CN109781883A - A method of based on QuEChERS- dispersive liquid-liquid microextraction trace detection 5 hydroxymethyl furfural - Google Patents
A method of based on QuEChERS- dispersive liquid-liquid microextraction trace detection 5 hydroxymethyl furfural Download PDFInfo
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Abstract
The invention discloses a kind of methods based on QuEChERS- dispersive liquid-liquid microextraction trace detection 5 hydroxymethyl furfural, belong to field of detection of food safety.The present invention carries out pre-treatment using QuEChERS joint dispersive liquid-liquid microextraction DLLME technology, carries out purification enrichment processing to detectable substance, can be used in the content of 5 hydroxymethyl furfural in determination of trace rice (solid sample).The present invention is effectively combined improved QuEChERS-DLLME pre-treatment with UPLC-MS/MS, and has carried out investigation and condition optimizing to pre-treating method, establishes optimal pre-treating method, and is successfully suitable for the measurement of actual sample.This method is easy to operate, consumption of organic solvent is few, environmental pollution is small, and it good purification, reduces matrix interference, improve extraction efficiency, with quick, accurate and effective, easy, stable, and the advantage for taking into account that high sensitivity, the rate of recovery are good, cheap, applicable object is wide etc., it is suitble to the determination of trace of 5 hydroxymethyl furfural in rice.
Description
Technical field
The invention belongs to analytical chemistry and technical field of food safety, and in particular to one kind is based on QuEChERS- dispersion liquid
The method of liquid micro-extraction trace detection 5 hydroxymethyl furfural.
Background technique
Rice is the Major Foods of most of populations in the world as a kind of amyloid product of richness.Rice is through electric cooker
After culinary art, keeping warm mode can be maintained under conditions of temperature is 60~80 DEG C.Meanwhile temperature in electric cooker and moisture content compared with
Therefore height can occur Maillard reaction in rice culinary art and insulating process and accelerate the formation of 5 hydroxymethyl furfural.According to report
Road, Maillard reaction and it is long when storage both contribute to the formation of 5 hydroxymethyl furfural in rice.5 hydroxymethyl furfural is food
The by-product of Maillard reaction generation occurs during heat treatment for product, is normally used as a kind of thermal damage's index, for commenting
Estimate the degree of rice Maillard reaction in hot procedure, and measures the finger containing confectionery in storage period mass loss
Mark.In addition to this, 5 hydroxymethyl furfural has potentially hazardous property as a kind of food contaminant in food.It is reported that 5-
Hydroxymethylfurfural is widely present in miscellaneous food rich in carbohydrate, such as cereal products, biscuit, jam, baby
Dairy produce, fruit juice etc..Specifically, Lamberts and Villanova et al. delivered about in rice contain 5- methylol
The document of furfural.The 5 hydroxymethyl furfural in food is analyzed there are many analysis method, wherein ultra performance liquid chromatography mass spectrometry
It is combined with the high sensitivity for efficiently separating ability and MS/MS of UPLC and the separation detection technique of extremely strong specificity, even if
Also have the characteristics that reproducible, high sensitivity, analysis speed are fast in complicated sample substrate, however since matrix is complex
And the content of 5 hydroxymethyl furfural is trace level, it is more difficult to directly adopt instrument analysis, it is therefore necessary in instrument point
Before analysis, analyte is purified, is enriched with pretreatment.The pre-treatment of sample is the key step for being related to analysis result accuracy
It suddenly, is the important step for removing complex matrices interference, improving the analyte rate of recovery.Currently, document report is many for extracting
The pre-treating method of 5 hydroxymethyl furfural in sample, such as traditional Solid Phase Extraction, liquid-phase extraction, but conventional method is time-consuming, consumption
Material is expensive and consumes a large amount of organic solvent.To solve this problem, dispersive liquid-liquid microextraction (DLLME) technology is answered extensively
With, which was put forward for the first time by Rezaee et al. in 2006, have it is easy to operate, the rate of recovery is high, extraction time is short,
Cheap, the advantages that enrichment factor is high, but it is not suitable for solid sample.So far, there is not yet about 5- hydroxyl first in rice
Effective pre-treatment purification of base furfural, enrichment method and trace detection report.Therefore, develop it is a kind of accurate, quickly, effectively,
Cheap, easy pre-treatment purification and enrichment method (QuEChERS- dispersive liquid-liquid microextraction) are for detecting and quantifying
5 hydroxymethyl furfural in rice is of great significance.
Summary of the invention
To solve the above-mentioned problems, the present invention provides a kind of quick, accurate and effective, easy, stable, and height is taken into account
Enrichment factor, environmental pollution be small, economic advantages pre-treating methods, to Maillard reaction by-product 5- in trace detection rice
Hydroxymethylfurfural.The present invention purifies skill as pre-treatment using improved QuEChERS combination dispersive liquid-liquid microextraction (DLLME)
Art, and be detection method using ultra performance liquid chromatography-mass spectrometry (UPLC-MS/MS), overcome existing method organic solvent
The problems such as consumption is big, time-consuming, matrix interference is more, complex steps, can rapidly detect the 5 hydroxymethyl furfural in rice, fit
It is wide with range, measurement result accuracy with higher, good sensitivity and the excellent rate of recovery.
