CN105911206A - Method for measuring amino acid in mainstream cigarette smoke - Google Patents
Method for measuring amino acid in mainstream cigarette smoke Download PDFInfo
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- CN105911206A CN105911206A CN201610419938.6A CN201610419938A CN105911206A CN 105911206 A CN105911206 A CN 105911206A CN 201610419938 A CN201610419938 A CN 201610419938A CN 105911206 A CN105911206 A CN 105911206A
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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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
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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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
- G01N2030/8818—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials involving amino acids
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Abstract
The invention discloses a method for measuring amino acid in mainstream cigarette smoke. The method includes the steps that amino acid in the mainstream cigarette smoke is captured by means of a Cambridge filter; after a cigarette is smoked, the Cambridge filter collecting a mainstream cigarette smoke sample is taken out, after an internal standard is added, ultrasonic extraction is conducted by means of a diluted hydrochloric acid solution, a filtering film enters an efficient liquid chromatogram-tandem mass spectrometer for measuring, and quantitation is conducted through the internal standard method. The method is characterized in that quantitation avoids influences of the matrix effect in the analysis process by means of the isotopic dilution internal standard method; direct sample injection is adopted so that the content of 17 amino acids in the mainstream cigarette smoke can be quickly and accurately detected, repeatability and the recovery rate are good, and the method is suitable for analysis of a large quantity of samples in industry.
Description
Technical field
The invention belongs to physical and chemical inspection technical field, be specifically related to the mensuration side of ammonia of main stream smoke of cigarette base acid content
Method.
Background technology
Aminoacid is to constitute the most basic material of organism protein, is one of indispensable composition in organism.It
Not only there is different physiological roles, be alternatively arranged as flavor agent and nutrient source and be widely used in food additive.Aminoacid is
A class important compound in Nicotiana tabacum L., at Nicotiana tabacum L. modulation, alcoholization, fermentation processing, baking silk until in combustion process, free amino acid
And there is enzyme catalysis and the browning reaction of non-enzymatic catalysis between reducing sugar (or carbonyl compound), generate multiple have roasting fragrant or quick-fried
The heterocyclic compounds such as the pyrans of popped rice fragrance characteristic, pyrazine, pyrroles, pyridines, some aminoacid such as phenylpropyl alcohol aminoacid also can be certainly
Body resolves into perfume compound, such as benzyl alcohol, phenethanol etc..
Aminoacid in cigarette smoke is to affect one of cigarette product key factor inhaling taste, and the aminoacid in Nicotiana tabacum L. also may be used
Process can be burnt and sucked by Medicated cigarette and enter human body, meet the metabolic needs of body.But with aminoacid in cigarette smoke it is at present
The detection method of object of study rarely has report.
For promote cigarette smoking quality, set up a kind of be suitable for ammonia of main stream smoke of cigarette base acidity test, analyze speed
Hurry up, repeatability and the response rate is good and the amino acid whose assay method of the analysis that is applicable to batch samples is significant.
Summary of the invention
The present invention compensate for the deficiency of existing analytical technology, it is provided that one is particularly suitable for ammonia of main stream smoke of cigarette base
The assay method of acid content.
The invention discloses the assay method of a kind of ammonia of main stream smoke of cigarette base acid, the method includes: use cambridge filter
Aminoacid in trapping cigarette mainstream flue gas；After cigarette smoking, the cambridge filter having collected cigarette mainstream flue gas sample is taken out,
After adding internal standard, use dilute hydrochloric acid solution to carry out ultrasonic extraction, enter high performance liquid chromatography-tandem mass instrument after filter membrane and carry out
Measure, inner mark method ration.
