CN108956804A - The quantitative detecting method of fluorescent whitening agent WS in a kind of edible mushroom - Google Patents
The quantitative detecting method of fluorescent whitening agent WS in a kind of edible mushroom Download PDFInfo
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- CN108956804A CN108956804A CN201810534026.2A CN201810534026A CN108956804A CN 108956804 A CN108956804 A CN 108956804A CN 201810534026 A CN201810534026 A CN 201810534026A CN 108956804 A CN108956804 A CN 108956804A
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- 239000006081 fluorescent whitening agent Substances 0.000 title claims abstract description 58
- 235000001674 Agaricus brunnescens Nutrition 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 15
- 239000007788 liquid Substances 0.000 claims abstract description 26
- 238000001514 detection method Methods 0.000 claims abstract description 15
- 238000001819 mass spectrum Methods 0.000 claims abstract description 8
- 238000000605 extraction Methods 0.000 claims abstract description 4
- 238000002360 preparation method Methods 0.000 claims abstract description 4
- 238000000746 purification Methods 0.000 claims abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 18
- 230000004044 response Effects 0.000 claims description 15
- 239000012224 working solution Substances 0.000 claims description 15
- 238000010792 warming Methods 0.000 claims description 12
- 239000007789 gas Substances 0.000 claims description 11
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 238000002414 normal-phase solid-phase extraction Methods 0.000 claims description 8
- 239000000126 substance Substances 0.000 claims description 8
- 125000000129 anionic group Chemical group 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- 239000003480 eluent Substances 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 6
- 239000000243 solution Substances 0.000 claims description 6
- 238000004458 analytical method Methods 0.000 claims description 5
- 238000004587 chromatography analysis Methods 0.000 claims description 5
- 230000001681 protective effect Effects 0.000 claims description 5
- 238000002098 selective ion monitoring Methods 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 238000005406 washing Methods 0.000 claims description 5
- 238000005282 brightening Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 3
- 239000005695 Ammonium acetate Substances 0.000 claims description 3
- NDAUXUAQIAJITI-UHFFFAOYSA-N albuterol Chemical compound CC(C)(C)NCC(O)C1=CC=C(O)C(CO)=C1 NDAUXUAQIAJITI-UHFFFAOYSA-N 0.000 claims description 3
- 229940043376 ammonium acetate Drugs 0.000 claims description 3
- 235000019257 ammonium acetate Nutrition 0.000 claims description 3
- GHQPBDDZGPAVJP-UHFFFAOYSA-N azanium;methanol;hydroxide Chemical compound N.O.OC GHQPBDDZGPAVJP-UHFFFAOYSA-N 0.000 claims description 3
- 239000012159 carrier gas Substances 0.000 claims description 3
- 239000001307 helium Substances 0.000 claims description 3
- 229910052734 helium Inorganic materials 0.000 claims description 3
- SWQJXJOGLNCZEY-UHFFFAOYSA-N helium atom Chemical compound [He] SWQJXJOGLNCZEY-UHFFFAOYSA-N 0.000 claims description 3
- 238000012544 monitoring process Methods 0.000 claims description 3
- 238000002390 rotary evaporation Methods 0.000 claims description 3
- 229960002052 salbutamol Drugs 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 1
- 238000005360 mashing Methods 0.000 claims 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 6
- 244000234623 Coprinus comatus Species 0.000 description 5
- 235000004439 Coprinus comatus Nutrition 0.000 description 5
- 241000233866 Fungi Species 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 239000012086 standard solution Substances 0.000 description 4
- 235000013305 food Nutrition 0.000 description 3
- IANQTJSKSUMEQM-UHFFFAOYSA-N 1-benzofuran Chemical compound C1=CC=C2OC=CC2=C1 IANQTJSKSUMEQM-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000123 paper Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000005464 sample preparation method Methods 0.000 description 2
- 239000003053 toxin Substances 0.000 description 2
- 231100000765 toxin Toxicity 0.000 description 2
- 230000002087 whitening effect Effects 0.000 description 2
- 241000222519 Agaricus bisporus Species 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 235000006521 Pleurotus eryngii var ferulae Nutrition 0.000 description 1
- 244000088486 Pleurotus eryngii var. ferulae Species 0.000 description 1
- 241000318836 Pleurotus nebrodensis Species 0.000 description 1
- 241001558929 Sclerotium <basidiomycota> Species 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 229960000956 coumarin Drugs 0.000 description 1
- 235000001671 coumarin Nutrition 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 231100000175 potential carcinogenicity Toxicity 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000004383 yellowing Methods 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The present invention provides the quantitative detecting methods of fluorescent whitening agent WS in edible mushroom a kind of, comprising: sample preparation；Sample extraction；Sample purification；GC-MS Analysis detection；The peak area that gas chromatography-mass spectrum detects obtained fluorescent whitening agent WS is substituted into following formula to calculate, to obtain the measured value of fluorescent whitening agent WS in prepare liquid；
Description
Technical field
Present invention relates particularly to the quantitative detecting methods of fluorescent whitening agent WS in edible mushroom a kind of.
