CN106353156A - Standard mix based on rapid detection of ewe milk composition and preparation process thereof - Google Patents
Standard mix based on rapid detection of ewe milk composition and preparation process thereof Download PDFInfo
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- CN106353156A CN106353156A CN201610654367.4A CN201610654367A CN106353156A CN 106353156 A CN106353156 A CN 106353156A CN 201610654367 A CN201610654367 A CN 201610654367A CN 106353156 A CN106353156 A CN 106353156A
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- 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
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/02—Food
- G01N33/04—Dairy products
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- 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
- G01N2001/2893—Preparing calibration standards
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Abstract
The invention discloses a standard mix based on rapid detection of ewe milk composition and a preparation process thereof. The standard mix is prepared from, by weight, 2.6-6.2% of proteins in 10 levels, 2.4-6.0% of fat in 10 levels, and 3.4-5.0% of lactose in 5 levels; by integrating early stage of the present study and experimental verification analysis, mass fractions of proteins, fat and lactose in ewe milk are acquired, the proteins and the fat are set to 10 gradient levels respectively, the lactose is set to 5 gradient levels, a set of 10 standard mix samples is prepared by quasi-level uniform design method, and each standard weighs 50 g and is about 50 ml; the standard mix is suitable for reducing errors in determination of ewe milk composition with a milk composition analyzer and may provide individual performance measurement to monitor milk production of sheep groups on a sheep farm, scientific data support is provided for genetic breeding of high-producing sheep groups, and scientific efficient guidance and management is provided for the production of the sheep farm.
Description
Technical field
The present invention relates to a kind of the developing and in particular to mixing mark based on quick detection Lac caprae seu ovis milk composition of a kind of standard substance
Quasi- product and its preparation technology, belong to field of food science.
Background technology
Breast in mainly contain the compositions such as moisture, fat, protein, Lactose, inorganic salt and vitamin, wherein with protein,
Based on fat and Lactose, i.e. usually said milk composition.No matter which kind of newborn source, component content in milk within the specific limits all can be
Fluctuation, fat content fluctuation is larger, and protein takes second place, and in most of the cases difference is not notable for the content of Lactose.Milk composition contains
The change of amount is affected by factors, as the kind of mammal, lactation period, parity, season, feedstuff, health status etc. because
Element.This laboratory has carried out comprehensive analysis to the Lac caprae seu ovis milk composition in Laoshan dairy goat difference lactogenic period, and result shows, Laoshan milk
In goat Lac caprae seu ovis, the content of milk composition is respectively protein 3.03%～6.05%, fat 2.45%～5.91%, Lactose
3.57%～4.62%, ash is 0.8%-1.04%, and calcium is 0.78～1.29mg/g.Lac caprae seu ovis replace after Lac Bovis seu Bubali as one kind
The emerging breast source of godmother's breast, its component content in milk granule is all little and uniform than Lac Bovis seu Bubali, and more conducively human consumption absorbs.Wherein, sheep
Lactoprotein granule is less than Lac Bovis seu Bubali, and uniformly, digestibility is up to 98% to granule；Fat is mainly made up of short-chain fatty acid, Oil globule
Diameter is less than Lac Bovis seu Bubali, and uniform in size, digestibility is up to 95%；Lactose is mainly made up of alpha-lactose and beta lactose, and the two ratio is closed
Fit and granule is little, more conducively human consumption absorbs, contain substantial amounts of adenosine triphosphate in Lac caprae seu ovis in addition, can fully decompose Lac caprae seu ovis
In Lactose, therefore seldom there are lactose intolerance and anaphylaxiss in diet Lac caprae seu ovis.
The heat stability of different breast sources breast forms directly related with it, and the heat stability of Lac caprae seu ovis is lower than Lac Bovis seu Bubali, milk composition
Change and milk composition between interaction key effect is played to milk heat stability.Processing in feature Lac caprae seu ovis
In, milk composition is also an important technology index and the economic indicator of examination milk goat production performance, the analysis of milk composition of Lac caprae seu ovis
Directly affects the quality index of high-end goat milk product.With regard to the detection method of milk composition, at present mostly according in GB to list
The detection method of one composition is detected one by one, slow compared to GB chemical method detection speed, the drawbacks of composition is single, utilize
Analysis of milk composition instrument fades in advantage to the method that milk composition is used for quickly detecting.This assay method of analysis of milk composition instrument is not only examined
Degree of testing the speed is fast, can also realize detecting protein, fat and three kinds of compositions of Lactose simultaneously, but the key of quick detection milk composition exists
Preparation and demarcation in hybrid standard product.Standard substance be a kind of have determined there are one or more sufficiently uniform characteristics
Value and can be used for calibration instrument, evaluates measuring method or the material to material assignment or material, and can be used as ginseng during measurement
Compare standard.Dhi (dairy herd improvement) is China's breeding milk animal breeding of method and dairy products' quality safety ensures body
The important component part of system, dhi standard substance be during dhi measures to Cow Milk fast analyzing constituent when the Lac Bovis seu Bubali mark calibrated
Quasi- product.Because the milk composition of Lac caprae seu ovis and its proportion of composing and structure are all different from Lac Bovis seu Bubali, so to Lac caprae seu ovis in production and processing
Inspection and its fabrication process condition are all not quite similar with Lac Bovis seu Bubali, using Lac Bovis seu Bubali standards calibration analysis of milk composition instrument come quick detection
Lac caprae seu ovis are also inaccurate.Standard substance with regard to Lac caprae seu ovis milk composition quick detection also do not occur at home or even in the world at present.Therefore,
Develop a kind of hybrid standard product of quick detection Lac caprae seu ovis milk composition for the unification effectively management of Yang Chang and to the high-end sheep of promotion
The deep processing of milk product all has great importance.