The assay method optimal technical scheme of a kind of ammonia of main stream smoke of cigarette base acid content that the present invention provides is:
1. the preparation of extract: with dilute HCl solution of concentrated hydrochloric acid preparation 0.01-0.5mol/L, as extract；
2. inner mark solution is prepared: weigh a certain amount of leucine-d3, it is 0.01-0.5 μm ol/mL with extract compound concentration
Inner mark solution；
3. prepare mixed standard solution: be solvent with extract, 17 kinds of aminoacid are mixed standard configuration and becomes different series concentration
Standard solution, proline, alanine, isoleucine, leucine, arginine, histidine, methionine, phenylalanine, serine,
Threonine, tyrosine, valine, aspartic acid, glutamic acid, lysine, the concentration of glycine are 0.01-0.1 μm ol/mL, Guang
The concentration of propylhomoserin is 0.01-0.1 μm ol/mL；
4. the suction of Medicated cigarette: the standard bar that the cigarette sample smoking machine to be measured balanced is specified according to GB/T19609
Part aspirates, and suction 10-30 props up Medicated cigarette, with 17 kinds of aminoacid in cambridge filter trapping cigarette mainstream flue gas；
5. sample pre-treatments and analysis: after cigarette smoking, takes off cambridge filter and puts in triangular flask, being sequentially added into internal standard
Solution and extract, ultrasonic extraction, take the supernatant after aqueous phase membrane filtration, measure for LC-MS/MS；
6. the drafting of standard curve: take mixed standard solution 0.02ml, 0.05ml, 0.1mL, 0.25mL, 0.5mL,
1.0mL, 5.0mL, each addition inner mark solution 100 μ L, it is settled to 50mL with extract, shakes up, obtain 6 level work standard solution；Right
Above-mentioned series standard solution carries out LC-MS/MS analysis, with object with interior target peak area ratio to the two corresponding concentration ratio
Value carries out linear regression analysis, tries to achieve 17 kinds of amino acid whose standard curves；
7. data process with detection: carried out inner mark method ration by the ratio of component peaks area with internal standard peak area.
The preferred technical scheme of assay method of a kind of ammonia of main stream smoke of cigarette base acid content that the present invention provides is:
1. the preparation of extract: with dilute HCl solution of concentrated hydrochloric acid preparation 0.05mol/L, as extract；
2. inner mark solution is prepared: weigh a certain amount of leucine-d3, by the internal standard that extract compound concentration is 0.1 μm ol/mL
Solution；
3. prepare mixed standard solution: be solvent with extract, 17 kinds of aminoacid are mixed standard configuration and becomes different series concentration
Standard solution, proline, alanine, isoleucine, leucine, arginine, histidine, methionine, phenylalanine, serine,
Threonine, tyrosine, valine, aspartic acid, glutamic acid, lysine, the concentration of glycine are 0.025 μm ol/mL, cystine
Concentration be 0.0125 μm ol/mL；
4. the suction of Medicated cigarette: the standard bar that the cigarette sample smoking machine to be measured balanced is specified according to GB/T19609
Part aspirates, and aspirates 20 Medicated cigarette, with 17 kinds of aminoacid in cambridge filter trapping cigarette mainstream flue gas；
5. sample pre-treatments and analysis: after cigarette smoking, takes off cambridge filter and puts in triangular flask, being sequentially added into internal standard
Solution and extract, ultrasonic extraction, take the supernatant after aqueous phase membrane filtration, measure for LC-MS/MS；
6. the drafting of standard curve: take mixed standard solution 0.02ml, 0.05ml, 0.1mL, 0.25mL, 0.5mL,
1.0mL, 5.0mL, each addition inner mark solution 100 μ L, it is settled to 50mL with extract, shakes up, obtain 6 level work standard solution；Right
Above-mentioned series standard solution carries out LC-MS/MS analysis, with object with interior target peak area ratio to the two corresponding concentration ratio
Value carries out linear regression analysis, tries to achieve 17 kinds of amino acid whose standard curves；
7. data process with detection: carried out inner mark method ration by the ratio of component peaks area with internal standard peak area.
Further, step 5. in, the addition of inner mark solution and extract is respectively 50-150 μ L and 30-100mL；
The preferably addition of inner mark solution and extract is respectively 100 μ L and 50mL.
Further, step 5. in, the time of ultrasonic extraction is 20-60min, and power is 200-500W；The most ultrasonic
The time of extraction is 30min, and power is 300W.
Further, step 5. in, aqueous phase filter membrane is 0.22-1.2 μm cellulose acetate sheets；Preferably aqueous phase filter membrane is
0.22 μm cellulose acetate sheets.
