CN101806682A - Preparation method of benzoic acid standard solution, method for determining standard curve of benzoic acid standard solution and method for detecting content of benzoic acid standard solution in cheese - Google Patents
Preparation method of benzoic acid standard solution, method for determining standard curve of benzoic acid standard solution and method for detecting content of benzoic acid standard solution in cheese Download PDFInfo
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Abstract
The invention provides a preparation method of a benzoic acid standard solution, a method for determining standard curve of the benzoic acid standard solution and a method for detecting content of the benzoic acid standard solution in cheese. The method for detecting the content of the benzoic acid standard solution in cheese comprises the steps of: adding sea sand and zinc acetate into the cheese for grinding, then adding 20mL of distilled water, uniformly mixing by using a vortex mixer and then centrifuging, filtering, taking 5mL of filtrate, regulating pH value of the filtrate by using 1mL of 6mol/L of hydrochloric acid, extracting by using dichloromethane, and determining the content of benzoic acid by using a gas chromatography-mass spectrometer (GC-MS/MS), wherein detection requirements can be met by using a capillary chromatographic column and temperature programming. The method for detecting the content of the benzoic acid in the cheese by using a gas chromatography-mass spectrometric method has short extraction time, simple and convenient operation, high detection accuracy and strong capacity of resisting disturbance, and can meet the requirement for large detection load in a laboratory.
Description
Technical field
The present invention relates to the technical field of the detection of chemical analysis, specifically, it relates to a kind of preparation method of benzoic acid standard solution, a kind of usefulness has a series of benzoic acid standard solution of different benzoic acid content and determines that the method for benzoic acid standard solution typical curve and the benzoic acid standard solution typical curve that a kind of basis is determined detect the method for benzoic acid content in the cheese.
Background technology
Benzoic acid (claiming benzoic acid again) is a kind of food preservative commonly used, identical with the antiseptic effect of its sodium salt, it has inhibiting effect to the activity of the respiratory enzyme system of multiple microbial cell, and growth has inhibition to the cells of microorganisms film, thereby reaches the breeding that suppresses microorganism.Because it can suppress the breeding of multiple microorganism, has the good sterilization effect again, is widely used in can, jam, soy sauce, vinegar, carbonated drink and all kinds of meat product.Benzoic acid is that various countries allow to use, and historical longer food preservative.
After benzoic acid enters human body, can react generation hippuric acid and glucosiduronate respectively with aminoacetic acid (glycocoll) in the human body and gluconic acid, the latter can excrete with urine, can not form harm to human body so absorb a small amount of benzoic acid every day.But during the benzoic acid of excess intake, but can work the mischief to the human liver.Because benzoic abstinence reaction is carried out in liver, even being absorbed a small amount of benzoic acid, the weak people of liver function also can aggravate disease, and benzoic acid also has certain pungency to skin, eyes and mucous membrane, and Sodium Benzoate can cause the enteron aisle discomfort.Regulation human body para Toluic Acid's intake every day is 5-10mg/kg after FAO (Food and Agriculture Organization of the United Nation) (FAO) and the The World Health Organization's experimental study, and China has also made regulation to benzoic maximum use amount in the varieties of food items.
Measure benzoic content with liquid phase chromatography in the national standard " mensuration of benzoic acid and sorbic acid in GB/T 21703-2008 breast and the dairy products ".The pre-treatment of this method perhaps takes by weighing solid sample 3g for taking by weighing fluid milk 20g earlier in the 150ml conical flask in the 150ml conical flask, add 10ml water and all dissolve.Add 25ml 0.1mol/L sodium hydroxide solution then, in 70 ℃ of water-baths, handled 15 minutes.After the cooling, add an amount of methyl alcohol and be transferred in the 100ml volumetric flask.With pH regulator to 8, add people 2ml 92g/L potassium ferrocyanide solution and 2ml 183g/l acetic acid zinc solution with the 0.5moll sulfuric acid solution then.Violent jolting was left standstill 15 minutes, mixed the back cool to room temperature, used methanol constant volume again, left standstill 15 minutes, and supernatant is membrane filtration after filtration.Collect filtrate as sample solution, be used for high performance liquid chromatograph and measure.