Content of the invention
It is an object of the invention to provide a kind of standard substance of Lac caprae seu ovis milk composition mensure and its preparation technology, according to this laboratory
Early-stage Study result, the mass fraction scope of main for Lac caprae seu ovis milk composition is defined as protein 2.60%～6.20% by the present invention,
Fat 2.40%～6.00%, Lactose 3.40%～5.00%, substantially covers the changing value of the general content of Lac caprae seu ovis milk composition.This
In invention, protein and fat are set to 10 levels, Lactose is set to 5 levels, according to the Uniform ity Design Method of pseudo level, if
Count out a set of 10 standard substance, the quality of each standard substance is 50g, about 50ml.
For achieving the above object, the present invention adopts following technical proposals:
First purpose of the present invention is a kind of preparation technology of the standard substance providing Lac caprae seu ovis milk composition to measure, including following
Step:
(1) Lac caprae seu ovis are carried out centrifugal degreasing, obtain fat standby；
(2) Lac caprae seu ovis after defat are heated, ultrafiltration, obtain the trapped fluid based on albumen and the transmission based on Lactose
Liquid, standby；
(3) by fat, albumen and Lactose according to the horizontal gradient setting, Lac caprae seu ovis sample is designed according to the method for uniform Design
Product uniform designs table；
(4) the standby milk composition obtaining described step (1) and (2) is according to Lac caprae seu ovis sample uniform designs table in step (3)
Carry out configuring, dissolve, filter, obtain a set of 10 samples standby；
(5) ready sample in step (4) is respectively adopted analysis of milk composition instrument (foss ft+) and chemical analysis method
Carry out the mensure of albumen, fat and Lactose, obtain two groups of measured values；
(6) two groups of measured values in step (5) are analyzed comparing, if the chemical definite value of butter oil, lactoprotein, Lactose and
Between instrument value two class value, there was no significant difference (p > 0.05), and the chemical definite value of each index in same sample and instrument value
Difference in the prescribed limit of analytical chemistry method detection error ± 0.2%, then the sample passes prepared in step (4), as
Hybrid standard product based on quick detection Lac caprae seu ovis milk composition.
In step (1), described Lac caprae seu ovis can be from fresh Lac caprae seu ovis or freezing Lac caprae seu ovis.Preferably, the present invention is from freezing
Lac caprae seu ovis, the refrigerator temperature storing during freezing be -22～-18 DEG C (preferably -20 DEG C), this temperature range both will not be destroyed
Component content in milk in Lac caprae seu ovis, also will not make milk composition go bad, and the holding time is longer；When raw Lac caprae seu ovis thaw, first by milk container from
Take out in refrigerator with cold water flush one time, then thaw under room temperature condition；After thawing completely, filtered using double gauze, filter
The impurity such as foreign body.
During centrifugation, Lac caprae seu ovis are sub-packed in centrifuge tube, and volume does not exceed the 2/3 of centrifugation pipe volume, in case Lac caprae seu ovis splash
Go out；Carry out adopting counter balance during trim, you can reach centrifugation and require.
Centrifugation adopts frozen centrifugation, and the condition of refrigerated centrifuger work is: 2～6 DEG C, 5000～6000r/min, and it is centrifuged 12
～18min, repeats two～tri- times；Preferably, the working condition of refrigerated centrifuger is: 4 DEG C, 5500r/min, and it is centrifuged 15min, weight
Again twice, this condition is conducive to the removing of fat and defat is as complete as possible；Under the conditions of obtained fat will be housed in 4 DEG C,
Sealed with preservative film, cold preservation is standby.
Preferably, after Lac caprae seu ovis carry out described centrifugal degreasing, need the Lac caprae seu ovis after defat stir, recycle gauze
Filter, obtain the Lac caprae seu ovis after at utmost defat, the Lac caprae seu ovis after this defat are in order to carry out the operation of step (2).Further preferably
Be to be filtered using double gauze.Lac caprae seu ovis after defat will stir, and is because during centrifugation, centrifuge tube bottom
Portion has a small amount of casein precipitate；In addition filtered again, on the one hand can be removed some fat of remnants, on the other hand also
The impurity such as a small amount of foreign body can be removed further.
In step (2), the temperature of heating is 40 DEG C～42 DEG C (preferably heating in water bath), time 15～20min, here heating
During to observe to have or not and addle or abnormal milk sample, if having, being given up and being recorded；Heat normal milk sample before the assay,
Should be sufficiently mixed uniformly, to reduce the evaluated error of parallel test.
Preferably, described ultrafiltration uses doughnut membrane ultrafiltration, and the condition of ultrafiltration is: 38～42 DEG C, 0.18～
0.22mpa, membrane molecule amount is 6000da, further preferred ultra-filtration conditions: 40 DEG C, 0.20mpa, treats Lactose permeate substantially not
Flow out, and when the now obtained trapped fluid volume based on albumen is of substantially equal with the fatty volume that defat obtains, stop
Ultrafiltration.Note: the Lac caprae seu ovis after defat are carried out before ultrafiltration, the cleaning of ultrafiltration apparatus to be carried out.During ultrafiltration apparatus cleaning, will transmit through
Liquid outlet is blocked, and pressure drops to 0, using isobaric cleaning, first cleans 2-3h with the sodium hydroxide solution that mass fraction is 2%, then
Clean 1-2h with the hydrochloric acid that mass fraction is 1%, then rinse 2-3h (can also be 40 DEG C of hot water) with clear water, finally with ultrapure
Twice of water rinse, is optionally replaced and cleaned liquid in cleaning process.
In step (3), described fat, albumen and lactose level gradient are as follows:
The mass fraction scope of protein is 2.6%～6.2%, be set as 2.6%, 3.0%, 3.4%, 3.8%, 4.2%,
4.6%th, 5.0%, 5.4%, 5.8%, 6.2%, 10 levels；The mass fraction scope of fat is 2.4%～6.0%, is set as
2.4%th, 2.8%, 3.2%, 3.6%, 4.0%, 4.4%, 4.8%, 5.2%, 5.6%, 6.0%, 10 levels；The matter of Lactose
Amount fraction range is 3.4%～5.0%, is set as 3.4%, 3.8%, 4.2%, 4.6%, 5.0%, 5 levels.
