CN104408314A - Prediction method for lead content fading in lactation of dairy cow and application - Google Patents
Prediction method for lead content fading in lactation of dairy cow and application Download PDFInfo
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Abstract
The invention discloses a prediction method for lead content fading in lactation of a dairy cow and application, and belongs to the technical field of food safety. The lead content of cow milk lactated by a lead exceeded dairy cow is measured, the obtained data is counted and analyzed to obtain a fitting equation, then the equation is verified to obtain the prediction method, and the obtained prediction equation is used for predicting the lead content fading time in lactation of the lead exceeded dairy cow. According to the method, the relative error of the predicted value of the lead concentration in the cow milk is within 5 percent, and the accuracy reaches 95.13-98.73 percent; the method is applicable to predicting the lead concentration fading level in the lactation process of the lead exceeded dairy cow and the time required for fading to be below limited concentration, so the investment in manpower and material resources of enterprises is reduced, the cost is saved, the loss is reduced, and meanwhile powerful and reliable guarantee is also provided for safety of dairy products.
Description
Technical field
The present invention relates to Forecasting Methodology and application that in a kind of Lactation of Dairy Cow, lead content disappears, belong to technical field of food safety.
Background technology
Milk is a kind of rich in nutritive value, close to perfect healthy food, be also one of very important raw material in food processing.In recent years, along with improving constantly of people's living standard, the consumption per head of milk also increases considerably, and people more and more pay attention to the safety problem of milk.And heavy-metal residual is one of safety factor affecting milk, heavy metal ion, not easily by bioprocess metabolism, thus can cause heavy metal ion content in milk to exceed standard.Heavy metal element, mainly comprise copper, lead, iron, zinc, cadmium, parent, chromium, gold, silver, violent, nickel, crane, molybdenum etc. about 45 kinds, in China's agricultural production, livestock products field, the distribution range of lead, cadmium, mercury pollution and the extent of injury are comparatively serious.
Plumbous widely distributed at occurring in nature.Owing to having good ductility, high density and LMP, lead is just used on a large scale in the early stage daily life of the mankind.At present, the production field such as various metal smelt, accumulator, paint, plating and chemical industry is the main source of lead contamination.In addition, also extensively adopt lead compound as anti-knock agent in gasoline, in recent years along with a large amount of growths of vehicles number, vehicle exhaust also becomes an important channel of lead contamination physical environment.Can combine with a series of protein, organized enzyme and amino acid after lead ion enters human body, affect the nerve of human body, blood circulation, the systems such as cardiovascular, digestion, uropoiesis, cause hypertension, arrhythmia cordis, ataxia and the symptom such as anaemia, stupor.In addition, children are responsive especially to the toxicity of lead, and International Medical organization prescribed just can be diagnosed as children ' s lead poisoning when Pb-B exceedes or equals 100 micrograms per litre.Saturnism can have a strong impact on Physique growth and the intelligent development of children, causes the serious harms such as growth retardation, immunity degradation and attention deficient disorder.
Country variant all limits lead content in milk and milk products with organizing, in National Standard of the People's Republic of China GB2762-2012 " pollutants in food limitation ", the Limited Doses of regulation lactogenesis, pasteurization milk, sterile milk, acidified milk, modulation Ruzhong lead is 0.05mg/kg, Limited Doses plumbous in milk powder, non-desalted whey powder is 0.5mg/kg, and Limited Doses plumbous in other dairy products is 0.3mg/kg; Limited Doses plumbous in regulation milk and milk products in Codex Alimentary Commission (CAC) CODEX STAN 193-1995 is 0.02mg/kg; EU Committee's regulations (EC) No 629/2008 specifies as in raw milk, thermal treatment breast and other dairy products, plumbous Limited Doses is 0.02mg/kg.
