CN107356538A - Using the method for vitamin A content in spectrophotometry animal's liver - Google Patents

Using the method for vitamin A content in spectrophotometry animal's liver Download PDF

Info

Publication number
CN107356538A
CN107356538A CN201710569178.1A CN201710569178A CN107356538A CN 107356538 A CN107356538 A CN 107356538A CN 201710569178 A CN201710569178 A CN 201710569178A CN 107356538 A CN107356538 A CN 107356538A
Authority
CN
China
Prior art keywords
vitamin
sample
animal
absorbance
liver
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201710569178.1A
Other languages
Chinese (zh)
Inventor
祝海珍
吕新河
吕慧
向芳
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Tourism Professional School
Original Assignee
Nanjing Tourism Professional School
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing Tourism Professional School filed Critical Nanjing Tourism Professional School
Priority to CN201710569178.1A priority Critical patent/CN107356538A/en
Publication of CN107356538A publication Critical patent/CN107356538A/en
Pending legal-status Critical Current

Links

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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4022—Concentrating samples by thermal techniques; Phase changes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
    • G01N2001/2866—Grinding or homogeneising
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4022—Concentrating samples by thermal techniques; Phase changes
    • G01N2001/4027—Concentrating samples by thermal techniques; Phase changes evaporation leaving a concentrated sample

Landscapes

  • Physics & Mathematics (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)
  • Spectroscopy & Molecular Physics (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The invention provides a kind of method of vitamin A in quick measure animal's liver.It is characterized in that：It includes following measure：First, vitamin A standard liquid is prepared；Secondly, 0mL, 0.05mL, 0.1mL, 0.2mL, 0.5mL and 1.0mL vitamin A titer accurately being measured respectively in 7 10mL color-comparison tubes, being settled to 5mL with chloroform respectively, each pipe adds acetic anhydride 1 and dripped；Before 7 colorimetric cylinders are moved into spectrophotometer, at 650nm wavelength, absorbance is adjusted to zero point with blank solution, 5mL antimony trichloride chloroform solns is rapidly added, determines its absorbance at once, draw standard curve, calculate its equation of linear regression；Then, the absorbance for determining vitamin A in animal's liver calculates recovery of standard addition.The method of vitamin A has simple to operate in this kind of quick measure animal's liver, and detection speed is than very fast, reliable results, high repeatability and other advantages.