Described fat, albumen and Lactose are respectively the fat that centrifugal degreasing obtains, the trapped fluid based on albumen and with breast
Permeate based on sugar adds lactose assay pure (the pure interpolation of lactose assay is depending on the content of Lactose in Lactose permeate)；This
Invention, when carrying out the configuration of standard substance, each graded levels of Lactose is repeated once, takes the uniform Design of pseudo level
Method.
Lactose is set to 5 horizontal gradients, is mixed-level, takes pseudo level Uniform ity Design Method, as shown in table 1.
The uniform designs table (g/100g) of table 1 pseudo level
In step (4), carry out sample configuration when, first, before configuration, the permeate based on Lactose is measured,
Probably to obtain the content of Lactose in every milliliter of Lactose permeate, albumen quality percentage composition is the trapped fluid based on albumen
Quality, the quality of the fat that fat mass percentage composition obtains for defat, Lactose weight/mass percentage composition is saturating based on Lactose
Cross contained Lactose in liquid and additional analysis pure lactose sum, in the configuration process of whole sample, added ultra-pure water regards
Depending on the content of Lactose in Lactose permeate, more few better, added preservative is 0.1%；40 DEG C of heating in water bath of sample are used in combination
Glass rod stirring and dissolving, then recycles double gauze to filter, after cooling completely, is placed in cold preservation in 4 DEG C of cold closetes.
In step (5), when subsequent measurements are carried out to the milk composition in the sample of configuration, the chemical analysis method being adopted
It is: lactoprotein is Kjeldahl's method, with reference to gb 5009.5-2010；Butter oil be sieve purple Gothic in (rose gottlieb)
Method, with reference to gb 5413.3-2010；Lactose is spectrophotography, with reference to sn/t 0871-2012.The analysis of milk composition being adopted
Instrument model foss ft+, FOX company of Denmark produces；Clean instrument three times with fixing cleanout fluid before mensure, school zero point, so
Afterwards with controlling sample inspection apparatus twice, determine that instrument property indices normally can start test sample.
In step (6), using two groups of measured values of excel Form Handle, soft using ibm spss statistics 19.0
Part is compared the paired-sample t test in Mean Method, butter oil, lactoprotein, the chemical definite value of Lactose and two groups of instrument value
The standard that between value, there was no significant difference (p > 0.05) comes from Wang Qinde, Yang Jian. food test design and statistical analysiss [m]. the 2nd
Version. Beijing: China Agricultyre University Press, 2009.；Described chemistry definite value is detected by mistake in analytical chemistry method with the difference of instrument value
In the prescribed limit of difference ± 0.2%, this standard comes from Li Weibin. analytical chemistry [m]. Beijing: and Higher Education Publishing House, 2005..
If the standard substance that the present invention prepares meet two above standard then it is assumed that this hybrid standard product is qualified simultaneously.The present invention
In in view of instrument and the artificial error measuring operation, through lot of experiment validation and analysis, do not affecting to calibrate the premise of effect
Under, can widen to ± 0.3% prescribed limit.
The Lac caprae seu ovis milk composition bioassay standard product of the present invention are the fat that obtains of trapped fluid based on lactoprotein, defat with
And the additional proper amount of lactose assay of the permeate with Lactose based on pure for main milk composition, and add 0.1% preservative with
And a small amount of ultra-pure water is configured.Wherein, lactoprotein, butter oil and Lactose are the milk composition obtaining roughly, do not enter
The strict purification process of row, it is also less that such milk composition loses.Additional preservative does not only have to the result of final products
Impact, and microbial activitiess can be suppressed, prevent product putrid and deteriorated, thus extending the pot-life of standard substance.
Second object of the present invention is to provide the standard substance that obtain using above-mentioned preparation technology, described standard substance be according to
Following mass fractions and respective horizontal setting: the mass fraction scope of protein is 2.6%～6.2%, be set as 2.6%,
3.0%th, 3.4%, 3.8%, 4.2%, 4.6%, 5.0%, 5.4%, 5.8%, 6.2%, 10 levels；The mass fraction of fat
Scope is 2.4%～6.0%, be set as 2.4%, 2.8%, 3.2%, 3.6%, 4.0%, 4.4%, 4.8%, 5.2%, 5.6%,
6.0%, 10 levels；The mass fraction scope of Lactose is 3.4%～5.0%, be set as 3.4%, 3.8%, 4.2%, 4.6%,
5.0%, 5 levels, the method according to pseudo level uniform Design is designed to a set of 10 samples, and this sample is preferably as table 1 institute
Show.
Preferably, this standard substance is the method making according to uniform Design: take the uniform designs table of pseudo level to configure,
By 5 horizontal gradients of Lactose, each gradient is repeated once.
The mass fraction scope of described milk composition covers the mass fraction scope of Lac caprae seu ovis milk composition.The water of described milk composition
Flat setting, its gradient difference draws according to single factor experiment result, so more meets the chemical definite value of milk composition and instrument is examined
Measured value is in the prescribed limit of analytical chemistry methods detection error.
The using method of this standard substance is: is quickly measured to demarcate during milk composition using analysis of milk composition instrument and uses.
