CN107525820A - A kind of method based on low-field nuclear magnetic resonance measure Mice Body composition - Google Patents
A kind of method based on low-field nuclear magnetic resonance measure Mice Body composition Download PDFInfo
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Abstract
The invention discloses a kind of method based on low-field nuclear magnetic resonance measure Mice Body composition, comprise the following steps：(1) Mice Body composition muscles, fat, the standard specimen of moisture are made；(2) standard specimen is tested, simulant standard samples are subjected to constant temperature processing, the NMR signal of each standard specimen, and extracted valid data are tested by body fat test pulse sequence；(3) model is established, the qualitative data at the significant figure strong point of three kinds of standard specimens and each standard specimen is established to the equation of linear regression group of muscles, fat, moisture；(4) graticule amendment, using test graticule test mixing sample, the dependency relation between test value and actual value is obtained according to the method for asking for optimal solution, carries out the amendment of model；(5) test sample, the nuclear magnetic resonance data of mouse to be measured is tested, test data is analyzed using the equation of linear regression group corrected, obtain the muscles, fat, the predicted value of moisture of Mice Body to be measured.Method provided by the invention, quickly it can determine Mice Body composition under mouse waking state.
Description
Technical field
The present invention relates to body composition of animal detection field, it is more particularly to a kind of based on low-field nuclear magnetic resonance determine Mice Body into
The method divided.
Background technology
Toy muscles, fat content test can embody research of the health status of animal especially in terms of obesity at present
Very valuable such as fat and feed, fat and gene etc.；There are CCA (whole bodies in the method for testing of existing body composition of animal
Chemical composition analysis method), BIA (Bioelectrical impedance analysis) and DXA (Dual energy X ray absorptiometry), computed tomography and
MR imaging method etc..Whole body chemical composition analytic approach, testing result is accurate, but needs to use chemical reagent to carry out fatty extraction
Take, it is time-consuming it is very long, animal need to be put to death.Bioelectrical impedance analysis, principle are that fat is substantially non-conductive, pass through test organism
Electric conductivity obtains muscles content indirectly, then fat content is calculated, and error is larger.Dual energy X ray absorptiometry, X ray pass through
There is different attenuation rates when the different bone of density, lean tissue mass (LTM), adipose tissue (FTM), being then calculated needs fiber crops
Liquor-saturated, time-consuming, to human body, animal pest.
These methods are required for anaesthetizing animal or calmness, experimental animal is kept absolutely motionless state, still
Anesthesia or calmness will cause the side effects such as animal heat reduction, and have the risk of death.In addition with isotope-dilution analysis
With whole body electrical conductivity method etc., the numerical value that these methods measure is not accurate enough, and the change of body composition can not be tracked
Research.
The content of the invention
Based on above mentioned problem, it is an object of the present invention to provide a kind of side based on low-field nuclear magnetic resonance measure Mice Body composition
Method, it can quickly determine Mice Body composition.
In order to solve the problems of the prior art, technical scheme provided by the invention is：
A kind of method based on low-field nuclear magnetic resonance measure Mice Body composition, comprises the following steps：
(1) standard specimen is made, muscles, the internal organ of mouse are simulated with fresh Fresh Grade Breast, the fat of mouse is simulated with rapeseed oil,
Free water in mouse body fluid is simulated with physiological saline, different quality is chosen and makes multiple standard specimens；
(2) standard specimen is tested, the simulant standard samples in step (1) are subjected to constant temperature processing, surveyed by body fat test pulse sequence
Try the NMR signal of each standard specimen, and extracted valid data；
(3) model is established, by the significant figure strong point of the three kinds of standard specimens obtained in step (2) and the qualitative data of each standard specimen
The equation of linear regression group of muscles, fat, moisture is established by meterological software；
(4) graticule amendment, the test graticule test mixing sample obtained using step (3), three components in biased sample
Content carries out the amendment of model, it is known that according to the dependency relation asked between the method acquisition test value of optimal solution and actual value；
(5) test sample, the nuclear magnetic resonance data of mouse to be measured is tested, utilizes the linear regression corrected in step (4)
Equation group is analyzed test data, obtains the muscles, fat, the predicted value of moisture of Mice Body to be measured.
The fresh grade breast slaughtered in three days, and ice are used in some embodiments wherein, in the step (1)
Fresh preservation；The rapeseed oil used is longitudinal relaxation time, the T2 sample close with the relaxation time of mouse adipose
Product, and it is less than 10% with mouse adipose relaxation time error.
In some embodiments wherein, the T2 of the rapeseed oil is 110.68 ± 10ms, and longitudinal direction is relaxed
The Henan time is 481.12 ± 10ms.