This method need be used and use methyl alcohol 45ml-65ml, can cause certain pollution to environment, and it is long to handle the sample time simultaneously, and operation is comparatively complicated.
This method is normally demarcated the working fluid with definite benzoic acid content earlier, and then uses the same method and handle and the test sample, calculates benzoic acid content in the sample according to the result of this twice measurement.But this method measuring accuracy is not high, because the numerical value of one-shot measurement may be subjected to the interference of some uncertain factors, therefore the sample benzoic acid content of determining according to the numerical value of this working fluid may be inaccurate.
In addition, at " GC-MS(gas chromatography-mass spectrography) is measured benzoic acid (sodium) content in the milk powder " (" chemistry in detecting ", 2008 the 18th the 1st phases of volume, 75-76, Gan Pingsheng, Huang Cong, in letter, disclose a kind of method Hou Jianrong), but this method only detects powdered milk sample, used qualitative ion, quota ion when not mentioning experiment with gas chromatography-mass spectrography detection benzoic acid content.
Summary of the invention
The preparation method who the purpose of this invention is to provide a kind of benzoic acid standard solution, this method is easy and simple to handle and have a degree of precision.
The present invention also provides a kind of definite method of benzoic acid standard solution typical curve, it can be equipped with the standard operation liquid with different benzoic acid content with the preparation method of above-mentioned benzoic acid standard solution, and adopts gas chromatography-mass spectrography accurately to determine the corresponding relation between benzoic acid content and the peak area.
The present invention provides a kind of method that detects benzoic acid content in breast and the cheese again, it adopts the benzoic acid in gas chromatography-mass spectrum/mass spectroscopy detection cheese sample, determine benzoic acid content in the cheese according to above-mentioned benzoic acid typical curve, and improve the antijamming capability when detecting.
For achieving the above object, the invention provides a kind of preparation method of benzoic acid standard solution, this method comprises the steps:
1) gets in the centrifuge tube of 1mL to 10-15ml of benzoic acid standard operation liquid of 20.0 μ g/ml;
2) in this centrifuge tube, add distilled water to 5ml, add 1mL hydrochloric acid solution (6mol/L) and 2ml methylene chloride again, mixed 1 minute, usefulness hydro-extractor under the 3000rpm-4000rpm condition centrifugal 5-10 minute with vortex mixer;
3) be filled into the sample introduction bottle with clear liquid under the absorption of 1ml syringe through filtering membrane, this is and makes benzoic acid standard solution to be measured.
The invention allows for and a kind ofly determine the method for benzoic acid standard solution typical curve according to above-mentioned benzoic acid standard solution preparation method, this method comprises:
1) prepares benzoic acid standard solution with different benzoic acid content;
2) demarcate above-mentioned steps 1 respectively with gas chromatography-mass spectrum/mass spectroscopy) the benzoic acid standard solution of variable concentrations of preparation;
3) determine the typical curve that benzoic acid content is corresponding with peak area.
According to of the present invention another determine the method for benzoic acid standard solution typical curve, wherein, the amount of benzoic acid standard solution is respectively: the benzoic acid mark liquid 0.1ml, 0.2ml, 0.5ml, 0.6ml, 0.8ml and the 1.0ml that get 20.0 μ g/ml.
Another kind according to the present invention is determined the method for benzoic acid standard solution typical curve, and wherein, the condition of gas chromatography is:
Chromatographic column: capillary chromatographic column, stationary phase are nitroso-terephthalic acid (TPA) modified poly (ethylene glycol) or suitable person, and the chromatographic column specification is 30m * 0.25mm * 0.2 μ m or 30m * 0.32mm * 0.2 μ m;
Injector temperature: 200-300 ℃;
Input mode: shunting, split ratio is 40: 1-50: 1;
Carrier gas: high-purity helium;
Flow velocity: the capillary chromatography column internal diameter is that the flow velocity of 0.25 μ m is 1.0-1.4ml/min, and the capillary chromatography column internal diameter is that the flow velocity of 0.32 μ m is 1.8-2.5ml/min;
Heating schedule is as follows: initial temperature is 140 ℃ to be kept 1-2 minute, be raised to 175-190 ℃ with 4-7 ℃/min then after, kept program 10-20.5 minute altogether 4-7 minute.