Process of industrialization makes Heavy Metals In Environment day by day serious, and the probability that heavy metal is moved in Contents in Cows and in cow's milk by environment increases, and in daily life, milk cow is taken in because of unexpected or after contact lead contamination input, lead content in produced cow's milk can be caused to exceed standard.When milk cow makes lead content in secreted cow's milk exceed national limit standard (0.05mg/kg) because of unexpected absorption or contact lead contamination input, cow's milk that milk cow secretes cannot be produced as raw milk, but milk cow cost is larger, by larger for its superseded enterprises' loss, lead content in this lactation is exceeded standard, but the milk cow not reaching saturnism level still has its commercial value, can by its isolated rearing, wait for that lead is discharged by own metabolism by milk cow, until lead content could be produced as raw milk again lower than after national limit standard in cow's milk secreted by it, but in order to monitor the fade level being isolated lead content in milk cow cow's milk and the time of disappearing to national Limited Doses, just must the cow's milk secreted by milk cow be detected in real time, add manpower, material resources and enterprise operation cost, increase enterprises' loss.Therefore, prior art is badly in need of a kind of method can predicting that in cow's milk, lead content disappears, to alleviate the input of business manpower, material resources, cost-saving, reduce the loss, and also ensures reliably for the safety of dairy products provides strong simultaneously.
Summary of the invention
For solving the deficiencies in the prior art, the invention provides Forecasting Methodology and application that in a kind of Lactation of Dairy Cow, lead content disappears, the technical scheme of employing is as follows:
The object of the present invention is to provide the Forecasting Methodology that in a kind of Lactation of Dairy Cow, lead content disappears, the method collects the cow's milk secreted by lead doping milk cow, detect initial concentration plumbous in cow's milk, management of feeding is isolated to lead doping milk cow, according to the cow's milk secreted by sampling plan collection isolation milk cow, gathered cow's milk is carried out to the mensuration of lead content, statistical study is carried out to gained lead content data and obtains fit equation, obtain Forecasting Methodology after again fit equation being verified, utilize the lead content regression time in gained predictive equation prediction lead doping Lactation of Dairy Cow; Described predictive equation is:: Y=X/ (2.0289+2.2300X
2); Wherein, X is the rear number of days of isolation, and Y is mass concentration plumbous in cow's milk.
The step of described method is as follows:
1) collect the cow's milk secreted by lead doping milk cow, detect initial concentration plumbous in cow's milk;
2) management of feeding is isolated to lead doping milk cow, according to the cow's milk secreted by sampling plan collection isolation milk cow;
3) sample pre-treatments is carried out to gathered cow's milk, detect content plumbous in cow's milk;
4) statistical study is carried out to detection data, obtain fit equation, fit equation is verified that described in rear acquisition predictive equation, predictive equation is: Y=X/ (2.0289+2.2300X
2); Wherein X is number of days after isolation, and Y is mass concentration plumbous in cow's milk;
6) predictive equation is utilized to predict lead concentration regression time in Lactation of Dairy Cow.
Described step 1) in lead doping milk cow be in secreted cow's milk lead content more than the non-poisoning milk cow of 0.05mg/kg.
Described step 1) in dairy bread be holstein cow.
Described step 1) in initial concentration not higher than 0.2315mg/kg.
Step 2) described sampling plan, be designated as the 1st day by just isolating the milk sample that milk cow gathers, the 1st day to the 19th day every day, sampling should be carried out, each institute sample thief from the same day 7 point, that the newborn mixing obtained is squeezed by 15 and 23 institutes is newborn; 20th day to the 47th day, sampling should be carried out every other day, each institute sample thief from the same day 7 point, 15 and 23 squeeze the mixing breast that breast obtains.
Preferably, step 3) lead content in described detection cow's milk adopts GFAAS (graphite furnace atomic absorption spectrometry) to detect.
Step 4) described statistical study carries out nonlinear fitting to gained lead content data.
The concrete steps of described method are:
1) collect the exceed standard milk sample of lead content at 0.05-0.2315mg/kg, and measure initial concentration;
2) management of feeding is isolated to lead doping milk cow, according to the cow's milk secreted by sampling plan collection isolation milk cow;
3) described sampling plan is: the milk sample that just isolation milk cow has gathered is designated as the 1st day, and the 1st day to the 19th day every day, sampling should be carried out, each institute sample thief from the same day 7 point, that the newborn mixing obtained is squeezed by 15 and 23 institutes is newborn; 20th day to the 47th day, sampling should be carried out every other day, each institute sample thief from the same day 7 point, 15 and 23 squeeze the mixing breast that breast obtains;
4) sample pre-treatments is carried out to gathered cow's milk, adopt GFAAS (graphite furnace atomic absorption spectrometry) to detect content plumbous in cow's milk;
5) nonlinear fitting is carried out to detection data and obtain fit equation, then checking acquisition predictive equation is carried out to fit equation;
6) described predictive equation is: Y=X/ (2.0289+2.2300X
2), wherein, X is the rear number of days of isolation, and Y is mass concentration plumbous in cow's milk; Utilize step 4) fade level of the predictive equation of gained prediction Ruzhong lead doping milk cow lead concentration in lactation process and time.
