CN106770729B - A kind of method for detecting free state and bound state trace cyanide - Google Patents
A kind of method for detecting free state and bound state trace cyanide Download PDFInfo
- Publication number
- CN106770729B CN106770729B CN201611075836.3A CN201611075836A CN106770729B CN 106770729 B CN106770729 B CN 106770729B CN 201611075836 A CN201611075836 A CN 201611075836A CN 106770729 B CN106770729 B CN 106770729B
- Authority
- CN
- China
- Prior art keywords
- cyanide
- derivative
- bottle
- free state
- pretreatment
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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
Landscapes
- 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)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention discloses a kind of detection free state and the methods of reference state micro cyanide, belong to technical field of analysis and detection, are mainly used for alcoholic drink and its feedstock analysis.The present invention can handle free state or reference state cyanide sample using a kind of specific pretreatment and derivatization device respectively.For free state cyanide, it is directly that cyanogen root is derivative to discharge hydrogen cyanide；Free hydrogen cyanide is first changed into for reference state cyanide, then derivative with derivative reagent.Qualitative and quantitative analysis is carried out using liquid chromatography-mass spectrometry for derivative, mark song is established using internal standard method, calculates separately free state and reference state cyanide content.The detection method detection limit that the present invention establishes can quantitatively be limited to 0.05 μ g/L down to 0.01 μ g/L, and linearly dependent coefficient is greater than 0.99, and recovery of standard addition is 93.8%~106%, and relative standard deviation is less than 10%.The method of the present invention detection sensitivity is high, and required time is short, and interference is small, can meet the requirement of reference state and the measurement of free state cyanide respectively.
Description
Technical field
The present invention relates to a kind of detection free state and the methods of reference state micro cyanide, belong to analysis and detection technology neck
Domain.It is mainly used for alcoholic drink and its feedstock analysis.
Background technique
Cyanide category extremely toxic substance can be entered in human body by respiratory tract, alimentary canal and skin, in cell mitochondrial
The ferric iron of oxidized form cytochrome oxidase combines, and prevents the ferric iron back in oxidizing ferment, interferes cell eupnea, makes
At body anoxic so that generating poisoning effect.In addition, cyanide is also possible to form other hazardous compounds under certain condition,
There are security risks in food, beverage.Cyanide existing for nature includes free state and reference state.Reference state cyanide is usual
Exist in the form of cyanogenic glycoside, by hydrolyzing to form free state cyanide, therefore is also important risk factor.Cyanide is
One important foodstuffs safety detection index of drinks, cyanide is also the important precursor to form urethanes.Beverage
Free state and reference state cyanide in wine and its raw material, can form free state, then form urethanes with alcohol.
There is a problem of that urethanes is exceeded in the alcoholic drink of part, for the health with the people that ensures food safety, beverage
Cyanide research in wine is very necessary.Accurate Determining cyanide is of great significance for the control of related hazardous material.
Currently, both at home and abroad to the measurement of cyanide apply more medical treatment detection, in terms of.Measuring method mainly has colorimetric method, glimmering
Light method and headspace gas chromatography.And the measurement of reference state cyanide is less common, generally requires more complex hydrolytic process.Than
Color method mainly has isonicotinic acid-pyrazolone, isonicotinic acid -- barbituric acid, Pyridine-barbituric acid method.Wherein, isonicotinic acid -- barbituric acid
Method is the method that GB/T5009.48-2003 is used to detect cyanide content in white wine.This method is cumbersome, it is muddy white occur
Turbid and color interference problem, there are false positive interference in qualitative, detection limit measures micro cyanide tired in 3mg/L
It is difficult.Headspace gas chromatography is to be performed the derivatization using reagent to cyanide, is detected by headspace extraction combination GC-ECD.
The cyanide that the method detects is free state cyanide, and detection limit is in 2.5 μ g/L.Detection of the existing method to cyanide, one
As that there are detection limits is higher, be easy to be interfered by sample substrate, reference state cyanide measures difficult or cumbersome, equipment
Complexity interferes the problems such as big.
The detection of cyanide is typically necessary by pretreatment, cyanogen root is acidified and is detected again for hydrogen cyanide, and combines
The pretreatment of state cyanide is then increasingly complex.Current general pre-processing device is frequently present of that conversion ratio is low or hydrogen cyanide loss,
Or the problems such as cumbersome, keep accuracy in detection low, it cannot actual content in test sample well.For micro free cyaniding
The measurement of object, it is more with gas phase or liquid chromatography both at home and abroad at present, it generally requires and free cyanide is first acidified and is spread out
Biochemistry, then carry out gas phase or liquid chromatogram measuring.And for micro reference state cyanide at present still without direct method for measuring.
Summary of the invention
The first purpose of the invention is to provide a kind of detection free state and the methods of reference state micro cyanide, solve existing
There are technology free state and reference state determination methods of cyanide detection limit for height, cumbersome, detection sensitivity is low to ask vulnerable to interference etc.
Topic.
