CN109187400B - Method for detecting pigment of duck egg shell - Google Patents
Method for detecting pigment of duck egg shell Download PDFInfo
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- CN109187400B CN109187400B CN201810979793.4A CN201810979793A CN109187400B CN 109187400 B CN109187400 B CN 109187400B CN 201810979793 A CN201810979793 A CN 201810979793A CN 109187400 B CN109187400 B CN 109187400B
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- 241000272525 Anas platyrhynchos Species 0.000 title claims abstract description 55
- 210000003278 egg shell Anatomy 0.000 title claims abstract description 52
- 102000002322 Egg Proteins Human genes 0.000 title claims abstract description 40
- 108010000912 Egg Proteins Proteins 0.000 title claims abstract description 40
- 239000000049 pigment Substances 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 20
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 31
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 26
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 20
- RCNSAJSGRJSBKK-NSQVQWHSSA-N Biliverdin IX Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(\C=C/2C(=C(C)C(=C/C=3C(=C(C=C)C(=O)N=3)C)/N\2)CCC(O)=O)N1 RCNSAJSGRJSBKK-NSQVQWHSSA-N 0.000 claims abstract description 18
- QBUVFDKTZJNUPP-UHFFFAOYSA-N biliverdin-IXalpha Natural products N1C(=O)C(C)=C(C=C)C1=CC1=C(C)C(CCC(O)=O)=C(C=C2C(=C(C)C(C=C3C(=C(C=C)C(=O)N3)C)=N2)CCC(O)=O)N1 QBUVFDKTZJNUPP-UHFFFAOYSA-N 0.000 claims abstract description 18
- 238000001556 precipitation Methods 0.000 claims abstract description 18
- GWZYPXHJIZCRAJ-UHFFFAOYSA-N Biliverdin Natural products CC1=C(C=C)C(=C/C2=NC(=Cc3[nH]c(C=C/4NC(=O)C(=C4C)C=C)c(C)c3CCC(=O)O)C(=C2C)CCC(=O)O)NC1=O GWZYPXHJIZCRAJ-UHFFFAOYSA-N 0.000 claims abstract description 17
- 238000001514 detection method Methods 0.000 claims abstract description 17
- 230000031700 light absorption Effects 0.000 claims abstract description 14
- 239000000843 powder Substances 0.000 claims abstract description 11
- 230000020477 pH reduction Effects 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 9
- 238000001035 drying Methods 0.000 claims abstract description 8
- 230000010355 oscillation Effects 0.000 claims abstract description 7
- 238000005303 weighing Methods 0.000 claims abstract description 7
- 238000004140 cleaning Methods 0.000 claims abstract description 4
- 239000012528 membrane Substances 0.000 claims abstract description 4
- 238000007781 pre-processing Methods 0.000 claims abstract description 4
- 239000006228 supernatant Substances 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 17
- 239000011550 stock solution Substances 0.000 claims description 9
- 238000007873 sieving Methods 0.000 claims description 7
- 238000002835 absorbance Methods 0.000 claims description 4
- 238000007602 hot air drying Methods 0.000 claims description 4
- KSFOVUSSGSKXFI-GAQDCDSVSA-N CC1=C/2NC(\C=C3/N=C(/C=C4\N\C(=C/C5=N/C(=C\2)/C(C=C)=C5C)C(C=C)=C4C)C(C)=C3CCC(O)=O)=C1CCC(O)=O Chemical compound CC1=C/2NC(\C=C3/N=C(/C=C4\N\C(=C/C5=N/C(=C\2)/C(C=C)=C5C)C(C=C)=C4C)C(C)=C3CCC(O)=O)=C1CCC(O)=O KSFOVUSSGSKXFI-GAQDCDSVSA-N 0.000 claims description 3
- 229950003776 protoporphyrin Drugs 0.000 claims description 3
- 238000002798 spectrophotometry method Methods 0.000 claims description 3
- 238000007865 diluting Methods 0.000 claims description 2
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 235000013601 eggs Nutrition 0.000 abstract description 17
- 238000009395 breeding Methods 0.000 abstract description 4
- 230000001488 breeding effect Effects 0.000 abstract description 4
- 230000000052 comparative effect Effects 0.000 description 9
- 244000144977 poultry Species 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 241000257159 Musca domestica Species 0.000 description 1
- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000011669 selenium Substances 0.000 description 1
- 229910052711 selenium Inorganic materials 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a method for detecting duck egg shell pigment, which comprises the following steps: s1, preprocessing: removing eggshell membranes from duck eggshells, cleaning and drying the eggshells to prepare powder, weighing the duck eggshell powder, adding mixed acidification precipitation liquid, performing vortex oscillation, and ultrasonically dissolving for 8-12 h in a dark environment, wherein the mixed acidification precipitation liquid comprises acetone, concentrated hydrochloric acid and acetic acid, and the volume ratio of the acetone to the concentrated hydrochloric acid to the acetic acid is 3: 1: 0.5-0.8; s2, drawing a standard curve; s3, detection and analysis: centrifuging the dissolved eggshell solution, taking supernatant, measuring the primary light absorption value at the wavelength of 300-760 nm at the interval of 2nm, calculating the protoporphyrin-IV content according to the light absorption value at 412nm, and calculating the biliverdin content according to the light absorption value at 680 nm. Compared with the prior art, the scheme of the invention has the advantages of accurate and reliable detection result and the like, and has good application prospect in seed selection and breeding of green-shell duck eggs.