Third object of the present invention is the application providing described standard substance in calibration analysis of milk composition instrument, specifically such as
Under:
Milk composition refers to the main component based on lactoprotein, butter oil and Lactose, the system of milk composition bioassay standard product
To reduce, for can be used for calibration analysis of milk composition instrument, the error that analysis of milk composition instrument measures milk composition, and this standard substance is one kind
Hybrid standard product.Analysis of milk composition instrument is carried out after orientation ratio Lac Bovis seu Bubali standards calibration using the Lac caprae seu ovis standard substance of the present invention, uses
In the detection of Lac caprae seu ovis, the detected value obtaining is more accurate, error is less.It is mainly reflected in following several respects:
(1) the mass fraction scope of the main milk composition of Lac caprae seu ovis that the Lac caprae seu ovis standard substance invented in the present invention determine is respectively
Protein 2.60%～6.20%, fat 2.40%～6.00%, Lactose 3.40%～5.00%, compared to of the prior art
Lac Bovis seu Bubali standard substance, substantially covers Lac caprae seu ovis and the changing value of the general content of Lac Bovis seu Bubali milk composition, with Lac Bovis seu Bubali standard of the prior art
Condition compares, calibration when the more accurate and suitable Lac caprae seu ovis milk composition of the standard substance in the present invention measures.
(2) Goat milk protein content is higher than Lac Bovis seu Bubali, and with beta-casein content at most, α s1- casein content is minimum,
And α s1- casein content highest in Lac Bovis seu Bubali, α s2- casein content is minimum；Lactalbumin and casein ratio value are higher than Lac Bovis seu Bubali
Middle lactalbumin and caseic ratio value.Compared to Lac Bovis seu Bubali standard substance of the prior art, the Lac caprae seu ovis mark that the present invention is invented
Quasi- product are content and proportions for protein in Lac caprae seu ovis for the calibration of protein, solve with of the prior art
Calibration for dhi compares, when the more accurate and suitable milk protein of the standard substance in the present invention measures.
(3) Goat milk fat content is higher than Lac Bovis seu Bubali, short chain (c2～c6) and middle chain (c8～c12) fatty acid proportion
It is significantly higher than Lac Bovis seu Bubali, based on unsaturated fatty acid, and Lac Bovis seu Bubali is based on satisfied fatty acid.Compared to Lac Bovis seu Bubali of the prior art
Standard substance, the Lac caprae seu ovis standard substance that the present invention is invented are for fat content in Lac caprae seu ovis and proportions for fatty calibration
, for solving compared with Lac Bovis seu Bubali standard substance of the prior art, the more accurate and suitable sheep of the standard substance in the present invention
Calibration during Determination of Milk Fat.
(4) lactose content of Goat milk is less than Lac Bovis seu Bubali, and alpha-lactose and beta lactose ratio are suitable, compared in prior art
Lac Bovis seu Bubali standard substance, the Lac caprae seu ovis standard substance that the present invention is invented are content and ratios for Lactose in Lac caprae seu ovis for the calibration of Lactose
Example is prepared, for solving compared with Lac Bovis seu Bubali standard substance of the prior art, standard substance in the present invention more accurately and
Calibration during suitable Lac caprae seu ovis Determination of lactose.
(5) in sum, the milk composition of goat dairy is different from Lac Bovis seu Bubali except percentage composition and ratio, its corresponding structure
Be different from Lac Bovis seu Bubali, that is, the particle diameter of milk composition little and uniform in size all than Lac Bovis seu Bubali so that the digestion rate of goat dairy and digestion
Rate is above Lac Bovis seu Bubali, and lactose intolerance and anaphylaxiss seldom diet Lac caprae seu ovis occur so that goat dairy becomes one kind continues Lac Bovis seu Bubali
The emerging breast source of breast milk, therefore, the inspection to Lac caprae seu ovis and its fabrication process condition and cattle in production and processing can be substituted afterwards
Breast is compared and is all not quite similar, and detects Lac caprae seu ovis using Lac Bovis seu Bubali standards calibration analysis of milk composition instrument also inaccurate.So with existing
Lac Bovis seu Bubali standard substance in technology are compared, and the standard substance using the present invention are more accurately and reliable when rectifying an instrument detection Lac caprae seu ovis,
And between parallel test measure data stability good, using during Lac Bovis seu Bubali standard substance calibration analysis of milk composition instrument detect Lac caprae seu ovis exist
Error is larger.
The invention has the beneficial effects as follows:
(1) present invention develops a kind of domestic at present using pseudo level Uniform ity Design Method or even had not occurred in the world
The hybrid standard product of Lac caprae seu ovis milk composition quick detection, not only the exploitation to Lac caprae seu ovis and its product has for the preparation of Plays product of the present invention
There is certain directive significance, more fully, further investigation Lac caprae seu ovis milk composition and the development further of whole Lac caprae seu ovis industry establish
Good basis, provides reliable theoretical foundation.
(2) as one kind preferred embodiment, the present invention adopts 2～6 DEG C, 5000～6000r/min, 12～18min
Carry out frozen centrifugation, and be repeated twice, then filtered, obtain skimmed milk, this condition not only make fat farthest on
Floating, the fat obtaining is purer and loses few, also can obtain the pure skimmed milk of comparison simultaneously and the foreign bodies such as some impurity sink
In centrifugation bottom of the tube, and lay a good foundation for subsequently carrying out the separation and Extraction of Lactose.
(3) present invention is based on when carrying out the configuration of standard sample, utilize as much as possible the milk composition of original Lac caprae seu ovis according to
Given mass percent carries out proportioning.As one kind preferred embodiment, the present invention utilizes Hollow Fiber Ultrafiltration dense first
Contracting technology, skimmed milk is preheating to 40 DEG C about, and being adjusted to pressure is 0.20mpa, and choosing membrane molecule amount is in 6000da
Fibre ultrafiltration film is circulated ultrafiltration, respectively obtains the trapped fluid based on albumen and the permeate based on Lactose.This
Although the quality of the bright albumen obtaining and Lactose, with solution as representative, does not carry out strict purification process, so can also protect
Card milk composition will not lose substantially, and final result confirms using this technology it is rational.
(4) as one kind preferred embodiment, the present invention is when carrying out the configuration of standard substance, by each gradient of Lactose
Level is repeated once, takes the Uniform ity Design Method of pseudo level, the butter oil of the standard substance of final gained, lactoprotein, Lactose
Chemical definite value and instrument detected value two class value between there was no significant difference (p > 0.05), and the difference of the two is in analytical chemistry method
In the prescribed limit of detection error ± 0.2%.