In the present invention, milk cow is holstein cow, and body weight is 550 ~ 620kg, one full year of life at age 2 ~ 4, and except milk cow to produce in cow's milk except lead content exceeds standard, without other diseases, body condition is good, and saturnism symptom does not appear in milk cow.
Method of the present invention can be applied to prediction lead doping milk cow lead concentration fade level and disappearing to the following required time of limitation concentration in lactation process.
The present invention carries out Isolation Management to the Ruzhong lead doping milk cow found, the sample squeezing breast every day is gathered, storage etc., in order to reach expection good result, construct a large amount of sample informations, ensure that institute's sample thief accuracy, prevent non-monitored breast to be mixed into or the generation of cross pollution situation of sample room.The accuracy in detection of the present invention to adopted Ruzhong lead is high, and method standard is consistent, and same people has detected all samples, to reduce systematic error.The present invention utilizes 1stOpt mathematical analysis software to carry out statistical study to the result gathering rear sample detection, nonlinear fitting can be carried out to result curve, need effectively to screen assessment to the curve simulated, and carry out a large amount of detections and calculate checking, to filter out optimum Fitting curve equation.
Beneficial effect of the present invention:
The method of the invention only needs the initial concentration of lead doping in the galactopoiesis of known milk cow institute, namely by concentration plumbous in prediction equation after this milk cow institute galactopoiesis every day, without the need to carrying out collection, the detection of milk sample every day to the milk cow of Ruzhong lead doping, easy and simple to handle, accuracy is high.Decrease because needing monitoring to lead concentration in the galactopoiesis of milk cow institute and the human cost consumed, comprise the collection of milk sample, storage, detection etc., material resources cost comprises collects milk sample consumables cost used, detects many costs such as reagent, consumptive material, instrumental analysis of Ruzhong heavy metal experimental technique used.Simultaneously the method for the invention is a kind of cow's milk lead concentration method of real-time of very time-saving and efficiency, and forecasting accuracy is high, solves the problems such as conventional monitoring methods used time long, monitoring result hysteresis quality.In addition, the prediction that the method for the invention disappears to lead concentration in the galactopoiesis of milk cow institute, for the monitoring of risks and assumptions plumbous in raw milk provides effective means, for dairy safety provides reliable guarantee.
Accompanying drawing explanation
Fig. 1 is best fit equation curve.
Fig. 2 calculated value and actual value residual error column diagram.
Embodiment
Below in conjunction with specific embodiment, the present invention will be further described, but the present invention is not by the restriction of embodiment.
Embodiment 1: case finds
Pasture, the Northeast has found that a collection of Ruzhong lead content exceeds the holstein cow of national limit standard (0.05mg/kg) after testing, is owing to having eaten lead doping in the cow's milk that causes of green grass or young crops storage corn that the accumulator that goes out of use pollutes by mistake through investigation confirmation milk cow.
Embodiment 2: the collection of Cow-feeding management and sample
Lead doping milk cow is chosen with reference to embodiment 1.
From the holstein cow that the Ruzhong lead content embodiment 1 exceeds standard, choose 40 carry out isolation and feed, side by side except the absorption of the storage of lead contamination input green grass or young crops corn, feed, water etc.Carry out feeding and squeezing breast, every day squeezes breast three times, squeezes 7 points, 15 points, 23 points that the breast time is every day.