A method of detection free state and reference state micro cyanide use not the cyanide of free state and reference state
Same pretreatment mode；Pretreatment to free state cyanide is after being directly acidified cyanogen root in pretreatment and derivatization device
Perform the derivatization reaction；Pretreatment to reference state cyanide is first heated using boiling water bath in pretreatment and derivatization device
Way of distillation decomposition releases after free state cyanide derivative reaction again；Derivative is detected, free state is calculated separately out
The content of cyanide and total cyanide, it is reference state cyanide content that total cyanide content, which subtracts free state cyanide content,.
The pretreatment and derivatization device include example reaction bottle and derivative receiving bottle, and acid is carried out in example reaction bottle
Solution reaction releases free state cyanide, and the free state cyanide released is imported into the derivative added with derivating agent by conduit
Receiving bottle performs the derivatization reaction in derivative receiving bottle.Detection free state and reference state cyanide method specifically include with
Lower step:
(1) pretreatment of free state and reference state cyanide sample and derivatization
A. free state cyanide measurement sample pretreatment and derivatization steps are as follows: accurately weigh appropriate amount of fluid sample or
Solid sample leaching liquor is added to the example reaction bottle of pretreatment and derivatization device, opens acid-adding port, is rapidly added appropriate amount of acid
Acid-adding port is closed immediately after change reagent to be reacted.It opens controllable switch and is passed through nitrogen, in the item for being passed through nitrogen in reaction process
Under part, the hydrogen cyanide of generation is transferred to derivative receiving bottle.Derivative receiving flask is previously added a certain amount of derivatization reagent and interior
Mark solution.Derivative to be measured is obtained after reaction, is used for liquid phase or gas chromatographic analysis.
B. reference state cyanide measurement sample pretreatment and derivatization steps are as follows: accurately weigh appropriate amount of fluid sample or
Solid sample is added to the example reaction bottle of pretreatment and derivatization device, and a certain proportion of distilled water and anti-interference is added
Agent.For containing ethanol sample, appropriate sodium hydroxide solution alkaline hydrolysis 10min is first added.Then acid-adding port is opened, excess is rapidly added
Acid-adding port is closed after acidizing reagent immediately.The heating of example reaction bottle is reacted and is distilled.It condenses and communicating pipe connects cooling water, will distillate
The hydrogen cyanide generated in liquid is transferred to derivative receiving bottle.Derivative receiving flask is previously added a certain amount of derivatization reagent and internal standard is molten
Liquid.Distillate is collected in derivative receiving bottle and carries out derivative reaction, after reaction constant volume and derivative to be measured, for liquid
Phase or gas chromatographic analysis.
(2) detection point is carried out to cyanide derivative products and internal standard 13C15N- cyanide derivatives using LC-MS instrument
Analysis；
(3) cyanide standard curve is made using LC-MS/MS-MRM method, and according in the use of cyanide standard curve
Mark method in sample free state and reference state cyanide content calculate.Calculate separately out free state cyanide and total cyaniding
The content of object, it is reference state cyanide content that total cyanide content, which subtracts free state cyanide content,.
The ratio of a certain proportion of distilled water of addition described in step (1) b, sample and distilled water is 1:(5-10) g/mL,
Soaking time is 0-5h；The acidizing reagent is tartaric acid, and additive amount is the 10-20% of sample quality；The anti-interference agent is
The acetic acid zinc solution of 100g/L, additive amount are sample: zinc acetate is equal to 1:(1-3) g/mL；The acidification examination added in step (1) a
Agent is lactic acid, reaction time 15-35min.
Agent derived from step (1) a and b be include 2.71mmol/L methanol dissolution two acetal of 2,3- naphthalene (NDA) it is molten
Liquid, the taurine solution of 50mmol/L, formaldehyde and ammonium hydroxide volume ratio are the mixed solution of 25:25:45:5；The inner mark solution
The 13C15N- cyanide standard solution for being 1-5mg/L for concentration.
LC-MS in step (2), chromatographic condition are as follows: chromatographic column: Acquity UPLC C18,2.1 × 100mm, 1.7 μ
m；40 DEG C of column temperature；2 μ L of sample volume；Flow velocity 0.3mL/min；Mobile phase: A is 2mM ammonium acetate, and B is acetonitrile, and gradient elution program is such as
Under:
Mass Spectrometry Conditions are as follows: ionization mode ESI (-)；Multiple reaction monitors mass spectrum mode；Ion source temperature is 150 DEG C；Collision
Energy 30eV；Orifice potential 25V；Capillary pressure 2500V.
A kind of measured a second object of the present invention is to provide free state and reference state micro cyanide pretreatment and spread out
Biochemical device.
The pretreatment and derivatization device include example reaction bottle and derivative receiving bottle, and acid is carried out in example reaction bottle
Solution reaction releases free state cyanide, and the free state cyanide released is imported into the derivative that derivating agent is added by conduit
Receiving bottle performs the derivatization reaction in derivative receiving bottle.