QuEChERS extraction and separation technology is improved in recent years, for measuring the differences such as vegetables, soil, tealeaves
The persticide residue of matrix.But so far, there is not yet handling rice (solid-like with QuEChERS extraction and separation technology
Product) in 5 hydroxymethyl furfural document report, also have no QuEChERS is combined with DLLME be used to purify, be enriched with concentration rice
The document of middle 5 hydroxymethyl furfural.
The first purpose of the invention is to provide a kind of pre-treating method of 5 hydroxymethyl furfural in rice, the method packets
It includes:
(1) in organic solvent, by the octadecylsilane chemically bonded silica of every 8~12g rice and 40mg~80mg
(C18), N- propyl ethylenediamine (PSA) effect of 40mg~80mg, is purified, is purified liquid；
(2) step (1) resulting scavenging solution is mixed with extractant, water is then added, realize enrichment to get pre-treatment sample
Product.
In one embodiment of the invention, the carbon black (GCB) of 0mg~15mg can also be added in the step (1).
In one embodiment of the invention, GCB is preferably added without in the step (1).
It in one embodiment of the invention, further include the magnesium sulfate that 0.5g~2g is added in the step (1)
(MgSO4) and 0.5g~1.5g sodium chloride (NaCl) be used as deicer.
In one embodiment of the invention, the organic solvent in the step (1) includes acetonitrile.
In one embodiment of the invention, the additive amount of organic solvent is the 0.5 of rice quality in the step (1)
~1.5 times of volumes.
In one embodiment of the invention, the volume ratio of the scavenging solution in the step (2) and extractant be (2~
8):1。
In one embodiment of the invention, extractant includes chloroform, methylene chloride, four in the step (2)
Chlorination carbon, chlorobenzene；It is preferred that chloroform.
In one embodiment of the invention, the pH of water is 7~10 in the step (2).
In one embodiment of the invention, in the step (2) the sum of water and scavenging solution and extractant volume body
Product is than being (0.5~2): 1.
In one embodiment of the invention, the preparation method of the rice includes: 1~1.5 times of body of addition in polished rice
Long-pending water cooking and heat-preservation 0h~42h is to get the rice for keeping the temperature different time, and the length of soaking time is to the contents of the present invention
It has an impact.
A second object of the present invention is to provide a kind of method of 5 hydroxymethyl furfural in quantitative detection rice, the methods
Include: the preparatory pre-treatment for carrying out rice, obtains pre-treatment sample, then detected；The method of the pre-treatment is above-mentioned
Pre-treating method.
In one embodiment of the invention, the detection includes high performance liquid chromatography (HPLC) detection, liquid chromatogram-
Mass spectrometry (LC-MS) detection, gas-chromatography (GC) detection, gas chromatography-mass spectrography (GC-MS) detection, Micellar Electrokinetic Capillary Chromatography hair
Tubule chromatography (MEKC) detection, high performance anion exchange chromatography (HPAEC) etc..
Third object of the present invention is to provide a kind of ultra performance liquid chromatography-mass spectrometry (UPLC-MS) trace detections
The method of 5 hydroxymethyl furfural, which comprises the pre-treatment for carrying out rice in advance obtains pre-treatment sample, then carries out
Liquid chromatograph mass spectrography detection；The method of the pre-treatment is above-mentioned pre-treating method.
In one embodiment of the invention, the condition of the liquid chromatogram are as follows: chromatographic column is reverse phase C18 chromatographic column
(including ACQUITY UPLC BEH reverse phase C18 column, PhenomenexSynergi Fusion reverse phase C18 column, SunFire
Waters reverse phase C18 column etc.), column temperature is 25~40 DEG C；The ultra-pure water solution that mobile phase A is 100%, Mobile phase B are 100%
Methanol takes gradient elution program, and flow velocity is 0.2~1mL/min, and sample volume is 10~15 μ L.