Background technique
Edible mushroom broadly refers to have large-scale fructification or the histioid fungi of sclerotium, including edible, pharmaceutically acceptable
, food medicine dual-purpose and itself containing certain toxin but through processing after edible a variety of macro fungis.People are traditionally
To can be formed in macro fungi with loose fleshiness organ of multiplication, degree of lignification is low, eats without toxin, for people,
Referred to as edible mushroom.Due to being increasingly subject to the attention of people with nutrition abundant and medical care effect, the mankind take the photograph edible mushroom
The source of protein is taken just gradually to be changed from animal protein to vegetable protein, mycoprotein, the demand to edible mushroom is increasing.
But in recent years, part dealer is under the tending to act of interests, and in order to keep edible mushroom appearance whiter, brighter, freshness date extends, commodity valence
Value increases, circulate field before sales to the edible mushrooms such as agaricus bisporus, coprinus comatus using fluorescent whitening agent or containing this at
The antistaling agent divided carries out processing rinsing, and edible mushroom is caused seriously to pollute.In addition, Pleurotus ferulae (also known as Pleurotus nebrodensis) has used fluorescence increasing
After the exceeded wrapping paper of white substance is packed, fluorescent brightening substance severe overweight is caused, is caused to consumer's health
It seriously endangers.
Fluorescent whitening agent WS is a kind of coumarin type fluorescent dye, has coumarone basic structure, there is excellent light resistance,
It is one of the fluorescent whitening agent that people have found earliest, it is water-soluble stronger, it can be easy to be adsorbed by food in aqueous solution, and cannot
It rinses out at once, there is excellent level-dyeing property and permeability, it is easy to use.Its whitening effect is former using complementary color optically
Reason, makes yellowing substance after fluorescent whitening agent is handled, can not only reflect visible light, moreover it is possible to absorb ultraviolet light other than visible light simultaneously
It is changed into the visible reflectance with hyacinthine or cyan to come out.Yellow and blue complementary color each other, counteract the original Huang of substance
Color is allowed to seem pure white.Fluorescent whitening agent has potential carcinogenicity on toxicity, may be to human body after being absorbed by the body
Liver, kidney etc. cause seriously to damage.
Therefore the method for quantitatively determining for studying and formulating fluorescent whitening agent WS in edible mushroom is of great significance.Currently, state
The report of inside and outside related fluorescent whitening agent detection, is concentrated mainly in paper, detergent and plastic material, SN/T 4396-2015
Define the measurement liquid chromatography-mass spectrography of fluorescent whitening agent 85, fluorescent whitening agent 71 and fluorescent whitening agent 113 in export food/
Mass spectrography, but this method does not include fluorescent whitening agent WS, and NY/T 1257-2006 defines the detection of fluorescent material in edible mushroom,
But this method is only qualitative, and accuracy is not high, has no the side of gas chromatograph-mass spectrometer (GC-MS) measurement edible fungus fluorescent whitening agent WS
Method report.In addition, ASTM, GB, the standards such as SN, which are looked into, newly has no that relevant criterion is reported via ISO, phase is also had no through patent network inquiry
Close patent report.Therefore it is badly in need of developing the gas chromatography-mass spectrum detection side of fluorescent whitening agent WS in accurately and rapidly edible mushroom
Method.
Summary of the invention
The technical problem to be solved in the present invention is to provide the quantitative detection side of fluorescent whitening agent WS in edible mushroom a kind of
Method.