The pretreatment of the cyanide measurement and derivatization device specifically include example reaction bottle (1), derivative receiving bottle
(2), example reaction bottle long duct (3), example reaction bottle tubulature (4), derivative receiving bottle long duct (5), atmospheric communicating pipe
(6), communicating pipe (7), controllable switch (8), acid-adding port (9) bottle stopper (10), cooling water outlet (11) and cooling water inlet are condensed
(12)；Described example reaction bottle long duct (3) one end is extend into below example reaction bottle bottom of bottle liquid level, one end and controllable switch
(8) it connects；Described derivative receiving bottle long duct (5) one end is extend into below derivative receiving bottle bottom of bottle liquid level；The atmosphere
Pipe (6) one end is extend into derivative receiving bottle, and one end is exposed to outside bottle；The condensation communicating pipe (7) is short by example reaction bottle
Conduit (4) is connected to example reaction bottle and derivative receiving bottle with derivative receiving bottle long duct (5), and the hydrogen cyanide that reaction flask generates is led
Enter to derivative receiving bottle；The controllable switch (8) connects logical nitrogen.
Third object of the present invention is to provide a kind of reference state and the pretreatment of free state micro cyanide and derivatization sides
Method.
Steps are as follows for sample pretreatment and derivative:
A. free state cyanide measurement sample pretreatment and derivatization steps are as follows: accurately weigh appropriate amount of fluid sample or
Solid sample leaching liquor is added to the example reaction bottle of pretreatment and derivatization device, opens acid-adding port, is rapidly added appropriate amount of acid
Acid-adding port is closed immediately after change reagent to be reacted.It opens controllable switch and is passed through nitrogen, in the item for being passed through nitrogen in reaction process
Under part, the hydrogen cyanide of generation is transferred to derivative receiving bottle.Derivative receiving flask is previously added a certain amount of derivatization reagent and interior
Mark solution.Derivative to be measured is obtained after reaction, is used for liquid phase or gas chromatographic analysis.
B. reference state cyanide measurement sample pretreatment and derivatization steps are as follows: accurately weigh appropriate amount of fluid sample or
Solid sample is added to the example reaction bottle of pretreatment and derivatization device, and a certain proportion of distilled water and anti-interference is added
Agent.For containing ethanol sample, appropriate sodium hydroxide solution alkaline hydrolysis 10min is added.Then acid-adding port is opened, excess acid is rapidly added
Acid-adding port is closed immediately after changing reagent.The heating of example reaction bottle is reacted and is distilled.It condenses and communicating pipe connects cooling water, by distillate
The hydrogen cyanide of middle generation is transferred to derivative receiving bottle.Derivative receiving flask is previously added a certain amount of derivatization reagent and internal standard is molten
Liquid.Distillate is collected in derivative receiving bottle and carries out derivative reaction, after reaction constant volume and derivative to be measured, for liquid
Phase or gas chromatographic analysis.
The pretreatment and derivatization device include example reaction bottle and derivative receiving bottle, and acid is carried out in example reaction bottle
Solution reaction releases free state cyanide, and the free state cyanide released is imported into the derivative that derivating agent is added by conduit
Receiving bottle performs the derivatization reaction in derivative receiving bottle.
The acidizing reagent added in step a is lactic acid, reaction time 15-35min.
Agent derived from step a and b be include 2.71mmol/L methanol dissolution two acetal of 2,3- naphthalene (NDA) solution,
Taurine solution, formaldehyde and the ammonium hydroxide volume ratio of 50mmol/L is the mixed solution of 25:25:45:5；The inner mark solution is
Concentration is 5mg/L's13C15N- cyanide standard solution.
The ratio of a certain proportion of distilled water of addition described in step b, sample and distilled water is 1:(5-10) g/mL, impregnates
Time is 1-5h；The acidizing reagent is tartaric acid, and additive amount is the 10-20% of sample quality；The anti-interference agent is 100g/
The acetic acid zinc solution of L, additive amount are sample: zinc acetate is equal to 1:(1-3) g/mL；In one embodiment of the invention, it walks
Sample is weighed in rapid b and is added distilled water immersion, and the ratio of sample and distilled water is 1:5g/mL, impregnates 2h；The addition tartaric acid
And zinc acetate, the additive amount of tartaric acid are the 10-20% of sample quality, the additive amount of the acetic acid zinc solution of 100g/L is sample:
Zinc acetate is equal to 1:2g/mL；The volume ratio of the additive amount of step a sample and lactic acid is 2:1, and the derivative reaction is at room temperature
It carries out, time 20min.
Beneficial effects of the present invention:
After being first acidified by the way of derivatization, the interference of other compositions in alcoholic drink is excluded, the inspection of cyanide is improved
Survey accuracy.Qualitative and quantitative analysis is carried out to cyanide using liquid chromatography-mass spectrometry.The present invention is using special pre-
Processing and derivatization device collect the hydrogen cyanide that reaction obtains using conduit degassing method, and whole device air-tightness is good, reduces because of hydrogen
Caused by the loss of cyanic acid detect inaccuracy, it can be achieved that micro cyanide detection.The quantitative Huang of the method that the present invention establishes
The detection limit of cyanide can be down to 0.01 μ g/L in wine, and linearly dependent coefficient is greater than 0.99, and recovery of standard addition is 93.8%~
106%, relative standard deviation is less than 10%.
