CN110132955A - The method of perchlorate in dispersive liquid-liquid microextraction-smart phone colorimetric estimation food - Google Patents
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Abstract
The invention discloses a kind of method using the perchlorate in dispersive liquid-liquid microextraction-smart phone colorimetric estimation food, be using methylene blue as color developing agent, in the acid system of pH=4.5 with ClO4 ‑Reaction generates hydrophobic green ionic associate；Methanol is reused as dispersing agent, methylene chloride makees extractant, and ion association extraction is enriched in organic phase methylene chloride using dispersive liquid-liquid microextraction technology；Then the RGB color intensity value of organic phase is obtained using the shooting of Android mobile phone Color Grab software, and according to ClO4 ‑Working curve determine the RGB color intensity and ClO of organic phase4 ‑Quantitative relationship between concentration, to realize ClO4 ‑Quantitative detection.This method accuracy is good, high sensitivity, organic reagent dosage are few, and the instrument and equipment being related to is cheap, is easy to field deployment and of less demanding to the technical level of operator, is expected to promote the use of as the quick detection device of perchlorate in food.
Description
Technical field
The present invention relates to the detection method of high chloro acid ion a kind of more particularly to a kind of dispersive liquid-liquid microextraction-intelligence hands
The method of perchlorate in machine colorimetric estimation food, belongs to technical field of food detection.
Background technique
Perchlorate has high stability (difficult to degrade), highly dissoluble, high diffusibility and holds as a kind of environmental contaminants
Continuous migration, exposure will lead to the long-term pollution to water source, soil in the environment, by being enriched with or converting, by breathing, eating
The approach such as object chain enter human body.Drinking Water, vegetable and fruit, meat, tealeaves, milk and its product and beverage etc. are China
The food of the daily intake of resident, wherein have different degrees of content perchlorate detect, Drinking Water, tealeaves, spinach and
The content of perchlorate reaches as high as ppm grades in the food such as "Hami" melon, and food-safe there are greater risks with human health.It grinds
Study carefully and show that the absorption of iodine can be hindered after perchlorate enters human body, and then interfere thyroid normal function, influences human body
Metabolism and nervous system growth and development, this adverse effect shows especially prominent between infant and teenager.
Since the content of perchlorate in food is at ppb-ppm grades, quantitative survey is difficult to using general chemical detection method
It is fixed.Ppb grade of quantitative detecting method is concentrated mainly on and uses ion chromatography, ion chromatography-mass spectrometry, efficient both at home and abroad at present
The technologies such as liquid chromatogram-matter combination.Ion chromatograph conductance detection method only depends on retention time to the qualitative of object, vulnerable to it
The interference of his coexisting ion and complex matrices, therefore the simple matrix, the lesser sample of interference such as be mainly used in water sample.Mass spectrum connection
With technology can the very harsh sample of Accurate Determining content, but operating cost is high, device-specific is strong, occupation rate of market is low, right
Laboratory hardware condition and personnel qualifications are higher, cannot achieve on-site test.
Visual retrieval technology based on portable equipments such as smart phone camera, digital camera, scanners is to utilize three
Primary colours principle establishes itself and target after grabbing the color space numerical value of image under conditions of keeping background color almost the same
The quantitative relationship of object concentration, these methods have many advantages, such as simple quick response, analytic process, real time communication and on-site test,
Have become more popular research field.Free smart phone takes color software Color Grab to can be obtained by once shooting
Numerical value including HSV, RGB, CMYK etc. including ten several color spaces, these numerical value do not need to carry out a series of complex computer image
As processing and mathematic(al) manipulation, the quantitative determination of object can be used to.
Summary of the invention
The object of the present invention is to provide the perchlorate in a kind of dispersive liquid-liquid microextraction-smart phone colorimetric estimation food
Method.
The present invention with methylene blue (MB) be color developing agent, in the acid system of pH=4.5 with ClO4 -Reaction generates hydrophobic
Green ionic associate；Methanol is reused as dispersing agent, methylene chloride makees extractant, utilizes dispersive liquid-liquid microextraction
(DLLME) ion association extraction is enriched in organic phase methylene chloride by technology；Then Android mobile phone Color Grab is used
Software shooting obtains the RGB color intensity value of organic phase, and according to ClO4 -Working curve determine the RGB color intensity of organic phase
With ClO4 -Quantitative relationship between concentration, to realize ClO4 -Quantitative detection.Specific method is described in detail below.