Description

Using the method for vitamin A content in spectrophotometry animal's liver
Technical field
The present invention relates to chemical composition detection technique field, and in particular to the detection method of vitamin A content, it is especially dynamic The detection method of vitamin A in thing liver.
Background technology
Vitamin A (vitamin A, VA) also name retinol, it is growth in humans's development and maintains body vital movement institute not The liposoluble vitamin that can lack.Vitamin A can not only maintain normal visual performance to prevent yctalopia, but also with dimension Hold complete and healthy, promotion body normal growth and development, the hematopoiesis function for strengthening body and the pre- anti-cancer of epithelial cell structure Etc. highly important function.Discovered in recent years vitamin A has different degrees of influence to brain growth and learning and memory.
Animal's liver is described as " king of vitamin A " in food.In recent years, in animal's liver vitamin A detection side Method is mainly high performance liquid chromatography (HPLC) and HPLC MS (HPLC-MS).
HPLC and HPLC-MS separating degrees are high, favorable reproducibility, and high sensitivity, accuracy is good, but its instrument is costly, general Logical physics ＆ chemistry lab does not possess instrument condition, and the instrumentation time is long, generally requires half an hour to one hour.
Spectrophotometer is physics ＆ chemistry lab underlying instrument, and instrumentation is simple, and detection speed is than very fast, reliable results, Favorable reproducibility.
The content of the invention
Goal of the invention：In order to overcome the deficiencies in the prior art, the present invention provides a kind of simple and quick Animal Liver The assay method of dirty middle vitamin A.
Technical scheme：
Using the method for vitamin A content in spectrophotometry animal's liver, including step：
(1) vitamin A standard liquid is configured：Retinyl acetate 100.0mg accurately is weighed in 500mL saponification flasks, is added 60mL absolute ethyl alcohols, 40mL30%KOH methanol solutions, flowed back saponification 30min in 100 DEG C, and 3 are extracted repeatedly with ether after cooling It is secondary, and obtain the standard liquid of vitamin A into 100mL volumetric flasks with methanol constant volume；
(2) 0.05-1.0mL vitamin A titer is measured respectively in colorimetric cylinder, is settled to respectively with chloroform 5mL, each pipe add acetic anhydride 1 and dripped；Before colorimetric cylinder is moved into spectrophotometer, at 650nm wavelength, adjusted and inhaled with blank solution Luminosity adds 5mL antimony trichlorides-chloroform soln, determines its absorbance at once to zero point；Using absorbance as ordinate, dimension Raw plain A contents draw canonical plotting for abscissa；
(3) to pre-treatment of the animal's liver sample by grinding, extraction and concentration, animal's liver extract solution is obtained；Measure The absorbance of resulting animal's liver extract solution, the content of vitamin A in sample is calculated according to following equation；
Wherein, in X- samples vitamin A content, unit mg/100g；C- is according to the animal's liver extract solution measured For absorbance by checking in the content of vitamin A in sample on canonical plotting, unit is μ g/mL；M- sample masses, unit g；V- Add the quantitative volume of chloroform, unit mL after extraction；100- is in terms of every hectogram sample.
The pre-treatment of the sample includes：
Grinding：Accurate weighing 5g samples, it is put into the mortar for filling 15g anhydrous sodium sulfates, it is complete is ground to moisture in sample Absorbed, and homogenized entirely；
Extraction：The sample that carefully will all homogenize is moved into triangular flask with cover, accurate to add 50mL ether, friction top Son, 5min is firmly shaken, vitamin A in sample is dissolved in ether, it is voluntarily clarified；
Concentration：The ether extracted liquid 5mL of clarification is taken, is put into colorimetric cylinder, is evacuated and is evaporated in 80 DEG C of water-baths, be added immediately 1mL chloroform dissolved residues.
The drafting of the standard curve is as follows：Accurately measure respectively 0mL, 0.05mL, 0.1mL, 0.2mL, 0.5mL and 1.0mL vitamin A titer is settled to 5mL with chloroform respectively in 7 colorimetric cylinders, and each pipe adds acetic anhydride 1 and dripped； Before 7 colorimetric cylinders are moved into spectrophotometer, at 650nm wavelength, absorbance is adjusted to zero point with blank solution, is rapidly added 5mL antimony trichlorides-chloroform soln, determine its absorbance at once；Using absorbance as ordinate, vitamin A content is horizontal seat Plotting canonical plotting.
A certain amount of vitamin A is added in the sample specimens after preceding processing, is then surveyed according to standard curve step The concentration of vitamin A in random sample product, so as to obtain the recovery of standard addition of sample.