This method can directly measure free state cyanide in sample, measure the content of reference state cyanide indirectly.Using pre-
Processing and derivatization device according to circumstances can carry out pretreatment and derivatization to free state and reference state cyanide respectively, be used for
Gas phase or liquid chromatogram measuring.The device can meet reference state and the pretreated different requirements of free state cyanide measurement respectively,
The respective content of accurate detection reference state and free state cyanide.
The pretreatment for the cyanide measurement that this method provides and derivatization device are easy to operate, and safety will especially combine
The more complicated pretreatment of state cyanide measurement and derivatization process merge in set of device, significantly simplify operation, subject to
Really measurement sample cyanide content is laid a good foundation.
Detailed description of the invention
Fig. 1: pretreatment and derivatization device figure
Fig. 2: cyanide standard items and internal standard13C15N- cyanide liquid chromatogram
Fig. 3: cyanide standard quality spectrogram
Fig. 4: cyanide measures standard curve
Specific embodiment
The preparation and preservation of reagent:
Potassium cyanide (KCN) stock solution of 10g/L is prepared, 4 DEG C of refrigerators save；Prepare the cyaniding potassium isotope of 5mg/L
(K13C15N) stock solution, 4 DEG C of refrigerators save；Two acetal of 2,3- naphthalene (NDA) of 2.71mM: 100mgNDA is dissolved in 20mL methanol
In, and make 10 times of dilutions, it is kept in dark place at 4 DEG C；50mM taurine: taking 313mg taurine to be settled to 50mL with ultrapure water, and 4 DEG C
Lower preservation.Mix derivating agent, including 2, two acetal of 3- naphthalene (NDA), taurine, formaldehyde, ammonium hydroxide, ratio 25:25:45:5.
The foundation of standard curve:
10mg/L potassium cyanide standard items stock solution is diluted to a series of titer of concentration, each 2mL of above-mentioned solution is taken, adds
Enter in reaction unit example reaction bottle, 2ml derivating agent and 20 μ L 5mg/L are added in derivative receiving bottle13C15In N- cyanide
Mark.1mL lactic acid is added into example reaction bottle to start to react, so that its acidification is generated hydrogen cyanide, and be transferred to derivative receiving bottle quilt
Derivating agent is derivative to be absorbed.20min is reacted at room temperature, then carries out LC-MS analysis.By measured peak area with
For the ratio of isotopic peak area as ordinate, concentration is that abscissa carries out linear regression analysis, draws standard curve.
Embodiment 1: the measurement of free state and reference state cyanide in yellow rice wine
(1) pretreatment of yellow rice wine sample and derivatization:
A. the sample pretreatment and derivatization that free state cyanide measures: in pretreatment and the example reaction of derivatization device
The yellow rice wine sample of 2mL is added in bottle, adds 1mL lactic acid, cyanogen root acidification is transformed into hydrogen cyanide.In the condition for being passed through nitrogen
Under, hydrogen cyanide is transferred in derivative receiving flask.Accurate addition 2mL derivating agent and 10 μ L concentration are in advance in derivative receiving flask
1mg/L internal standard13C15N- cyanide standard solution.Derivative reaction, time 20min, room temperature occur for derivating agent and hydrogen cyanide
Lower progress.Product after being derived, sample detection free cyanide content.
B. the sample pretreatment and derivatization that reference state cyanide measures: in pretreatment and the example reaction of derivatization device
The yellow rice wine sample of 2mL and the sodium hydroxide of 0.5mL 2g/L are added in bottle, 10ml is added after alkaline hydrolysis 10min and is saturated tartaric acid solution
With the zinc acetate of 2mL 100g/L, boiling water bath heating distillation is carried out, the derivative receiving flask of distillate is received.In derivative receiving flask
In accurate addition 2mL derivating agent and 100 μ L concentration are 5mg/L internal standard in advance13C15N- cyanide standard solution.Work as distillate
Stop distillation when close to 10mL.By distillate constant volume, sample detection total cyanide content.
(2) it is produced using the silent winged TSQ Quantum Ultra EMR liquid chromatography-tandedm mass spectro-metry instrument of match is derivative to cyanide
Object and internal standard13C15N- cyanide derivatives are tested and analyzed.
Chromatographic condition: chromatographic column: Acquity UPLC C18,2.1 × 100mm, 1.7 μm；40 DEG C of column temperature；2 μ L of sample volume；
Flow velocity 0.3mL/min；Mobile phase: A is 2mM ammonium acetate, and B is acetonitrile, and gradient elution program is as follows:
Mass Spectrometry Conditions: ionization mode ESI (-)；Multiple reaction monitors mass spectrum mode；Ion source temperature is 150 DEG C；Impact energy
Measure 30eV；Orifice potential 25V；Capillary pressure 2500V.
(3) LC-MS/MS-MRM method is established, carries out quantitative calculating respectively by cyanide standard curve using internal standard method
The content of free state and total cyanide in yellow rice wine, the reference state cyanide that obtains that total cyanide subtracts the calculating of free state cyanide contain
Amount.As a result as shown in the table:
Embodiment 2: the measurement of total cyanide in water
(1) pretreatment of sample and derivatization:
The sample pretreatment of total cyanide determination and derivatization: add in the example reaction bottle of pretreatment and derivatization device
Enter the water sample of 2mL, about 10mL distilled water is added, be rapidly added the zinc acetate of 1g tartaric acid and 2mL 100g/L, carries out boiling water
Bath heating distillation receives the derivative receiving flask of distillate.It is accurate in advance in derivative receiving flask to add 2mL derivating agent and 100 μ
L concentration is 5mg/L internal standard13C15N- cyanide standard solution.When distillate stops distillation when close to 10mL.By distillate
Constant volume, sample detection total cyanide content.