According to of the present invention another determine the method for benzoic acid standard solution typical curve, wherein, mass spectral condition is:
Ionization mode: electricity bombardment ionization (EI);
Ionizing energy: 70eV;
Ion source temperature: 200-220 ℃;
Transmission line temperature: 150-200 ℃;
Solvent delay time: 3.0-5.0 minute;
Sample size: 1-2 μ L;
Scan pattern: multiple-reaction monitoring (MRM), qualitative ion mass-to-charge ratio (m/z) 122＞105,122＞77, quota ion mass-to-charge ratio (m/z) 122＞105;
Collision energy: mass-to-charge ratio (m/z) 122＞105 is 8V, and 122＞77 is 20V.
The present invention has proposed the method that a kind of benzoic acid standard solution typical curve of determining with said method detects benzoic acid content in the cheese again, and this method comprises:
1) take by weighing 3-5g sea sand to the 50ml centrifuge tube, in centrifuge tube, take by weighing 1-3g cheese sample then,
Add the 1-2g zinc acetate, accurately add 20ml distilled water;
2) fully stir with glass bar, cheese evenly is blended in the extra large sand;
3) fully mix with vortex mixer;
4) with hydro-extractor under the 3000rpm-4000rpm condition centrifugal 5-10 minute;
5) filter water layer, draw 5ml filtrate in the centrifuge tube of a 10-15ml, add 6mol/L hydrochloric acid solution 1ml, the 2ml methylene chloride;
6) mixed 1 minute with vortex mixer, with hydro-extractor under the 3000rpm-4000rpm condition centrifugal 5-10 minute;
7) membrane filtration is to the sample introduction bottle after filtration with clear liquid under the absorption of 1ml syringe, and this is and makes benzoic acid sample solution to be measured;
8) with gas chromatography-mass spectrum/mass spectrometer described benzoic acid sample solution to be measured is measured;
9) measurement result and the determined benzoic acid content typical curve of step 8) gained are compared, determine the benzoic acid content of described benzoic acid sample solution to be measured;
10) determine benzoic acid content in the described cheese according to the benzoic acid content of described benzoic acid sample solution to be measured.
Gas chromatography-mass spectrography of the present invention detects the method for benzoic acid content in breast and the cheese, adopt the benzoic acid in gas chromatography-mass spectrum/mass spectrometric determination cheese, characteristics such as have that the organic solvent of consumption is few, the running time short, simple to operation, detection accuracy rate height, antijamming capability are strong.
Description of drawings
The following drawings only is intended to the present invention done and schematically illustrates and explain, not delimit the scope of the invention.Wherein,
Fig. 1 is when gas chromatography-mass spectrum/mass spectrometer is in the SCAN pattern, and benzoic acid content is the total ions chromatogram that the benzoic acid standard solution 1.0ml of 20.0 μ g/ml obtains;
Fig. 2 is when gas chromatography-mass spectrum/mass spectrometer is in the SCAN pattern, and benzoic acid content is the mass spectrogram that the benzoic acid standard solution 1.0ml of 20.0 μ g/ml obtains;
Fig. 3 is when gas chromatography-mass spectrum/mass spectrometer is in the MRM pattern, and benzoic acid content is the total ions chromatogram that the benzoic acid standard solution 1.0ml of 20.0 μ g/ml obtains;
Fig. 4 is when gas chromatography-mass spectrum/mass spectrometer is in the MRM pattern, and benzoic acid content is the mass spectrogram that the benzoic acid standard solution 1.0ml of 20.0 μ g/ml obtains;
Fig. 5 is a kind of benzoic acid standard solution canonical plotting when gas chromatography-mass spectrum/mass spectrometer is in the MRM pattern, horizontal ordinate is that benzoic quality contained in the benzoic acid standard solution of being drawn (is got the benzoic acid mark liquid 0.1ml of 20.0 μ g/ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml and 1.0ml), be respectively and contain benzoic acid 1.0 μ g in every milliliter of methylene chloride, 2.0 μ g, 4.0 μ g, 6.0 μ g, 8.0 μ g, 10.0 being respective quality, μ g, ordinate (promptly get the benzoic acid mark liquid 0.1ml of 20.0 μ g/ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml Dui Ying peak area and 1.0ml).
