
The mezzanine level of green, the content of blue rich anthocyanidin are 0.11mg/g；The rich anthocyanidin content of indigo plant of second stage increases more
For 1.02mg/g；In 2nd stage to the 4th stage, fruit color is red by green turn, then turns blue, and anthocyanidin content significantly rises；6th rank
Duan Guoshi full maturity.
C) plant abscisic acid (ABA) changes of contents
The detection of the plant ABA content is as follows:
The Blueberry for weighing 3g differing maturity respectively is placed in mortar, and the PBS of 0.01mol/L is used to grind as homogenate
Fruit is ground, and the homogenate after grinding is transferred completely into 50mL centrifuge tube with the PBS of 0.01mol/L, every group of use
The total amount of 0.01mol/LPBS is 27mL；Homogenate is centrifuged 30min in the case where 4000r/min, takes supernatant to be measured；According to
The application method of plant abscisic acid (ABA) enzyme-linked immunoassay kit is operated, according to obtained standard curve and sample
OD value, calculate every gram of fruit contained by ABA amount；ABA is one of the important hormone for accelerating fruit abscission, it can inhibit fruit
Growth increases resistance, stomata can be caused to close in fruit water shortage, reduce the transpiration of fruit.As shown in Figure 9, blue
Certain kind of berries fruit ABA contained by the 1st initial stage is 0.56 μ g/g, and ABA contained by every gram of Blueberry of second stage is increased rapidly
1.82 μ g/g, in maturation later, ABA content keeps steady growth, and the 3rd stage rose to 2.31 μ g/g, the 4th stage
ABA contained by every gram of Blueberry is 2.55 μ g/g, and the 5th stage continued to rise to 3.04 μ g/g, after fruit full maturity, ABA
Rise to the 3.89 μ g/g of maximum value of whole process.
(3) it according to the curve of the detection data of step (2) and production, is counted using Excel2010 and SPSS22 software
According to statistics with analysis, mapping comparison is carried out with origin8.1 software, carries out color levels figure with Photoshop2018 software
It draws；As a result it is indicated with average value ± standard deviation (x ± s)；
(4) content further according to step (3) develops the colour atla of Blueberry respectively.
The development of blue rich colour atla
Figure 11 is the blueberry colour atla according to blue rich fruit color variation production.1st stage fruit is generally green, volume with
Quality is smaller, and the hardness of fruit is higher with flesh firmness, and titratable acid content is higher, is fruit most sour period, soluble solid
The content of the indexs such as object, anthocyanidin and ABA content is lower, and total phenol substance and Flavonoid Content highest, fruit are in prematurity
State.There is the red of small area by bottom in 2nd stage, and volume and the complete green fruit of quality versus have obvious increasing
Add, fruit and flesh firmness are decreased obviously, and titratable acid content is also remarkably decreased, and soluble solid content is the same as green fruit ratio
It relatively has increased slightly, minimum occur in total phenol substance and Flavonoid Content, and anthocyanidin rises more significant with ABA content.3rd stage
Red area increases, and volume and quality increase smaller compared to the 2nd stage, and it is smaller that the hardness of fruit reduces amplitude, flesh firmness with
The reduction of titratable acid content is more significant, and soluble solid, total phenol substance, flavonoids and anthocyanidin content increase by a fairly big margin,
ABA content increases more significant.4th stage fruit becomes red completely, and the growth of volume and quality is still smaller, and fruit is hard
Degree, flesh firmness, titratable acid, total phenol substance and Flavonoid Content reduction are more significant, and soluble solid growth slows down, flower
Green element goes up with ABA content held stationary.5th phases red is deepened, and kermesinus is presented in fruit, and bottom has the portion of small area to separate
Begin to become blue, fruit is indulged diameter and more dramatically increased, and transverse diameter and quality growth conspicuousness are lower, and the hardness of fruit reduces amplitude and is greater than pulp
Hardness reduces amplitude, and titratable acid, Flavonoid Content have significant decrease, and soluble solid content has more significant growth,
Total phenol substance, anthocyanidin and ABA content rise more significant.6th stage fruit becomes blue completely, and volume and quality have conspicuousness increasing
Long, hardness of fruit reduction is more significant, and flesh firmness reduction amplitude is smaller, and titratable acid, total phenol content of material have conspicuousness drop
Low, soluble solid, anthocyanidin, flavonoids and ABA content have conspicuousness rise.