Simulant standard samples are placed in insulating box in some embodiments wherein, in the step (2) half an hour with
On, treatment temperature is 36~38 DEG C, standard specimen temperature is reached mouse temperature.
In some embodiments wherein, body fat test pulse sequence parameter is in the step (2)：Repeated sampling etc.
Treat the time：TW=100ms, RFD=0.7ms, analog gain RG1：[10~20, be integer], digital gain DRG1：[1~3,
It is integer], repeated sampling times N S：[2~4, be integer], echo number 1：NECH1=800, echo number 2：NECH2
=50,90 ° of upset times N TI=28, and VDL1 number, half echo time：DL2=0.523ms, pre-amp gain
PRG：[0~3, be integer],
VDL1=20,24,29,35,42,51,61,73,88,106,128,154,186,224,269,324,390,470,
566,681,820,988,1189,1432,1724,2076,2500,100.
The selection mode of data is：In 1350 data that each sample test obtains, the relatively good point of signal to noise ratio is chosen,
Preceding every 50 points of 500 points only take point 2-31, and valid data points are 1170.
In some embodiments wherein, the equation of linear regression of the step (3) is each component same test time
Significant figure strong point and each standard specimen quality obtain linear fit equation by least square regression.
In some embodiments wherein, modification method is by the slope of three components in master mould in the step (4)
Correction factor k corresponding to being multiplied by respectivelyMuscles、kFat、kMoisture。
Compared with prior art, it is an advantage of the invention that：
Using technical scheme, by establishing Mice Body composition test model, to small under mouse waking state
Mouse is tested, and the muscles of mouse, fat and moisture are predicted with the nuclear magnetic resonance data of mouse, simple to operate, analysis
Time is short, and without anesthetized mice, to mouse not damaged, test accuracy is high, reproducible；Ground in diabetes, obesity, metabolism
Study carefully has important application with biomedical sectors such as nutrition.
Brief description of the drawings
Fig. 1 is the body composition test curve (1350 points of total data) that the present invention measures mouse samples；
Fig. 2 a, the graticule slope figure (1170 points of valid data) that 2b, 2c are three component standard specimens；
Fig. 3 a, the graticule intercept figure (1170 points of valid data) that 3b, 3c are three component standard specimens；
Fig. 4 is simulant standard samples by asking for the muscles calculated value of optimal solution acquisition and the recurrence spectrogram of actual value；
Fig. 5 is the recurrence spectrogram of fatty calculated value and actual value that simulant standard samples are obtained by asking for optimal solution；
Fig. 6 is simulant standard samples by asking for the moisture calculated value of optimal solution acquisition and the recurrence spectrogram of actual value；
Fig. 7 is that mouse muscles content nuclear-magnetism calculates calculated value and the recurrence spectrogram of CCA measured values；
Fig. 8 is that mouse adipose content nuclear-magnetism calculates calculated value and the recurrence spectrogram of CCA measured values.
Embodiment
Such scheme is described further below in conjunction with specific embodiment.It should be understood that these embodiments are to be used to illustrate
The present invention and be not limited to limit the scope of the present invention.The implementation condition used in embodiment can be done according to the condition of specific producer
Further adjustment, unreceipted implementation condition is usually the condition in normal experiment.
Embodiment 1
Mouse weight used in embodiment is 10-50g.
A kind of method based on low-field nuclear magnetic resonance measure Mice Body composition, comprises the following steps：
(1) standard specimen is made：
With muscles, the internal organ of fresh Fresh Grade Breast simulation mouse, with the fat of rapeseed oil simulation mouse, with physiological saline mould
Intend the Free water in mouse body fluid.Fresh grade breast is within 3 days to slaughter the time, and chilled preservation；Rapeseed oil is longitudinal direction
Relaxation time (T1), T2 (T2) sample close with the relaxation time of mouse adipose, and relaxed with mouse adipose
Henan time error is less than 10%, in this example, the rapeseed oil T2=110.68 ± 10ms, T1=481.12 ± 10ms of selection.
Choose different quality and make multiple standard specimens, each standard specimen composition is as shown in table 1；
Each standard specimen constituent table of table 1
(2) standard specimen is tested：
Simulant standard samples in step (1) are subjected to constant temperature processing, placed more than half an hour in 37 DEG C of insulating box, are ensured
Sample temperature is up to standard.The NMR signal of each standard specimen, and extracted valid data are tested by body fat test pulse sequence.
Test equipment：The MesoMR23-060H-I produced using Suzhou Niu Mai analytical instrument limited company, toy
Body composition analysis equipment, resonant frequency 23.346MHz, magnet strength 0.54T, probe coil diameter 70mm, experimental temperature control
At 31.99~32.01 DEG C.