(5) standard substance of the quick detection Lac caprae seu ovis milk composition prepared by the present invention are a kind of hybrid standard product, and it is right to can be used to
Determining instrument carries out regular calibration, so not only can ensure that accuracy to milk composition determination data, reliability and concordance,
And determination data can be made to have comparability in China or even worldwide.
(6) the hybrid standard product of the quick detection Lac caprae seu ovis milk composition prepared by the present invention can realize individual performance measurement,
Implement monitoring sheep field flock of sheep to give milk situation, and can routinely improve flock of sheep quality, to obtain the original Lac caprae seu ovis sample of high-quality, real
Now flock of sheep pasture is produced and carry out scientific and efficient ground guidance management.In addition, monitoring is implemented by the individual milk composition in sheep field, for height
The genetic breeding producing flock of sheep provides the data of science to support, it is possible to achieve the production performance comparation and assessment between sheep field and comparison in difference,
And then by relatively to realize international joint compared with external milk sheep field.
Specific embodiment
Below by instantiation the present invention will be further elaborated it should explanation, the description below be only for
The explanation present invention, is not defined to its content.
Heretofore described Lac caprae seu ovis take from 200 healthy adult Laoshan milk that milk goat plant is liked in Shandong Province Tai'an three
The fresh mixing breast of goat ewe, milking parlour is concentrated and is milked.
Heretofore described lactose assay is pure, and its concentration reaches more than 99.9%, and fine chemistry industry institute is recovered in Tianjin.
Heretofore described preservative is that potassium sorbate chemistry is pure, Tianjin Bo Di Chemical Co., Ltd..
Heretofore described ultra-pure water is made by oneself by Qilu University of Technology.
Embodiment 1: pseudo level Uniform ity Design Method prepares Lac caprae seu ovis milk composition hybrid standard product
1) take out 6 bottles of milk (about 1300ml) from deep freezer, first use twice of cold water flush, then solve under room temperature condition
Freeze, be filled in large beaker followed by double gauze, filter off the impurity such as foreign body；
2) Lac caprae seu ovis after filtering are sub-packed in centrifuge tube, the volume of subpackage is less than the 2/3, Ran Houli of centrifugation pipe volume
Carry out trim with counter balance；
3) centrifuge tube after trim respectively is put in refrigerated centrifuger, 4 DEG C, 5500r/min, is centrifuged 15min, repeat two
Secondary, carry out frozen centrifugation, and cut out, with key, the fat that upper strata freezes, the fat obtaining is put in beaker, is sealed with preservative film
Beaker mouth, is put in cold preservation in 4 DEG C of refrigerators standby；
4) the skimmed milk Glass rod after fat will be gone to stir, reuse double gauze and be filled in beaker, enter one
Step removing partial impurities and on a small quantity fat, obtain skimmed milk, using preservative film sealing, are put in cold preservation in 4 DEG C of refrigerators standby；
5) skimmed milk obtaining is taken out from cold closet, in room temperature placement 10min about, heating in water bath to 40 DEG C, choose
Pressure 0.20mpa, membrane molecule amount 6000da, it is circulated ultrafiltration, treat that the permeate based on Lactose does not flow out and substantially with egg
When fatty volume that trapped fluid volume and the defat of Bai Weizhu obtains is basically identical, stops ultrafiltration, respectively obtain based on Lactose
Permeate and the trapped fluid based on albumen, with preservative film sealing, standby；
6) permeate obtaining based on Lactose is taken 10 μ l, carry out the mensure of Lactose using spectrophotography, obtain
In this time testing, every milliliter of Lactose permeate is containing about Lactose 0.05g.
7) obtained standby milk composition is configured with reference to table 1, wherein, albumen quality percentage composition is to be with albumen
The quality of main trapped fluid, the quality of the fat that fat mass percentage composition obtains for defat, Lactose weight/mass percentage composition be with
Contained Lactose and additional analysis pure lactose sum in permeate based on Lactose, in the configuration process of whole sample, institute
Plus ultra-pure water be less than 10%, added potassium sorbate preservative is 0.1%；Concrete configuration table is as shown in table 2:
The preparation (/ 50g) of table 2 pseudo level uniform Design hybrid standard product
8) by 40 DEG C of heating in water bath of the sample having configured and use Glass rod stirring and dissolving, then recycle double gauze mistake
Filter, is sub-packed in sterilized centrifuge tube, after cooling completely, is placed in cold preservation in 4 DEG C of cold closetes, carries out the survey of follow-up indices
Fixed.
