CN106290442B - Utilize the method for body component content in low-field nuclear magnetic resonance technology for detection mouse - Google Patents
Utilize the method for body component content in low-field nuclear magnetic resonance technology for detection mouse Download PDFInfo
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- G01N24/00—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects
- G01N24/08—Investigating or analyzing materials by the use of nuclear magnetic resonance, electron paramagnetic resonance or other spin effects by using nuclear magnetic resonance
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Abstract
The invention provides a kind of method of body component content in technology for detection mouse using low-field nuclear magnetic resonance, comprise the following steps：Choose experiment mice and carry out low-field nuclear magnetic resonance analysis, gather nmr echo signal using CPMG pulse sequence method, obtain echo attenutation curve data；The body fluid of every experiment mice sample, fat and lean meat content are measured respectively, obtain body fluid, fat and lean meat content data；It is fitted by meterological software, establishes the regression model of body fluid, fat and lean meat content；Regression model is assessed；Determine the echo attenutation curve data of testing sample.The method of the present invention, can be under waking state, while detects body fluid in mouse, fat, lean meat content, and quick and precisely, and measurement process need not be anaesthetized, and mouse is not destroyed in itself.
Description
Technical field
It is more particularly to a kind of to utilize low-field nuclear magnetic resonance technology for detection the present invention relates to body composition of animal content detection field
The method of body fluid, fat and lean meat content in mouse.
Background technology
Body fluid, fat, lean meat content in animal body can often embody the health status of animal.In existing animal body
In body composition measurement method, traditional whole body chemical composition analytic approach is to measure the goldstandard of body composition, but this method
Process is long, workload is big, and needs to kill animal, therefore measurement repeatedly can not be carried out to animal.
New measure body composition of animal technology, such as Bioelectrical impedance analysis, dual energy x-ray absorption process, computerized tomography
Scanning and MR imaging method etc..These methods are required for anaesthetizing animal or calmness, experimental animal is kept absolute
Motionless state, but anesthesia or calmness will bring animal ingestion amount to reduce, the side effect such as hypothermia, and have death
Risk.In addition with isotope-dilution analysis and whole body electrical conductivity method etc., the numerical value that these methods measure is not accurate enough, and can not
Change to body composition is tracked research.
The content of the invention
It is an object of the present invention to for prior art in the blank of body composition of animal context of detection, a kind of utilization of proposition
The method of body fluid, fat and lean meat content in low-field nuclear magnetic resonance technology Fast nondestructive evaluation mouse, meanwhile, ensuring method letter
Single, quick, accurate, environmental protection.
To reach above-mentioned purpose, the invention provides one kind to utilize body composition in low-field nuclear magnetic resonance technology for detection mouse to contain
The method of amount, comprises the following steps：
S1, some of 10~30g experiment mices are chosen, as detection sample；
S2, low-field nuclear magnetic resonance analysis is carried out to mouse samples described in step S1, core is gathered using CPMG pulse sequence method
Magnetic resonance echo signals, obtain echo attenutation curve data；
The CPMG pulse sequence parameter is：90 degree of pulsewidth P1：13 μ s, 180 degree pulsewidth P2：26 μ s, repeated sampling wait
Time Tw：1000-10000ms, analog gain RG1：[10 to 20, be integer], digital gain DRG1：[2 to 5, it is whole
Number], pre-amp gain PRG：[1,2,3], NS：4,8,16, NECH：1000-10000, receiver bandwidth SW：100,200,
300KHz, start the control parameter RFD in sampling time：0.002-0.05ms, time delay D L1：0.1-0.5ms；
S3, mouse measurement：Mouse sample described in step S1 is dissected, the lean meat of mouse systemic is isolated, as small
Mouse lean meat detects sample, and other parts are as body fluid and fat detection sample；
The body fluid and fat detection sample and lean meat detection sample of the every mouse sample obtained respectively to dissection carry out body
The measurement of liquid parameter, fatty parameter and lean meat parameter, obtain body fluid, fat and lean meat content data；
The body fluid and fat detection sample and lean meat detection sample of the mouse sample are determined using 105 DEG C of drying constant weight methods
This, obtains the body fluid content of mouse sample described in step S1；The body fluid and fat detection sample are determined using soxhlet extraction,
Obtain the fat content of mouse sample described in step S1；The lean meat content is equal to protein in lean meat detection sample and contained
The summation of amount, moisture and content of ashes, wherein protein content are by Kjeldahl nitrogen determination, and moisture is by 105 DEG C of bakings
Dry constant weight method measure, content of ashes are determined by 550 DEG C of calcination methods；
105 DEG C of drying constant weight methods of moisture in the lean meat of the measurement mouse sample body fluid content and mouse sample
Concrete operations are：
(1) weigh with scale the body fluid and the weight of fat detection sample and lean meat detection sample of every mouse sample；
(2) body fluid and fat detection sample or lean meat detection sample are respectively placed in 105 DEG C of baking ovens, per 2h days
The flat weight for checking the body fluid and fat detection sample or lean meat detection sample, weigh with scale the body again after constant weight
The weight of liquid and fat detection sample or lean meat detection sample；The body fluid and fat detection sample or lean meat detection is calculated
The moisture of sample；
(3) moisture that the body fluid of same mouse sample and fat detection sample are detected to sample with lean meat is added
The body fluid content of as described mouse sample.