One, ClO4 -Working curve drafting
The preparation of perchlorate standard solution: perchlorate standard substance concentration is 1000 μ g/mL: directly being measured after dilution；
The perchlorate standard solution of various concentration is sequentially added in 15mL centrifuge tube, 550 μ L, 50 μ g/L methylene blue is molten
Liquid, the acetic acid-sodium acetate buffer solution of pH=4.5 1mL, is settled to 10mL, shakes up, and stands 2min, is separately added into 70 μ L methanol,
350 μ L methylene chloride, be vortexed extraction 60s, and 4000r/min is centrifuged 5min, and centrifugation bottom of the tube obtains enriched sample organic phase；It places
The color of 30min(organic phase is in more stable state), using Android mobile phone software Color Grab to centrifugation bottom of the tube
Organic phase take pictures and take color, obtain organic phase RGB color numerical value；With various concentration ClO4 -The color relative intensity value of standard solution
I maps to corresponding concentration, and (I is ordinate, ClO4 -Concentration establishes standard curve as abscissa), data model result is shown in figure
1:ClO4 -Corresponding color relative intensity value I is in a linear relationship within the scope of 20 ~ 500 μ g/L, related coefficient square R2=
0.9996, equation of linear regression are as follows:
I=0.2950×C+3.5906
The color relative intensity value I of I --- standard solution
C——ClO4 -Concentration, unit are μ g/L
Color relative intensity value I is defined as follows:
I=|IR-IRblank |+|IG-IGblank |+|IB-IBblank|
I in formulaR、IG、IBIndicate R, G, the channel B color value of sample solution, IRblank、IGblank、IBblankIndicate that reagent blank is molten
The color value of liquid R, G, channel B.
The color intensity value for measuring 11 groups of standard blank solutions simultaneously, obtains its standard deviation=0.5045, according to LOD=3
σ/k, LOQ=10 σ/k acquire this method detection and are limited to 5.13 μ g/L, are quantitatively limited to 17.50 μ g/L.
Two, ClO in sample4 -The measurement of concentration
1、ClO4 -The measurement of concentration
(1) preparation of sample solution: food samples and acetic acid solution being vortexed after mixing, 10 ~ 20min of ultrasound；With being covered with activity
The filter paper of charcoal filters, and filtrate is through C18Solid phase extraction column purification crosses 0.22 μm of filter membrane to get sample purification liquid；
(2) 5 ~ 8mL sample purification liquid, 550 μ L methylene blue solutions (50 μ g/L), 1mL are sequentially added in 15mL centrifuge tube
The acetic acid-sodium acetate buffer solution of pH=4.5, is settled to 10mL, shakes up, and stands 2min, is separately added into 70 μ L methanol, 350 μ L dichloros
Methane, be vortexed extraction 60s, and 4000r/min is centrifuged 5min, and centrifugation bottom of the tube obtains enriched sample organic phase；30min is placed, is made
It is taken pictures with organic phase of the Android mobile phone software Color Grab to centrifugation bottom of the tube, obtains organic phase RGB color numerical value；It will enrichment
The color relative intensity value I of sample organic phase is brought into the equation of linear regression of standard curve, obtains ClO in sample purification liquid4 -
Concentration, by can be calculated ClO in sample4 -Content.
2, anti-interference measurement
Under these experimental conditions, some common zwitterions have been investigated to ClO4 -The influence of measurement.For 200 μ g/L's
ClO4 -, think noiseless as relative error≤± 5 %, the results are shown in Table 1.The results show that I-And ClO3 -Ion pair measurement is done
Disturb it is relatively high, if answering pre-separation to remove in the presence of a large amount of, reducing its interference effect to measurement.
。
3, analytical characteristics
Under these experimental conditions, ClO is chosen4 -Concentration is the standard working solution of 100 μ g/L and 300 μ g/L, investigates the weight of method
Existing property, in a few days the RSD of the color relative intensity value I of five measurements is 1.37%, in the daytime the color relative intensity value I of five measurements
RSD be 1.99%.Show that the reproducibility of this method is very good, method is reliable and stable.