In one embodiment of the invention, the method specifically includes:
(1) it takes 8~12g of rice to be put into centrifuge tube, the acetonitrile solution of 0.5~1.5 times of volume is added and is cut by high speed
Cut disperser be homogenized under 5000~8000rpm revolving speed 2~5min progress slightly propose operation, obtain homogenate；
(2) purification process:
Magnesium sulfate (the MgSO of 0.5g~2g is added in the homogenate of step (1)4) and 0.5g~1.5g sodium chloride
(NaCl) it is used as deicer, be then vortexed rapidly 0.5~1.5min, then is centrifuged 4~8min at 7000~9000rpm to precipitate
Rice sample obtains the clear acetonitrile layer containing 5 hydroxymethyl furfural；Then adsorbent is added in clear solution includes 40mg
The octadecylsilane chemically bonded silica (C18) of~80mg, the lipid for being mainly used for removing in sample and other nonpolar chaff interferents；
The N- propyl ethylenediamine (PSA) of 40mg~80mg is used as weak anionic exchange adsorbing substance, commonly used in removal fatty acid, organic
Sour, various carbohydrates and matrix coextraction object etc.；The carbon black (GCB) of 0mg~15mg removes toner as a kind of, is mainly used for removing sample
Pigment in product.Then be vortexed rapidly 0.5~1.5min and 4~8min of centrifugation at 7000~9000rpm, by clear upper layer
Clear liquid is placed in centrifuge tube, as scavenging solution；
(3) the further purified treatment of dispersive liquid-liquid microextraction and enrichment concentration process:
The extractant of acetonitrile layer (scavenging solution is as dispersing agent) and certain volume in the step of taking certain volume (2) is mixed
Close, make the volume ratio (2~8) of scavenging solution and extractant: 1, then to addition pH range in mixed liquor 7~10 it is ultrapure
Water, makes the volume ratio (0.5~2) of the sum of ultrapure water and scavenging solution and extractant volume: 1；Mixed solution shows cloud immediately
Mist is centrifuged 4~8min at 7000~9000rpm, then collects undermost organic phase and be dried with nitrogen, it is standby to obtain residue
With；
(4) 5 hydroxymethyl furfural in ultra performance liquid chromatography-mass spectrometry rice:
Ultra performance liquid chromatography condition are as follows: chromatographic column is reverse phase C18 chromatographic column (including ACQUITY UPLC BEH reverse phase
C18 column, PhenomenexSynergi Fusion reverse phase C18 column, SunFire Waters reverse phase C18 column etc.), column temperature be 25~
40℃；The ultra-pure water solution that mobile phase A is 100%, the methanol that Mobile phase B is 100%, takes gradient elution program, flow velocity is
0.2~1mL/min, sample volume are 10~15 μ L；Mass Spectrometry Conditions are as follows: ion source is electric spray ion source (ESI), source parameter are as follows:
Entrance potential (EP) is 10V, and electron spray voltage is 5500V, and collision energy (CE) is 15V, and ion source temperature is 600 DEG C, source gas
Body and auxiliary gas are respectively set as 15L/min and 18L/min, and scan pattern is positive ion scan, and acquisition mode is more reactions
It monitors (MRM)；
(5) calculating of the 5 hydroxymethyl furfural assay result in rice:
The 5 hydroxymethyl furfural standard solution for pipetting certain concentration is added in rice, carries out sample pre-treatments-to it
Ultra performance liquid chromatography-mass spectrometry (improved QuEChERS- dispersive liquid-liquid microextraction pre-treatment purification method-UPLC-MS/
MS it) measures, obtains standard curve；It brings the measurement result in step (5) into standard curve and acquires 5- hydroxyl first in rice to be analyzed
The content of base furfural.
Advantages of the present invention and effect:
Improved QuEChERS and dispersive liquid-liquid microextraction technology are combined by the present invention, and effectively UPLC-MS/MS are combined to examine
Survey method establishes the Maillard reaction by-product 5- hydroxyl in QuEChERS-DLLME-UPLC-MS/MS method determination of trace rice
The detection method of methyl furfural.This method slightly mentions the 5 hydroxymethyl furfural in rice with acetonitrile solvent, is adsorbed using C18, PSA etc.
Agent purification reaches further purification then by dispersive liquid-liquid microextraction step (conditions such as Optimized Extraction agent, dispersing agent, pH)
With the effect of enrichment concentration, UPLC-MS/MS analysis measurement is then carried out.The method increase to determinand 5 hydroxymethyl furfural
Selectivity, also considerably increase enrichment times, new approaches provided for the development and application of pretreatment technology, also to trace detection
The extracting and developing of Maillard reaction by-product 5 hydroxymethyl furfural, enrichment concentration increase new method in rice (solid sample).
The method of the present invention effectively shortens the pre-treatment time, and the preceding place of 6 samples can be at least completed at the same time in 30min
Reason, and organic reagent dosage is few, waste is few, environmental pollution is small；In addition, by deicer, adsorbent, extractant,
The condition optimizing of dispersing agent etc. reduces the interference of complex matrices, improves purification efficiency and the rate of recovery.The result shows that the party
Method not only realizes highly selective and highly sensitive effective combination to 5 hydroxymethyl furfural, but also has accurate, quick, letter
Just, effectively, it is stable, inexpensive, reproducible, the rate of recovery is high the advantages that, be suitble to rice (solid sample) in 5 hydroxymethyl furfural
Determination of trace.Wherein detection limit and quantitative limit are respectively 0.1,0.3 μ g/L, sensitivity with higher；It is in a few days and smart in the daytime
The relative standard deviation of density is within 9%, and bat is between 93.95%~114.22%；Average recovery rate exists
Between 82.87%~104.86%, relative standard deviation is within 9%.
Detailed description of the invention
Fig. 1 is the QuEChERS-DLLME pre-treating method of embodiment 1-2 Optimal improvements, wherein A: improved QuEChERS
Method condition optimizing；B:DLLME optimization；
Fig. 2 is rice in embodiment 3 without pre-treatment and improved QuEChERS-DLLME pre-treating method contrastive colours
Wherein a, c: spectrogram adds the 5 hydroxymethyl furfural mark product of 50,500 μ g/L, the chromatography without pre-treatment respectively in rice sample
Figure；B, the 5 hydroxymethyl furfural mark product of 50,500 μ g/L, improved QuEChERS-DLLME d: are added in rice sample respectively
The chromatogram of pre-treating method；
Fig. 3 is the only improved QuEChERS processing and only through DLLME pre-treatment and improved of 4 rice of embodiment
QuEChERS-DLLME pre-treating method compares chromatogram.
Specific embodiment
For a better understanding of the present invention, the content of invention is furtherd elucidate below with reference to embodiment, but of the invention interior
Appearance is not limited solely to the following examples.
The measuring method of enrichment factor:
The enrichment factor of improved QuEChERS-DLLME pre-treatment is calculated according to following formula: enrichment factor EF=CA/
CB, CA are that 5 hydroxymethyl furfural standard items are added in rice to pass through the peak face that obtains after improved QuEChERS-DLLME pre-treatment
Product, CB are the peak area that 5 hydroxymethyl furfural standard items are added in rice and obtain without the above pre-treating method.
Embodiment 1
One, sample pre-treatments:
(1) rice is prepared:
The water of 1.5 times of volumes is added in polished rice, is then cooked in electric cooker and obtains rice.
The thick of 5 hydroxymethyl furfural proposes process in rice:
It weighs rice 10g to be put into centrifuge tube, the acetonitrile solution of 10mL is added and is existed by high-speed shearing disperser
3min is homogenized under 8000rpm revolving speed to carry out slightly proposing operation.
(2) improved QuEChERS pre-treatment purification process:
Magnesium sulfate (the MgSO of 1g is added in the rice in step (1)4) and 0.5g sodium chloride (NaCl) be used as deicer,
Then rapid vortex 1min, then centrifugation 5min is clear containing 5- methylol chaff to precipitate the acquisition of rice sample at 8000rpm
The acetonitrile layer of aldehyde.Then the octadecylsilane chemically bonded silica that a certain amount of adsorbent includes 60mg is added in clear solution
(C18), the N- propyl ethylenediamine (PSA) of 50mg.Then rapid vortex 1min, then it is centrifuged 5min at 8000rpm, it will be clear
Supernatant liquor is placed in centrifuge tube, as scavenging solution.
(3) the further purified treatment of dispersive liquid-liquid microextraction and enrichment concentration process:
The extractant of acetonitrile layer (scavenging solution is as dispersing agent) and certain volume in the step of taking certain volume (3) is mixed
It closes, makes the volume ratio 4:1 of scavenging solution and extractant, the extractant is optimized for chloroform.Then to being added in mixed liquor
The ultrapure water that the pH of certain volume is 7, makes the ratio 1:1 of the sum of ultrapure water and scavenging solution extractant volume.Mixed solution is vertical
It is formed and shows cloud, 5min is centrifuged at 8000rpm, undermost organic phase is then collected and is dried with nitrogen, obtained
Pre-treatment sample.
Two, the 5 hydroxymethyl furfural in ultra performance liquid chromatography-mass spectrometry rice is utilized:
Ultra performance liquid chromatography condition are as follows: chromatographic column: ACQUITY UPLC BEH-RP C18 chromatographic column (2.1mm ×
100mm, 1.7 μm), column temperature is 40 DEG C.The ultra-pure water solution that mobile phase A is 100%, the methanol that Mobile phase B is 100%, takes
Gradient elution program, flow velocity 0.3mL/min, sample volume are 10 μ L.Mass Spectrometry Conditions are as follows: ion source is electric spray ion source
(ESI), source parameter are as follows: entrance potential (EP) is 10V, and electron spray voltage is 5500V, and collision energy (CE) is 15V, ion source temperature
Degree is 600 DEG C, and source gas and auxiliary gas are respectively set as 15L/min and 18L/min, and scan pattern is positive ion scan, is adopted
Integrated mode is multiple-reaction monitoring (MRM).
Calculate the 5 hydroxymethyl furfural assay result in rice:
The 5 hydroxymethyl furfural standard solution for pipetting certain concentration is added in rice, carries out sample pre-treatments-to it
Ultra performance liquid chromatography-mass spectrometry (improved QuEChERS-DLLME-UPLC-MS/MS) measurement, obtains standard curve；It will
Testing result brings the content that standard curve acquires 5 hydroxymethyl furfural in rice to be analyzed into.
The efficiency evaluation of quantitative analysis and method:
(1) linear relationship, detection limit, quantitative limit: suitable 5 hydroxymethyl furfural standard solution is accurately pipetted, is added to
In rice sample (sample of no 5 hydroxymethyl furfural), analyzed after pre-treatment through UPLC-MS/MS.With target components peak area
For ordinate, adding concentration (μ g/L) accordingly with component is abscissa, draws standard curve, the results are shown in Table 1.5- hydroxyl first
Base furfural has preferable linear relationship in the range of 0.5~50.0 μ g/L, and regression coefficient is greater than 0.999, and detection limits and determines
Amount limit is respectively 0.1,0.3 μ g/L, sensitivity with higher.
The standard curve range of 5 hydroxymethyl furfural, regression equation, detection limit and quantitative limit in 1 rice of table
Note: subscript a and b value respectively indicates 3 times and 10 times of signal-to-noise ratio.
(2) in rice sample, 2.0,5.0,10.0,20.0 and 50.0 μ g/L, tetra- concentration accuracy and precision: are done
Horizontal addition experiment, each concentration level do 6 parallel samples, carry out pre-treatment and instrument analysis respectively, in a few days and in the daytime
Preci-sion and accuracy is as shown in table 2.As shown in Table 2, in a few days with the relative standard deviation of day to day precision within 9%,
Bat meets domestic and international analytical standard and requirement between 93.95%~114.22%.
The preci-sion and accuracy (n=6) of 5 hydroxymethyl furfural in 2 rice of table
(3) rate of recovery: being respectively measured actual sample by basic, normal, high three concentration mark-on level, i.e., 2.0,
5.0,10.0 μ g/L, tri- concentration levels do recovery testu, and each concentration level does 3 parallel samples, place before carrying out respectively
Reason and instrument analysis, and the rate of recovery is calculated by scalar quantity and measured value, the results are shown in Table 3.As shown in Table 3, average recovery rate
Between 82.87%~104.86%, relative standard deviation meets domestic and international analytical standard and requirement within 9%.
The rate of recovery (n=3) of 5 hydroxymethyl furfural in 3 rice of table
(4) actual sample measures:
The rice (i.e. different 5 hydroxymethyl furfural contents) of different soaking times is handled simultaneously with the method that the present invention establishes
5 hydroxymethyl furfural detection is carried out, the results are shown in Table 4, and this method is easy, quick, accurate and effective, has good applicability
And operability.
Table 4 keeps the temperature the measurement of 5 hydroxymethyl furfural in the rice sample of different time
Superscript letters (a-f) indicate statistical significant difference (p < 0.05).
Embodiment 2:
Using experiment of single factor, the method for 5 hydroxymethyl furfural in improved QuEChERS-DLLME extraction rice is carried out
Condition is investigated and optimization, the type including deicer, adsorbent species and content, dispersive liquid-liquid microextraction extractant and dispersing agent
With the conditions such as dosage, the range of pH, select optimal condition, obtain in optimal conditions to 5 hydroxymethyl furfural in rice most
Big extraction yield (highest enrichment factor).
QuEChERS condition optimizing:
1) optimize MgSO4With the dosage of NaCl: fixed C18 20mg, PSA 20mg, volume ratio 1 are 4:1, volume ratio 2 is
2:1, pH 7, by MgSO4Dosage replace with 1g, 2g, 3g, 4g respectively, by the dosage of NaCl replace with respectively 0g, 0.5g,
1g, 1.5g, specific experiment parameter is as shown in table 5, and treated shown in concentration effect such as Fig. 1 (A), as addition 2g MgSO4When,
Enrichment factor is 0.6, the results showed that 1g MgSO is added4When with 0.5g NaCl, the enrichment factor of 5 hydroxymethyl furfural reaches
0.90。
Table 5 optimizes MgSO4With the experiment parameter of NaCl dosage
"-" represents the experiment parameter of application in table 5.
2) optimize the component and dosage of adsorbent:
According to 1) optimum results, 1g MgSO is selected4With 0.5g NaCl, optimize adsorbent condition, measures concentration effect, tool
Body experiment parameter is as shown in table 6, shown in treated concentration effect such as Fig. 1 (A), as a result, it has been found that: as the dosage of C18 is from 20mg
Increase to 60mg, the enrichment factor of 5 hydroxymethyl furfural also increases.C18 amount continues growing the enrichment for but leading to 5 hydroxymethyl furfural
The factor is substantially reduced, and when adding to 200mg, enrichment factor only has 0.1.Therefore, the dosage of C18 is that 60mg can get highest richness
Collect the factor 1.12；When adding 50mg PSA, the maximum enrichment factor 1.56 of 5 hydroxymethyl furfural can get, as addition 100mg
When PSA, enrichment factor only has 0.6.In addition, by shown in Fig. 1 (A), with the increase of GCB amount, the enrichment of 5 hydroxymethyl furfural because
Son is reducing always, and when 10mg GCB is added, enrichment factor only has 0.7, to ensure to have preferable sensitivity, in sample of the present invention
GCB is not added in product pre-treatment.
The experiment parameter of the optimization adsorbent amount of table 6
"-" represents the experiment parameter of application in table 6.
The result shows that 1g MgSO4, the combination of 0.5g NaCl, 60mg C18,50mg PSA and 0mg GCB can be rice
The extraction of 5 hydroxymethyl furfural provides preferable purified treatment effect, enrichment factor 1.98 in meal.
DLLME condition optimizing:
1) selection of Optimized Extraction agent: choosing the optimal conditions of QuEChERS, changes the type of extractant, including dichloromethane
Alkane, chloroform, carbon tetrachloride, chlorobenzene, other conditions are constant, and specific experiment parameter is as shown in table 7, measurement treated enrichment
Intensity selects chloroform that maximum 5 hydroxymethyl furfural enrichment factor can be obtained as a result as shown in Fig. 1 (B) in extractant
1.56。
2) dosage of Optimized Extraction agent: choosing the optimal conditions of QuEChERS, and solvent selection chloroform changes purification
The dosage (volume ratio 1) of liquid and extractant, respectively 0.25:1,1:1,4:1,8:1, other conditions are constant, specific experiment parameter
As shown in table 7, measurement treated enrichment intensity, as a result as shown in Fig. 1 (B), when volume ratio is from 0.25:1~4:1,5- hydroxyl first
The enrichment factor of base furfural is consequently increased, and volume ratio 1 continues increase enrichment factor and downward trend occurs.Therefore, best body
Value of the product than 1 is 4:1, obtains 1.56 enrichment factor.
The experiment parameter of table 7 Optimized Extraction agent type and dosage
"-" represents the experiment parameter of application in table 7.
3) optimize the process conditions of water: choosing the optimal conditions of QuEChERS, solvent selection chloroform, volume ratio 1
For 4:1, changes the ratio (volume ratio 2) of the sum of water and scavenging solution extractant volume, be respectively as follows: 0.5:1,1:1,2:1,4:1, specifically
Experiment parameter is as shown in table 8, and treated that enrichment intensity obtains maximum as a result as shown in Fig. 1 (B) when ratio is 1:1 for measurement
Enrichment factor 1.98；Meanwhile it is 3~10 that ultrapure water, which adjusts pH range, pH value increases to 10 from 3,5 hydroxymethyl furfural
Maximum enrichment factor 2.0 is obtained in pH 10, but when pH value is from 7 to 10, the enrichment factor of 5 hydroxymethyl furfural increases slow
Slowly, for the convenience of experimental implementation, pH, which may be selected 7, can be obtained the 5 hydroxymethyl furfural extraction effect that enrichment factor is 1.98
Rate.
The process condition experiment parameter of the optimization water of table 8
"-" represents the experiment parameter of application in table 8.
Embodiment 3:
Rice is tested and analyzed without pre-treatment: the water of 1.5 times of volumes being added in polished rice, then cooks in electric cooker
And keeping warm mode 0h (heat preservation 0h is generated without 5 hydroxymethyl furfural, noiseless to addition mark product) is maintained, and add 50,500 μ g/L
5 hydroxymethyl furfural mark product.It weighs rice 10g to be put into centrifuge tube, the acetonitrile solution of 1 times of volume is added and is cut by high speed
Disperser even 3min under 8000rpm revolving speed is cut to carry out slightly proposing operation.Then, homogenate is by 0.22 μm of film filtering, by super
High performance liquid chromatography separation and Mass Spectrometer Method.
The improved QuEChERS-DLLME pre-treating method of rice is tested and analyzed: being selected under 1 conditional of embodiment
QuEChERS-DLLME pre-treating method the rice of 5 hydroxymethyl furfural mark product of 50,500 μ g/L of addition is carried out at purification
Reason then passes through ultra performance liquid chromatography separation and Mass Spectrometer Method.
The chromatogram of 5 hydroxymethyl furfural is as shown in Fig. 2, by Fig. 2 b, d in the rice obtained under both the above treatment conditions
It is found that improved QuEChERS-DLLME pre-treating method, gained spectrogram is noiseless peak, peak type is well-symbolized and without dragging
Tail, furthermore the 5 hydroxymethyl furfural of same concentrations has significantly enrichment inspissation.However, the spectrogram without pre-treatment is miscellaneous
Peak is more, and peak type is asymmetric, and trail serious (Fig. 2 a, c).Therefore, improved QuEChERS-DLLME pre-treating method reduces
The interference of matrix, strong operability, enrichment concentrated effect is good, the measurement of 5 hydroxymethyl furfural suitable for rice (solid sample).
Embodiment 4:
Only improved QuEChERS pre-treatment is tested and analyzed rice: the water of 1.5 times of volumes being added in polished rice, then
It is cooked in electric cooker and maintains keeping warm mode 30h.The rice 10g for weighing heat preservation different time is put into centrifuge tube, is added 1 times
The acetonitrile solution of volume simultaneously be homogenized 3min by high-speed shearing disperser under 8000rpm revolving speed and carries out thick proposing operation.Then exist
1gMgSO is added in homogenate4, 0.5gNaCl as deicer, then vortex 1min is simultaneously centrifuged 5min at 8000rpm rapidly
The clear acetonitrile layer containing 5 hydroxymethyl furfural is obtained to precipitate rice sample.Then in clear solution be added 60mgC18,
50mg PSA, subsequent vortex 1min rapidly are simultaneously centrifuged 5min at 8000rpm, and clear supernatant liquor is passed through 0.22 μm of film mistake
Filter passes through ultra performance liquid chromatography separation and Mass Spectrometer Method.
Rice is only tested and analyzed through DLLME pre-treatment: the water of 1.5 times of volumes being added in polished rice, then in electric cooker
Middle cooking simultaneously maintains keeping warm mode 30h.The rice 10g for weighing heat preservation different time is put into centrifuge tube, and the second of 1 times of volume is added
Nitrile solution simultaneously be homogenized 5min by high-speed shearing disperser under 8000rpm revolving speed and carries out thick proposing operation.Then, certain volume is taken
Scavenging solution (dispersing agent) mixed with the chloroform (extractant) of certain volume, make the volume ratio (body of scavenging solution and extractant
1) product is than being 4:1.The ultrapure water for being 7 then to the pH value that certain volume is added in mixed liquor extracts ultrapure water and scavenging solution
The ratio (volume ratio 2) of the sum of agent volume is 1:1.Mixed solution shows cloud immediately, 5min is centrifuged at 8000rpm, so
After collect undermost organic phase through being dried with nitrogen, the filtering of 0.22 μm of film is redissolved and passed through with ultrapure water, then passes through ultra high efficiency
Liquid chromatogram separation and Mass Spectrometer Method.
The improved QuEChERS-DLLME pre-treating method of rice is tested and analyzed: selecting optimal conditions
QuEChERS-DLLME pre-treating method carries out purified treatment to rice, is then examined by ultra performance liquid chromatography separation and mass spectrum
It surveys.
The chromatogram of 5 hydroxymethyl furfural is as shown in Figure 3 in the rice obtained under the conditions of three of the above processing mode.By Fig. 3
It is found that the chromatography peak intensity for the 5 hydroxymethyl furfural that only improved QuEChERS pre-treatment obtains is minimum and noise is more,
Matrix interference is more serious；The chromatogram only obtained through DLLME pre-treatment makes moderate progress, but still has more noise, peak type
There are hangover and asymmetry；In contrast, the 5 hydroxymethyl furfural spectrogram base that improved QuEChERS-DLLME pre-treatment obtains
Line is more steady, noise is less, matrix interference is small and peak type is symmetrical, peak intensity is higher.Also, only improved QuEChERS
Pre-treatment and the detection only obtained through DLLME pre-treatment limit are respectively 5.0 μ g/L, 2.0 μ g/L；The rate of recovery is respectively 25.86%
~42.94%, 44.02%~68.55%.
Therefore, improved QuEChERS-DLLME pre-treating method eliminate significantly the interference of matrix, catharsis it is obvious,
It is good to be enriched with concentrated effect, the measurement of 5 hydroxymethyl furfural suitable for rice.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all in essence of the invention
Within mind and principle, any modification, equivalent substitution, improvement and etc. done be should be included within the scope of the present invention.
Claims (10)
1. the pre-treating method of 5 hydroxymethyl furfural in a kind of rice, which is characterized in that the described method includes:
(1) in organic solvent, by the octadecylsilane chemically bonded silica of every 8~12g rice and 40mg~80mg, 40mg~
The N- propyl ethylenediamine of 80mg acts on, and is purified, is purified liquid；
(2) step (1) resulting scavenging solution is mixed with extractant, water is then added, realize enrichment to get pre-treatment sample.
2. the method according to claim 1, wherein further including the sulphur that 0.5g~2g is added in the step (1)
Sour magnesium and 0.5g~1.5g sodium chloride are as deicer.
3. method according to claim 1 or 2, which is characterized in that 0mg~15mg can also be added in the step (1)
Carbon black.
4. method according to claim 1 to 3, which is characterized in that the organic solvent in the step (1) includes second
Nitrile.
5. method according to claim 1 to 3, which is characterized in that the additive amount of organic solvent in the step (1)
For 0.5~1.5 times of volume of rice quality.
6. the method according to claim 1, wherein the volume of scavenging solution and extractant in the step (2)
Than for (2~8): 1.
7. the method according to claim 1, wherein water consumption and scavenging solution and extractant in the step (2)
Total volume volume ratio be (0.5~2): 1.
8. a kind of method of 5 hydroxymethyl furfural in quantitative detection rice, which is characterized in that the described method includes: carrying out rice in advance
The pre-treatment of meal obtains pre-treatment sample liquid, is then detected；The method of the pre-treatment is claim 1-7 any described
Pre-treating method.
9. according to the method described in claim 8, it is characterized in that, the detection includes high performance liquid chromatography detection, liquid phase color
Spectrum-mass spectrometry detection, gas chromatographic detection, gas chromatography-mass spectrography detection, Micellar Electrokinetic Chromatography detection, height
Imitate anion-exchange chromatography.
10. a kind of ultra performance liquid chromatography-mass spectrometry trace detection 5 hydroxymethyl furfural method, which is characterized in that described
Method includes: the preparatory pre-treatment for carrying out rice, obtains pre-treatment sample, then carries out liquid chromatograph mass spectrography detection；Institute
The method for stating pre-treatment is pre-treating method as claimed in claim 1 to 7.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112858508A (en) * | 2021-01-13 | 2021-05-28 | 浙江省疾病预防控制中心 | Method for determining furfural compounds in coffee product |
| CN113009002A (en) * | 2019-12-20 | 2021-06-22 | 中粮营养健康研究院有限公司 | Sample pretreatment method containing furfural substances and method for detecting content of furfural substances |
| CN113325119A (en) * | 2021-06-11 | 2021-08-31 | 中国热带农业科学院南亚热带作物研究所 | Pesticide residue sample pretreatment concentration method |
| CN115389642A (en) * | 2021-05-25 | 2022-11-25 | 中粮营养健康研究院有限公司 | Method for judging Maillard reaction degree in dairy product |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010013225A1 (en) * | 2008-08-01 | 2010-02-04 | Universidade Nova De Lisboa | Method for the analysis, identification and quantification of furfurals as food quality markers |
| CN108414634A (en) * | 2018-02-12 | 2018-08-17 | 淮阴工学院 | The detection method of 5 hydroxymethyl furfural content in a kind of root of Chinese clematis |
-
2019
- 2019-01-17 CN CN201910043814.6A patent/CN109781883A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010013225A1 (en) * | 2008-08-01 | 2010-02-04 | Universidade Nova De Lisboa | Method for the analysis, identification and quantification of furfurals as food quality markers |
| CN108414634A (en) * | 2018-02-12 | 2018-08-17 | 淮阴工学院 | The detection method of 5 hydroxymethyl furfural content in a kind of root of Chinese clematis |
Non-Patent Citations (6)
| Title |
|---|
| MICHAELA POLOVKOV 等: "Determination and occurrence of 5-hydroxymethyl-2-furaldehyde in white and brown sugar by high performance liquid chromatography", 《FOOD CONTROL》 * |
| TING-TING FENG 等: "Isotope dilution quantification of 5-hydroxymethyl-2-furaldehyde in beverages using vortex-assisted liquid–liquid microextraction coupled with ESI-HPLC-MS/MS", 《ANALYTICAL METHODS》 * |
| XIANBING XU 等: "A rapid clean-up method for the quantitation of 5-hydroxymethyl-2-furaldehyde in thermally treated abalone (Haliotis discus) muscle by HPLC-MS/MS", 《ANAL. METHODS》 * |
| 刘学芝 等: "固相萃取-超高效液相色谱法测定浓缩石榴汁中羟甲基糠醛含量", 《食品工业科技》 * |
| 刘潇潇 等: "HPLC法测定参芪扶正注射液中5-羟甲基糠醛的含量及其来源的初步探讨", 《药物分析杂志》 * |
| 蔡玮红 等: "UPLC-MS/MS法同时测定焙烤食品中的丙烯酰胺、4-甲基咪唑与5-羟甲基糠醛", 《现代食品科技》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113009002A (en) * | 2019-12-20 | 2021-06-22 | 中粮营养健康研究院有限公司 | Sample pretreatment method containing furfural substances and method for detecting content of furfural substances |
| CN112858508A (en) * | 2021-01-13 | 2021-05-28 | 浙江省疾病预防控制中心 | Method for determining furfural compounds in coffee product |
| CN112858508B (en) * | 2021-01-13 | 2022-10-18 | 浙江省疾病预防控制中心 | Method for determining furfural compounds in coffee product |
| CN115389642A (en) * | 2021-05-25 | 2022-11-25 | 中粮营养健康研究院有限公司 | Method for judging Maillard reaction degree in dairy product |
| CN113325119A (en) * | 2021-06-11 | 2021-08-31 | 中国热带农业科学院南亚热带作物研究所 | Pesticide residue sample pretreatment concentration method |
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