The concrete operations of soxhlet extraction of the measurement mouse sample fat content are：
(1) body fluid for having measured body fluid content and fat detection sample are ground into powder and is placed in petroleum ether, added
Heat backflow, extract the fat of the body fluid and fat detection sample；
(2) rotary evaporator evaporated in vacuo petroleum ether is used, extracted fat weight, as mouse are weighed after constant weight
The fat content of sample.
The concrete operations of Kjeldahl's method of protein content are in the lean meat of the measurement mouse sample：
(1) take part to measure the lean meat detection sample of moisture in lean meat to be digested, while enter line blank
Experiment；
(2) distilled with half kjeldahl apparatus to neutrality；
(3) it is in lavender with HCI solution, records salt acid consumption and blank assay salt acid consumption, pass through calculating
Obtain the protein content in the mouse sample lean meat.
The concrete operations of 550 DEG C of calcination methods of content of ashes are in the measurement mouse lean meat：
Take part to measure the lean meat detection sample of moisture in lean meat to be placed in crucible and weigh, place into 550
DEG C Muffle furnace in heat 6~8h, weighed after cooling, obtain the content of ashes in the mouse sample lean meat.
S4, by the echo attenutation curve data and the body fluid, fat and lean meat content data pass through meterological software
It is fitted using partial least-square regression method (PLSR) and principal component regression method (PCR), establishes body fluid, fat and lean meat contains
The regression model of amount；
S5, model evaluation：According to the forecast of regression model value and the coefficient R of actual valuecal 2And Rcv 2, it is square
Root error RMSEC and prediction standard difference SEP are assessed the regression model；
S6, the echo attenutation curve data for determining testing sample, the evaluated body fluid of utilization, fat and lean meat contain
The regression model of amount, the echo attenutation curve data to testing sample are analyzed, and obtain corresponding body fluid, and fat and lean meat contain
The predicted value of amount.
Under preferred embodiment, experiment mice described in step S1 is kunming mice, ICR mouse, LACA mouse, NIH mouse；Measurement
When, the mouse for selecting to vary in weight is as model sample.
The technological innovation of the present invention is：
1st, detection method operating process of the present invention is simple, and mouse samples to be measured are reproducible without pre-treatment, point
It is short to analyse the time, to mouse without destruction, after establishing for the regression model of prediction to every other mouse samples to be measured only
Need measurement echo attenutation curve data fat and lean meat content, can be surveyed by forecast of regression model body fluid for non-intrusion type
Amount method, and the body fluid of mouse can be measured simultaneously, fat and lean meat content, the numerical value of detection accurately, stably improve survey
Amount efficiency, it can meet that animal to live body, not anaesthetizing carries out quick body composition detection.
2nd, the inventive method need not be anaesthetized or calm to animal, and animal is not damaged, keeps experimental animal
Absolutely motionless state, will not come to examined lead for animal because anesthesia or it is calm and caused by food ration reduction, hypothermia,
The problem of even dead.
3rd, the inventive method is to the conventional foundation done the mouse strain tested and carry out body component content model, to study from now on
Experiment on the change of Mice Body component content provides conveniently.
4th, isotope-dilution analysis determines the amount without adipose tissue by the calculating to whole body total Water, is obtained so as between
Fat mass, the key of this method assume that about 73% non-fat tissue is all water, brought to measurement very big not true
Qualitative and error.Whole body electrical conductivity method, principle is substantially non-conductive using adipose tissue, and water and dielectric then turn into good and led
Isoelectric substance, lean meat content is measured indirectly by determining electrical conductivity, then by the way that fat mass is calculated, but this method measures
Numerical value it is not accurate enough, the atomic small error of cutability can cause the very big error of fat mass, and can not be to body composition
Change be tracked research.The inventive method is relative to existing isotope-dilution analysis, whole body electrical conductivity method etc., the numerical value measured
It is more accurate, and research can be tracked to the change of body composition.