Embodiment 2: wait horizontal homogeneous method for designing to prepare Lac caprae seu ovis milk composition hybrid standard product
Method in order to preferably determine uniform Design, the horizontal homogeneous method for designing preparation mixing mark such as embodiment 2 employing
Quasi- product.Lactose is decreased gradient difference by this test, be set to 3.4%, 3.6%, 3.8%, 4.0%, 4.2%, 4.4%,
4.6%th, 4.8%, 5.0%, 5.2%, 10 horizontal gradients, the Uniform ity Design Method of level such as take, as shown in table 3.This reality
When carrying out sample configuration in testing, configured with reference to table 3, with embodiment 1, concrete configuration table is as shown in table 4 for remaining step:
The uniform designs table (g/100g) of the levels such as table 3
Table 4 grade horizontal homogeneous design the preparation (/ 50g) of hybrid standard product
Embodiment 3: add the hybrid standard product of emulsifying agent group preparation and the comparison of matched group
Add emulsifying agent sucrose ester when carrying out the configuration of standard substance (because sample needs 40 DEG C of water-baths to add after configuring
Thermal agitation is dissolved, and sucrose ester is soluble in warm water), addition is 0.2%.This every milliliter of Lactose permeate of test is containing about Lactose
0.065g, according to quality 50g of the final constant volume of standard substance, can meet the requirement of Lactose weight/mass percentage composition it is not necessary to add
Lactose assay is pure.Carry out sample configuration when, configured with reference to table 1, remaining step with embodiment 1, concrete configuration table such as table 5 institute
Show:
Table 5 adds the preparation (/ 50g) of the hybrid standard product of emulsifying agent
Test example 1: the chemical definite value of milk composition and the mensure of instrument value in the hybrid standard product that pseudo level design method is prepared
(1) mensure of milk composition chemistry definite value
The mensure of a Lactose
Take out testing sample, first 40 DEG C of heating in water bath 15～20min, and fully shaking is uniform.Weigh about 0.4～0.7g examination
Sample (being accurate to 0.0001g) is placed in 10ml tool plug graduated cylinder (or 10ml volumetric flask), is diluted with water to scale, mixes.And accurately
Pipette 2.5ml in centrifuge tube, addition 1.25ml solution of zinc sulfate, 1.25ml potassium ferrocyanide solution, with little Glass rod gently
After stirring evenly, 2min is centrifuged with the rotating speed of 2000r/min, takes supernatant liquid as sample liquid, for measuring.Remove and do not weigh sample
Outward, all carry out by said determination step, obtain sample blank liquid.Accurately pipette 0.5ml sample liquid and commensurability sample blank liquid, point
It is not placed in 10ml color comparison tube, add 2.5ml colour former.Accurately pipette Lactose standard working solution 0.00,0.20,0.40,0.60,
0.80 and 1.00ml, it is respectively placed in the color comparison tube that 10ml contains 2.5ml colour former.By sample liquid, sample blank liquid and Lactose standard
The color comparison tube of liquid, stoppers plastic plug, moves into accurately heating 6min, taking-up in boiling water bath, after cold water cooling, is diluted with water to
10ml.After shaking up, mensuration absorbance at wavelength 520nm.During mensure, Lactose titer is with without Lactose standard working solution
Blank solution adjusts zero point, draws canonical plotting, and sample liquid adjusts spectrophotometer zero point with sample blank liquid, measures its extinction
Degree, and substitute into the content calculating Lactose in sample liquid in calibration curve equation.
The mensure of b lactoprotein
Take out testing sample, first 40 DEG C of heating in water bath 15～20min, and fully shaking is uniform.Weigh 2g～6g sample (essence
Really to 0.01g) be placed in dry 100ml nitrogen fixing bottle, add 0.1g copper sulfate, 3g potassium sulfate and 10ml sulphuric acid, jog after
Bottleneck puts a little funnel, and bottle is tiltedly propped up on foraminate asbestos gauge with 45° angle.Careful heating, treats the whole carbonization of content, bubble
After foam stops completely, strengthen firepower, and keep liquid micro-boiling in bottle, after being in aeruginouss clear to liquid, be further continued for adding
Hot 0.5h～1h.Take off and let cool, be carefully added into 20ml water.After letting cool, move in 50ml volumetric flask, and be washed with a small amount and determine nitrogen
Bottle, washing liquid is incorporated in volumetric flask, adds water to scale, mixes standby.Do reagent blank test simultaneously.Then occur to vapor
The built-in water of device, at 2/3, adds several beades, methylate red ethanol solution few drops and several milliliters of sulphuric acid, to keep water to be in acid
Property, the water in heated and boiled steam evaporator simultaneously keeps seething with excitement.20.0ml boric acid solution and 2～3 are added into receiving bottle
Mixing indicator solution (the interim mixed liquors of a C.I. 13020. ethanol solution and 5 points of bromocresol green ethanol solution), and make under condensing tube
Under end insertion liquid level, according to nitrogen content in sample, accurately draw 2.0ml～10.0ml sample treatment solution and reaction is injected by little funnel
Room, with the little funnel of 10ml water washing and to be allowed to flow into reaction indoor, subsequently clamps screw clamp.10.0ml sodium hydroxide solution is fallen
Enter little funnel, unclamping screw clamp makes it slowly flow into reative cell, clamps screw clamp immediately, and adds water in little funnel with leakproof
Gas.Clamping screw clamp, starts to distill.The indoor liquid of question response comes to life, and after redistillation 10min, mobile distillate receives
Bottle, liquid level leaves condensing tube lower end, redistillation 1～2min.Then rinsed outside condensing tube lower end with a small amount of water, take off distillate
Receiving bottle.Titrate to terminal with 0.01mol/l Hydrochloric Standard Titration, the volume of record quota of expenditure hydrochloric acid solution.Simultaneously
Make reagent blank.
The mensure of c butter oil
Take out testing sample, 40 DEG C of heating in water bath 15～20min, and fully shaking is uniform.Weigh 1g～2g (to be accurate to
Testing sample 0.0001g) is placed in 10ml tool plug graduated cylinder, adds 0.2ml ammonia, shakes up.Cover stopper, graduated cylinder is placed in 65
In ± 5 DEG C of water-bath, heating 15～20min (shaking once every 5min), takes out standing, is cooled to room temperature.After standing 30s
Carry out next step operation.Add 1ml dehydrated alcohol, relax but thoroughly carry out mixing and shake up, standing, it is cooled to room temperature.Add
2.5ml absolute ether, fully vibration 1min (about 60 times), then stand 1min.Add 2.5ml petroleum ether, fully vibrate 1min
(about 60 times), then stand at least 30min, until supernatant clarification, and occur substantially being layered, read ether layer volume v.To weigh
Bottle constant weight, weighs (being accurate to 0.0001g).Accurately draw 1ml ether liquid to be placed in the weighing botle of constant weight, allow ether in fume hood
Liquid evaporates into substantially completely, then by weighing botle constant weight, weighs (being accurate to 0.0001g), and record, thus calculate the matter of butter oil
Amount.
(2) mensure of analysis of milk composition instrument (model foss ft+, FOX company of Denmark produces) detected value
1) check whether cleanout fluid and zeroing liquid reach master scale line, whether each part of stream has milk leakage phenomenon, power supply
And whether data wire connection is firm.