(2) it is produced using the silent winged TSQ Quantum Ultra EMR liquid chromatography-tandedm mass spectro-metry instrument of match is derivative to cyanide
Object and internal standard13C15N- cyanide derivatives are tested and analyzed.
Chromatographic condition: chromatographic column: Acquity UPLC C18,2.1 × 100mm, 1.7 μm；40 DEG C of column temperature；2 μ L of sample volume；
Flow velocity 0.3mL/min；Mobile phase: A is 2mM ammonium acetate, and B is acetonitrile, and gradient elution program is as follows:
Mass Spectrometry Conditions: ionization mode ESI (-)；Multiple reaction monitors mass spectrum mode；Ion source temperature is 150 DEG C；Impact energy
Measure 30eV；Orifice potential 25V；Capillary pressure 2500V.
(3) LC-MS/MS-MRM method is established, quantitative calculate in water is carried out by cyanide standard curve using internal standard method
The content of cyanide, the content for measuring total cyanide is 7.49 μ g/L.
Embodiment 3: the evaluation of cyaniding object detecting method
(1) repeated --- precision test
By 6 parts of standard solution that concentration is 10 μ g/L, detected respectively, final result is as shown in table 1.Test result table
Bright, this method has good reproducibility.
1 precision of table
(2) rate of recovery --- accuracy test
Taking free state concentration of cyanide is 6 parts of yellow rice wine sample of 4.71 μ g/L, and the accurate concentration that is added is 2.5,10 μ g/ respectively
The standard items of L, are measured according to above-mentioned steps.The rate of recovery is respectively 93.8%, 106%.Details are shown in Table 2.
2. rate of recovery of table
(3) detection limit and quantitative limit
When sample volume is 2 μ L, S/N=3, the LOD of cyanide measurement is 0.01 μ g/L；When sample volume is 2 μ L, S/N=
When 10, LOQ is 0.05 μ g/L.
Embodiment 4: the measurement of free state and reference state cyanide in yellow rice wine raw material
(1) pretreatment of yellow rice wine material sample and derivatization:
A. the sample pretreatment and derivatization that free state cyanide measures: in pretreatment and the example reaction of derivatization device
The material sample of 1g is accurately added in bottle, about 5mL distilled water immersion 2h is added, adds 2.5mL lactic acid, cyanogen root acidification is converted to
For hydrogen cyanide.Under conditions of being passed through nitrogen, hydrogen cyanide is transferred in derivative receiving flask.It is added in advance in derivative receiving flask
2mL derivating agent and 10 μ L concentration are 1mg/L internal standard13C15N- cyanide standard solution.Derivatization occurs for derivating agent and hydrogen cyanide
Reaction, time 20min are carried out at room temperature.Product after being derived, sample detection free cyanide content.
B. the sample pretreatment and derivatization that reference state cyanide measures: in pretreatment and the example reaction of derivatization device
The material sample of 1g is accurately added in bottle, about 10mL distilled water immersion 2h is added, is rapidly added 1g tartaric acid and 2mL 100g/L
Zinc acetate, carry out boiling water bath heating distillation, by distillate with 2mL derivating agent receive.2mL is added in advance in derivative receiving flask
Derivating agent and 100 μ L concentration are 5mg/L internal standard13C15N- cyanide standard solution.When distillate stops steaming when close to 10mL
It evaporates.By distillate constant volume, internal standard after constant volume13C15The final concentration of 50 μ g/L of N- cyanide, sample detection total cyanide content.
(2) it is produced using the silent winged TSQ Quantum Ultra EMR liquid chromatography-tandedm mass spectro-metry instrument of match is derivative to cyanide
Object and internal standard13C15N- cyanide derivatives are tested and analyzed.
Chromatographic condition: chromatographic column: Acquity UPLC C18,2.1 × 100mm, 1.7 μm；40 DEG C of column temperature；2 μ L of sample volume；
Flow velocity 0.3mL/min；Mobile phase: A is 2mM ammonium acetate, and B is acetonitrile, and gradient elution program is as follows:
Mass Spectrometry Conditions: ionization mode ESI (-)；Multiple reaction monitors mass spectrum mode；Ion source temperature is 150 DEG C；Impact energy
Measure 30eV；Orifice potential 25V；Capillary pressure 2500V.
(3) LC-MS/MS-MRM method is established, carries out quantitative calculating respectively by cyanide standard curve using internal standard method
The content of free state and total cyanide in yellow rice wine, the reference state cyanide that obtains that total cyanide subtracts the calculating of free state cyanide contain
Amount.As a result as shown in the table:
The above preferred embodiment is merely to illustrate the contents of the present invention, in addition to this, the present invention there are also other embodiments,
In every case those skilled in the art are because of technical inspiration involved in the present invention, and equivalent replacement or equivalent deformation mode is used to be formed
Technical solution is fallen within the scope of protection of the present invention.