Under these experimental conditions, ClO is used4 -Concentration is the standard working solution of 200 μ g/L, investigate by 0.22 μm of filter membrane and
By 0.5g active carbon, C18To the adsorption capacity of object, obtaining the rate of recovery is respectively for solid-phase extraction column, 0.22 μm of filter membrane
96.85% and 97.06%, illustrate active carbon, C18Solid phase extraction column and 0.22 μm of filter membrane are to the ClO in range of linearity concentration4 -Base
This does not generate adsorptive hindrance.
Under these experimental conditions, it chooses and ClO is housed4 -Concentration is respectively three centrifuge tubes of 0,50,500 μ g/L, and every
Centrifuge tube rotates one week in ten times, and the rgb value of 36 degree of reading organic phases of rotation, is calculated RSD points of color intensity value every time
Not Wei 0.35%, 0.84% and 1.10%, illustrate mobile phone take color measurement substantially not by nuance existing between centrifuge tube and centrifugation
The influence of the factors such as pipe sample introduction position, reproducibility are preferable.
In conclusion the present invention extracts the ClO in sample with acetic acid solution4 -Afterwards, by active carbon and C18Solid Phase Extraction is small
Column remove interfering substance, using methanol as dispersing agent, methylene chloride as extractant, be enriched with using dispersive liquid-liquid microextraction
ClO4 -Ionic associate, then with cell phone software Color Grab take color obtain methylene chloride phase RGB color intensity value, really
Determine color relative intensity value and ClO4 -There are quantitative linearity relationships for concentration.This method is used successfully to perchloric acid in measurement numerous food
The content of salt, and using the test of the mark-on reclaims of various concentration, to measure the use ion chromatography that different Reagent Companies provide legal
The standard solution and liquid chromatogram of value are combined the mass spectroscopy accuracy and reliability of method.This method accuracy is good, clever
For sensitivity compared with high, organic reagent dosage is few, the instrument and equipment being related to is cheap, is easy to field deployment and to the skill of operator
Art level requirement is not high, is expected to promote the use of as the quick detection device of perchlorate in food.
Detailed description of the invention
Fig. 1 is ClO4 -Working curve.
Specific embodiment
It is described further below by measuring method of the specific embodiment to perchlorate of the present invention.
The measurement of perchlorate in embodiment 1, drinking water and bottled Drinking Mineral Water
The preparation of sample: it directly measures.
The measurement of sample: sequentially adding 8mL sample in 15mL centrifuge tube, 550 μ L methylene blue solutions (50 μ g/L),
The acetic acid-sodium acetate buffer solution of pH=4.5 1mL, is settled to 10mL, shakes up, and stands 2min, is separately added into 70 μ L methanol, 350 μ L
Methylene chloride, be vortexed extraction 60s, and 4000r/min is centrifuged 5min, and centrifugation bottom of the tube obtains enriched sample organic phase；It places
30min is taken pictures using organic phase of the Android mobile phone software Color Grab to centrifugation bottom of the tube, obtains organic phase RGB color number
Value；The color relative intensity value I of enriched sample organic phase is brought into the equation of linear regression of standard curve, sample purification is obtained
ClO in liquid4 -Concentration, by can be calculated ClO in sample4 -Content.Measurement result is shown in Table 2.
The measurement of perchlorate in embodiment 2, tea beverage
The preparation of sample: taking 40mL tea beverage in 50mL centrifuge tube, and 40 μ L glacial acetic acids are added, and is vortexed and mixes, ultrasonic 10min,
Sample solution is after the filter paper for being covered with 0.5g active carbon in advance filters, and the filtrate collected is through C18Solid phase extraction column purification,
Cross 0.22 μm of filter membrane.