Further, step 5. and 6. in, high-efficient liquid phase chromatogram condition is:
Chromatographic column: Agilent ZORBAX Eclipse AAA chromatographic column, column parameter is 150mm × 4.6mm × 5 μm；
Sample size 5 μ L；
Column temperature is 30 DEG C；
Flow velocity is 0.4mL/min, and gradient elution program is:
Further, step 5. and 6. in, Mass Spectrometry Conditions is: Mass Spectrometer Method uses ESI ion source, scans at cation
Under mode, selecting MRM mode of operation to carry out second mass analysis, the main Mass Spectrometry Conditions after optimization is: electron spray voltage
5000V；Atomization gas 50psi, auxiliary adds steam 50psi, gas curtain gas 10psi, baking temperature 600 DEG C；The feature of each compound from
Son and parameter are respectively as follows:
The present invention, relative to prior art, achieves the most progressive:
The instrument and equipment that the present invention uses is laboratory conventional analysis equipment, and this makes the detection side that the present invention provides
Method testing cost is low, it is easier to promote；
The present invention use Isotope internal standard dilution standard measure avoid COMPLEX MIXED objects system Quantitative Separation, purification tired
Difficulty, preferably overcomes the impact of matrix effect, and testing result accuracy is high；
The present invention has filled up and has used high performance liquid chromatography-tandem mass combined instrument to measure the acid of ammonia of main stream smoke of cigarette base
The blank of detection method, takes the lead in establishing employing high performance liquid chromatography-tandem mass combined instrument and measures in cigarette mainstream flue gas 17
Plant amino acid whose analysis method；It is fast that the detection method that the present invention provides analyzes speed, and repeatability and the response rate are good, are particularly well-suited to
The detection analysis of batch samples in industry.
Accompanying drawing explanation
Fig. 1 is the extraction ion flow graph of glycine, alanine, serine, proline；
Fig. 2 is the extraction ion flow graph of valine, threonine, aspartic acid, leucine, isoleucine；
Fig. 3 is the extraction ion flow graph of internal standard, lysine, glutamic acid, methionine.
Fig. 4 is the extraction ion flow graph of histidine, phenylalanine, arginine, tyrosine, cystine.
Detailed description of the invention
For ease of being better understood from the present invention, the present invention further illustrates in conjunction with specific embodiments:
Embodiment 1
Instrument and reagent, material: American AB SCIEX 5500LC-MS/MS；U.S. Agilent 1290HPLC；
Agilent ZORBAX Eclipse AAA chromatographic column (150mm × 4.6mm, particle diameter 5 μm), 0.22 μm micropore filtering film, ultrasonic
Washer；Ultra-pure water (18M Ω), acetonitrile (chromatographically pure, lark prestige chemical reagents corporation), 17 kinds of mixed marks of aminoacid.
The preparation of extract: with dilute HCl solution of concentrated hydrochloric acid preparation 0.05mol/L, as extract；
Preparation inner mark solution: weigh a certain amount of leucine-d3, molten by the internal standard that extract compound concentration is 0.1 μm ol/mL
Liquid；
Preparation mixed standard solution: be solvent with extract, 17 kinds of aminoacid mixed standard configuration and becomes the mark of different series concentration
Quasi-solution, proline, alanine, isoleucine, leucine, arginine, histidine, methionine, phenylalanine, serine, Soviet Union
Propylhomoserin, tyrosine, valine, aspartic acid, glutamic acid, lysine, the concentration of glycine are 0.025 μm ol/mL, cystine
Concentration is 0.0125 μm ol/mL；
The suction of Medicated cigarette: the standard conditions that the cigarette sample smoking machine to be measured balanced is specified according to GB/T19609
Aspirate, aspirate 20 Medicated cigarette, with 17 kinds of aminoacid in cambridge filter trapping cigarette mainstream flue gas；
Sample pre-treatments: after cigarette smoking, takes off cambridge filter and puts in triangular flask, is sequentially added into 100 μ L internal standards molten
Liquid and 50mL extract, ultrasonic (power is 300W) extraction 30min, take the supernatant and filter through 0.22 μm cellulose acetate sheets
After, measure for LC-MS/MS；
High-efficient liquid phase chromatogram condition:
Chromatographic column: Agilent ZORBAX Eclipse AAA chromatographic column, column parameter is 150mm × 4.6mm × 5 μm；
Sample size 5 μ L；
Column temperature is 30 DEG C；
Flow velocity is 0.4mL/min, and gradient elution program is as shown in table 1:
Table 1 Gradient Elution condition
Mass Spectrometry Conditions: Mass Spectrometer Method uses ESI ion source, under cation scan mode, selects MRM mode of operation to carry out
Second mass analysis, the main Mass Spectrometry Conditions after optimization is: electron spray voltage 5000V；Atomization gas 50psi, auxiliary adds steam
50psi, gas curtain gas 20psi, baking temperature 600 DEG C；Characteristic ion and the parameter of each compound are as shown in table 2:
2 17 kinds of aminoacid of table and interior target characteristic ion and parameter
The drafting of standard curve: take mixed standard solution 0.02ml, 0.05ml, 0.1mL, 0.25mL, 0.5mL, 1.0mL,
5.0mL, each addition inner mark solution 100 μ L, be settled to 50mL with extract, shake up, obtain 6 level work standard solution；To above-mentioned system
Row standard solution carries out LC-MS/MS analysis, carries out the two corresponding concentration proportion with interior target peak area ratio with object
Linear regression analysis, tries to achieve 17 kinds of amino acid whose standard curves；17 kinds of amino acid whose equations of linear regression, correlation coefficient, detections
Limit and quantitative limit are as shown in table 3.
3 17 kinds of amino acid whose equations of linear regression of table, correlation coefficient, detection limit and quantitative limit
Precision: in order to investigate the precision of method, carries out 5 times with certain cigarette sample and in a few days repeats experiment, measurement result
Show that the relative standard deviation of 17 kinds of aminoacid in a few days measurement results, between 0.93%～7.29%, shows the in a few days essence of method
Density is good.Meanwhile, having carried out sample 5 days repeating experiment in the daytime, measurement result shows 17 kinds of aminoacid measurement results in the daytime
Relative standard deviation between 2.33%～9.64%, result shows that the repeatability in the daytime of the method is good.
The response rate: use sample mark-on method that on high, medium and low three contents levels, certain cigarette sample has been carried out the response rate
Mensuration, result shows, the result of 17 kinds of amino acid whose average recovery rates is between 89.1%-103.8%, and the response rate is higher,
Meet and analyze requirement.
The mensuration of actual sample: the method using the present invention to set up determines the amino in 6 cigarette sample main flumes
Acid (table 3), the content range of ammonia of main stream smoke of cigarette base acid is that 1.2-1539.7ng/ props up.
Amino acid whose testing result (ng/ props up) in table 3 cigarette sample main flume
As can be seen from the above experimental data, the present invention uses Isotope internal standard dilution standard measure to avoid complex mixture
System Quantitative Separation, the difficulty of purification, preferably overcome the impact of matrix effect, testing result precision is high, reproducible,
The response rate is high, and it is fast to analyze speed, is particularly well-suited to the detection analysis of batch samples in industry.The method pair that the present invention provides
In promoting, cigarette smoking quality is significant.
Claims (12)
1. the assay method of an ammonia of main stream smoke of cigarette base acid, it is characterised in that: the method comprises the following steps:
With the aminoacid in cambridge filter trapping cigarette mainstream flue gas；After cigarette smoking, cigarette mainstream flue gas sample will be collected
Cambridge filter take out, add after internal standard, use dilute hydrochloric acid solution to carry out ultrasonic extraction, after filter membrane, enter high-efficient liquid phase color
Spectrum-tandem mass spectrometer is measured, inner mark method ration.
The assay method of a kind of ammonia of main stream smoke of cigarette base the most according to claim 1 acid, it is characterised in that the method
Comprise the following steps:
1. the preparation of extract: with dilute HCl solution of concentrated hydrochloric acid preparation 0.01-0.5mol/L, as extract；
2. inner mark solution is prepared: weigh a certain amount of leucine-d3, by the internal standard that extract compound concentration is 0.01-0.5 μm ol/mL
Solution；
3. prepare mixed standard solution: be solvent with extract, 17 kinds of aminoacid are mixed standard configuration and becomes the standard of different series concentration
Solution, proline, alanine, isoleucine, leucine, arginine, histidine, methionine, phenylalanine, serine, Soviet Union's ammonia
Acid, tyrosine, valine, aspartic acid, glutamic acid, lysine, the concentration of glycine are 0.01-0.1 μm ol/mL, cystine
Concentration be 0.01-0.1 μm ol/mL；
4. the suction of Medicated cigarette: the standard conditions that the cigarette sample smoking machine to be measured balanced specifies according to GB/T19609 are entered
Row suction, suction 10-30 props up Medicated cigarette, with 17 kinds of aminoacid in cambridge filter trapping cigarette mainstream flue gas；
5. sample pre-treatments and analysis: after cigarette smoking, takes off cambridge filter and puts in triangular flask, being sequentially added into inner mark solution
And extract, ultrasonic extraction, take the supernatant after aqueous phase membrane filtration, measure for LC-MS/MS；
6. the drafting of standard curve: take mixed standard solution 0.02ml, 0.05ml, 0.1mL, 0.25mL, 0.5mL, 1.0mL,
5.0mL, respectively adds inner mark solution, uses extract constant volume, shakes up, and obtains 6 level work standard solution；Above-mentioned series standard solution is entered
Row LC-MS/MS analyzes, and with interior target peak area ratio, the two corresponding concentration proportion is carried out linear regression with object and divides
Analysis, tries to achieve 17 kinds of amino acid whose standard curves；
7. data process with detection: carried out inner mark method ration by the ratio of component peaks area with internal standard peak area.
The assay method of a kind of ammonia of main stream smoke of cigarette base the most according to claim 1 acid, it is characterised in that the method
Comprise the following steps:
1. the preparation of extract: with dilute HCl solution of concentrated hydrochloric acid preparation 0.05mol/L, as extract；
2. inner mark solution is prepared: weigh a certain amount of leucine-d3, with the inner mark solution that extract compound concentration is 0.1 μm ol/mL；
3. prepare mixed standard solution: be solvent with extract, 17 kinds of aminoacid are mixed standard configuration and becomes the standard of different series concentration
Solution, proline, alanine, isoleucine, leucine, arginine, histidine, methionine, phenylalanine, serine, Soviet Union's ammonia
Acid, tyrosine, valine, aspartic acid, glutamic acid, lysine, the concentration of glycine are 0.025 μm ol/mL, cystine dense
Degree is 0.0125 μm ol/mL；
4. the suction of Medicated cigarette: the standard conditions that the cigarette sample smoking machine to be measured balanced specifies according to GB/T19609 are entered
Row suction, aspirates 20 Medicated cigarette, with 17 kinds of aminoacid in cambridge filter trapping cigarette mainstream flue gas；
5. sample pre-treatments and analysis: after cigarette smoking, takes off cambridge filter and puts in triangular flask, being sequentially added into inner mark solution
And extract, ultrasonic extraction, take the supernatant after aqueous phase membrane filtration, measure for LC-MS/MS；
6. the drafting of standard curve: take mixed standard solution 0.02ml, 0.05ml, 0.1mL, 0.25mL, 0.5mL, 1.0mL,
5.0mL, each addition inner mark solution 100 μ L, be settled to 50mL with extract, shake up, obtain 6 level work standard solution；To above-mentioned system
Row standard solution carries out LC-MS/MS analysis, carries out the two corresponding concentration proportion with interior target peak area ratio with object
Linear regression analysis, tries to achieve 17 kinds of amino acid whose standard curves；
7. data process with detection: carried out inner mark method ration by the ratio of component peaks area with internal standard peak area.
The assay method of ammonia of main stream smoke of cigarette base the most according to claim 3 acid, it is characterised in that: in step 5.
In, inner mark solution addition is 50-150 μ L, and extract addition is 30-100mL.
The assay method of ammonia of main stream smoke of cigarette base the most according to claim 4 acid, it is characterised in that: in step 5.
In, inner mark solution addition is 100 μ L, and extract addition is 50mL.
The assay method of ammonia of main stream smoke of cigarette base the most according to claim 3 acid, it is characterised in that: in step 5.
In, the time of ultrasonic extraction is 20-60min, and power is 200-500W.
The assay method of ammonia of main stream smoke of cigarette base the most according to claim 6 acid, it is characterised in that: in step 5.
In, the time of ultrasonic extraction is 30min, and power is 300W.
The assay method of ammonia of main stream smoke of cigarette base the most according to claim 3 acid, it is characterised in that: in step 5.
In, aqueous phase filter membrane is 0.22-1.2 μm cellulose acetate sheets.
The assay method of ammonia of main stream smoke of cigarette base the most according to claim 8 acid, it is characterised in that: in step 5.
In, aqueous phase filter membrane is 0.22 μm cellulose acetate sheets.