Description
Technical Field
The invention relates to the technical field of poultry breeding, in particular to a method for detecting a duck egg shell pigment.
Background
The quality of the duck egg shells has important significance for the transportation, storage and processing of poultry eggs, and the color of the duck egg shells directly influences the sale of the duck eggs and indirectly influences the economic benefits of poultry breeders. The color of the egg shell of the duck egg is closely related to the quality of the egg shell, the green duck egg is fresh and natural, and the quality of the egg shell is obviously superior to that of a white-shell egg. In the course of processing, transporting and storing, the better eggshell quality is favorable for reducing the economic loss caused by breakage, and the green-shell duck egg, because the protein content is higher than the white-shell duck egg by 4.23 percent, except that the selenium and lysine content is slightly lower than the white-shell egg, the other 15 kinds of amino acid and zinc content are all increased to different extent than the white-shell duck egg, and because the green-shell duck egg has natural and bright-colored appearance, the green-shell duck egg is deeply loved by consumers, and the consumption is increased day by day. Eggshell pigment is a composite pigment mainly comprising protoporphyrin-IV and biliverdin, wherein the protoporphyrin-IV forms yellow, pink, light yellow or brown, while the biliverdin causes the eggshell to appear blue and green, so that the color of the eggshell of the duck depends on the proportion of the three pigments. Therefore, the accurate detection of protoporphyrin-IV and biliverdin-IX in the egg shell of the duck is of great significance for breeding.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: provides a method for detecting the pigment of the duck egg shell with accurate and reliable result.
In order to solve the technical problems, the invention adopts the technical scheme that: a method for detecting duck egg shell pigment comprises the following steps:
s1, preprocessing: removing eggshell membranes from duck eggshells, cleaning and drying the eggshells to prepare powder, weighing the duck eggshell powder, adding mixed acidification precipitation liquid, performing vortex oscillation, and ultrasonically dissolving for 8-12 h in a dark environment, wherein the mixed acidification precipitation liquid comprises acetone, concentrated hydrochloric acid and acetic acid, and the volume ratio of the acetone to the concentrated hydrochloric acid to the acetic acid is 3: 1: 0.5-0.8;
s2, drawing a standard curve: weighing 0.36mg of protoporphyrin-IV and 0.26mg of biliverdin, respectively dissolving in 6ml of the mixed acidified precipitation solution, carrying out vortex oscillation, keeping for 8-12 h in a dark place by ultrasonic, respectively preparing into protoporphyrin standard stock solution and biliverdin standard stock solution, respectively gradually diluting the two standard stock solutions into a series of solutions by using the mixed acidified precipitation solution, measuring an absorbance value at intervals of 2nm from the wavelength of 300-760 nm by using ultraviolet spectrophotometry, and respectively drawing standard working curves of the protoporphyrin-IV and the biliverdin according to the absorbance values and corresponding concentrations at 412nm and 680 nm;
s3, detection and analysis: centrifuging the dissolved eggshell solution, taking supernatant, measuring the primary light absorption value at the wavelength of 300-760 nm at the interval of 2nm, calculating the protoporphyrin-IV content according to the light absorption value at 412nm, and calculating the biliverdin content according to the light absorption value at 680 nm.
Further, the drying is natural drying or hot air drying, and preferably, the temperature of the hot air drying is 35-45 ℃.
Further, in the step S1, the mass-to-volume ratio of the duck egg shell powder to the mixed acidified precipitation solution is 0.15-0.4 g:6 ml.