In summary, mouse body composition analysis is carried out using nuclear magnetic resonance technique, is a kind of very potential quick inspection
Survey new technology.
Brief description of the drawings
Fig. 1 is the echo attenutation curve of the body component content of Kunming mouse sample of the present invention；
Fig. 2 be the predicted value of regression model established by principal component regression method (PCR) of Kunming mouse body fluid content with truly
Value returns spectrogram；
Fig. 3 be the predicted value of regression model established by principal component regression method (PCR) of Kunming mouse fat content with truly
Value returns spectrogram；
Fig. 4 be the predicted value of regression model established by principal component regression method (PCR) of Kunming mouse lean meat content with truly
Value returns spectrogram；
Fig. 5 is the predicted value for the regression model that Kunming mouse body fluid content is established by partial least-square regression method (PLSR)
Spectrogram is returned with actual value；
Fig. 6 is the predicted value for the regression model that Kunming mouse fat content is established by partial least-square regression method (PLSR)
Spectrogram is returned with actual value；
Fig. 7 is the predicted value for the regression model that Kunming mouse lean meat content is established by partial least-square regression method (PLSR)
Spectrogram is returned with actual value.
Embodiment
In order that those skilled in the art are better understood from the present invention, the present invention is entered with reference to embodiment
One step explanation.
Embodiment 1
Mouse used in following embodiments, by taking Kunming mouse as an example, but Kunming mouse is not limited to, as long as body weight is in 10-
30 grams can be applicable.
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material used, reagent etc., unless otherwise specified, are commercially obtained in following embodiments.
Specific implementation step is as follows：
Instrumental correction：Parameter is arranged to：90 degree of pulsewidth P1：13 μ s, 180 degree pulsewidth P2：26 μ s, repeated sampling stand-by period
Tw：3000ms, analog gain RG1：15, digital gain DRG1：3, pre-amp gain PRG：1, NS：8, NECH：5000, receive
Machine bandwidth SW：200KHz, start the control parameter RFD in sampling time：0.002ms, time delay D L1：0.5ms.
Sample measures：Sample low field nmr analysis：Using Mini MR-Rat magnetic resonance imaging analysis instrument to 30 Kunming mouses
Sample carries out low field nmr analysis, using CPMG pulse sequence, measures Kunming mouse T2 T2, it is bent to obtain echo attenutation
Line.(representative curve for typical sample provided in figure) as shown in Figure 1.
The present embodiment will measure the body fluid, fat, lean meat content of each mouse sample, respectively with the nuclear-magnetism of every mouse
Data are corresponding.Each mouse sample all need complete to its body fluid, fat, lean meat content measure.First carry out body fluid and thin
The measure of moisture in meat, then carry out determination of fat；Then the lean meat sample Jing Guo determination of moisture is divided into
Two parts, the measure of protein content and content of ashes in lean meat is carried out respectively, then empirically sample accounts for mouse muscle total amount
Ratio, converse protein content and content of ashes in the lean meat of every mouse；Finally carry out can be calculated every mouse
Body fluid, fat, lean meat content.
The assay method of body fluid content：105 DEG C drying constant weight methods concrete operations be：
Sample is detected as Kunming mouse body lean meat by the use of the muscle of scalpel separation Kunming mouse body whole body, other parts are as body
Liquid and fat detection sample, the body fluid and fat detection sample and lean meat detection sample of the every Kunming mouse body that weighs with scale respectively
Weight；The body fluid and fat detection sample or lean meat detection sample are respectively placed in 105 DEG C of baking ovens, examined per 2h with balance
Look into the body fluid and fat detection sample or lean meat detection sample weight, weighed with scale again after constant weight the body fluid and
The weight of fat detection sample or lean meat detection sample；The body fluid and fat detection sample or lean meat detection sample is calculated
Moisture；By the moisture of the body fluid of same Kunming mouse body sample and fat detection sample and lean meat detection sample
It is added, as the body fluid content of Kunming mouse body sample.Measurement result is as shown in table 1.