Get about 150mL milk sample with the cleaned tygon sampling jar of acid during each squeezing breast and be put in 4 DEG C of refrigerator storage, treat three times milk sample gather after, the sample of three sampled points is mixed with the volume ratio of morning, noon and afternoon 4:3:3, be dispensed in the cleaned vinyon centrifuge tube of 50mL acid (in triplicate), the sample of collection is numbered, puts into-20 DEG C of refrigerators immediately and preserve.The milk sample that firm isolation milk cow gathers is designated as the 1st day, and the 1st day to the 19th day every day, sampling should be carried out, each institute sample thief from the same day 7 point, 15 and 23 squeeze and newbornly must mix breast; 20th day to the 47th day, sampling should be carried out every other day, each institute sample thief from the same day 7 point, 15 and 23 squeeze and newbornly must mix breast;
Embodiment 3: the detection of lead content in milk sample
Sample collection is carried out with reference to embodiment 2.
(1) sample pre-treatments
Milk sample is placed in room temperature (25 DEG C) by-20 DEG C of refrigerators taking-ups thaw, concussion mixing in vortex oscillator.About 5g milk sample (being accurate to 0.001g) is taken in 50mL glass digestion tube with disposable plastic tube, add 10mL 9:1 nitration mixture (nitric acid: perchloric acid), add a cover soaked overnight to clear up in advance, spend the night after clearing up and again digestion tube is put into elder generation's heating and decompose at 100 DEG C on electric heating counteract appearance, when digestion solution is yellow transparent state, temperature is progressively promoted (120 DEG C-150 DEG C), finally be elevated to more than 180 DEG C heating and decomposes and catch up with acid, if sample becomes black, then dripping several red fuming nitric acid (RFNA)s continues to clear up until black disappears, to be heated to sample closely dry after, take off digestion tube cooling after with 2% nitric acid constant volume to 50ml, fully shake up and treat machine testing.Do 3 parallel reagent blanks simultaneously.
(2) GFAAS (graphite furnace atomic absorption spectrometry) detects lead content
A. the drafting of typical curve
Plumbous for 40 μ g/L standard working solution, the cup position of the sample disc that the system of being placed on is edited, instrument carries out gradient dilution according to the concentration gradient editted and automatically introduces sample standard solution, and dilution is ultrapure water.Plumbous concentration of standard solution is respectively 0 μ g/L, 4 μ g/L, 10 μ g/L, 20 μ g/L, 30 μ g/L, 40 μ g/L.Apparatus measures is by low concentration to high concentration, and sample size is 20 μ L.Calibration curve type of calibration is Linear, Calculated Intercept, external standard method is adopted to measure, with the concentration of standard substance for independent variable, element absorbance is dependent variable, utilizes the process of instrument AA WinLab software to do linear regression, draws out typical curve, and obtain related coefficient, for the measurement of sample concentration.
B. the condition of instrument
Table 1 graphite furnace atomic absorption spectrophotometer running parameter
| Instrument condition | Plumbous Pb |
| Hollow cathode lamp (nm) | 283.31 |
| Lamp current (mA) | 10 |
| Slit width (nm) | 0.7 |
| Sample size (μ L) | 20 |
| Background correction | Deuterium lamp |
| Matrix modifier (μ L) | Nothing |
The graphite furnace heating schedule that table 2 lead detects
Embodiment 4: the foundation of model
Collect sample with reference to embodiment 2, detect lead content in sample with reference to embodiment 3.
The present invention chooses 30 lead doping milk cows for Modling model, the plumbous initial concentration in Ruzhong is 0.2315mg/kg, the 1stOpt mathematical analysis software of 7 degree of freedom high-tech company limited (7D-Soft High Technology Inc.) stand-alone development is used to carry out statistical study to its milk sample testing result, to isolate rear number of days (d) for horizontal ordinate, mass concentration (mg/kg) plumbous in cow's milk carries out non-linear curve fitting for ordinate.Analytic process is as follows:
The sample concentration of 30 cow head every days to be averaged mass concentration plumbous in every day cow's milk, code book is opened in 1stOpt, edit code, mass concentration (mg/kg) plumbous in number of days (d) and cow's milk is input in 1stOpt code book respectively as independent variable and dependent variable, carry out formula fitting, draw the function model equation that 10 degrees of fitting are higher, in table 3, this matching adopts standard difference evolution algorithm Differential Evolution (DE) to carry out non-linear curve fitting.