It is concluded that after comparative example 1 and embodiment 2
As shown in Figure 1, the quality of two kind Blueberries presented with the increase of maturity ascendant trend (p <
0.05).In fruit mezzanine level, two kind Blueberry mass differences are little, and fruit mass is about 0.85~0.887g.
Subsequent indigo plant Feng Bidu gram mass growth is slightly faster, and fruit mass reaches 1.2g or so.3rd stage and the 4th stage indigo plant Feng Guoshi
Quality growth rate slows down, and fruit mass is about 1.27g.In growth after the 3rd stage, the rich quality of Du Ke BELAND increases
Fastly, blue rich fruit mass of the final stage of ripeness is 1.95g, and Du Ke fruit mass is close to 2.5g.
As shown in Figure 2, it is measured as fruit belt leather hardness, the i.e. hardness of fruit for the first time.It is measured as fruit not belt leather for the second time
Hardness, i.e. flesh firmness.With the continuous maturation of fruit, Blueberry hardness and flesh firmness present decreasing trend (p <
0.05) as can be seen from Figure 2, immature green fruit hardness is very big, and when measurement when probe penetrates fruit, audible pericarp is clear
Crisp rupture sound, the hardness of fruit are up to 4.34N, about more than twice of ripening fruits hardness (1.25N)；Immature fruit not belt leather
The hardness of measurement is 1.81N, is ripening fruits not more than 8 times of belt leather hardness (0.22N).4th stage and the 5th stage Du Ke fruit
The a little higher than indigo plant of hardness is rich, remaining stage, the blue rich hardness of fruit was greater than Du Ke.
As seen from Figure 3, two kind Blueberry titratable acid contents are reduced with fruit maturation degree, and difference is aobvious
It writes (p < 0.05).Wherein, Lan Feng with fruit maturation degree raising, the percentage of titratable acid from initial 3.3217% by
Decrescence as little as 0.5438%, and Du Ke, in the 4th stage and the 5th phase, the percentage of titratable acid reduces more slow,
2.4% or so floating is about rested on, then declines suddenly in the percentage of last stage of ripeness titratable acid, is reduced to
0.4313%.
Soluble solid refers mainly to soluble carbohydrate material.As shown in Figure 4, as fruit is constantly mature, solubility is solid
Shape object content is gradually increasing, significant difference (p < 0.05).Two kind blueberries are in the case where initial green state completely, Du Ke
Soluble solid content is 6.5% or so, and blue rich soluble solid content is 5.2% or so.Du Ke is solvable in the 3rd stage
Property solid content, which increases sharply, reaches 9%, and being less about in the 4th stage increment is 9.2%, the 6th stage soluble solid
Content is up to 11.2%.Lan Feng is 6.7% in the 3rd stage soluble solid content, and the 4th stage amount of increase is smaller, and the 5th stage was
7.5% or so, the 6th stage amount of increase is larger, and soluble solid content is 9.2% or so.Compare in figure that two curves, Du Ke can
Dissolubility solid content is all larger than Lan Feng in the entire fruit maturation stage.
Polyphenol in Blueberry has inoxidizability, can be antitumor.According to Fig. 6 as can be seen that Blueberry total phenol
Content is changed significantly (p < 0.05) in maturation.Du Ke fruit total phenol content rises steadily.In immature green stage,
Fruit contains less total phenol, and content is 185.99mg/g or so.After red occurs in fruit, total phenol substance is increased, and contains
Amount is in 198.76mg/g or so.3rd stage slightly slightly slowed down to the 5th stage total phenol content growth trend, when fruit was in for the 6th stage
When, total phenol content of material increases comparatively fast, and content is up to 228.74mg/g or so.And indigo plant Feng Guoshi total phenol in entire maturation
Content of material fluctuation is larger.Total phenol content of material maximum value appears in the 1st stage of fruit maturation, the 2nd very fast drop of stage content
Down to 176.76mg/g, with fruit maturation, total phenol content of material gradually goes up again in upper and lower fluctuation, and the 5th stage is reachable
209.89mg/g, after the 3rd stage Du Ke BELAND it is rich containing total phenol substance it is some more.