Embodiment
Understand for technical characterictic of the present invention, purpose and effect being had more clearly, now contrast description of drawings the specific embodiment of the present invention.Each the concrete numerical value that it is pointed out that among the present invention to be mentioned not is the numeral on the complete mathematical meaning, and every numerical value that meets on the chemical engineering meaning all is suitable for.
Experimental apparatus and reagent that each embodiment adopts are as follows:
450GC-300MS gas chromatography instrument: available from U.S.'s Varian (VARIAN) company;
Chromatographic column: capillary chromatographic column, stationary phase are nitroso-terephthalic acid (TPA) modified poly (ethylene glycol) or suitable person, and the chromatographic column specification is 30m * 0.25mm * 0.2 μ m or 30m * 0.32mm * 0.2 μ m;
Centrifuge tube: specification is 10-15ml and 50ml;
Benzoic acid standard solution: take from State Standard Matter Research Centre, concentration 1.0g/L, numbering: GBW (E) 100006;
Benzoic acid standard operation liquid: concentration is 20.0 μ g/ml.The preparation method gets the benzoic acid titer 1ml that concentration is 1.0g/L, uses the distilled water constant volume to 50ml;
Hydrochloric acid solution: concentration is 6mol/L.The preparation method gets the 50ml hydrochloric acid solution, uses the distilled water constant volume in the 100mL volumetric flask;
Sea sand: it is pure to belong to analysis;
Zinc acetate: it is pure to belong to analysis;
Methylene chloride: belong to chromatographically pure.
The instrument condition that each embodiment adopts is as follows:
GC conditions:
Injector temperature: 200-300 ℃;
Input mode: shunting, split ratio is 40: 1-50: 1;
Carrier gas: high-purity helium;
Flow velocity: the capillary chromatography column internal diameter is that the flow velocity of 0.25 μ m is 1.0-1.4ml/min, and the capillary chromatography column internal diameter is that the flow velocity of 0.32 μ m is 1.8-2.5ml/min;
Heating schedule is as follows: initial temperature is 140 ℃ to be kept 1-2 minute, be raised to 175-190 ℃ with 4-7 ℃/min then after, kept program 10-20.5 minute altogether 4-7 minute.
The mass spectrum condition:
Ionization mode: electricity bombardment ionization (EI);
Ionizing energy: 70eV;
Ion source temperature: 200-220 ℃;
Transmission line temperature: 150-200 ℃;
Solvent delay time: 3.0-5.0 minute;
Sample size: 1-2 μ L;
Scan pattern: multiple-reaction monitoring (MRM), qualitative ion mass-to-charge ratio (m/z) 122＞105,122＞77, quota ion mass-to-charge ratio (m/z) 122＞105;
Collision energy: mass-to-charge ratio (m/z) 122＞105 is 8V, and 122＞77 is 20V.
The preparation of embodiment 1 benzoic acid standard solution
Illustrate as benzoic acid content to be the preparation method of the benzoic acid standard solution of 10.0 μ g/ml.
Get earlier in 1ml to 15ml centrifuge tube of benzoic acid standard operation liquid of 20.0 μ g/ml, in this centrifuge tube, add water to 5ml, add 1mL hydrochloric acid solution (concentration 6mol/L) and 2ml methylene chloride again, mixed 1 minute, with hydro-extractor under the 3000rpm condition centrifugal 5 minutes with vortex mixer; Membrane filtration is to the sample introduction bottle after filtration with clear liquid under the absorption of 1ml syringe, and this is and makes benzoic acid standard solution to be measured.
Though top compound method of having described a kind of standard solution of benzoic acid content, but it will be appreciated by those skilled in the art that, when as long as preparation process begins, in some centrifuge tubes of same capacity, add the benzoic acid standard operation liquid of different amounts, so just can obtain a series of benzoic acid standard solution with different benzoic acid content.Here can different condition carry out centrifugal treating, be that 3500rpm and 4000rpm etc., centrifugation time are 8 minutes and 10 minutes as rotating speed.