The measurement of sample: sequentially adding 8mL sample in 15mL centrifuge tube, 550 μ L methylene blue solutions (50 μ g/L),
The acetic acid-sodium acetate buffer solution of pH=4.5 1mL, is settled to 10mL, shakes up, and stands 2min, is separately added into 70 μ L methanol, 350 μ L
Methylene chloride, be vortexed extraction 60s, and 4000r/min is centrifuged 5min, and centrifugation bottom of the tube obtains enriched sample organic phase；It places
30min is taken pictures using organic phase of the Android mobile phone software Color Grab to centrifugation bottom of the tube, obtains organic phase RGB color number
Value；The color relative intensity value I of enriched sample organic phase is brought into the equation of linear regression of standard curve, sample purification is obtained
ClO in liquid4 -Concentration, by can be calculated ClO in sample4 -Content.Measurement result is shown in Table 2.
Embodiment 3, tealeaves, the measurement for being dehydrated perchlorate in spinach
Tealeaves, the preparation for being dehydrated spinach samples: son is sieved with 100 mesh sieve after tealeaves and dehydration spinach are crushed, weighs 0.25 ~ 3g sample
The glacial acetic acid solution of 25mL0.1% is added in 50mL centrifuge tube in powder, is vortexed and mixes, and 60 DEG C of ultrasonic extraction 20min are put to room
Temperature, 10000r/min are centrifuged 15min, take supernatant to cross the filter paper for being covered with 0.5g active carbon, the filtrate collected is through C18Solid phase
Small column purification is extracted, 0.22 μm of filter membrane is crossed.
The measurement of sample: 8mL sample solution, 550 μ L methylene blue solution (50 μ g/ are sequentially added in 15mL centrifuge tube
L), the acetic acid-sodium acetate buffer solution of pH=4.5 1mL, is settled to 10mL, shakes up, stand 2min, be separately added into 70 μ L methanol,
350 μ L methylene chloride, be vortexed extraction 60s, and 4000r/min is centrifuged 5min, and centrifugation bottom of the tube obtains enriched sample organic phase；It places
30min is taken pictures using organic phase of the Android mobile phone software Color Grab to centrifugation bottom of the tube, obtains organic phase RGB color number
Value；The color relative intensity value I of enriched sample organic phase is brought into the equation of linear regression of standard curve, sample purification is obtained
ClO in liquid4 -Concentration, by can be calculated ClO in sample4 -Content.Measurement result is shown in Table 2.
Embodiment 4, apple, in panax ginseng fruit perchlorate measurement
The preparation of sample: taking edible part 200g or more, and sampling machine is smashed, and takes 10 ~ 15g sample in 50mL centrifuge tube after homogeneous
In, add 0.1% acetic acid solution to 25mL, is vortexed and mixes, ultrasonic extraction 20min crosses the filter paper for being covered with 0.5g active carbon, collects
Filtrate is through C18Solid phase extraction column purification, crosses 0.22 μm of filter membrane.
The measurement of sample: sequentially adding 8mL sample in 15mL centrifuge tube, 550 μ L methylene blue solutions (50 μ g/L),
The acetic acid-sodium acetate buffer solution of pH=4.5 1mL, is settled to 10mL, shakes up, and stands 2min, is separately added into 70 μ L methanol, 350 μ L
Methylene chloride, be vortexed extraction 60s, and 4000r/min is centrifuged 5min, and centrifugation bottom of the tube obtains enriched sample organic phase；It places
30min is taken pictures using organic phase of the Android mobile phone software Color Grab to centrifugation bottom of the tube, obtains organic phase RGB color number
Value；The color relative intensity value I of enriched sample organic phase is brought into the equation of linear regression of standard curve, sample purification is obtained
ClO in liquid4 -Concentration, by can be calculated ClO in sample4 -Content.Measurement result is shown in Table 2.
In order to verify the reliability of detection method, with this method determine that an other Reagent Company provides by
The perchlorate standard substance of ion chromatography Par value, and choose drinking water, dehydration spinach and jasmine tea and give to CNAS
The laboratory of qualification uses SN/T 4089-2015 " measurement liquid chromatography-mass spectrography/mass spectrography of perchlorate in export food "
Specified in liquid chromatography-mass spectrography/mass spectrometric determination its perchlorate content, the results are shown in tables 3.Obtained result is used
T inspection is statisticallyd analyze, and the t value being calculated is less than its tabulated value t0.95,2=4.303, show the confidence level water 95%
Under flat, there was no significant difference for measurement result.