Claims (9)
1. a kind of method of detection free state and reference state micro cyanide, which is characterized in that using identical pretreatment and spread out
Biochemical device uses different pretreatment modes to the cyanide of free state and reference state；Pretreatment to free state cyanide
It is to carry out derivative reaction after being directly acidified cyanogen root in pretreatment and derivatization device；Pretreatment to reference state cyanide
It is to derive again after first releasing free state cyanide using boiling water bath heating way of distillation decomposition in pretreatment and derivatization device
Change reaction；LC-MS detection is carried out to derivative and calculates the content of cyanide；
The pretreatment and derivatization device include example reaction bottle and derivative receiving bottle, and it is anti-that acidolysis is carried out in example reaction bottle
Free state cyanide should be released, the free state cyanide released is imported into the derivative reception that derivating agent is added by conduit
Bottle performs the derivatization reaction in derivative receiving bottle；
Comprising the following steps:
(1) pretreatment of free state and reference state cyanide sample and derivatization
A. steps are as follows for the sample pretreatment of free state cyanide measurement and derivatization: accurately weighing appropriate amount of fluid sample or solid
Sample leaching liquor is added to the example reaction bottle of pretreatment and derivatization device, opens acid-adding port, is rapidly added appropriate acidification examination
Acid-adding port is closed after agent immediately to be reacted, controllable switch is opened and is passed through nitrogen, in reaction process under conditions of being passed through nitrogen,
The hydrogen cyanide of generation is transferred to derivative receiving bottle, derivative receiving flask is previously added a certain amount of derivatization reagent and internal standard is molten
Liquid obtains derivative to be measured after reaction, analyzes for LC-MS instrument；
B. steps are as follows for the sample pretreatment of reference state cyanide measurement and derivatization: accurately weighing appropriate amount of fluid sample or solid
Sample is added to the example reaction bottle of pretreatment and derivatization device, and a certain proportion of distilled water and anti-interference agent is added, right
In containing ethanol sample, appropriate sodium hydroxide solution alkaline hydrolysis 10min is first added, then opens acid-adding port, is rapidly added excessive acidification
Acid-adding port is closed after reagent immediately, the heating of example reaction bottle is reacted and distilled, and condensation communicating pipe connects cooling water, will be in distillate
The hydrogen cyanide of generation is transferred to derivative receiving bottle, and derivative receiving flask is previously added a certain amount of derivatization reagent and inner mark solution,
Distillate is collected in derivative receiving bottle and carries out derivative reaction, after reaction constant volume and derivative to be measured, for liquid matter
Combined instrument analysis；
(2) using LC-MS instrument to cyanide derivative products and internal standard13C15N- cyanide derivatives are tested and analyzed；
(3) cyanide standard curve is made using LC-MS/MS-MRM method, and uses internal standard method according to cyanide standard curve
To in sample free state and reference state cyanide content calculate.
2. the method for a kind of detection free state according to claim 1 and reference state micro cyanide, which is characterized in that step
Suddenly the acidizing reagent added in (1) a is lactic acid, reaction time 15-35min.
3. the method for a kind of detection free state according to claim 1 and reference state micro cyanide, which is characterized in that step
Suddenly the ratio of a certain proportion of distilled water of addition described in (1) b, sample and distilled water is 1:(5-10) g/mL, and soaking time is
0-5h；The acidizing reagent is tartaric acid, and additive amount is the 10-20% of sample quality；The anti-interference agent is acetic acid zinc solution,
The concentration of the acetic acid zinc solution is 100g/L, and additive amount is sample: zinc acetate 1:(1-3) g/mL；The alkaline hydrolysis hydroxide
Sodium solution is the sodium hydroxide solution of 0.5-1mL 2g/L, alkaline hydrolysis 10min.
4. the method for a kind of detection free state according to claim 1 and reference state micro cyanide, which is characterized in that step
Suddenly agent derived from (1) a and b is two acetal of 2,3- naphthalene (NDA) solution for including the dissolution of 2.71mmol/L methanol, 50mmol/L
Taurine solution, formaldehyde and ammonium hydroxide volume ratio be 25:25:45:5 mixed solution；The inner mark solution is that concentration is 1-
5mg/L's13C15N- cyanide standard solution.