Test parameter：90 degree of pulsewidth P1=12us, 180 degree pulsewidth P2=25us, repeated sampling stand-by period：TW=
100ms, RFD=0.7ms, analog gain RG1：[10~20, be integer], digital gain DRG1：[1~3, be integer],
Repeated sampling times N S：[2~4, be integer], echo number 1：NECH1=800, echo number 2：NECH2=50,90 ° are turned over
Turn times N TI=28, half echo time：DL2=0.523ms, pre-amp gain PRG：[0~3, be integer].
The extracting method of valid data is：In 1350 data points that each sample test obtains, winning the confidence, it is relatively good to make an uproar
Point, preceding every 50 points of 500 points only take a little 2~31, number of effective points totally 1170.
(3) model is established：
The significant figure strong point of the three kinds of standard specimens obtained in step (2) and the qualitative data of each standard specimen is soft by meterological
Part establishes the equation of linear regression group of muscles, fat, moisture by simulated annealing.
Equation of linear regression group passes through minimum for the significant figure strong point of each component same test time with each standard specimen quality
Square law returns to obtain linear fit equation, and each component has the equation of linear regression group that 1170 test graticules form standard specimen,
There are 3 × 1170 graticules altogether, the slope of each 1170 linear equation each points of three components of foundation is cut as shown in Fig. 2 a, 2b, 2c
Away from distribution as shown in Fig. 3 a, 3b, 3c；
Simulated annealing：Muscles quality, fat mass, the number range of biodiversity are set as 0~50g, initial value is equal
For 0g, iterations 10000, regressed by simulated annealing rule and change the numerical value of muscles, fat, moisture, obtained and calculate signal
1170 points and test signal 1170 point tolerance sum minimums when muscles quality, fat mass, biodiversity is most
Excellent solution is the model calculation.
(4) Modifying model：
The test graticule test mixing sample obtained using step (3), test value is obtained according to the method for asking for optimal solution
Dependency relation between actual value, with correction model.Biased sample test result is shown in Table 2.
The mixed sample test result of table 2
The update equation of muscles quality is：Y=0.9698*x, R2=0.998；
The update equation of fat mass is：Y=0.9607*x, R2=0.999；
The update equation of biodiversity is：Y=1.071*x, R2=0.994；
Wherein x represents that sample quality is true, and y represents sample measurement quality, and 0.9698,0.9607,1.071 correspond to muscle respectively
Meat, fat, the correction factor k of moistureMuscles、kFat、kMoisture。
The slope of three composition equation groups in step (3) is modified, modification method is three groups of respective slopes
Data are multiplied by corresponding correction factor, i.e. the 1170 of muscles system of linear equations slope data is multiplied by 0.9698 respectively, fat line
1170 slope datas of property equation group are multiplied by 0.9607 respectively, and 1170 slope datas of moisture system of linear equations are multiplied by respectively
1.071 complete Modifying model.
Tested after amendment, three component assays respectively as shown in Fig. 4, Fig. 5, Fig. 6,
The correction equation of simulated fat：Y=0.9958*x+0.0093, R2=0.9993；
Simulate the correction equation of muscles：Y=0.9992*x-0.1368, R2=0.9977；
The correction equation of moisture is：Y=0.998*x+0.0112, R2=0.9997.
(5) sample test：Model carries out body composition test after establishing to 25 mouse samples, and it is bent to obtain nuclear magnetic resonance test
Line, Fig. 1 are the test curve of a mouse test sample, and test data is divided using the equation of linear regression group corrected
Analysis, carry out equation optimal solution using simulated annealing and ask for, obtain the predicted value of corresponding muscles, fat, free moisture, its
In middle muscles and fat content compared with CCA methods, be shown in Table 3.
(6) data verification：Mouse is killed, the fat mass of mouse is calculated by chemical composition analysis method, is total to nuclear-magnetism
The mouse compositional data that the method for shaking tests to obtain is analyzed, and muscles, fat, moisture are analyzed by regression analysis model
Accuracy, data are shown in Table 3：
Compared with the nuclear-magnetism method of table 3 dissects (CCA) muscles and fat content with mouse
Wherein, the mean error of muscles is 0.41g, and fatty mean error is 0.49g.
Mouse muscles content nuclear-magnetism calculated value and the recurrence collection of illustrative plates of CCA measured values are as shown in fig. 7, mouse adipose content nuclear-magnetism
Calculated value and the recurrence collection of illustrative plates of CCA measured values are as shown in Figure 8.
It can be seen that being tested by low field nuclear-magnetism, three composition regression equation groups determine calibration method, can accurately use
It is simple to operate to mouse samples not damaged in mouse muscles, fat and determination of moisture, fast.
The foregoing examples are merely illustrative of the technical concept and features of the invention, its object is to allow the person skilled in the art to be
Present disclosure can be understood and implemented according to this, it is not intended to limit the scope of the present invention.It is all smart according to the present invention
The equivalent transformation or modification that refreshing essence is done, should all be included within the scope of the present invention.
Claims (7)
- A kind of 1. method based on low-field nuclear magnetic resonance measure Mice Body composition, it is characterised in that comprise the following steps：(1) standard specimen is made, muscles, the internal organ of mouse are simulated with fresh Fresh Grade Breast, the fat of mouse is simulated with rapeseed oil, with life The Free water in salt water modeling mouse body fluid is managed, chooses the standard specimen for making multiple different qualities；(2) standard specimen is tested, the simulant standard samples in step (1) are subjected to constant temperature processing, is tested by body fat test pulse sequence each The NMR signal of standard specimen, and extracted valid data；(3) model is established, the significant figure strong point of the three kinds of standard specimens obtained in step (2) and the qualitative data of each standard specimen are passed through Meterological software establishes the equation of linear regression group of muscles, fat, moisture；(4) Modifying model, the test graticule test mixing sample obtained using step (3), the content of three components in biased sample , it is known that the dependency relation between test value and actual value is obtained according to the method for asking for optimal solution, according to correction factor k to former mould The slope of type is modified, wherein, k is coefficient using actual value as x, using calculated value as y function, y=k*x；(5) test sample, the nuclear magnetic resonance data of mouse to be measured is tested, utilizes the equation of linear regression corrected in step (4) Group is analyzed test data, obtains the muscles, fat, the predicted value of moisture of Mice Body to be measured.
- 2. the method according to claim 1 based on low-field nuclear magnetic resonance measure Mice Body composition, it is characterised in that：It is described The fresh grade breast slaughtered in three days, and chilled preservation are used in step (1)；The rapeseed oil used for longitudinal relaxation time, The T2 sample close with the relaxation time of mouse adipose, and be less than with mouse adipose relaxation time error 10%.
- 3. the method according to claim 2 based on low-field nuclear magnetic resonance measure Mice Body composition, it is characterised in that：It is described The T2 of rapeseed oil is 110.68 ± 10ms, and longitudinal relaxation time is 481.12 ± 10ms.
- 4. the method according to claim 1 based on low-field nuclear magnetic resonance measure Mice Body composition, it is characterised in that：It is described Simulant standard samples are placed more than half an hour in insulating box in step (2), treatment temperature is 36~38 DEG C, reaches standard specimen temperature Mouse temperature.
- 5. the method according to claim 1 based on low-field nuclear magnetic resonance measure Mice Body composition, it is characterised in that：It is described Body fat test pulse sequence parameter is in step (2)：The repeated sampling stand-by period：TW=100ms, RFD=0.7ms, simulation increase Beneficial RG1：[10~20, be integer], digital gain DRG1：[1~3, be integer], repeated sampling times N S：[2~4, For integer], echo number 1：NECH1=800, echo number 2：NECH2=50,90 ° of upset times N TI=28, during half echo Between：DL2=0.523ms, pre-amp gain PRG：[0~3, be integer].
- 6. the method according to claim 1 based on low-field nuclear magnetic resonance measure Mice Body composition, it is characterised in that：It is described The equation of linear regression of step (3) passes through a most young waiter in a wineshop or an inn for the significant figure strong point of each component same test time with each standard specimen quality Multiply recurrence and obtain linear fit equation.
- 7. the method according to claim 1 based on low-field nuclear magnetic resonance measure Mice Body composition, it is characterised in that：It is described Modification method is that the slope of three components in master mould is multiplied by into corresponding correction factor k respectively in step (4)Muscles、kFat、kMoisture。
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111272799A (en) * | 2019-10-22 | 2020-06-12 | 浙江大学 | Method for detecting water migration of frozen portunid based on low-field nuclear magnetic resonance |
| CN112683940A (en) * | 2020-12-24 | 2021-04-20 | 恒天海龙（潍坊）新材料有限责任公司 | Method for rapidly determining content of flame retardant in inorganic flame-retardant cellulose fiber |
| CN112881450A (en) * | 2020-05-09 | 2021-06-01 | 上海纽迈电子科技有限公司 | Quantitative analysis model construction and quantitative analysis method and system for tissue components |
| CN112946003A (en) * | 2019-12-11 | 2021-06-11 | 上海纽迈电子科技有限公司 | Method for detecting fat content in milk powder |
| CN114460121A (en) * | 2022-01-28 | 2022-05-10 | 中国农业科学院农业质量标准与检测技术研究所 | Method for detecting water content and fat content of livestock meat by using low-field nuclear magnetic resonance technology |
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