Beneficial effect：Compared with prior art, this method instrumentation is simple, and detection speed is fast, reliable results by the present invention, Favorable reproducibility.
Brief description of the drawings
Fig. 1 is the standard curve and regression equation schematic diagram of the present invention.
Embodiment
The present invention is further described below in conjunction with the accompanying drawings.
It is permanent that used instrument includes UV-4802H types ultraviolet-uisible spectrophotometer, AL204 assay balances, HH-6 digital displays Warm water bath and reflux condensate device.
Used reagent includes antimony trichloride, chloroform, ether；Retinyl acetate (chromatographically pure 99.99%)； Antimony trichloride-chloroform soln (50g/L)；Vitamin A standard liquid.
Unless otherwise indicated, the reagent used in the present invention is that analysis is pure, and water used is distilled water.
Wherein, antimony trichloride-chloroform soln (50g/L) prepares antimony trichloride solution with chloroform, is stored in palm fibre In color bottle.
The preparation of vitamin A standard liquid：Retinyl acetate 100.0mg accurately is weighed in 500mL saponification flasks, is added 60mL absolute ethyl alcohols, 40mL30%KOH methanol solutions, flowed back saponification 30min in 100 DEG C, and 3 are extracted repeatedly with ether after cooling It is secondary, and with methanol constant volume into 100mL volumetric flasks, obtain the stock solution that concentration is 1.0mg/mL.
Experimental method：
The processing of sample：
(1) grind：Accurate weighing 5g animal's livers, it is put into the mortar for filling 15g anhydrous sodium sulfates, is ground in sample Moisture is absorbed completely, and is homogenized.
(2) extract：The sample that carefully will all homogenize is moved into triangular flask with cover, accurate to add 50mL ether, pressure Tight lid, firmly shakes 5min, vitamin A in sample is dissolved in ether, it is voluntarily clarified.
(3) concentrate：The ether extracted liquid 5mL of clarification is taken, is put into colorimetric cylinder, is evacuated and is evaporated in 80 DEG C of water-baths, at once 1mL chloroform dissolved residues are added, obtain animal's liver extract solution.
The drafting of standard curve：
0mL, 0.05mL, 0.1mL, 0.2mL, 0.5mL and 1.0mL vitamin A titer are accurately measured respectively in 7 ratios In colour tube, 5mL is settled to chloroform respectively, each pipe adds acetic anhydride 1 and dripped.Before 7 colorimetric cylinders are moved into spectrophotometer, At 650nm wavelength, absorbance is adjusted to zero point with blank solution, 5mL antimony trichlorides-chloroform soln is rapidly added, stands Carve and determine its absorbance.Using absorbance as ordinate, vitamin A content is that abscissa draws canonical plotting.
The measure of sample：
Using the method for the processing and analysis of sample, three kinds of processing chicken gizzard, duck liver and pork liver animal's livers, then according to mark The plot step determination sample of directrix curve, the content of vitamin A in sample is calculated according to following equation；
Wherein, in X- samples vitamin A content, unit is milligram per hectogram (mg/100g)；C- is by canonical plotting The content of vitamin A in sample is checked in, unit is micrograms per millilitre (μ g/mL)；M- sample masses, unit are gram (g)；V- is extracted Afterwards plus the quantitative volume of chloroform, unit is milliliter (mL)；100- is in terms of every hectogram sample.
Three kinds of chicken gizzard, duck liver and pork liver animal's livers are have chosen in experiment, according to containing for its vitamin A of determination of experimental method Amount.
The precision of the method for the present invention is the relative standard deviation (RSD) by determination sample) come what is investigated, survey respectively Determine the content 6 times of vitamin A in three kinds of chicken gizzard, duck liver and pork liver animal's livers, calculate relative standard deviation (RSD).
The content of vitamin A is shown in Table 1 in animal's liver：
Table 1
The measure of recovery of standard addition：
Using the processing method of sample, three kinds of processing chicken gizzard, duck liver and pork liver animal's livers, while toward adding one in sample Quantitative vitamin A, then according to the concentration of vitamin A in standard curve step measurements sample, so as to obtain the mark-on of sample Recovery.
The degree of accuracy of the method for the present invention is to be determined by the recovery of standard addition of sample to investigate.According to recovery of standard addition Experimental method, carry out the recovery of standard addition measure of vitamin A in animal's liver.
Recovery of standard addition experimental result is shown in Table 2.
Table 2
Described above is only the preferred embodiment of the present invention, it should be pointed out that：For the ordinary skill people of the art For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should It is considered as protection scope of the present invention.

Claims (4)

1. using the method for vitamin A content in spectrophotometry animal's liver, it is characterised in that：Including step：
(1) vitamin A standard liquid is configured：Retinyl acetate 100.0mg accurately is weighed in 500mL saponification flasks, adds 60mL Absolute ethyl alcohol, 40mL30%KOH methanol solutions, flow back saponification 30min in 100 DEG C, is extracted repeatedly with ether 3 times after cooling, and With methanol constant volume into 100mL volumetric flasks, the standard liquid of vitamin A is obtained；
(2) 0.05-1.0mL vitamin A titer is measured respectively in colorimetric cylinder, is settled to 5mL with chloroform respectively, respectively Pipe adds acetic anhydride 1 and dripped；Before colorimetric cylinder is moved into spectrophotometer, at 650nm wavelength, with blank solution adjust absorbance to Zero point, 5mL antimony trichlorides-chloroform soln is added, determines its absorbance at once；Using absorbance as ordinate, vitamin A contains Measure and draw canonical plotting for abscissa；
(3) to pre-treatment of the animal's liver sample by grinding, extraction and concentration, animal's liver extract solution is obtained；Measure gained The absorbance of the animal's liver extract solution arrived, the content of vitamin A in sample is calculated according to following equation；
<mrow> <mi>X</mi> <mo>=</mo> <mfrac> <mi>c</mi> <mi>m</mi> </mfrac> <mo>&amp;times;</mo> <mi>V</mi> <mo>&amp;times;</mo> <mfrac> <mn>100</mn> <mn>1000</mn> </mfrac> </mrow>
Wherein, in X- samples vitamin A content, unit mg/100g；C- is according to the extinction of the animal's liver extract solution measured The content by checking in vitamin A in sample on canonical plotting is spent, unit is μ g/mL；M- sample masses, unit g；V- is extracted Afterwards plus the quantitative volume of chloroform, unit mL；100- is in terms of every hectogram sample.
2. according to the method for claim 1, it is characterised in that：The pre-treatment of the sample includes：
Grinding：Accurate weighing 5g samples, it is put into the mortar for filling 15g anhydrous sodium sulfates, is ground to moisture quilt completely in sample Absorb, and homogenize；
Extraction：The sample that carefully will all homogenize is moved into triangular flask with cover, accurate to add 50mL ether, compresses lid, 5min is firmly shaken, vitamin A in sample is dissolved in ether, it is voluntarily clarified；
Concentration：The ether extracted liquid 5mL of clarification is taken, is put into colorimetric cylinder, is evacuated and is evaporated in 80 DEG C of water-baths, be added immediately 1mL Chloroform dissolved residue.
3. according to the method for claim 1, it is characterised in that：The drafting of the standard curve is as follows：Accurately measure respectively 0mL, 0.05mL, 0.1mL, 0.2mL, 0.5mL and 1.0mL vitamin A titer are in 7 colorimetric cylinders, respectively with three chloromethanes Alkane is settled to 5mL, and each pipe adds acetic anhydride 1 and dripped；Before 7 colorimetric cylinders are moved into spectrophotometer, at 650nm wavelength, with sky White solution adjusts absorbance to zero point, is rapidly added 5mL antimony trichlorides-chloroform soln, determines its absorbance at once；To inhale Luminosity is ordinate, and vitamin A content is that abscissa draws canonical plotting.
4. according to the method for claim 1, it is characterised in that：Added in the sample specimens after preceding processing a certain amount of Vitamin A, then according to the concentration of vitamin A in standard curve step measurements sample, so as to obtain the mark-on reclaims of sample Rate.
CN201710569178.1A 2017-07-13 2017-07-13 Using the method for vitamin A content in spectrophotometry animal's liver Pending CN107356538A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710569178.1A CN107356538A (en) 2017-07-13 2017-07-13 Using the method for vitamin A content in spectrophotometry animal's liver

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710569178.1A CN107356538A (en) 2017-07-13 2017-07-13 Using the method for vitamin A content in spectrophotometry animal's liver

Publications (1)

Publication Number Publication Date
CN107356538A true CN107356538A (en) 2017-11-17

Family

ID=60293545

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710569178.1A Pending CN107356538A (en) 2017-07-13 2017-07-13 Using the method for vitamin A content in spectrophotometry animal's liver

Country Status (1)

Country Link
CN (1) CN107356538A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114230501A (en) * 2021-12-24 2022-03-25 河南省奥林特药业有限公司 Preparation method of pork liver extract rich in vitamin A

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101676719A (en) * 2008-09-19 2010-03-24 河南瑞驰生物科技有限公司 Method for measuring diastatic enzyme activity using a spectrophotometer
CN105372374A (en) * 2015-12-11 2016-03-02 厦门出入境检验检疫局检验检疫技术中心 Method for detecting three fat-soluble vitamins A, E and K1 in milk powder
CN105866309A (en) * 2016-05-20 2016-08-17 芜湖宝瓶智能化服务外包有限公司 Determination method of vitamin A and vitamin E

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101676719A (en) * 2008-09-19 2010-03-24 河南瑞驰生物科技有限公司 Method for measuring diastatic enzyme activity using a spectrophotometer
CN105372374A (en) * 2015-12-11 2016-03-02 厦门出入境检验检疫局检验检疫技术中心 Method for detecting three fat-soluble vitamins A, E and K1 in milk powder
CN105866309A (en) * 2016-05-20 2016-08-17 芜湖宝瓶智能化服务外包有限公司 Determination method of vitamin A and vitamin E

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
张应年: "三氯化锑比色法测定牛肝视黄醇", 《甘肃农业大学学报》 *
祝海珍等: "高校液相色谱法测定动物肝脏中维生素A的含量", 《中国营养学会第十三届全国营养科学大会暨全球华人营养科学家大会》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114230501A (en) * 2021-12-24 2022-03-25 河南省奥林特药业有限公司 Preparation method of pork liver extract rich in vitamin A

Similar Documents

Publication Publication Date Title
CN109142564A (en) Method that is a kind of while detecting vitamin A. D. E
CN101893612A (en) Method for determining content of astaxanthin in antarctic krill oil by chromatography
CN107255685A (en) The high performance liquid chromatography of ultraviolet absorber in a kind of detection cosmetics
CN109030658B (en) Method for detecting fructo-oligosaccharide and raffinose in infant milk powder
CN101526509A (en) Method for rapidly determining content of preservatives in condiment
CN105842379A (en) Method for measuring phenolic estrogen by means of derivation
CN106841457B (en) The measuring method of methaqualone and diazepam residual quantity in a kind of animal derived food
Huang et al. Determination of Vitamins D2 and D3 in Edible Fungus by Reversed‐Phase Two‐Dimensional Liquid Chromatography
CN106153785A (en) A kind of online sample introduction of aflatoxin analyzes method
CN102735772B (en) Quality controlling method of ganoderma aqueous extract
CN107356538A (en) Using the method for vitamin A content in spectrophotometry animal&#39;s liver
CN108020627B (en) Method for determining residual quantity of three phenoxy carboxylic acid pesticides in tobacco by ultra-high performance synthetic phase chromatography-tandem mass spectrometry
CN104359900B (en) Kit for quickly detecting content of proline in honey
CN105572242A (en) Non-target quick screening and detecting method for PDE-5 inhibitor banned drugs in health foods
CN109254088A (en) A kind of method of the chloro- 1,2- propylene glycol of 3- in measurement flavouring
CN106198810A (en) A kind of quality determining method of the Chinese medicine composition with treatment tumor chemoradiotherapy bone marrow depression
CN107703080A (en) A kind of assay method of dandelion extract general flavone content
CN105866309A (en) Determination method of vitamin A and vitamin E
CN110068643A (en) A method of based on vitamin in Liquid Chromatography-Tandem Mass Spectrometry measurement medical food
CN108709942B (en) Method for determining vitamin A and vitamin E in milk powder
CN106680062A (en) Method for determining content of anionic surfactant by utilizing resonance rayleigh scattering method
CN107748211A (en) A kind of method using 5 kinds of macamides in deep co-melting solvent extraction measure maca
CN104345104B (en) Standard sample for headspace analysis of trace benzene in water, preparation method thereof and method for determining trace benzene in water by using standard sample
CN103278498A (en) Detecting method and kit of sibutramine hydrochloride
CN109507306B (en) Method for determining acetyl tributyl citrate in water-based adhesive

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20171117