5. pretreatment and the derivatization device of a kind of measurement of cyanide, it is characterised in that: the pretreatment of the cyanide measurement and
Derivatization device includes example reaction bottle and derivative receiving bottle, carries out acidolysis reaction in example reaction bottle and releases free state cyanogen
The free state cyanide released is imported into derivative receiving bottle by conduit by compound, and derivating agent is added in derivative receiving bottle
For receiving the free state cyanide generated in reaction flask and performing the derivatization；
The pretreatment of the described cyanide measurement and derivatization device specifically include example reaction bottle (1), derivative receiving bottle (2),
Example reaction bottle long duct (3), example reaction bottle tubulature (4), derivative receiving bottle long duct (5), atmospheric communicating pipe (6), cold
Solidifying communicating pipe (7), controllable switch (8), acid-adding port (9) bottle stopper (10), cooling water outlet (11) and cooling water inlet (12)；It is described
Example reaction bottle long duct (3) one end is extend into below example reaction bottle bottom of bottle liquid level, and one end is connect with controllable switch (8)；Institute
Derivative receiving bottle long duct (5) one end is stated to extend into below derivative receiving bottle bottom of bottle liquid level；It stretches described atmospheric communicating pipe (6) one end
Enter into derivative receiving bottle, one end is exposed to outside bottle；The condensation communicating pipe (7) is by example reaction bottle tubulature (4) and spreads out
Raw receiving bottle long duct (5) connection example reaction bottle and derivative receiving bottle, imported into derivative for the hydrogen cyanide that reaction flask generates and connect
Receive bottle；The controllable switch (8) connects logical nitrogen.
6. a kind of free state and the preprocess method of reference state micro cyanide measurement, which is characterized in that steps are as follows:
A. steps are as follows for the sample pretreatment of free state cyanide measurement and derivatization: accurately weighing appropriate amount of fluid sample or solid
Sample leaching liquor is added to the example reaction bottle of pretreatment and derivatization device, opens acid-adding port, is rapidly added appropriate acidification examination
Acid-adding port is closed after agent immediately to be reacted, controllable switch is opened and is passed through nitrogen, in reaction process under conditions of being passed through nitrogen,
The hydrogen cyanide of generation is transferred to derivative receiving bottle, derivative receiving flask is previously added a certain amount of derivatization reagent and internal standard is molten
Liquid obtains derivative to be measured after reaction, is used for liquid phase or gas chromatographic analysis；
B. steps are as follows for the sample pretreatment of reference state cyanide measurement and derivatization: accurately weighing appropriate amount of fluid sample or solid
Sample is added to the example reaction bottle of pretreatment and derivatization device, and a certain proportion of distilled water and anti-interference agent is added, right
In containing ethanol sample, appropriate sodium hydroxide solution alkaline hydrolysis 10min is first added, then opens acid-adding port, is rapidly added excessive acidification
Acid-adding port is closed after reagent immediately, the heating of example reaction bottle is reacted and distilled, and condensation communicating pipe connects cooling water, will be in distillate
The hydrogen cyanide of generation is transferred to derivative receiving bottle, and derivative receiving flask is previously added a certain amount of derivatization reagent and inner mark solution,
Distillate is collected in derivative receiving bottle and carries out derivative reaction, after reaction constant volume and derivative to be measured, for liquid phase
Or gas chromatographic analysis.
7. a kind of free state and the preprocess method of reference state micro cyanide measurement, feature exist according to claim 6
In the pretreatment of the free state and reference state cyanide carries out in pretreatment and derivatization device, described to pre-process and spread out
Biochemical device includes example reaction bottle and derivative receiving bottle, carries out acidolysis reaction in example reaction bottle and releases free state cyaniding
The free state cyanide released is imported into derivative receiving bottle by conduit by object, in the derivative receiving bottle that derivating agent is added
It receives the free state cyanide generated in reaction flask and performs the derivatization.
8. a kind of free state according to claim 6 and the preprocess method of reference state micro cyanide measurement, feature
It is,
The ratio of a certain proportion of distilled water of addition described in step b, sample and distilled water is 1:(5-10) g/mL, soaking time
For 0-5h；The acidizing reagent is tartaric acid, and additive amount is the 10-20% of sample quality；The anti-interference agent is 100g/L's
Acetic acid zinc solution, additive amount are sample: zinc acetate is equal to 1:(1-3) g/mL；
The acidizing reagent added in step a is lactic acid, reaction time 15-35min；
Agent derived from step a and b be include 2.71mmol/L methanol dissolution two acetal of 2,3- naphthalene (NDA) solution,
Taurine solution, formaldehyde and the ammonium hydroxide volume ratio of 50mmol/L is the mixed solution of 25:25:45:5；The inner mark solution is
Concentration is 1-5mg/L's13C15N- cyanide standard solution.