In definite method of benzoic acid content typical curve of the present invention, can adopt said method earlier, preparing benzoic acid content respectively is the benzoic acid standard solution of 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml and 1.0ml.
Determining of embodiment 2 benzoic acid standard solution typical curves
Owing to use equipment provided by the present invention (as 300-MS gas chromatography instrument) to use above condition all can draw the typical curve of benzoic acid standard solution, so used condition is as follows in the present embodiment:
GC conditions:
Temperature is 200 ℃,
Input mode: shunting, split ratio is 40: 1;
Flow velocity: the capillary chromatography column internal diameter is that the flow velocity of 0.25 μ m is 1.0ml/min, and the capillary chromatography column internal diameter is that the flow velocity of 0.32 μ m is 1.8ml/min;
Heating schedule is as follows: initial temperature is 140 ℃ and keeps 1min, be raised to 175 ℃ with 4 ℃/min then after, keep 4min, program is 10min altogether;
Ion source temperature: 200 ℃;
Transmission line temperature: 150 ℃;
Mass spectral condition is:
Ionization mode: electricity bombardment ionization (EI);
Ionizing energy: 70eV;
Ion source temperature: 200 ℃;
Transmission line temperature: 150 ℃;
The solvent delay time: 3.0 minutes;
Sample size: 1 μ L;
Scan pattern: multiple-reaction monitoring (MRM), qualitative ion mass-to-charge ratio (m/z) 122＞105,122＞77, quota ion mass-to-charge ratio (m/z) 122＞105;
Collision energy: mass-to-charge ratio (m/z) 122＞105 is 8V, and 122＞77 is 20V.
Condition according to above-mentioned preparation benzoic acid standard solution, regulate instrument according to the instrument condition that adopts, treat in stable condition after, with gas chromatography-mass spectrography the benzoic acid standard solution of preparing with different benzoic acid content is analyzed respectively, determine the typical curve that benzoic acid content is corresponding with peak area according to the numerical value of being demarcated again, this is benzoic acid standard solution typical curve, wherein, the amount of benzoic acid standard solution is respectively: the benzoic acid mark liquid 0.1ml, 0.2ml, 0.5ml, 0.6ml, 0.8ml and the 1.0ml that get 20.0 μ g/ml.
Fig. 1 is when gas chromatography-mass spectrum/mass spectrometer is in the SCAN pattern, and benzoic acid content is to get the total ions chromatogram that the benzoic acid standard solution 1.0ml of 20.0 μ g/ml obtains; Fig. 2 is when gas chromatography-mass spectrum/mass spectrometer is in the SCAN pattern, and benzoic acid content is to get the mass spectrogram that the benzoic acid standard solution 1.0ml of 20.0 μ g/ml obtains; Fig. 3 is when gas chromatography-mass spectrum/mass spectrometer is in the MRM pattern, and benzoic acid content is to get the total ions chromatogram that the benzoic acid standard solution 1.0ml of 20.0 μ g/ml obtains; Fig. 4 is when gas chromatography-mass spectrum/mass spectrometer is in the MRM pattern, and benzoic acid content is to get the mass spectrogram that the benzoic acid standard solution 1.0ml of 20.0 μ g/ml obtains;
Fig. 5 is a kind of benzoic acid standard solution canonical plotting when gas chromatography-mass spectrum/mass spectrometer is in the MRM pattern.Horizontal ordinate is a quality (getting benzoic acid mark liquid 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml and the 1.0ml of 20.0 μ g/ml) contained in the benzoic acid standard solution of being drawn, be respectively and contain benzoic acid 1.0 μ g, 2.0 μ g, 4.0 μ g, 6.0 μ g, 8.0 μ g, 10.0 μ g in every milliliter of methylene chloride, ordinate is the corresponding peak area of respective quality (promptly getting benzoic acid mark liquid 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml and the 1.0ml of 20.0 μ g/ml).