Table 3 fitting function equation and major parameter
Note: R value: Correlation Coef.; R
2: R-Square; RMSE:Root of Mean Square ErrorSSE:Sum of Square Error; DC:Determination Coef.
Consider R value, the R of fit equation
2value, residual error scope, RMSE root-mean-square error, SSE average error summation, DC determine the parameter values such as coefficient, and the distribution situation of data point in matched curve, show that y=x/ (p1+p2*x^2) equation is optimum fit curve, wherein p1=2.0289, p2=2.2300.As shown in Figure 1, calculated value and actual value residual error column diagram are as shown in Figure 2 for optimum fit curve.
Embodiment 5: modelling verification
Remain milk cow that in 10 lactations, lead content exceeds standard as modelling verification group, they are isolated and ensures do not have lead contamination input to contact, gather milk sample every day and detect the content of Ruzhong lead, the initial lead content of 10 cow heads is respectively 0.0254, 0.0271, 0.0260, 0.0218, 0.0179, 0.0156, 0.0192, 0.0233, 0.0265, 0.0182mg/kg, the time point a corresponding to initial concentration is obtained according to set up models fitting Solving Equations, then a+n (n=1, 2, 3 ...) the plumbous mass concentration corresponding in model curve in sky be isolation after n days institute predict the mass concentration of lead in Lactation of Dairy Cow.Prediction concentrations and actual detectable concentration are carried out the accuracy of contrast verification institute of the present invention Modling model equation, carry out accuracy of judgement degree with the relative error of prediction concentrations,
the accuracy rate of prediction concentrations is calculated simultaneously,
concrete model the result, in table 4.
Table 4 plan-validation result
As shown in Table 4, carry out prediction checking by models fitting equation to Acetochlor concentration in the galactopoiesis of 10 cow head institutes, obtain the relative error of prediction concentrations and actual measurement concentration, relative error range is 2.14 ~ 4.24%, within the scope of reasonable error; Rate of accuracy reached 95.13 ~ 98.73%, accuracy rate is in good scope.The Forecasting Methodology that the Ruzhong Acetochlor concentration that the present invention sets up disappears has very high accuracy.
This proof scheme simultaneously to 10 cow head Ruzhong lead concentrations disappear to national limit standard (0.05mg/kg) below time paracmasis carry out prediction checking.Initial concentration x plumbous in cow's milk is obtained according to model equation
0corresponding number of days y
0, the number of days that limitation concentration 0.05mg/kg is corresponding is 8.866d, uses 8.866-y
0the value obtained rounds up to count and is time of day required when lead concentration in Lactation of Dairy Cow disappears to limitation concentration.Calculate in this proof scheme numbering 1 ~ No. 10 milk cow to produce in cow's milk the plumbous time point corresponding to initial concentration value and be respectively 1.3602, 2.1366, 3.2817, 2.4488, 2.7393, 2.2681, 2.3667, 1.6614, 4.4170, 3.7620 my god, calculate the time of disappearing to Limited Doses with this and be respectively 8, 7, 6, 7, 7, 7, 7, 8, 5, 6 days, actual lead concentration in 10 checking milk cow institutes galactopoiesis of predicted regression time point is detected, show that actual detectable concentration is 0.0452, 0.0466, 0.0438, 0.0422, 0.0432, 0.0455, 0.0463, 0.0451, 0.0422, 0.0417mg/kg, actual detectable concentration has been up to state standards below limitation concentration all, show the present invention set up equation model can to Ruzhong lead concentration reach country limitation concentration below time paracmasis carry out Accurate Prediction.
Embodiment 6: utilize model to predict
In Dec, 2013 obtains in somewhere the milk cow sample that in 5 lactations, lead content exceeds standard, and after isolating, collects milk sample every day and detects the content of Ruzhong lead, utilizing equation of the present invention to produce lead concentration in cow's milk to this 5 cow head simultaneously and predict it.Result shows, in this prediction equation cow's milk, lead concentration accuracy rate is up in the good range of 95.27 ~ 98.12%, has good practicality to the monitoring of lead concentration in cow's milk.Although the present invention with preferred embodiment openly as above; but it is also not used to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; can do various change and modification, what therefore protection scope of the present invention should define with claims is as the criterion.