As shown in Figure 7, Du Ke flavonoids in maturation content totally present ascendant trend, but ascensional range compared with
Small, only final stage of ripeness content goes up by more significant (p < 0.05).In prematurity period in the 1st stage, the content of flavonoids
It just can reach 2.22mg/g, be 2.31mg/g in the content of final stage of ripeness flavonoids.And in final stage of ripeness fruit
Flavonoid Content is relatively stable, standard deviation very little.There is maximum value in the 1st stage in blue Feng Guoshi Flavonoid Content, about
2.45mg/g, the 2nd minimum 2.17mg/g of stage content or so, the 3rd stage rapid Flavonoid Content rise rapidly, and the 4th
Stage and the decline of the 5th stage, final stage of ripeness content are 2.28mg/g.
As shown in Figure 8, in fruit anthocyanidin content with fruit maturation present ascendant trend (p < 0.05).It is in fruit color
The mezzanine level of green, the content of Du Ke anthocyanidin are 0.22mg/g, and the content of blue rich anthocyanidin is 0.11mg/g；Second-order
It is 0.24mg/g that the Du Ke anthocyanidin content of section, which increases less, and it is relatively mostly 1.02mg/g that blue rich anthocyanidin content, which increases,；2nd stage
To the 4th stage, fruit color is red by green turn, then turns blue, and anthocyanidin content significantly rises, and blue rich anthocyanidin content is higher than Du Ke,
Anthocyanidin content differs smaller in the 5th stage during the blueberry of two kinds is mature, and Du Ke content is slightly above blue rich；6th stage
Fruit full maturity, anthocyanidin content are blue rich higher than Du Ke.The light absorption value of anthocyanidin is green when Du Ke fruit final full maturity
13.5 times of color fruit anthocyanidin light absorption value, blue Feng Guoshi variation is bigger, about increases 29 times.
Figure 10 and Figure 11 is comprehensively compared, develops the colour atla of reaction Blueberry maturation variation, as shown in figure 12.
Fruit is difficult in the 1st stage that color is full green to the 2nd transition stage；The fruit red area in the 2nd stage compared with
It is few, it is also difficult to change to the phase III；3rd stage of ripeness switch to the 4th stage of ripeness only need to store in room temperature a d when
Between；4th stage of ripeness is more of short duration to two transition stage times later, and storing 12h fruit color in room temperature can
Become completely blue.But changes the fruit of color its size, acid content, pectin equal size by storage and change smaller, mouthfeel is not yet
Such as the mature fruit picked again is grown in end of the branch.Blueberry is not easily to fall off in mezzanine level carpopodium, holds very much in picking
Easily fruit is followed to fall off from tree；Base of fruit is in closed state in mezzanine level, and with fruit maturation, base of fruit is gradually opened, most
It is fully deployed eventually in the stage of ripeness.
It is sold it is recommended that the fruit in the 4th or the 5th stage of picking carries out transport, it is ensured that consumer can be used for the 5th or the 6th stage
Fruit obtains more nutriments and better mouthfeel.
The above is only presently preferred embodiments of the present invention, not does limitation in any form to the present invention, it is all according to
According to any simple modification to the above embodiments in technical spirit of the invention, equivalent variations, guarantor of the invention is each fallen within
Within the scope of shield.
Claims (1)
1. a kind of colorimetric card for determining Blueberry maturity grade, which is characterized in that be the color and fruit according to Blueberry
The relationship of quality, by outside the change of fruit internal physiological biochemistry and fruit color and structure feature contact established can
Reflect the colour atla of Blueberry difference mature period, comprising the following steps:
(1) no disease and pests harm picked the 4-7 month and the Blueberry without machinery wound are chosen, classifies according to maturity, is divided into 6 classes,
Respectively immature green fruit, start the micro- green fruit of mature greenbelt blush fruit, half-mature red tape, half-mature red fruit,
Most of mature red tape blue fruit and all mature blue fruit；It is to be measured that above-mentioned Blueberry is put into 20 DEG C of insulating boxs；Institute
The blueberry stated is two kinds of Du Ke and Lan Feng；
(2) it is detected as follows respectively, and then obtains the indices feature of the Blueberry in the different stages of ripeness:
A) fruit growth index, the fruit growth index include: transverse diameter, vertical diameter, quality and hardness；
With the transverse diameter and vertical diameter of the vernier caliper measurement fruit that precision is 0.02mm；Wherein: six kinds of stages of ripeness of blue rich kind
The vertical diameter of Blueberry respectively be: 9.928 ± 0.312a, 10.41 ± 0.536ab, 10.548 ± 0.289ab,
10.588 ± 0.322ab, 10.936 ± 0.788bc, 11.376 ± 0.825c；Transverse diameter respectively is 12.986 ± 0.222a,
14.282 ± 0.811b, 14.337 ± 0.221b, 14.364 ± 0.563b, 14.896 ± 1.022b, 16.636 ± 1.245c；Du
The vertical diameter of the Blueberry in six kinds of stages of ripeness of gram kind respectively is: 9.365 ± 0.386a, 10.036 ± 0.254ab,
10.787 ± 0.525bc, 11.241 ± 0.249c, 11.770 ± 0.683c, 13.092 ± 1.182d；Transverse diameter respectively is:
13.313 ± 1.209a, 13.793 ± 0.050ab, 14.147 ± 0.500abc, 15.107 ± 0.508bc, 15.347 ±
0.480c, 17.227 ± 1.148d；The unit of the transverse diameter and vertical diameter is mm；
Fruit quality is weighed using a ten thousandth balance；The different stages of ripeness are produced to the influence curve of Blueberry quality；
The hardness of fruit is measured using Texture instrument, probe diameter 8mm, traverse distance 7mm, finding speed 0.5mm/s；
The different stages of ripeness are produced to the influence curve of Blueberry hardness；
B) nutrition content change, the nutrition content include: titratable acid content, soluble solid content,
Pectin content, total phenol content of material, Flavonoid Content and anthocyanidin content；
The detection method of the titratable acid content is as follows:
Each 5g of different times fruit is chosen, is placed in mortar and is fully ground, fruit homogenate is transferred completely into 100mL capacity
In bottle, and it is settled to scale with distilled water, shaken up, filtered after standing 30 minutes；
It takes 20.00mL filtrate in triangular flask, two drop phenolphthalein indicators is added, are titrated with calibrated sodium hydroxide solution, note
Data, in triplicate parallel laboratory test are recorded, and does blank control group；
In formula: V --- sample extracting solution total volume, mL；
V2--- taken filtrate volume, mL when titration；
C --- sodium hydroxide titration liquid concentration, mol/L；
V1--- the sodium hydroxide solution volume of titration filtrate consumption, mL；
V0--- the sodium hydroxide solution volume of titration blank control group consumption, mL；
The quality of m --- sample, g；
F --- conversion factor, the most citric acid conversion factor of content is 0.064g/mmoL in blueberry；
The detection method of the soluble solid content is as follows:
It takes 3g Blueberry to be placed in mortar to be fully ground, detects mirror in WYT Refractometer with 4 layers of filtered through gauze homogenate drop
On, record reading indicates soluble solid content in fruit with mass fraction；Parallel laboratory test in triplicate；
The detection method of the pectin content is as follows:
Pectin substance content is measured using colorimetry of carbazole, the Blueberry of 5g differing maturity is ground in mortar, after filtering
Filtrate is transferred in 50mL centrifuge tube, each ethyl alcohol that 25mL95% is added, through heating 30min on boiling water bath, taking-up is cooled to