Further, in the steps S1 and S2, the ultrasonic power and the ultrasonic time are consistent, and the ultrasonic power is 750-900W.
Further, in the step S2, the dilution ratio of the diluted serial solution is 2 times, 4 times, 8 times, 16 times and 32 times in sequence.
Further, in the step S3, the centrifugation speed is 3000-4000 r/min, and the time is 40-50 min.
Further, the step S1 includes sieving the powdered duck egg shells, wherein the sieving operation is 80-mesh sieving.
Further, the mass percent of HCl in the concentrated hydrochloric acid is 36-38%.
Further, CH in the acetic acid3The mass percent of COOH is 36-38%.
The invention has the beneficial effects that: the method has the advantages that the pigment is better extracted in the dissolving process by skillfully designing the proportion of the organic acid to the inorganic acid, the detection of the duck eggshell pigment by adopting the scheme of the invention has accurate and reliable determination result, better stability of the method, high detection efficiency, ultrasonic dissolution, shorter time consumption and low cost, the detection can be accurately finished by an ultraviolet spectrophotometer, expensive precise detection equipment is not required, and the application range is wide.
Detailed Description
In order to explain the technical content, the objects and the effects of the present invention in detail, the following description will be given with reference to the embodiments.
The embodiment of the invention is as follows: a method for detecting duck egg shell pigment comprises the following steps:
s1, preprocessing: respectively taking 10 commercially available Maka duck green shell duck eggs (the egg weight is 65.82 +/-0.32 g, the color is basically consistent) and flying duck green shell duck eggs (the egg weight is 69.63 +/-0.94 g, the color is basically consistent), removing egg shell membranes from duck egg shells, cleaning, naturally drying, preparing into powder, sieving with a 80-mesh sieve, weighing 0.25g of duck egg shell powder, adding 6ml of mixed acidification precipitation liquid, carrying out vortex oscillation, and ultrasonically dissolving for 8 hours at 800W in a dark environment, wherein the mixed acidification precipitation liquid comprises acetone, concentrated hydrochloric acid and acetic acid, and the volume ratio of the acetone to the concentrated hydrochloric acid to the acetic acid is 3: 1: 0.5;
s2, drawing a standard curve: weighing 0.36mg of protoporphyrin-IV and 0.26mg of biliverdin, respectively dissolving in 6ml of the mixed acidification precipitation solution, carrying out vortex oscillation, carrying out ultrasonic preservation for 8h in the dark at 800W, respectively preparing a protoporphyrin standard stock solution and a biliverdin standard stock solution, respectively using the mixed acidification precipitation solution to gradually dilute the two standard stock solutions into a series of solutions which are 2 times, 4 times, 8 times, 16 times and 32 times in sequence, measuring the light absorption value once at an interval of 2nm from the wavelength of 300-760 nm outside ultraviolet spectrophotometry, and respectively drawing standard working curves of the protoporphyrin-IV and the biliverdin according to the light absorption values and corresponding concentrations at 412nm and 680 nm;
s3, detection and analysis: centrifuging the dissolved eggshell solution at 4000r/min for 40min, taking supernatant, measuring the light absorption value once at the wavelength of 300-760 nm at intervals of 2nm, calculating the content of protoporphyrin-IV according to the light absorption value at 412nm, and calculating the content of biliverdin according to the light absorption value at 680 nm.
The first comparative example is: the difference from the first embodiment is that: comparative example a mixed acidified precipitation solution was used comprising acetone and concentrated hydrochloric acid in a 3:1.5 volume ratio.
The second comparative example is: the difference from the first embodiment is that: comparative example a mixed acidified precipitation solution was used comprising acetone and concentrated hydrochloric acid in a 3:1.5 volume ratio and without sonication when dissolved.
The average values of the detection results of the pigments in the sheldrake and the flying duck are respectively shown in tables 1 and 2:
TABLE 1 Duck eggshell pigment detection result table of Musca domestica
| Examples | Comparison example 1 | Comparative example two | |
| protoporphyrin-IV (mg) | 12.45 | 11.38 | 10.25 |
| Biliverdin (mg) | 3.55 | 2.95 | 2.53 |
TABLE 2 Duck eggshell pigment detection result table of flying duck
| Examples | Comparison example 1 | Comparative example two | |
| protoporphyrin-IV (mg) | 11.82 | 11.24 | 9.83 |
| Biliverdin (mg) | 2.73 | 2.12 | 2.03 |
The detection results of the comparative example and the comparative examples I and II show that the content of the pigment extracted by the scheme of the invention is higher, so that the pretreatment operation effect of the invention is better.