The present invention is implemented as follows:
The quantitative detecting method of fluorescent whitening agent WS in a kind of edible mushroom, comprising the following steps:
(1), sample preparation: taking representative sample, after its edible portion is directly shredded without washing, is smash with tissue
It is broken to be machined to pulpous state, mix well, acquisition prepares sample, then sealing and in -18 DEG C or less freezen protectives, it is spare；
(2), sample extraction: weighing 5g sample from preparing in sample, is accurate to 0.01g, 10mL acetonitrile is added, 5mL pH is 10
50mmol/L ammonium acetate solution, in mixing 3min on liquid blending device, 40 DEG C of water bath sonicators extract 10min, 4000r/min from
Heart 5min collects supernatant；
(3), sample purification: weak anionic solid-phase extraction column is activated using preceding with 3mL methanol, 3mL deionized water, is drawn
10mL supernatant is all extracted by solid phase after the weak anionic solid-phase extraction column activated, and with the flow velocity lower than 1.0mL/min
Column is taken, eluent is discarded, then is eluted respectively with 3mL deionized water and 3mL methanol, eluent is discarded, finally uses 2% (body of 10mL
Product) the elution of ammonia water-methanol solution, collect eluent, 50 DEG C of rotary evaporations are settled to 1mL with n-hexane, cross film to close dry,
Obtain prepare liquid；
(4), analysis detection: carrying out analysis detection using gas chromatography-mass spectrography, i.e. glimmering to obtain measured object in prepare liquid
The response of optical brightener WS is chosen the corresponding standard working solution of response according to the response condition of measured object in prepare liquid and is carried out
Chromatography, standard working solution are equipped with comprising five concentration gradients including zero point, and fluorescence in standard working solution and prepare liquid
The response of whitening substance should all be in instrument linear response range, and blank control is arranged simultaneously；
The condition of the gas chromatography-mass spectrum are as follows:
1) chromatographic column: HP-5MS, specification 30m × 0.25mm × 0.25 μm or other columns imitate comparable chromatographic column；
2) chromatographic column temperature program: 60 DEG C of initial column temperature, 1min is kept；Then 220 DEG C are warming up to 20 DEG C/min, kept
1min；250 DEG C are warming up to 5 DEG C/min again, keeps 1min；280 DEG C are warming up to 10 DEG C/min again, keeps 1min；Again with 20
DEG C/min is warming up to 300 DEG C, keep 2min；
3) carrier gas: helium purity >=99.999%, constant current mode, flow velocity 1.0mL/min；
4) injector temperature: 250 DEG C；
5) sample volume: 1 μ L；
6) input mode: Splitless injecting samples；Solvent delay: 6.0min；
7) ion source: the source EI, 70eV；
8) ion source temperature: 230 DEG C, 150 DEG C of level four bars temperature；
9) chromatography and mass spectrometer interface temperature: 280 DEG C；
10) mass scanning mode: Salbutamol Selected Ion Monitoring SIM, monitoring ion are 216,231 and 188；Its relative abundance ratio:
216:231:188=100:38:25；
(5), result calculates: the peak area that gas chromatography-mass spectrum detects fluorescent whitening agent WS obtained is substituted into following formula
(1) it is calculated, to obtain the measured value of fluorescent whitening agent WS in prepare liquid；
Wherein:
Fluorescent whitening agent WS residual content in X-sample, μ g/kg；
The peak area of fluorescent whitening agent WS in A-prepare liquid；
The peak area of fluorescent whitening agent WS in As-standard working solution；
Fluorescent whitening agent WS concentration in c-standard working solution, μ g/L；
The final constant volume of V-prepare liquid, mL；
M-sample quality, g.
The present invention has the advantages that having filled up sky of the China in edible mushroom on the quantitative measurement technology of fluorescent whitening agent WS
It is white, and have many advantages, such as high easily operated and detection accuracy, the low, high sensitivity of detection limit, reproducible.
Specific embodiment
The quantitative detecting method of fluorescent whitening agent WS in a kind of edible mushroom, comprising the following steps:
1, sample preparation: taking representative sample about 500g, after the chopping of its edible portion (unavailable washing), uses tissue
Sample is processed pulp by bruisher, is mixed well, and test is divided into two parts, is packed into clean container and is sealed and carry out mark, and -18 DEG C
Following freezen protective, it is spare；
2, sample extraction: about 5g sample (being accurate to 0.01g) is weighed in sample in 50mL tool plug centrifuge tube from preparing, is added
10mL acetonitrile, the 50mmol/L ammonium acetate solution (pH 10) of 5mL, in mixing 3min, 40 DEG C of water bath sonicators on liquid blending device
10min is extracted, 4000r/min is centrifuged 5min, and it is to be measured to collect supernatant.
3, sample purification: weak anionic solid-phase extraction column is activated using preceding with 3mL methanol, 3mL deionized water, and 10mL is drawn
Supernatant all passes through solid-phase extraction column after the weak anionic solid-phase extraction column activated, and with the flow velocity lower than 1.0mL/min,
It is eluted with 3mL deionized water and 3mL methanol, is finally eluted with the ammonium hydroxide methanol of 10mL 2% respectively again, collect eluent in chicken
In heart bottle, 50 DEG C of rotary evaporations are done to close, are settled to 1mL with n-hexane, cross film, to be measured.
4, instrument test: analysis detection is carried out using gas chromatography-mass spectrography, to obtain measured object i.e. fluorescence in prepare liquid
The response of brightening agent WS chooses the corresponding standard working solution of response according to the response condition of measured object in prepare liquid and carries out color
Spectrum analysis, standard working solution are equipped with comprising five concentration gradients including zero point, and fluorescence increases in standard working solution and prepare liquid
The response of white substance should all be in instrument linear response range, and blank control is arranged simultaneously；
The condition of the gas chromatography-mass spectrum are as follows:
1) chromatographic column: HP-5MS (30m × 0.25mm × 0.25 μm) or other columns imitate comparable chromatographic column；
2) chromatographic column temperature program: 60 DEG C of initial column temperature, 1min is kept；Then 220 DEG C are warming up to 20 DEG C/min, kept
1min；250 DEG C are warming up to 5 DEG C/min again, keeps 1min；280 DEG C are warming up to 10 DEG C/min again, keeps 1min；Again with 20
DEG C/min is warming up to 300 DEG C, keep 2min；
3) carrier gas: helium (purity >=99.999%), constant current mode, flow velocity 1.0mL/min；
4) injector temperature: 250 DEG C；
5) sample volume: 1 μ L；
6) input mode: Splitless injecting samples；Solvent delay: 6.0min；
7) ion source: the source EI, 70eV；
8) ion source temperature: 230 DEG C, 150 DEG C of level four bars temperature；
9) chromatography and mass spectrometer interface temperature: 280 DEG C；
10) mass scanning mode: Salbutamol Selected Ion Monitoring (SIM), monitoring ion are 216,231 and 188；Its relative abundance
Than: 216:231:188=100:38:25；
5, result calculates: the peak area that gas chromatography-mass spectrum detects fluorescent whitening agent WS obtained is substituted into following formula (1)
It is calculated, to obtain the measured value of fluorescent whitening agent WS in prepare liquid；
Wherein:
Fluorescent brightening substance fluorescent whitening agent WS residual content, μ g/kg in X-sample；
The peak area of fluorescent whitening agent WS in A-prepare liquid；
The peak area of fluorescent whitening agent WS in As-standard working solution；
Fluorescent whitening agent WS concentration in c-standard working solution, μ g/L；
The final constant volume of V-prepare liquid, mL；
M-sample quality, g.
Embodiment 1: sample to be tested --- coprinus comatus
1.1 sample preparations: taking representative sample about 500g, after the chopping of its edible portion (unavailable washing), uses group
It knits bruisher and sample is processed into pulp, mix well, test is divided into two parts, and it is packed into clean container and seals and carry out mark, -18
DEG C or less freezen protective, it is spare；.
The test of 1.2 backgrounds: the examination in 5g (being accurate to 0.01g) 1.1 is respectively weighed into six 50mL tool plug centrifuge tubes
Sample is then successively operated according to step 2, step 3, step 4 in the specific embodiment of the invention, is finally counted according to step 5
It calculates resulting each actual measured value and average value is as shown in table 1, which is background values.
The background values determination data of fluorescent whitening agent WS in 1 coprinus comatus of table
1.3 verifyings one: 10 μ g/kg fluorescent whitening agent WS of addition
The sample in 5g (being accurate to 0.01g) 1.1, which is respectively weighed, into six 50mL tool plug centrifuge tubes (is specifically shown in Table 2
In sample weighting amount), then into each centrifuge tube add 0.5mg/L fluorescent whitening agent WS standard solution 0.1mL, then successively
It is operated according to step 2, step 3, step 4 in the present invention, resulting each actual measured value is finally calculated according to step 5 and is listed in
In table 2.
The related data of the fluorescent whitening agent WS of 10 μ g/kg is added in 2 coprinus comatus of table
1.4 verifyings two: 50 μ g/kg fluorescent whitening agent WS of addition
The sample in 5g (being accurate to 0.01g) 1.1, which is respectively weighed, into six 50mL tool plug centrifuge tubes (is specifically shown in Table 3
In sample weighting amount), then into each centrifuge tube add 1mg/L fluorescent whitening agent WS standard solution 0.25mL, then successively
It is operated according to step 2, step 3, step 4 in the present invention, and resulting each actual measured value finally will be calculated according to step 5
It is listed in Table 3 below.
The rate of recovery data of the fluorescent whitening agent WS of 50 μ g/kg are added in 3 coprinus comatus of table in addition
Embodiment 2: sample to be tested --- needle mushroom
2.1 sample preparations: taking representative sample about 500g, after the chopping of its edible portion (unavailable washing), uses group
It knits bruisher and sample is processed into pulp, mix well, test is divided into two parts, and it is packed into clean container and seals and carry out mark, -18
DEG C or less freezen protective, it is spare；.
The test of 2.2 backgrounds: the examination in 5g (being accurate to 0.01g) 1.1 is respectively weighed into six 50mL tool plug centrifuge tubes
Sample is then successively operated according to step 2, step 3, step 4 in the specific embodiment of the invention, is finally counted according to step 5
It calculates resulting each actual measured value and average value is as shown in table 4, which is background values.
The background values determination data of fluorescent whitening agent WS in 4 needle mushroom of table
2.3 verifyings one: 10 μ g/kg fluorescent whitening agent WS of addition
The sample in 5g (being accurate to 0.01g) 2.1, which is respectively weighed, into six 50mL tool plug centrifuge tubes (is specifically shown in Table 5
In sample weighting amount), then into each centrifuge tube add 0.5mg/L fluorescent whitening agent WS standard solution 0.1mL, then successively
It is operated according to step 2, step 3, step 4 in the present invention, resulting each actual measured value is finally calculated according to step 5 and is listed in
In table 5.
The related data of the fluorescent whitening agent WS of 10 μ g/kg is added in 5 needle mushroom of table
2.4 verifyings two: 50 μ g/kg fluorescent whitening agent WS of addition
The sample in 5g (being accurate to 0.01g) 2.1, which is respectively weighed, into six 50mL tool plug centrifuge tubes (is specifically shown in Table 4
In sample weighting amount), then into each centrifuge tube add 1mg/L fluorescent whitening agent WS standard solution 0.25mL, then successively
It is operated according to step 2, step 3, step 4 in the present invention, and resulting each actual measured value finally will be calculated according to step 5
It is listed in Table 6 below.
The 50 μ g/kg (rate of recovery data of fluorescent whitening agent WS are added in addition in 6 needle mushroom of table
By above-mentioned experimental result it is found that when the present invention is applied to fluorescent whitening agent WS content in edible mushroom, lower limit of measurement are as follows:
10 μ g/kg, rate of recovery 77.8-96.0%, correlation coefficient r good in 10~200 μ g/L range internal standard curve linear relationships
Greater than 0.999.
From the size of the rate of recovery and RSD in respective verification result can determine whether out detection method accuracy of the invention compared with
It is high.
Although specific embodiments of the present invention have been described above, those familiar with the art should be managed
Solution, we are merely exemplary described specific embodiment, rather than for the restriction to the scope of the present invention, it is familiar with this
The technical staff in field should be covered of the invention according to modification and variation equivalent made by spirit of the invention
In scope of the claimed protection.
Claims (1)
1. the quantitative detecting method of fluorescent whitening agent WS in a kind of edible mushroom, it is characterised in that: the following steps are included:
(1), sample preparation: taking representative sample, after its edible portion is directly shredded without washing, uses tissue mashing machine
Process pulp, mix well, acquisition prepares sample, then sealing and in -18 DEG C or less freezen protectives, it is spare；
(2), sample extraction: weighing 5g sample from preparing in sample, is accurate to 0.01g, 10mL acetonitrile is added, 5mLpH is 10
50mmol/L ammonium acetate solution, in mixing 3min on liquid blending device, 40 DEG C of water bath sonicators extract 10min, 4000r/min centrifugation
5min collects supernatant；
(3), sample purification: weak anionic solid-phase extraction column is activated using preceding with 3mL methanol, 3mL deionized water, is drawn on 10mL
Clear liquid all passes through solid-phase extraction column after the weak anionic solid-phase extraction column activated, and with the flow velocity lower than 1.0mL/min, then
It is eluted respectively with 3mL deionized water and 3mL methanol, discards eluent, finally eluted with the ammonia water-methanol solution of 10mL2%, received
Collect eluent, 50 DEG C of rotary evaporations are done to close, are settled to 1mL with n-hexane, are crossed film, obtain prepare liquid；
(4), analysis detection: carrying out analysis detection using gas chromatography-mass spectrography, is increased with obtaining measured object i.e. fluorescence in prepare liquid
The response of white dose of WS chooses the corresponding standard working solution of response according to the response condition of measured object in prepare liquid and carries out chromatography
Analysis, standard working solution are equipped with comprising five concentration gradients including zero point, and fluorescent brightening in standard working solution and prepare liquid
The response of substance should all be in instrument linear response range, and blank control is arranged simultaneously；
The condition of the gas chromatography-mass spectrum are as follows:
1) chromatographic column: HP-5MS, specification 30m × 0.25mm × 0.25 μm or other columns imitate comparable chromatographic column；
2) chromatographic column temperature program: 60 DEG C of initial column temperature, 1min is kept；Then 220 DEG C are warming up to 20 DEG C/min, kept
1min；250 DEG C are warming up to 5 DEG C/min again, keeps 1min；280 DEG C are warming up to 10 DEG C/min again, keeps 1min；Again with 20
DEG C/min is warming up to 300 DEG C, keep 2min；
3) carrier gas: helium purity >=99.999%, constant current mode, flow velocity 1.0mL/min；
4) injector temperature: 250 DEG C；
5) sample volume: 1 μ L；
6) input mode: Splitless injecting samples；Solvent delay: 6.0min；
7) ion source: the source EI, 70eV；
8) ion source temperature: 230 DEG C, 150 DEG C of level four bars temperature；
9) chromatography and mass spectrometer interface temperature: 280 DEG C；
10) mass scanning mode: Salbutamol Selected Ion Monitoring SIM, monitoring ion are 216,231 and 188；Its relative abundance ratio: 216:
231:188=100:38:25；
(5), result calculate: by gas chromatography-mass spectrum detect fluorescent whitening agent WS obtained peak area substitute into following formula (1) into
Row calculates, to obtain the measured value of fluorescent whitening agent WS in prepare liquid；
Wherein:
Fluorescent whitening agent WS residual content in X-sample, μ g/kg；
The peak area of fluorescent whitening agent WS in A-prepare liquid；
The peak area of fluorescent whitening agent WS in As-standard working solution；
Fluorescent whitening agent WS concentration in c-standard working solution, μ g/L；
The final constant volume of V-prepare liquid, mL；
M-sample quality, g.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001124750A (en) * | 1999-10-26 | 2001-05-11 | Ube Kagaku Bunseki Center:Kk | Measuring method for alkylphenols |
| CN105606430A (en) * | 2015-12-24 | 2016-05-25 | 上海微谱化工技术服务有限公司 | Separation and analysis methods of aromatic hydrocarbon additive in polymer with benzene ring structure |
| CN105628823A (en) * | 2016-02-04 | 2016-06-01 | 中山出入境检验检疫局检验检疫技术中心 | Method adopting high performance liquid chromatography to detect fluorescent whitener in flour |
-
2018
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001124750A (en) * | 1999-10-26 | 2001-05-11 | Ube Kagaku Bunseki Center:Kk | Measuring method for alkylphenols |
| CN105606430A (en) * | 2015-12-24 | 2016-05-25 | 上海微谱化工技术服务有限公司 | Separation and analysis methods of aromatic hydrocarbon additive in polymer with benzene ring structure |
| CN105628823A (en) * | 2016-02-04 | 2016-06-01 | 中山出入境检验检疫局检验检疫技术中心 | Method adopting high performance liquid chromatography to detect fluorescent whitener in flour |
Non-Patent Citations (4)
| Title |
|---|
| M. CASTILLO等: "Characterization of organic pollutants in industrial ef£uents by high-temperature gas chromatography-mass spectrometry", 《TRENDS IN ANALYTICAL CHEMISTRY》 * |
| 冼燕萍等: "固相萃取-超高效液相色谱分离测定洗涤用品中4种荧光增白剂", 《色谱》 * |
| 张建莹等: "超高效液相色谱串联质谱法检测食用菌中荧光增白剂", 《食品安全质量检测学报》 * |
| 黄薇等: "高效液相色谱法测定食用菌中10种荧光增白剂", 《理化检验-化学分册》 * |
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