Room temperature；It is put into centrifuge, discards supernatant liquid after 8000r/min centrifugation 15min；The ethyl alcohol of 25mL95% is respectively added again, in boiling water
It heats in bath, is so repeated 3 times, then precipitating is transferred in former test tube, 20 milliliters of distilled water are respectively added, are protected in 50 DEG C of water-baths
Warm 30min；Taking-up is put into centrifuge after being cooled to room temperature, 8000r/min is centrifuged 15min, supernatant is transferred to 100mL volumetric flask
In, with a small amount of distillation water washing precipitating, supernatant is gone in volumetric flask together after centrifugation, to equipped with heavy after distilling water washing
25mL0.5mol/L sulfuric acid solution is respectively added in the centrifuge tube of starch, 1h is heated in boiling water bath and is put after taking-up is cooled to room temperature
Enter centrifuge 8000r/min centrifugation 15min, in the 100mL volumetric flask before supernatant is transferred to, with distilled water constant volume and shakes
It is even；1mL extracting solution is drawn from each volumetric flask, is added separately in 25mL scale test tube, it is each that the 6.0mL concentrated sulfuric acid is added, in
20min is heated in boiling water bath, it is each that 0.2mL1.5g/L carbazole-ethanol solution is added after being cooled to room temperature, it shakes up；It places in the dark
After 30min, measuring solution in wavelength with ultraviolet specrophotometer is the light absorption value at 530nm；
The detection method of the total phenol substance, flavonoids and anthocyanidin content is as follows:
2g blueberry is weighed, the hydrochloric acid-methanol solution that the volume fraction by pre-cooling is 1% a little is added in mortar, in ice bath
Under the conditions of grinding homogenate after be all transferred in 25mL scale test tube, be settled to scale, mix, be protected from light in 4 DEG C extract 20min, the phase
Between shake for several times, after filtering, collect filtrate it is to be measured, with 1% hydrochloric acid-methanol solution make blank reference zeroing, take filtrate respectively at
The absorbance value that solution is measured at wavelength 280nm, 325nm, 600nm and 530nm, records data, as total phenol substance, flavonoids
With the content of anthocyanidin, parallel laboratory test in triplicate makes standard curve with various concentration gallic acid, calculates total phenol substance and contains
Amount indicates flavonoid substances content, i.e. ODxs/g in wavelength with every gram of Blueberry for the absorbance value at 325nm；In wavelength
Anthocyanidin content (U) is indicated for the difference of the absorbance value at 530nm and 600nm, i.e. U=(ODso-ODn)/g；
C) plant abscisic acid (ABA) changes of contents
The detection of the plant ABA content is as follows:
The Blueberry for weighing 3g differing maturity respectively is placed in mortar, and the PBS of 0.01mol/L is used to grind fruit as homogenate
It is real, and the homogenate after grinding is transferred completely into 50mL centrifuge tube with the PBS of 0.01mol/L, every group uses 0.01mol/
The total amount of LPBS is 27mL；Homogenate is centrifuged 30min in the case where 4000r/min, takes supernatant to be measured；It falls off according to plant
The application method of sour (ABA) enzyme-linked immunoassay kit is operated, according to the OD value of obtained standard curve and sample, meter
Calculate the amount of ABA contained by every gram of fruit；
(3) according to the curve of the detection data of step (2) and production, data system is carried out using Excel2010 and SPSS22 software
Meter and analysis, carry out mapping comparison with origin8.1 software, the drafting of color levels figure are carried out with Photoshop2018 software；
As a result it is indicated with average value ± standard deviation (x ± s)；
(4) content further according to step (3) develops the colour atla of Blueberry respectively.
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| CN110358660A (en) * | 2019-06-28 | 2019-10-22 | 四川农业大学 | A kind of preparation method of blueberry pulp |
| CN110823814A (en) * | 2019-11-27 | 2020-02-21 | 上海市农业科学院 | Simple method for evaluating flavonoid and total phenol content of eggplant peel |
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