The related data of the typical curve that draws is as follows:
The initial linear curve fitting, linear equation: y=+3.8717e+4x-3.5148e+4;
Deviation: 0.28%.
The benzoic acid content typical curve that utilizes said method of the present invention to determine can detect the benzoic acid content in the cheese easily.Below in conjunction with embodiment 3, describe the detection method of benzoic acid content in the cheese in detail.
The detection of benzoic acid content in embodiment 3 cheese
At first, take by weighing 3g sea sand to the 50ml centrifuge tube, in centrifuge tube, take by weighing 1g cheese sample, add the 1g zinc acetate, accurately add 20ml distilled water, fully stir, cheese evenly is blended in the extra large sand with glass bar, fully mix with vortex mixer, with hydro-extractor under the 3000rpm condition centrifugal 5 minutes.
Then, filter water layer, draw 5ml filtrate in the centrifuge tube of a 15ml, add 6mol/L hydrochloric acid solution 1ml, the 2ml methylene chloride mixed 1 minute with vortex mixer, with hydro-extractor under the 3000rpm condition centrifugal 5 minutes.
At last, membrane filtration is to the sample introduction bottle after filtration with clear liquid under the absorption of 1ml syringe, and this is and makes benzoic acid sample solution to be measured.Benzoic acid sample solution to be measured is injected into obtains peak area in gas chromatography-mass spectrum/mass spectrometer, from typical curve, check in benzoic content in the style (μ g).
Benzoic acid content X in the style with massfraction (mg/kg) expression, is calculated as follows:
In the formula:
Cs---from typical curve, check in benzoic content in the sample (μ g).
m
i---the quality (g) of sample.
In this embodiment, Cs is 0.324 μ g, m
iBe 1.0024g, calculate 1.36mg/kg by following formula.
With gas chromatograph-mass spectrometer (GCMS) above-mentioned liquid to be measured is measured, result that will measure and determined benzoic acid content typical curve are compared subsequently, determine the benzoic acid content of liquid to be measured.Like this, just can calculate the benzoic acid content in the cheese according to the benzoic acid content of liquid to be measured.
More than the gram number of the extra large sand of Xuan Zeing, zinc acetate can change according to the cheese sample of being got, such as when getting 3g cheese sample, extra large sand and zinc acetate are answered corresponding increase, as get extra large sand 5g and zinc acetate 2g.Hydro-extractor also can be selected different values in the condition that provides.These all it will be appreciated by those skilled in the art that and can realize, do not give unnecessary details at this.
As mentioned above, determine in the method that by preparing a series of benzoic acid standard solution with different benzoic acid content, the accuracy that can improve the benzoic acid content typical curve reduces the interference in measuring at benzoic acid content typical curve of the present invention.Equally in detecting cheese in the method for benzoic acid content, adopt the benzoic acid in gas chromatography-mass spectrum/mass spectrometric determination cheese, have characteristics such as consume that organic solvent is few, extraction time short, simple to operation, detection accuracy rate height, antijamming capability are strong, well satisfied the big needs of laboratory detection limit.
The above only is the schematic embodiment of the present invention, is not in order to limit scope of the present invention.Any those skilled in the art, equivalent variations, the modification of being done under the prerequisite that does not break away from design of the present invention and principle and combining all should belong to the scope of protection of the invention.
Claims (6)
1. benzoic acid standard solution preparation method is characterized in that this method comprises:
1) gets in the centrifuge tube of 1mL to 10ml-15ml of benzoic acid standard operation liquid of 20.0 μ g/ml;
2) in described centrifuge tube, add distilled water to 5ml, add 1mL hydrochloric acid solution (6mol/L) and 2ml methylene chloride again, mixed 1 minute, usefulness hydro-extractor under the 3000rpm-4000rpm condition centrifugal 5-10 minute with vortex mixer;
3) be filled into the sample introduction bottle with clear liquid under the absorption of 1ml syringe through filtering membrane, this is and makes benzoic acid standard solution to be measured.