Present invention test and material used in the examples, reagent, instrument and equipment are as follows:
1.1 materials and reagent
C18Agela company, the U.S. solid phase extraction column (500mg, 6mL)；Perchlorate standard solution (1000 μ g/mL) north
Capital north great achievement measurement technology research institute；It is limited that perchlorate standard substance (1.000 ± 0.001mg/mL) Jinan crowd marks science and technology
Company；Methylene blue (analysis is pure) Sigma Co., USA；Jasmine tea, bottled tea drinks, apple, life fruit, bottled natural drink
It is bought with mineral water from local supermarket；Drinking water is well water；Being dehydrated spinach is that complete sample is examined in laboratory；Other chemical reagent are
Domestic analysis is pure.
1.2 instrument and equipment
PHS-3C acidometer Shanghai Precision Scientific Apparatus Co., Ltd；IKA group company, IKA MS3 turbula shaker Germany；
TG16 centrifuge Changsha Ying Tai Instrument Ltd.；The new limited public affairs of sesame biotechnology in SB25-12DTD ultrasonic cleaner Ningbo
Department；The portable balance Mettler-Toledo Instrument (Shanghai) Co., Ltd. of PL-E；M651CY smart phone (camera pixel:
800 ten thousand) China Mobile；Color Grab mobile phone takes color software to download from the http://website www.loomatix. com.
Claims (3)
1. the method for the perchlorate in dispersive liquid-liquid microextraction-smart phone colorimetric estimation food is with methylene blue (MB)
As color developing agent, in the acid system of pH=4.5 with ClO4 -React the hydrophobic nonionic associated matter generated；Reuse methanol conduct
Dispersing agent, methylene chloride make extractant, and ion association extraction is enriched to organic phase two using dispersive liquid-liquid microextraction technology
In chloromethanes；Then using the RGB color intensity value of Android mobile phone Color Grab software shooting acquisition organic phase, and according to
ClO4 -Working curve determine the RGB color intensity and ClO of organic phase4 -Quantitative relationship between concentration, to realize ClO4 -'s
Quantitative detection.
2. the method for the perchlorate in dispersive liquid-liquid microextraction as described in claim 1-smart phone colorimetric estimation food,
It is characterized in that: the following steps are included:
(1) after food samples and glacial acetic acid solution vortex oscillation being mixed, 10 ~ 20min of ultrasound；With the filter paper mistake for being covered with active carbon
Filter, filtrate is through C18Solid phase extraction column purification crosses 0.22 μm of filter membrane to get sample purification liquid；
(2) 5 ~ 8mL sample purification liquid, 550 μ L methylene blue solutions (50 μ g/L), 1mL are sequentially added in 15mL centrifuge tube
The acetic acid-sodium acetate buffer solution of pH=4.5, is settled to 10mL, shakes up, and stands 2min, is separately added into 70 μ L methanol, 350 μ L dichloros
Methane, be vortexed extraction 60s, and 4000r/min is centrifuged 5min, and centrifugation bottom of the tube obtains enriched sample organic phase；30min is placed, is made
It is taken pictures with organic phase of the Android mobile phone software Color Grab to centrifugation bottom of the tube, obtains organic phase RGB color numerical value；It will enrichment
The color relative intensity value I of sample organic phase is brought into the equation of linear regression of standard curve, obtains ClO in sample purification liquid4 -
Concentration, by can be calculated ClO in sample4 -Content；
ClO4 -In 20 ~ 500 μ g/L ranges, the equation of linear regression of standard curve is I=0.2950 × C+3.5906
The color relative intensity value I of I --- standard solution
C——ClO4 -Concentration, unit are μ g/L
The color relative intensity value I is defined as follows:
I=|IR-IRblank |+|IG-IGblank |+|IB-IBblank|
I in formulaR、IG、IBIndicate R, G, the channel B color value of sample solution, IRblank、IGblank、IBblankIndicate blank reagent solution
R, G, channel B color value.
3. the method for the perchlorate in dispersive liquid-liquid microextraction as described in claim 1-smart phone colorimetric estimation food,
Be characterized in that: the sample is drinking water, mineral water, beverage, tealeaves, dehydration spinach, fruit.
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