A kind of quick detection of the fungimycin based on two color sensor dynamic white balancesMethodTechnical field
The present invention relates to a kind of rapid detection methods of fungimycin based on two color sensor dynamic white balances.
Background technique
Currently, food-safety problem is got worse, and the safety problem as caused by fungimycin substance is very universal.VeryVerticillium toxin (mycotoxins) is metabolite caused by fungi grows in food or feed, it seriously threatens the mankind and movesThe health and lives safety of object.Common mycotoxin mainly has aflatoxin (aflatoxins), ochratoxin A(ochratox A), fumarubicin (fumonisins) and zearalenone (zearalenone) etc..Due to most of trueMycomycin all has toxicity, and therefore, stringent limit has been formulated the content of fungimycin in food-related products in countries in the worldAmount standard.
Aflatoxin in field of food safety, every gram of food is with 10-9Gram for unit measure, have be detected ratioThe features such as high, sample distribution is unevenly low with there are content.Therefore, higher want is proposed to limit standard and detection techniqueIt asks.Currently, the conventional instrument detection method for mycotoxin mainly has thin-layered chromatography (TLC), high performance liquid chromatography(HPLC), the instrument analytical methods such as mass spectrometer (GCMS) and liquid chromatography mass spectrometric method (LCMS).Using a kind of high performance liquid chromatographyTandem mass spectrometry (HPLC-MS/MS) come measure analysis peanut and its product in five kinds of fungimycins, this method high sensitivity,Selectivity is good.But such methods are made a general survey of, generally existing sample pretreatment process is more complicated, instrument and equipment is expensive, need to specially graspThe problems such as making personnel and taking time and effort, to limit its scope of application, and conventional instrument detection method is not able to satisfy sceneThe demand of rapid screening analysis.Therefore, efficient mycotoxin detection pre-treatment and novel Fast Detection Technique and equipment are developedIt is particularly important.It is recently proposed a kind of quick detection side based on non-marked aptamer Visual retrieval aflatoxin B1Method, caused color change after being agglomerated under Salt treatment using nanogold are realized for the sensitive quick of aflatoxin B1Visual retrieval.But be based on colorimetric determination when, due to different batches nanogold 520 nanometer wave strong points absorbanceSlightly difference, when aptamers and salt ionic concentration are not fixed in solution, absorbance can all be affected, so having to testing resultCertain influence.Currently, antigen-antibody method, i.e. immunological method are also used for the quick detection to mycotoxin.But it is thisMethod needs to prepare the antibody of specificity, and antibody itself is due to unstability, so the cost that should not be saved and prepareIt is higher.These deficiencies constrain extensive use of the immunological method in mycotoxin field.
In above-described various methods, detection limit is universal higher, and unsuitable mycotoxin haves the characteristics that content is low.Therefore the sensitivity of detection method is continuously improved, enhancing anti-interference ability, improves detection speed, and reduces testing cost alwaysIt is all the driving source of mycotoxin Fast Detection Technique exploitation.
Summary of the invention
The present invention is studied for the detection of fungimycin content, is proposed a kind of yellow bent for field quick detectionThe technology of mould toxin.Herein for mycotoxin, a kind of detection method for determining solution concentration using solution colour is proposed, is led toThree primary colors (RGB) value for obtaining solution colour using color sensor is crossed, realizes the matching of three primary colors and various concentration solution, fromAnd establish the linear relationship between solution concentration and rgb value.This method improves traditional single-sensor color acquisition system simultaneouslySystem realizes the dynamic white balance of environment light using dual sensor, and detection system is made to have the adaptive ability of stronger environment light.Experiments have shown that the detection method speed is fast, high-efficient, versatile, measurement accuracy and robustness with higher.
A kind of rapid detection method of the fungimycin based on two color sensor dynamic white balances, it is characterised in that:
Color sensor I passes through three Color Channels of RGB, utilizes white light in timer/counter acquisition environmentInitial data；
Color sensor II utilizes the color data of timer/counter acquisition solution by tri- Color Channels of RGB；
Using Mean Filtering Algorithm, the initial data of white light in environment light is filtered, environment light is obtainedWhite balance parameter；
Sensor is in data acquisition, and there are pulse interference signals, carries out anti-arteries and veins to the solution colour data of acquisitionPunching interference average value filtering processing；
The white balance parameter obtained using sensor I is as the rgb value of benchmark solution colour；
Relationship between the two is established with corresponding rgb value according to sample concentration, by the linear relationship having built up, thenThe colouring information that detection unknown concentration solution is removed using two color sensor systems, finally extrapolates corresponding concentration value.
The correspondence reference frequency of RGB white in the detected environment light of sensor I in Fig. 1 is set to Fr、Fg、Fb, Tr、Tg、TbThe correspondence reference period of respectively white RGB, sample frequency difference when II test sample color of sensorIt is set as fr、fg、fb, tr、tg、tbSampling period when for test sample color utilizes formula according to the testing principle of color sensor(1), (2) and (3) respectively obtain the rgb value of solution example.