10. according to the assay method of described ammonia of main stream smoke of cigarette base acid arbitrary in claim 1-9, it is characterised in that:
Step 5. and 6. in, high-efficient liquid phase chromatogram condition is:
Chromatographic column: Agilent ZORBAX Eclipse AAA chromatographic column, column parameter is 150mm × 4.6mm × 5 μm；
Sample size 5 μ L；
Column temperature is 30 DEG C；
Flow velocity is 0.4mL/min, and gradient elution program is:
The assay method of 11. ammonia of main stream smoke of cigarette base according to claim 10 acid, it is characterised in that: in step 5.
6. in, Mass Spectrometry Conditions is: Mass Spectrometer Method uses ESI ion source, under cation scan mode, selects MRM mode of operation to enter
Row second mass analysis, the main Mass Spectrometry Conditions after optimization is: electron spray voltage 5000V；Atomization gas 50psi, auxiliary adds steam
50psi, gas curtain gas 10psi, baking temperature 600 DEG C；Characteristic ion and the parameter of each compound are respectively as follows:
The assay method of 12. ammonia of main stream smoke of cigarette base according to claim 11 acid, it is characterised in that:
1. the preparation of extract: with dilute HCl solution of concentrated hydrochloric acid preparation 0.05mol/L, as extract；
2. inner mark solution is prepared: weigh a certain amount of leucine-d3, with the inner mark solution that extract compound concentration is 0.1 μm ol/mL；
3. prepare mixed standard solution: be solvent with extract, 17 kinds of aminoacid are mixed standard configuration and becomes the standard of different series concentration
Solution, proline, alanine, isoleucine, leucine, arginine, histidine, methionine, phenylalanine, serine, Soviet Union's ammonia
Acid, tyrosine, valine, aspartic acid, glutamic acid, lysine, the concentration of glycine are 0.025 μm ol/mL, cystine dense
Degree is 0.0125 μm ol/mL；
4. the suction of Medicated cigarette: the standard conditions that the cigarette sample smoking machine to be measured balanced specifies according to GB/T19609 are entered
Row suction, aspirates 20 Medicated cigarette, with 17 kinds of aminoacid in cambridge filter trapping cigarette mainstream flue gas；
5. sample pre-treatments and analysis: after cigarette smoking, takes off cambridge filter and puts in triangular flask, being sequentially added into inner mark solution
And extract, ultrasonic extraction, take the supernatant after aqueous phase membrane filtration, measure for LC-MS/MS；
6. the drafting of standard curve: take mixed standard solution 0.02ml, 0.05ml, 0.1mL, 0.25mL, 0.5mL, 1.0mL,
5.0mL, each addition inner mark solution 100 μ L, be settled to 50mL with extract, shake up, obtain 6 level work standard solution；To above-mentioned system
Row standard solution carries out LC-MS/MS analysis, carries out the two corresponding concentration proportion with interior target peak area ratio with object
Linear regression analysis, tries to achieve 17 kinds of amino acid whose standard curves；
7. data process with detection: carried out inner mark method ration by the ratio of component peaks area with internal standard peak area；
Step 5. in, inner mark solution addition is 100 μ L, and extract addition is 50mL；
Step 5. in, the time of ultrasonic extraction is 30min, and power is 300W；
Step 5. in, aqueous phase filter membrane is 0.22 μm cellulose acetate sheets；
Step 5. and 6. in, high-efficient liquid phase chromatogram condition is:
Chromatographic column: Agilent ZORBAX Eclipse AAA chromatographic column, column parameter is 150mm × 4.6mm × 5 μm；
Sample size 5 μ L；
Column temperature is 30 DEG C；
Flow velocity is 0.4mL/min, and gradient elution program is:
Mass Spectrometry Conditions is: Mass Spectrometer Method uses ESI ion source, under cation scan mode, selects MRM mode of operation to carry out two
Level mass spectral analysis, the main Mass Spectrometry Conditions after optimization is: electron spray voltage 5000V；Atomization gas 50psi, auxiliary adds steam
50psi, gas curtain gas 10psi, baking temperature 600 DEG C；Characteristic ion and the parameter of each compound are respectively as follows:
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108709946A (en) * | 2018-06-13 | 2018-10-26 | 新疆维吾尔自治区分析测试研究院 | A kind of method of 17 kinds of amino acid in detection food |
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