Relative Standard Deviation (RSD) was calculated among 10 samples of the examples, the first control and the second control, and the results of the sheldrake and the flying duck are shown in the following table 2:
TABLE 2 RSD values of sheldrake and flying duck
| Examples | Comparison example 1 | Comparative example two | |
| Sheldrake (%) | 1.06 | 2.06 | 2.36 |
| Flying duck (%) | 1.55 | 2.12 | 2.69 |
As can be seen from Table 2, the relative standard deviation RSD of the detection of the sheldrake or the flying duck by the scheme of the invention is lower than 1.55 percent, which shows that the detection method of the invention has good repeatability and higher precision.
In conclusion, the method for detecting the pigment of the duck egg shells provided by the invention has the advantages of accurate and reliable detection result and the like, and has good application prospect in seed selection and breeding of the green-shell duck eggs.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all equivalent modifications made by the present invention in the specification or directly or indirectly applied to the related technical field are included in the scope of the present invention.
Claims (8)
1. A method for detecting duck eggshell pigment is characterized by comprising the following steps: the method comprises the following steps:
s1, preprocessing: removing eggshell membranes from duck eggshells, cleaning and drying the eggshells to prepare powder, weighing the duck eggshell powder, adding mixed acidification precipitation liquid, performing vortex oscillation, and ultrasonically dissolving for 8-12 h in a dark environment, wherein the mixed acidification precipitation liquid comprises acetone, concentrated hydrochloric acid and acetic acid, and the volume ratio of the acetone to the concentrated hydrochloric acid to the acetic acid is 3: 1: 0.5-0.8;
s2, drawing a standard curve: weighing 0.36mg of protoporphyrin-IV and 0.26mg of biliverdin, respectively dissolving in 6ml of the mixed acidified precipitation solution, carrying out vortex oscillation, keeping for 8-12 h in a dark place by ultrasonic, respectively preparing into protoporphyrin standard stock solution and biliverdin standard stock solution, respectively gradually diluting the two standard stock solutions into a series of solutions by using the mixed acidified precipitation solution, measuring an absorbance value at intervals of 2nm from the wavelength of 300-760 nm by using ultraviolet spectrophotometry, and respectively drawing standard working curves of the protoporphyrin-IV and the biliverdin according to the absorbance values and corresponding concentrations at 412nm and 680 nm;
s3, detection and analysis: centrifuging the dissolved eggshell solution, taking supernatant, measuring a primary light absorption value at an interval of 2nm from the wavelength of 300-760 nm, calculating protoporphyrin-IV content according to the light absorption value at 412nm, and calculating the biliverdin content according to the light absorption value at 680 nm;
the mass percent of HCl in the concentrated hydrochloric acid is 36-38%;
the mass percentage of CH3COOH in the acetic acid is 36-38%.
2. The method for detecting duck egg shell pigment according to claim 1, wherein: the drying is natural drying or hot air drying.
3. The method for detecting duck egg shell pigment according to claim 2, wherein: the temperature of the hot air drying is 35-45 ℃.
4. The method for detecting duck egg shell pigment according to claim 1, wherein: in the step S1, the mass-to-volume ratio of the duck egg shell powder to the mixed acidized precipitation liquid is 0.15-0.4 g:6 ml.
5. The method for detecting duck egg shell pigment according to claim 1, wherein: in the steps S1 and S2, the ultrasonic power and the ultrasonic time are consistent, and the ultrasonic power is 750-900W.
6. The method for detecting duck egg shell pigment according to claim 1, wherein: in the step S2, the dilution times of the diluted serial solutions are 2 times, 4 times, 8 times, 16 times and 32 times in sequence.
7. The method for detecting duck egg shell pigment according to claim 1, wherein: in the step S3, the centrifugal speed is 3000-4000 r/min, and the time is 40-50 min.
8. The method for detecting duck egg shell pigment according to claim 1, wherein: the step S1 further comprises the step of sieving the duck egg shells which are made into powder, wherein the sieving operation is 80-mesh sieving.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101122591A (en) * | 2007-08-16 | 2008-02-13 | 中国农业大学 | Method for detecting eggshell composition deposition rule |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101122591A (en) * | 2007-08-16 | 2008-02-13 | 中国农业大学 | Method for detecting eggshell composition deposition rule |
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