Wherein, since color sensor can not directly acquire measurement frequency in the detection process, so system passes through detectionCycle T and t calculate rgb value.It before calculating RGB, needs that cycle T and t is once filtered, wherein T, t distinguishTwo sensors corresponding period when for sampling pulse signal.N is sampling number, TiDuring acquisition environment light pulse signalA wherein period, tiTo obtain wherein period during sample pulse signal；
Using two color sensors during detecting color, the automation adjustment of detection data, tool can be realizedThere is environment self-adaption.Experimental data proof diagram is as follows:
The invention has the following beneficial effects:
(1) feature is automated.Two Color Detecting Systems that the present invention uses are able to achieve the automation feature of color detection,When environment light changes, it can adjust automatically, guarantee the accuracy of testing result, there is environment self-adaption sexual function；
(2) portability feature.Scene, detection on the spot, easy to use, popularization may be implemented in the detection device；
(3) easy to operate, quick.By established linear relationship, it can directly pass through detection unknown concentration solutionIts corresponding concentration can be obtained in colouring information；
(4) high sensitivity.By detecting unknown concentration, its relative error is obtained lower than 7%, meets the accurate of detection dataDegree requires；
(5) at low cost, profitable.This detection device production cost is low, greatly reduces testing cost.
For the detection of mycotoxin, the quick inspection proposed in this paper that dynamic white balance is realized using two color sensorsSurvey method does not need the antibody of preparation specificity yet, only needs independent of the sample pre-treatments of expensive instrument and equipment and complexityBy acquiring the color change information with analytical solution, the linear relationship between solution concentration and three primary colors (RGB) is established.TogetherWhen, due to realizing automatically correcting in real time for white balance, avoiding the change pair of environment light using two color sensor measurementsThe influence of testing result ensure that the stability of detection system and the accuracy of testing result.The experiment show detectionThe feasibility and validity of method.This method can be equally used for detecting other in relation to candex kind object matter.
Detailed description of the invention
The realization of Fig. 1 dynamic white balance
The variation of rgb value under Fig. 2 varying environment light
Scheming (a) and (b) is the situation of change for the R value that single, double sensor measures under two kinds of varying environment light.
Scheming (c) and (d) is the situation of change for the G value that single, double sensor measures under two kinds of varying environment light.
Scheming (e) and (f) is the situation of change for the B value that single, double sensor measures under two kinds of varying environment light.
Conclusion:
1, rgb value of the same sample under varying environment light can be fluctuated and be changed；
2, when environment light changes, the data fluctuations that Method for Single Sensor System collects are bigger；
3, when environment light changes, the data fluctuations that double-sensor system collects are smaller.
Fig. 3 is Color Detecting System circuit diagram, and wherein color sensor model TCS230, voltage conversion chip areThe power supply line of ULN2003, color sensor carry out decoupling using 0.01 μ of μ F -0.1 F, and the enable end OE pin of chip draws with GNDIt should be connected using Low ESR between foot, to improve noise resisting ability.Fig. 2 is solution example to be detected, is loaded in centrifuge tube, moltenLiquid product accounts for the 85% of centrifugation pipe volume.
Specific embodiment
The preparation of reagent mainly has the preparation of nanogold, AFB1 aptamers and nanogold under high salt concn in the present inventionEffect.
1, the preparation of nanogold
Experiment prepares colloidal gold using trisodium citrate reduction method, and 95.8mL ultrapure water is added in three neck round bottom flask,1% chlorauric acid solution of 4.2mL mass fraction is added, magnetic force is heated at reflux stirring, persistently boils 10min, 10mL is then added1% citric acid three sodium solution of mass fraction, solution colour redden in 1min, continue the 20min that boils, naturally cold under stirringBut to room temperature, obtained claret liquid, as nanogold.
2, the preparation of solution example
The concentration of aflatoxin B1 (AFB1) aptamers and the concentration of NaCl salting liquid are this detection method reaction systemsKey factor pass through the aptamers and NaCl to various concentration since the length of different aptamers, base composition are differentThe comparison of solution, has determined optimum reaction condition.The optium concentration that aptamers have been determined is 10 μm of ol/L, and concentration is 2mol/L'sNaCl solution.
Detecting step
Two innovative color sensor systems that this system uses during obtaining color sample information, are realizedAutomation in detection process, has the function of adaptivity, improves stability, the accuracy of detection data.It realizes simultaneouslyA method of new detection aflatoxin concentration, the specific steps are as follows:
Color sensor I passes through three Color Channels of RGB, utilizes white light in timer/counter acquisition environmentInitial data；
Color sensor II utilizes the color data of timer/counter acquisition solution by tri- Color Channels of RGB；
Using Mean Filtering Algorithm, the initial data of white light in environment light is filtered, environment light is obtainedWhite balance parameter；
Sensor is in data acquisition, and there are pulse interference signals, so the solution colour data to acquisition carry outAnti- impulse disturbances average value filtering processing；Remove maximum value and minimum value and carries out average value filtering processing again；
The white balance parameter obtained using sensor I is as the rgb value of benchmark solution colour；
Relationship between the two is established with corresponding rgb value according to sample concentration, can be obtained by established linear relationshipTo the solution of unknown concentration.
By the linear relationship having built up, detection unknown concentration solution then is removed using two color sensor systemsColouring information finally extrapolates corresponding concentration value.