Determination of fat method：Fat content is carried out using the body fluid and fat detection sample that have determined body fluid content
Measure, due to do not have during body fluid is determined fat loss, so for fat measure do not influence.Pass through survey
Surely the fat content of 30 mouse can be respectively obtained.Drying Kunming mouse is crushed, wrapped with filter paper, by round-bottomed flask in 105 DEG C
Dried in drying box to constant weight, filter paper bag is put into extractor, 150ml petroleum ethers are added into round-bottomed flask, connect extracting
Device, condensed water is connected, 9h is extracted at 90 DEG C of water bath with thermostatic control, evaporation removes petroleum ether, dried in 105 DEG C of driers of round-bottomed flask
2h, it is thoroughly volatilized, weigh flask weight, Kunming mouse fat content is calculated, measurement result is as shown in table 1.
The measure of lean meat：Kunming mouse lean meat content=protein content+content of ashes+moisture.
Protein content is determined, using Kjeldahl's method, laboratory sample amount is 0.2g or so；Determine ash content laboratory sample amount
For 3g or so.
Kjeldahl's method surveys protein content：0.1mol/L hydrochloric acid is demarcated, weighing part has determined the lean meat of body fluid content
Sample is detected, sample is digested, while carries out blank assay, adds 0.2g copper sulphate, 6g potassium sulfates, then adds the 20ml concentrated sulfuric acids,
200 DEG C of heating, then 420 DEG C are risen to, liquid is in blue-green and clear, is distilled with half kjeldahl apparatus to neutrality, uses hydrochloric acid
It is in lavender to be titrated to solution, records salt acid consumption and blank assay salt acid consumption, calculates to obtain Kunming mouse protein content.
Content of ashes：(550 DEG C of calcination methods) takes the sample drying after surname extraction fat to weigh, and takes part to weigh, crucible
Calcination in Muffle furnace is placed in weigh within 1 hour, sample is put into crucible, is placed in calcination 8h in 550 DEG C of Muffle furnaces, is weighed after cooling,
Calculate to obtain Kunming mouse content of ashes.
By Kunming mouse lean meat content=protein content+content of ashes+moisture, the lean meat of Kunming mouse is calculated
Content, as a result as shown in table 1.
The foundation of model：By the echo attenutation relaxation curve data and body fluid content of Kunming mouse sample, fat content and thin
Meat content is fitted by meterological software respectively, utilizes principal component regression method (PCR) and PLS algorithm
(PLSR) body fluid content, fat content and the PCR (calibration set, validation-cross collection) of lean meat and PLSR (calibration set, interaction, are established
Checking collection) regression model.Software used is unscrambler9.7, it is necessary to explanation, the metering in the present embodiment
Learn software can be it is any can carry out principal component regression method (PCR) and PLS algorithm (PLSR) analysis and establish
The software of regression model, it is not limited to the citing of the present embodiment.
Determine to establish the optimal main cause subnumber needed for model by prediction residue variance and principal component Figure of the quantitative relationship.Such as
PCR forecast of regression model body fluid contents shown in table 2, the optimal main cause subnumber difference needed for fat content and lean meat content model
For 7,8 and 9.
The Kunming mouse body component content of table 1
The parameter of the Kunming mouse body component content PCR models of table 2
30 Kunming mouse (6-8 weeks age, purchased from Dalian Medical Univ's Experimental Animal Center) sample fluids to be measured, fat and
The measure of lean meat content：Low-field nuclear magnetic resonance is carried out to Kunming mouse sample to be measured, obtains echo attenutation curve data, by using
The body fluid having built up, fat and lean meat content PCR and PLSR regression model, to the echo attenutation curve of Kunming mouse sample to be measured
Data are analyzed, and obtain the predicted value of corresponding body fluid, fat and lean meat content.
The PCR regression models of Kunming mouse body fluid content as shown in Figure 2, calibration set and validation-cross collection coefficient Rcal 2With
Rcv 2Respectively 0.9664,0.9406.Such as the PCR regression models of Fig. 3 Kunming mouse fat contents, calibration set and validation-cross collection phase
Close coefficients Rcal 2And Rcv 2Respectively 0.9707,0.9304.Such as the PCR regression models of Fig. 4 Kunming mouse lean meat contents, calibration set and
Validation-cross collection coefficient Rcal 2It is respectively 0.9937,0.9850 with Rcv2.PLSR predictions Kunming mouse body fluid as shown in table 3 contains
Optimal main cause subnumber needed for the regression model of amount, fat content and lean meat content is respectively 6,7 and 7.Kunming mouse as shown in Figure 5
The PLSR regression models of body fluid content, calibration set and validation-cross collection coefficient Rcal 2And Rcv 2Respectively 0.9769,
0.9429.The PLSR regression models of Kunming mouse fat content as shown in Figure 6, calibration set and validation-cross collection coefficient Rcal 2With
Rcv 2Respectively 0.9913,0.9544.The PLSR regression models of Kunming mouse lean meat content as shown in Figure 7, calibration set and validation-cross
Collect coefficient Rcal 2It is respectively 0.9973,0.9802 with Rcv2.
The parameter of the Kunming mouse body component content PLSR models of table 3
The evaluation of model：Table 2 shows the evaluation result of Kunming mouse body fluid, fat and lean meat content PCR regression models.Body
The PCR calibration sets regression model of liquid and the result of validation-cross collection are close, coefficient Rcal 2And Rcv 20.94 is all higher than, just
Root error RMSEC and prediction standard difference SEP is respectively 0.5275 and 0.6674, smaller, utilizes PCR model prediction Kunming mouse bodies
Liquid hold-up error is smaller (table 4), illustrates that low-field nuclear magnetic resonance method combination PCR regression models can predict Kunming mouse body exactly
Liquid hold-up.The PCR regression model coefficient Rs of fatcal 2And Rcv 20.93 is all higher than, root-mean-square error RMSEC and SEP difference
It is smaller for 0.1071 and 0.1576, it is smaller (table 5) using PCR model prediction Kunming mouse fat content errors, illustrate low field core
Magnetic resonance method combination PCR regression models can predict the fat content of Kunming mouse exactly.The PCR regression models of lean meat are related
Coefficients Rcal 2And Rcv 2It is all higher than 0.98, root-mean-square error RMSEC and SEP are respectively 0.1876 and 0.2803, smaller, are utilized
PCR model prediction Kunming mouse lean meat content errors are smaller (table 6), illustrate that low-field nuclear magnetic resonance method combination PCR regression models can
To predict the lean meat content of Kunming mouse exactly.Table 3 shows Kunming mouse body fluid, fat and lean meat content PLSR regression models
Evaluation result.The PLSR regression models calibration set of body fluid and the result of validation-cross collection are close, coefficient Rcal 2And Rcv 2It is big
In 0.94, root-mean-square error RMSEC and prediction standard difference SEP are respectively 0.4436 and 0.6604, smaller, utilize PLSR models
Predict that Kunming mouse body fluid content error is smaller (table 7), illustrate that low-field nuclear magnetic resonance combination PLSR regression models can be pre- exactly
Survey the body fluid content of Kunming mouse.The PLSR forecast model coefficient Rs of fatcal 2And Rcv 20.95 is all higher than, root-mean-square error
RMSEC and SEP is respectively 0.0583 and 0.1283, smaller, smaller using PLSR model prediction Kunming mouse fat content errors
(table 8), illustrate that low-field nuclear magnetic resonance method combination PLSR regression models can predict the fat content of Kunming mouse exactly.Lean meat
PLSR forecast model coefficient Rscal 2And Rcv 20.98 is all higher than, root-mean-square error RMSEC and SEP are respectively 0.1239 He
0.3264, it is smaller, it is smaller (table 9) using PLSR model prediction Kunming mouse lean meat content errors, illustrate low-field nuclear magnetic resonance side
Method combination PLSR regression models can predict the lean meat content of Kunming mouse exactly.
The Kunming mouse body fluid PCR model predictions of table 4
The Kunming mouse fat PCR model predictions of table 5
The Kunming mouse lean meat PCR model predictions of table 6
The Kunming mouse body fluid PLSR model predictions of table 7
The Kunming mouse fat PLSR model predictions of table 8
The Kunming mouse lean meat PLSR model predictions of table 9
To sum up, the checking to regression model is passed through, it can be seen that using method foundation of the invention for predicting Kunming
The regression model of mouse body fluid, fat and lean meat content, either using principal component regression method (PCR) or using offset minimum binary
Regression algorithm (PLSR) is fitted, and can be accurately used for predicting the body fluid of Kunming mouse, fat and lean meat content, to be measured
Kunming mouse sample is easy to operate without destruction, can improve detection speed.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited thereto,
Any one skilled in the art in the technical scope of present disclosure, technique according to the invention scheme and its
Inventive concept is subject to equivalent substitution or change, should all be included within the scope of the present invention.
Claims (1)
1. a kind of method of body component content in technology for detection mouse using low-field nuclear magnetic resonance, it is characterised in that including as follows
Step：
S1, some of 10~30g experiment mices are chosen, as detection sample；
S2, low-field nuclear magnetic resonance analysis is carried out to mouse samples described in step S1, be total to using CPMG pulse sequence method collection nuclear-magnetism
Shake echo-signal, obtain echo attenutation curve data；
The CPMG pulse sequence parameter is：90 degree of pulsewidth P1：13 μ s, 180 degree pulsewidth P2：26 μ s, repeated sampling stand-by period
Tw：1000-10000ms, analog gain RG1：[10 to 20, be integer], digital gain DRG1：[2 to 5, be integer], it is preceding
Put large gain PRG：[1,2,3], NS：4,8,16, NECH：1000-10000, receiver bandwidth SW：100,200,300KHz,
Start the control parameter RFD in sampling time：0.002-0.05ms, time delay D L1：0.1-0.5ms；
S3, mouse measurement：Mouse sample described in step S1 is dissected, isolates the lean meat of mouse systemic, it is thin as mouse
Meat detects sample, and other parts are as body fluid and fat detection sample；
The body fluid and fat detection sample and lean meat detection sample of the every mouse sample obtained respectively to dissection carry out body fluid ginseng
The measurement of several, fatty parameter and lean meat parameter, obtain body fluid, fat and lean meat content data；
The body fluid and fat detection sample and lean meat detection sample of the mouse sample are determined using 105 DEG C of drying constant weight methods, is obtained
Obtain the body fluid content of mouse sample described in step S1；The body fluid and fat detection sample are determined using soxhlet extraction, obtained
The fat content of mouse sample described in step S1；The lean meat content is equal to protein content, water in lean meat detection sample
Divide content and the summation of content of ashes, wherein protein content is by Kjeldahl nitrogen determination, and moisture is by 105 DEG C of drying constant weights
Method is determined, and content of ashes is determined by 550 DEG C of calcination methods；
105 DEG C of drying constant weight methods of moisture is specific in the lean meat of the measurement mouse sample body fluid content and mouse sample
Operate and be：
(1) weigh with scale the body fluid and the weight of fat detection sample and lean meat detection sample of every mouse sample；
(2) body fluid and fat detection sample or lean meat detection sample are respectively placed in 105 DEG C of baking ovens, examined per 2h with balance
Look into the body fluid and fat detection sample or lean meat detection sample weight, weighed with scale again after constant weight the body fluid and
The weight of fat detection sample or lean meat detection sample；The body fluid and fat detection sample or lean meat detection sample is calculated
Moisture；
(3) moisture that the body fluid of same mouse sample and fat detection sample and lean meat are detected to sample is added as
The body fluid content of the mouse sample；
The concrete operations of soxhlet extraction of the measurement mouse sample fat content are：
(1) body fluid for having measured body fluid content and fat detection sample are ground into powder and is placed in petroleum ether, heated back
Stream, extract the fat of the body fluid and fat detection sample；
(2) rotary evaporator evaporated in vacuo petroleum ether is used, extracted fat weight, as mouse sample are weighed after constant weight
Fat content；
The concrete operations of Kjeldahl's method of protein content are in the lean meat of the measurement mouse sample：
(1) take part to measure the lean meat detection sample of moisture in lean meat to be digested, while carry out blank assay；
(2) distilled with half kjeldahl apparatus to neutrality；
(3) it is in lavender with HCI solution, salt acid consumption and blank assay salt acid consumption is recorded, by being calculated
Protein content in the mouse sample lean meat；
The concrete operations of 550 DEG C of calcination methods of content of ashes are in the measurement mouse lean meat：
Take part to measure the lean meat detection sample of moisture in lean meat to be placed in crucible and weigh, place into 550 DEG C
6~8h is heated in Muffle furnace, is weighed after cooling, obtains the content of ashes in the mouse sample lean meat；
S4, by the echo attenutation curve data and the body fluid, fat and lean meat content data are used by meterological software
Partial least-square regression method and principal component regression method are fitted, and establish the regression model of body fluid, fat and lean meat content；
S5, model evaluation：According to the forecast of regression model value and the coefficient R of actual valuecal 2And Rcv 2, root-mean-square error
The regression model is assessed with prediction standard difference；
S6, the echo attenutation curve data for determining testing sample, the evaluated body fluid of utilization, fat and lean meat content
Regression model, the echo attenutation curve data to testing sample are analyzed, and obtain corresponding body fluid, fat and lean meat content
Predicted value.
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