2) check regulated power supply, whether firmly the power line grafting of ups, turns on the power switch, and checks corresponding control instruction
Lamp displays whether normally, to confirm that ups power supply is powered normally.
3) position moving out downwards instrument host switch to, waits preheating；Open and control computer monitor power supply, print
The external powers such as machine, finally open and control computer power.
4) open software program, wait each part warning to carry out 2～3 cleanings after disappearing, and carry out 2～3 zeroings, adjust
After zero is stable, ten made hybrid standard product are put into 42 DEG C of the temperature that 15～20min in water-bath can reach regulation simultaneously
± 1 DEG C, heat time heating time not can exceed that 45min.Heating process checking for addled or abnormal milk sample, if having, being moved back
Go out and record.The milk sample of heating is taken out from water-bath, should be sufficiently mixed uniformly, input sample is simultaneously placed directly in survey on machine
Fixed, to complete corresponding module is calibrated.In continuous mode, optical viewer has or not milk sample and overflows or leak and oozes phenomenon.
5), after mensure terminates, preserve the testing result that determining instrument exports automatically.
Test example 2: wait the chemical definite value of standard substance of horizontal homogeneous design and the mensure of instrument value
Method and step is with test example 1, hybrid standard product: prepare sheep using the horizontal homogeneous method for designing that waits in embodiment 2
Newborn milk composition hybrid standard product.
Test example 3: add the chemical definite value of standard substance of emulsifying agent and the mensure of instrument value
Method and step is with test example 1, hybrid standard product: using the mixing mark adding the preparation of emulsifying agent group in embodiment 3
Quasi- product.
(3) measurement result of the chemical definite value of milk composition and instrument value in the hybrid standard product prepared
1) pseudo level group and etc. horizontal group comparison
Change using pseudo level Uniform ity Design Method and Lactose in the hybrid standard product waiting horizontal homogeneous method for designing to prepare
Learn definite value and instrument value testing result is as shown in table 6, using two groups of measured values of excel Form Handle, using ibm spss
Statistics19.0 software is compared the paired-sample t test in Mean Method, obtains p value.
Table 6 pseudo level group and etc. the chemical definite value of Lactose and instrument value (%) in the hybrid standard product of horizontal group
Conclusion: in table 6, p=0.05 >=α=0.05 of the hybrid standard product that pseudo level is prepared, instrument value and chemical definite value
Two groups of measured value differences are not notable.And wait level prepare hybrid standard product p=0.04 < α=0.05, two groups of measured value differences
Significantly.Therefore the Uniform ity Design Method of pseudo level be better than etc. level Uniform ity Design Method, illustrate Lac caprae seu ovis milk composition measure mixing mark
The preparation of quasi- product preferably adopts the Uniform ity Design Method of pseudo level, and that is, the gradient of Lactose is set to five levels.
2) add the comparison of emulsifying agent group and matched group
Add the chemical definite value of fat and instrument value testing result such as table 7 in emulsifying agent group and the hybrid standard product of matched group
Shown, using two groups of measured values of excel Form Handle, it is compared average using ibm spss statistics 19.0 software
Paired-sample t test in method, obtains p value.
Table 7 emulsifying agent interpolation group and fatty chemical definite value in the standard substance of matched group and instrument value (%)
Conclusion: in table 7, matched group is not added with difference between the chemical definite value of the hybrid standard product of emulsifying agent and instrument value
Not notable (p=0.10 > 0.05).And the difference of the chemical definite value and instrument value adding emulsifying agent group has exceeded the inspection of analytical chemistry method
Survey the prescribed limit of error ± 0.2%, and significant difference (p=0.02 < 0.05) between two groups of measured values, therefore, add emulsifying
The hybrid standard product that agent is prepared are unqualified.It follows that the preparation of hybrid standard product based on quick detection Lac caprae seu ovis milk composition is not
Preferably add emulsifying agent.
3) the chemical definite value of milk composition and the comparison of instrument value in the hybrid standard product of present invention preparation
The present invention adopts the change of butter oil, lactoprotein and Lactose in hybrid standard product prepared by pseudo level Uniform ity Design Method
Learn definite value as shown in table 8 with the testing result of instrument value, using two groups of measured values of excel Form Handle, using ibm spss
Statistics 19.0 software is compared the paired-sample t test in Mean Method, obtains p value.
The chemical definite value of milk composition and the comparison (%) of instrument value in the hybrid standard product of table 8 present invention preparation
Conclusion: as can be seen from Table 8, the chemical definite value of the milk composition of hybrid standard product of the present invention and instrument detected value
Difference is all in the prescribed limit of analytical chemistry methods detection error ± 0.2%, and the chemical definite value of fat, protein, Lactose
Difference is not notable (p > 0.05) and instrument value between.Therefore, using a set of 10 marks of the Uniform ity Design Method preparation of pseudo level
Quasi- product can use as the hybrid standard product of quick detection Lac caprae seu ovis milk composition, and that is, the method for the present invention is feasible.
Test example 4: ratio analysis of milk composition instrument calibrated using hybrid standard product in the present invention and Lac Bovis seu Bubali standard substance
Relatively
Prepare Lac caprae seu ovis to be measured, the wherein mass fraction of protein is 2.60%～6.20%, and the mass fraction of fat is
2.40%～6.00%, the mass fraction of Lactose is 3.40%～5.00%.Lac Bovis seu Bubali standard substance are Shandong Province's livestock husbandry veterinarian general station
Breeding oxen performance measurement station provides.
The hybrid standard product being respectively adopted in the table 1 of the present invention and Lac Bovis seu Bubali standard substance carry out school to breast point analytical instrument
Standard, then detects Lac caprae seu ovis to be measured using breast point analytical instrument (foss ft+), the result obtaining is as shown in table 9.
Table 9 hybrid standard product of the present invention and Lac Bovis seu Bubali standard substance calibrate respectively after comparison to testing sample measured value
Note: yd: calibrate the milk composition measured value after analysis of milk composition instrument using hybrid standard product of the present invention；Hd: treat test sample
The chemical definite value of product.Nd: using the milk composition measured value after dhi standards calibration analysis of milk composition instrument；Yd-hd: for yd and hd
Difference；Nd-hd: for the difference of nd and hd.
Conclusion: table 9 shows, treat after the standards calibration instrument of the present invention fat of test sample, albumen, the instrument value of Lactose with
Between chemical definite value, there was no significant difference (p > 0.05), and its difference is all in the regulation model of analytical chemistry method detection error ± 0.2%
In enclosing, and better than the measured value after Lac Bovis seu Bubali standards calibration.Thus illustrate, carry out calibrating breast compared to Lac Bovis seu Bubali standard substance and be divided into point
Analyser, is more applicable for the quick detection of Lac caprae seu ovis milk composition after the Lac caprae seu ovis milk composition hybrid standard product calibration instrument of the present invention,
The detected value obtaining is more accurate, error is less.
Although above-mentioned binding tests form is described to the specific embodiment of the present invention, not the present invention is protected
The restriction of shield scope, on the basis of technical scheme, those skilled in the art do not need to pay creative work and are
The various modifications that can make or deformation are still within protection scope of the present invention.
Claims (10)
1. a kind of preparation technology of the hybrid standard product based on quick detection Lac caprae seu ovis milk composition, is characterized in that, comprise the following steps:
(1) Lac caprae seu ovis are carried out centrifugal degreasing, obtain fat standby；
(2) Lac caprae seu ovis after defat are heated, ultrafiltration, obtain the trapped fluid based on albumen and the permeate based on Lactose, standby
With；
(3) by fat, albumen and Lactose according to the horizontal gradient setting, Lac caprae seu ovis sample is designed according to the method for uniform Design equal
Even design table；
(4) the standby milk composition that described step (1) and (2) obtain is carried out according to Lac caprae seu ovis sample uniform designs table in step (3)
Configuration, dissolving, filtration, obtain a set of 10 samples standby；
(5) ready sample in step (4) is respectively adopted analysis of milk composition instrument and chemical analysis method carry out albumen, fat and
The mensure of Lactose, obtains two groups of measured values；
(6) two groups of measured values in step (5) are analyzed comparing, if the chemical definite value of butter oil, lactoprotein, Lactose and instrument
Between value two class value, there was no significant difference (p > 0.05), and the difference of the chemical definite value of each index in same sample and instrument value
In the prescribed limit of analytical chemistry method detection error ± 0.2%, then the middle sample passes prepared of step (4), are as based on value
The hybrid standard product of quick detection Lac caprae seu ovis milk composition.
2. technique as claimed in claim 1, is characterized in that: in step (1), centrifugal condition is: 2～6 DEG C, 5000～6000r/
Min, is centrifuged 12～18min.
3. technique as claimed in claim 1, is characterized in that: in step (1), after Lac caprae seu ovis carry out described centrifugal degreasing, needs
Lac caprae seu ovis after defat are stirred, recycles filtered through gauze, obtain the Lac caprae seu ovis after at utmost defat, the Lac caprae seu ovis after this defat
In order to carry out the operation of described step (2).
4. technique as claimed in claim 1, is characterized in that: in step (2), described ultrafiltration uses hollow-fibre membrane and surpasses
Filter, the condition of ultrafiltration is: 38～42 DEG C, 0.18～0.22mpa, membrane molecule amount is 6000da；Preferably, treat that Lactose passes through liquid-based
This does not flow out, and when the now obtained trapped fluid volume based on albumen is of substantially equal with the fatty volume that defat obtains,
Stop ultrafiltration.
5. technique as claimed in claim 1, is characterized in that: in step (3), described Uniform ity Design Method adopts the equal of pseudo level
Even method for designing.
6. technique as claimed in claim 1, is characterized in that: in step (4), carry out sample configuration when, albumen quality percentage
Content is the quality of the trapped fluid based on albumen, the quality of the fat that fat mass percentage composition obtains for defat, Lactose matter
Amount percentage composition is contained Lactose in permeate based on Lactose and additional analysis pure lactose sum.
7. technique as claimed in claim 1, is characterized in that: in step (6), using two groups of measured values of excel Form Handle, profit
It is compared the paired-sample t test in Mean Method with ibm spss statistics 19.0 software.
8. the standard substance being prepared using the technique any one of claim 1～7.
9. standard substance as claimed in claim 8, is characterized in that: be to be prepared into according to following mass fractions and respective horizontal gradient
Arrive:
The mass fraction scope of protein is 2.6%～6.2%, be set as 2.6%, 3.0%, 3.4%, 3.8%, 4.2%,
4.6%th, 5.0%, 5.4%, 5.8%, 6.2%, 10 levels；The mass fraction scope of fat is 2.4%～6.0%, is set as
2.4%th, 2.8%, 3.2%, 3.6%, 4.0%, 4.4%, 4.8%, 5.2%, 5.6%, 6.0%, 10 levels；The matter of Lactose
Amount fraction range is 3.4%～5.0%, is set as 3.4%, 3.8%, 4.2%, 4.6%, 5.0%, 5 levels.
10. the standard substance described in claim 8 or 9 are used for the calibration of analysis of milk composition instrument during quick detection Lac caprae seu ovis milk composition.
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| CN112683613A (en) * | 2020-12-09 | 2021-04-20 | 秦皇岛市农产品质量安全监督检验中心(秦皇岛市兽药监察所、秦皇岛市饲料监测站) | Preparation method of ivermectin matrix standard substance in goat milk |
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| CN114964944A (en) * | 2022-04-20 | 2022-08-30 | 中检科(北京)测试认证有限公司 | Nitrite-containing infant formula milk powder matrix standard sample and preparation method thereof |
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