2. determine to it is characterized in that the method for benzoic acid standard solution typical curve according to the described benzoic acid standard solution preparation of claim 1 method for one kind:
1) according to the described method of claim 1, the benzoic acid standard solution that preparation has different benzoic acid content;
2) demarcate above-mentioned steps 1 respectively with gas chromatography-mass spectrography) preparation described various benzoic acid standard solution;
3) determine the typical curve that benzoic acid content is corresponding with peak area, this typical curve is benzoic acid standard solution typical curve.
3. the method for definite benzoic acid standard solution typical curve as claimed in claim 2, wherein, the amount of described benzoic acid standard solution is respectively: the benzoic acid standard solution 0.1ml, 0.2ml, 0.5ml, 0.6ml, 0.8ml and the 1.0ml that get 20.0 μ g/ml.
4. the method for definite benzoic acid standard solution typical curve as claimed in claim 2, wherein, the condition of described gas chromatography is:
Chromatographic column: capillary chromatographic column, stationary phase are nitroso-terephthalic acid (TPA) modified poly (ethylene glycol) or suitable person, and the chromatographic column specification is 30m * 0.25mm * 0.2 μ m or 30m * 0.32mm * 0.2 μ m;
Injector temperature: 200-300 ℃;
Input mode: shunting, split ratio is 40: 1-50: 1;
Carrier gas: high-purity helium;
Flow velocity: the capillary chromatography column internal diameter is that the flow velocity of 0.25 μ m is 1.0-1.4ml/min, and the capillary chromatography column internal diameter is that the flow velocity of 0.32 μ m is 1.8-2.5ml/min;
Heating schedule is as follows: initial temperature is 140 ℃ to be kept 1-2 minute, be raised to 175-190 ℃ with 4-7 ℃/min then after, kept program 10-20.5 minute altogether 4-7 minute.
5. the method for definite benzoic acid standard solution typical curve as claimed in claim 4, wherein, described mass spectral condition is:
Ionization mode: electricity bombardment ionization (EI);
Ionizing energy: 70eV;
Ion source temperature: 200-220 ℃;
Transmission line temperature: 150-200 ℃;
Solvent delay time: 3.0-5.0 minute;
Sample size: 1-2 μ L;
Scan pattern: multiple-reaction monitoring (MRM), qualitative ion mass-to-charge ratio (m/z) 122＞105,122＞77, quota ion mass-to-charge ratio (m/z) 122＞105;
Collision energy: mass-to-charge ratio (m/z) 122＞105 is 8V, and 122＞77 is 20V.
6. a benzoic acid standard solution typical curve of determining with the described method of claim 2 detects the method for benzoic acid content in the cheese, it is characterized in that comprising:
1) takes by weighing 3-5g sea sand to the 50ml centrifuge tube, in centrifuge tube, take by weighing 1-3g cheese sample then, add the 1-2g zinc acetate, accurately add 20ml distilled water;
2) fully stir with glass bar, cheese evenly is blended in the extra large sand;
3) fully mix with vortex mixer;
4) with hydro-extractor under the 3000rpm-4000rpm condition centrifugal 5-10 minute;
5) filter water layer, draw 5ml filtrate in one centrifuge tube, add 6mol/L hydrochloric acid solution 1ml, the 2ml methylene chloride;
6) mixed 1 minute with vortex mixer, with hydro-extractor under the 3000rpm-4000rpm condition centrifugal 5-10 minute;
7) membrane filtration is to the sample introduction bottle after filtration with clear liquid under the absorption of 1ml syringe, and this is the benzoic acid standard solution to be measured that makes;
8) with gas chromatography-mass spectrum/mass spectrometer described benzoic acid standard solution to be measured is measured;
9) measurement result and the determined benzoic acid standard solution typical curve of step 8) gained are compared, determine the benzoic acid content of described benzoic acid standard solution to be measured;
10) determine benzoic acid content in the described cheese according to the benzoic acid content of described benzoic acid standard solution to be measured.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102495144A (en) * | 2011-11-11 | 2012-06-13 | 深圳天祥质量技术服务有限公司 | Detecting method of Benzene content |
| CN117310051A (en) * | 2023-11-28 | 2023-12-29 | 江西农业大学 | Method for detecting benzoic acid in gastrointestinal tract contents of monogastric animals |
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| CN101806682B (en) | 2012-05-30 |
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