Claims (10)
1. the Forecasting Methodology that in a Lactation of Dairy Cow, lead content disappears, it is characterized in that: collect the cow's milk secreted by lead doping milk cow, detect initial concentration plumbous in cow's milk, management of feeding is isolated to lead doping milk cow, according to the cow's milk secreted by sampling plan collection isolation milk cow, Determination of Pb is carried out to gathered cow's milk, statistical study is carried out to gained lead content data and obtains fit equation, obtain predictive equation after again fit equation being verified, utilize gained predictive equation to predict the regression time of lead content in lead doping Lactation of Dairy Cow;
Described predictive equation is: Y=X/ (2.0289+2.2300X
2); Wherein, X is the rear number of days of isolation, and Y is mass concentration plumbous in cow's milk.
2. method according to claim 1, it is characterized in that, step is as follows:
1) collect the cow's milk secreted by lead doping milk cow, detect initial concentration plumbous in cow's milk;
2) management of feeding is isolated to lead doping milk cow, according to the cow's milk secreted by sampling plan collection isolation milk cow;
3) sample pre-treatments is carried out to gathered cow's milk, detect content plumbous in cow's milk;
4) statistical study is carried out to detection data, obtain fit equation, after fit equation is verified, obtain predictive equation;
Described predictive equation is: Y=X/ (2.0289+2.2300X
2); Wherein, X is the rear number of days of isolation, and Y is mass concentration plumbous in cow's milk;
5) predictive equation is utilized to predict lead concentration regression time in Lactation of Dairy Cow.
3. method according to claim 2, is characterized in that, step 1) described lead doping milk cow, for lead content in secreted cow's milk is more than the non-poisoning milk cow of 0.05mg/kg.
4. method according to claim 2, is characterized in that, step 1) described dairy bread is holstein cow.
5. method according to claim 2, is characterized in that, step 1) described initial concentration is higher than 0.2315mg/kg.
6. method according to claim 2, it is characterized in that, step 2) described sampling plan is that the milk sample of just isolating milk cow collection is designated as the 1st day, 1st day to the 19th day every day, sampling should be carried out, each institute sample thief from the same day 7 point, 15 and 23 squeeze the mixing breast that breast obtains; 20th day to the 47th day, sampling should be carried out every other day, each institute sample thief from the same day 7 point, 15 and 23 squeeze the mixing breast that breast obtains.
7. method according to claim 2, is characterized in that, step 3) lead content in described detection cow's milk adopts GFAAS (graphite furnace atomic absorption spectrometry) to detect.
8. method according to claim 2, is characterized in that, step 4) described statistical study carries out nonlinear fitting to gained lead content data.
9. method according to claim 2, it is characterized in that, concrete steps are:
1) collect the exceed standard milk sample of lead content at 0.05-0.2315mg/kg, and measure initial concentration;
2) management of feeding is isolated to lead doping milk cow, according to the cow's milk secreted by sampling plan collection isolation milk cow;
Described sampling plan is: the milk sample that just isolation milk cow has gathered is designated as the 1st day, and the 1st day to the 19th day every day, sampling should be carried out, each institute sample thief from the same day 7 point, that the newborn mixing obtained is squeezed by 15 and 23 institutes is newborn; 20th day to the 47th day, sampling should be carried out every other day, each institute sample thief from the same day 7 point, 15 and 23 squeeze the mixing breast that breast obtains;
3) sample pre-treatments is carried out to gathered cow's milk, adopt GFAAS (graphite furnace atomic absorption spectrometry) to detect content plumbous in cow's milk;
4) nonlinear fitting is carried out to detection data and obtain fit equation, then checking acquisition predictive equation is carried out to fit equation;
Described predictive equation is: Y=X/ (20.289+2.2300X
2), wherein, X is the rear number of days of isolation, and Y is mass concentration plumbous in cow's milk;
5) step 4 is utilized) fade level of the predictive equation of gained prediction Ruzhong lead doping milk cow lead concentration in lactation process and time.
10. the method described in claim 1-9 is applied to prediction lead doping milk cow lead concentration fade level and disappearing to the following required time of limitation concentration in lactation process.
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