9. the method for a kind of detection free state according to claim 1 to 4 and reference state micro cyanide is yellow in detection
Application in wine and its raw material cyanide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611075836.3A CN106770729B (en) | 2016-11-30 | 2016-11-30 | A kind of method for detecting free state and bound state trace cyanide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611075836.3A CN106770729B (en) | 2016-11-30 | 2016-11-30 | A kind of method for detecting free state and bound state trace cyanide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106770729A CN106770729A (en) | 2017-05-31 |
| CN106770729B true CN106770729B (en) | 2019-06-21 |
Family
ID=58900916
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611075836.3A Active CN106770729B (en) | 2016-11-30 | 2016-11-30 | A kind of method for detecting free state and bound state trace cyanide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106770729B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107991429B (en) * | 2017-11-29 | 2020-04-24 | 长春黄金研究院 | Method for determining cyanide easily released in cyanide-containing wastewater containing sulfide |
| CN108303492B (en) * | 2018-01-21 | 2020-06-26 | 长春黄金研究院有限公司 | Method for determining cyanide easily released in cyanide-containing solid waste containing sulfide |
| CN108490061B (en) * | 2018-03-19 | 2020-07-28 | 广东华晟安全职业评价有限公司 | A kind of detection method for detecting cyanide content |
| CN112881548A (en) * | 2021-01-12 | 2021-06-01 | 济南趵突泉酿酒有限责任公司 | Method for rapidly detecting cyanide in sorghum |
| CN115166106A (en) * | 2021-11-26 | 2022-10-11 | 国家食品安全风险评估中心 | Method for detecting cyanide content in white spirit or white spirit fermentation process sample |
| CN115127889A (en) * | 2022-06-28 | 2022-09-30 | 贵州茅台酒股份有限公司 | Method for rapidly detecting nitrogen-containing compounds in white spirit |
| CN115420838B (en) * | 2022-08-25 | 2023-07-07 | 长沙晨辰医药科技有限公司 | Cyanide derivatization detection method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003098187A2 (en) * | 2002-05-15 | 2003-11-27 | Diversa Corporation | Assays and kits for detecting the presence of nitriles and/or cyanide |
| CN105699574A (en) * | 2016-04-05 | 2016-06-22 | 秦皇岛出入境检验检疫局检验检疫技术中心 | Method for detecting content of cyanide in milk and milk powder |
| CN206248619U (en) * | 2016-11-30 | 2017-06-13 | 江南大学 | Pretreatment and derivatization device that a kind of cyanide is determined |
-
2016
- 2016-11-30 CN CN201611075836.3A patent/CN106770729B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003098187A2 (en) * | 2002-05-15 | 2003-11-27 | Diversa Corporation | Assays and kits for detecting the presence of nitriles and/or cyanide |
| CN105699574A (en) * | 2016-04-05 | 2016-06-22 | 秦皇岛出入境检验检疫局检验检疫技术中心 | Method for detecting content of cyanide in milk and milk powder |
| CN206248619U (en) * | 2016-11-30 | 2017-06-13 | 江南大学 | Pretreatment and derivatization device that a kind of cyanide is determined |
Non-Patent Citations (1)
| Title |
|---|
| Determination of Blood Cyanide by HPLC-MS;A. Tracqui等;《Journal of Analytical Toxicology》;20020430;第26卷;第145页 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106770729A (en) | 2017-05-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106770729B (en) | A kind of method for detecting free state and bound state trace cyanide | |
| Smith et al. | New colorimetric reagent specific for cooper | |
| Xie et al. | Reverse flow injection spectrophotometric determination of iodate and iodide in table salt | |
| WO2009139656A1 (en) | The alcohol thermal dehydratation chamber, apparatus and method for determination of isotopic composition of non-exchangeable hydrogen and deuterium atoms in ethanol samples | |
| CN107255685A (en) | The high performance liquid chromatography of ultraviolet absorber in a kind of detection cosmetics | |
| CN103196849A (en) | Detection method of lead content in triacetin | |
| Varlet et al. | Innovative method for carbon dioxide determination in human postmortem cardiac gas samples using headspace-gas chromatography–mass spectrometry and stable labeled isotope as internal standard | |
| Chen et al. | Dynamic ultrasound-assisted extraction coupled on-line with solid support derivatization and high-performance liquid chromatography for the determination of formaldehyde in textiles | |
| Ribeiro et al. | Flow-injection spectrophotometric determination of methyldopa in pharmaceutical formulations | |
| CN110609100A (en) | Method for simultaneously detecting 6 organic acids in compound acidifier by gas chromatography | |
| Xie et al. | An efficient method for determining the acid value in edible oils by solvent-assisted headspace gas chromatography | |
| CN109001366A (en) | System and method for simultaneous determination of trivalent chromium and hexavalent chromium by ion chromatography-post-column derivatization | |
| CN103134870B (en) | Method for measuring sorbic acid and benzoic acid in foodstuff | |
| Cravey et al. | Current status of blood alcohol methods | |
| CN104198417B (en) | Method for testing heavy metal arsenic in compound food additive | |
| CN108593832A (en) | LC-MS-MS methods that are a kind of while measuring six kinds of mycotoxins in corn | |
| CN110954632A (en) | Method for determining content of 1,2-hexanediol in cosmetics | |
| CN104764850B (en) | A method for rapid quantification of urea in liquor by gas chromatography-mass spectrometry | |
| McLeod et al. | Rapid microstill determination of free and total sulfite in wine with conductimetric detection | |
| Li et al. | Direct automatic determination of the methanol content in red wines based on the temperature effect of the KMnO 4/K 2 S 2 O 5/fuchsin sodium sulfite reaction system | |
| Hu et al. | Rapid determination of formaldehyde in sanitary paper napkins for product quality control by headspace gas chromatography | |
| CN206248619U (en) | Pretreatment and derivatization device that a kind of cyanide is determined | |
| Cardeal et al. | New calibration method for gas chromatographic assay of carbon monoxide in blood | |
| Zhao-Fu et al. | Flow injection-chemiluminescence method for determination of hydrocortisone in human serum by using trivalent silver complex | |
| CN110954621A (en) | Method for determining hexavalent chromium in chromium yeast |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |