CN108195987B - Method for analyzing stearic acid content in stearate - Google Patents
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Abstract
The invention belongs to the technical field of analysis, and particularly relates to a method for analyzing the stearic acid content in stearate. The method comprises the steps of firstly adopting concentrated nitric acid and absolute ethyl alcohol to pretreat stearate to obtain stearic acid, then dissolving the stearic acid by the absolute ethyl alcohol, and then adopting special chromatographic and mass spectrum conditions to measure to obtain the content of the stearic acid. The method provided by the invention has the advantages that the sample is simply processed and then directly subjected to sample injection analysis, so that the content of stearic acid in stearate is simply, quickly and accurately analyzed, the accuracy is high, the precision is good, the recovery rate is high, and the detection limit is low.
Description
Technical Field
The invention belongs to the technical field of analysis, and particularly relates to a method for analyzing the stearic acid content in stearate.
Background
Stearic acid, mainly used for producing sodium stearate, magnesium stearate, calcium stearate, lead stearate, aluminium stearate, cadmium stearate, ferric stearate and potassium stearate. As an important raw material for synthesizing the stearate, the stearate is mainly from natural extracts, trace factors can have great influence on the product, and the uncertainty of the stearate product is high.
Stearates are a versatile class of additives used in many articles of manufacture and can provide many benefits to polymer products. Widely used for preparing cosmetics, plastic cold-resistant plasticizers, mold release agents, stabilizers, surfactants, rubber vulcanization accelerators, waterproofing agents, polishing agents, metal soaps, metal mineral flotation agents, softeners, pharmaceuticals and other organic chemicals. In addition, it can be used as solvent for oil-soluble pigment, crayon slip agent, stencil paper polishing agent, and emulsifier for stearin. The product can be used as lubricant, defoaming agent, and food additive such as stearin, sorbitan stearate, and sucrose ester in food industry. Used as raw materials of auxiliary agents and raw materials of daily chemical products.
At present, the content of stearic acid in stearate is generally analyzed by an acid-base titration method at home and abroad, and then the content of stearate in stearate is deduced. The method has the advantages of low analysis cost, short analysis period and the like, but the stearate generally contains palmitate and salts in other forms, but common acid-base titration cannot effectively distinguish the stearate from the palmitate, so that the accuracy of a detection result is influenced.
In view of this, it is necessary to develop a new detection method that can distinguish stearic acid from palmitic acid in a sample. The liquid chromatography-mass spectrometry (HPLC-MS) technology is a separation and analysis technology which takes high performance liquid chromatography as a separation means and mass spectrometry as an identification tool. At present, no report is available for analyzing the stearic acid content by using an HPLC-MS analysis means.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a method for analyzing the stearic acid content in stearate by liquid chromatography-mass spectrometry, which only needs to simply process a sample and directly perform sample injection analysis, thereby realizing simple, rapid and accurate quantitative analysis of stearic acid, and having high accuracy, good precision, high recovery rate and low detection limit.
The technical scheme adopted by the invention for solving the technical problem comprises the following steps:
chromatographic and mass spectrum conditions:
(1) the used instrument is a high performance liquid chromatography-quadrupole-time-of-flight mass spectrometer, and the chromatographic column is an ODS series liquid chromatographic column;
(2) liquid phase conditions: the mobile phase A is methanol; the mobile phase B is water; flow rate: 0.1-1.0 mL/min, elution program: contains A: 80-95% of the total weight of the composition, and the balance of B; the column temperature is 15-25 ℃, and the sample injection amount is 1-20 mu L;
(3) mass spectrum conditions: ESI ionization source, anion detection mode, ion source temperature 300 ℃, dry gas flow 5 ~ 15L/min, atomizer gas pressure 30 ~ 40psig, capillary voltage 2500 ~ 3000V, capillary exit voltage 100 ~ 120V, taper hole voltage 55 ~ 65V, octupole rod radio frequency voltage 650 ~ 750V, ration ion: 283.2650 (M-H); and (3) qualitative ion: m/z 329.2697(M + COOH).
Drawing of (II) standard curve
Weighing a stearic acid standard substance, preparing a series of standard solutions, determining by adopting the chromatographic and mass spectrum conditions, and drawing a peak area-concentration standard curve; carrying out regression analysis according to the ion peak area corresponding to the quantitative ions and the stearic acid concentration to obtain a linear equation of y-827095.7811 x-25950.0581, R20.9993x represents the concentration of stearic acid in the stearic acid standard solution, mg/L, and y represents the ion peak area corresponding to the quantitative ion;
(III) detection of sample to be detected
Get m1g, adding stearic acid salt, adding concentrated nitric acid and absolute ethyl alcohol, heating, stopping heating when an oil layer becomes clear, cooling, filtering, drying a filter cake to obtain pretreated stearic acid m2g；
Taking the stearic acid m3And g, adding absolute ethyl alcohol into a 100ml volumetric flask as a sample to be detected, fully and uniformly mixing, fixing the volume, diluting the sample according to the concentration range of a standard curve by the dilution factor of f times to obtain a liquid to be detected, and performing liquid quality analysis. Determining the liquid to be detected under the chromatographic and mass spectrum conditions, and recording the peak area of the quantitative ions; substituting the peak area-concentration standard curve or linear equation in the step (II) to obtain the concentration c of stearic acid in the solution to be detected in unit mg/L, and obtaining the content of stearic acid by adopting the following formula;
wherein:
x represents the content of stearic acid in a sample to be detected in mg/kg;
c, checking the concentration of stearic acid in the solution to be detected by a standard curve, wherein the unit is mg/L;
f is the dilution multiple of the sample;
m1-mass of stearate, g;
m2-mass of stearic acid after pretreatment, g;
m3-mass of stearic acid of the sample to be tested, g;
v-volume of constant volume.
In the invention, the volume ratio of the concentrated nitric acid to the ethanol is 3: 1.
The content of stearate in the stearate sample is deduced from the content of stearic acid in the stearate obtained by the above analysis.
The invention has the beneficial effects that:
the sample is directly injected and analyzed after being simply processed, so that the content of stearic acid in stearate is simply, quickly and accurately analyzed;
secondly, the method has high accuracy, good precision, high recovery rate and low detection limit;
the invention adopts the high performance liquid chromatography-quadrupole-time-of-flight mass spectrometer for the first time, and can realize the high-efficiency and accurate determination of the stearic acid content in the stearate by matching with special liquid chromatography and mass spectrum parameters.
Drawings
Fig. 1 shows a stearic acid standard use solution extraction ion flow of 40mg/L [ M/z: 283.2650(M-H) ];
fig. 2 is stearic acid extraction ion flow [ M/z ═ 283.2650(M-H) ] in a sample to be tested in example 1 of the present invention;
FIG. 3 is a linear regression equation of ion peak area and stearic acid concentration according to the present invention;
Detailed Description
Example one
(I) laboratory instruments and reagents
(1) The used instrument is a high performance liquid chromatography-quadrupole-time-of-flight mass spectrometer, and the chromatographic column is an ODS series liquid chromatographic column;
(2) experimental reagent: methanol, ultrapure water, ethanol, nitric acid and stearic acid standard substance
(II) chromatographic and mass spectrum conditions:
(1) liquid phase conditions: mobile phase: a: methanol; b, water; flow rate: 0.1-1.0 mL/min, elution program: contains A: 80-95% of the total weight of the composition, and the balance of B; the column temperature is 15-25 ℃, and the sample injection amount is 1-20 mu L;
(2) mass spectrum conditions: ESI ionization source, anion detection mode, ion source temperature 300 ℃, dry gas flow 5 ~ 15L/min, atomizer gas pressure 30 ~ 40psig, capillary voltage 2500 ~ 3000V, capillary exit voltage 100 ~ 120V, taper hole voltage 55 ~ 65V, octupole rod radio frequency voltage 650 ~ 750V, ration ion: 283.2650 (M-H); and (3) qualitative ion: m/z 329.2697(M + COOH).
(III) drawing of Standard Curve
Weighing 100mg of stearic acid standard substance in a 100ml volumetric flask, adding absolute ethyl alcohol, fixing the volume after the sample is fully dissolved to obtain a standard stock solution with the stearic acid concentration of 1000mg/L, respectively sucking 0ml, 1ml, 2ml, 3ml, 4ml and 5ml in the 100ml volumetric flask, fixing the volume with the absolute ethyl alcohol to obtain a series of standard solutions with the stearic acid concentration of 0, 10, 20, 30, 40 and 50mg/L, measuring under the conditions of the chromatogram and the mass spectrum, recording the peak area of quantitative ion M/z which is 283.2650(M-H), and drawing a peak area-concentration standard curve; regression analysis was performed based on the ion peak area corresponding to the quantitative ion M/z 283.2650(M-H) and the stearic acid concentration to obtain the linear equation: 827095.7811x-25950.0581, R20.9993(x is the concentration of stearic acid, mg/L)
(IV) detection of sample to be detected
Get m1g sodium stearate of a certain company, adding concentrated nitric acid and ethanol (the volume ratio of the concentrated nitric acid to the ethanol is 3:1), heating, stopping heating when an oil layer becomes clear, cooling, filtering, and drying a filter cake at 50 ℃ to obtain m2g stearic acid.
Respectively take m3g stearic acid is added into absolute ethyl alcohol in a volumetric flask of 100ml, and after the stearic acid is fully and uniformly mixed, the volume is constant, and the root is obtainedAnd (5) diluting the sample according to the concentration range of the standard curve by the dilution factor of f to obtain the solution to be detected, and performing liquid quality analysis. Determining the liquid to be detected under the chromatographic and mass spectrum conditions, and recording the peak area of the quantitative ions; substituting into the peak area-concentration standard curve or linear equation in step III to obtain stearic acid concentration c in the solution to be tested, wherein the stearic acid content is 12.32% by adopting the following formula.
Wherein:
x represents the content of stearic acid in a sample to be detected in mg/kg;
c, checking the concentration of stearic acid in the solution to be detected by a standard curve, wherein the unit is mg/L;
f is the dilution multiple of the sample;
m1-mass of stearate, g;
m2-mass of stearic acid after pretreatment, g;
m3-mass of sample to be tested stearic acid, g.
Example two
(I) laboratory instruments and reagents
(1) The used instrument is a high performance liquid chromatography-quadrupole-time-of-flight mass spectrometer, and the chromatographic column is an ODS series liquid chromatographic column;
(2) experimental reagent: methanol, ultrapure water, ethanol, nitric acid and stearic acid standard substance
(II) chromatographic and mass spectrum conditions:
(1) liquid phase conditions: mobile phase: a: methanol; b, water; flow rate: 0.1-1.0 mL/min, elution program: contains A: 80-95% of the total weight of the composition, and the balance of B; the column temperature is 15-25 ℃, and the sample injection amount is 1-20 mu L;
(2) mass spectrum conditions: ESI ionization source, anion detection mode, ion source temperature 300 ℃, dry gas flow 5 ~ 15L/min, atomizer gas pressure 30 ~ 40psig, capillary voltage 2500 ~ 3000V, capillary exit voltage 100 ~ 120V, taper hole voltage 55 ~ 65V, octupole rod radio frequency voltage 650 ~ 750V, ration ion: 283.2650 (M-H); and (3) qualitative ion: m/z 329.2697(M + COOH).
(III) drawing of Standard Curve
Weighing 100mg of stearic acid standard substance in a 100ml volumetric flask, adding absolute ethyl alcohol, fixing the volume after the sample is fully dissolved to obtain a standard stock solution with the stearic acid concentration of 1000mg/L, respectively sucking 0ml, 1ml, 2ml, 3ml, 4ml and 5ml in the 100ml volumetric flask, fixing the volume with the absolute ethyl alcohol to obtain a series of standard solutions with the stearic acid concentration of 0, 10, 20, 30, 40 and 50mg/L, measuring under the conditions of the chromatogram and the mass spectrum, recording the peak area of quantitative ion M/z which is 283.2650(M-H), and drawing a peak area-concentration standard curve; regression analysis was performed based on the ion peak area corresponding to the quantitative ion M/z 283.2650(M-H) and the stearic acid concentration to obtain the linear equation: 827095.7811x-25950.0581, R20.9993(x is the concentration of stearic acid, mg/L)
(IV) detection of sample to be detected
Get m1g potassium stearate of a company, adding concentrated nitric acid and ethanol (the volume ratio of the concentrated nitric acid to the ethanol is 3:1), heating, stopping heating when an oil layer becomes clear, cooling, filtering, and drying a filter cake at 50 ℃ to obtain m2g stearic acid.
Respectively take m3Adding absolute ethyl alcohol into stearic acid g in a volumetric flask of 100ml, fully and uniformly mixing, fixing the volume, diluting the sample according to the concentration range of a standard curve by f times to obtain a solution to be detected, and carrying out liquid quality analysis. Determining the liquid to be detected under the chromatographic and mass spectrum conditions, and recording the peak area of the quantitative ions; substituting into the peak area-concentration standard curve or linear equation in step III to obtain stearic acid concentration c in the solution to be tested, wherein the stearic acid content is 19.15% by adopting the following formula.
Wherein:
x represents the content of stearic acid in a sample to be detected in mg/kg;
c, checking the concentration of stearic acid in the solution to be detected by a standard curve, wherein the unit is mg/L;
f is the dilution multiple of the sample;
m1-mass of stearate, g;
m2-mass of stearic acid after pretreatment, g;
m3-mass of sample to be tested stearic acid, g.
EXAMPLE III
(I) laboratory instruments and reagents
(1) The used instrument is a high performance liquid chromatography-quadrupole-time-of-flight mass spectrometer, and the chromatographic column is an ODS series liquid chromatographic column;
(2) experimental reagent: methanol, ultrapure water, ethanol, nitric acid and stearic acid standard substance
(II) chromatographic and mass spectrum conditions:
(1) liquid phase conditions: mobile phase: a: methanol; b, water; flow rate: 0.1-1.0 mL/min, elution program: contains A: 80-95% of the total weight of the composition, and the balance of B; the column temperature is 15-25 ℃, and the sample injection amount is 1-20 mu L;
(2) mass spectrum conditions: ESI ionization source, anion detection mode, ion source temperature 300 ℃, dry gas flow 5 ~ 15L/min, atomizer gas pressure 30 ~ 40psig, capillary voltage 2500 ~ 3000V, capillary exit voltage 100 ~ 120V, taper hole voltage 55 ~ 65V, octupole rod radio frequency voltage 650 ~ 750V, ration ion: 283.2650 (M-H); and (3) qualitative ion: m/z 329.2697(M + COOH).
(III) drawing of Standard Curve
Weighing 100mg of stearic acid standard substance in a 100ml volumetric flask, adding absolute ethyl alcohol, fixing the volume after the sample is fully dissolved to obtain a standard stock solution with the stearic acid concentration of 1000mg/L, respectively sucking 0ml, 1ml, 2ml, 3ml, 4ml and 5ml in the 100ml volumetric flask, fixing the volume with the absolute ethyl alcohol to obtain a series of standard solutions with the stearic acid concentration of 0, 10, 20, 30, 40 and 50mg/L, measuring under the conditions of the chromatogram and the mass spectrum, recording the peak area of quantitative ion M/z which is 283.2650(M-H), and drawing a peak area-concentration standard curve; according to the ion peak area corresponding to quantitative ion M/z-283.2650 (M-H) and the concentration of stearic acidAnd (5) carrying out regression analysis to obtain a linear equation: 827095.7811x-25950.0581, R20.9993(x is the concentration of stearic acid, mg/L)
(IV) detection of sample to be detected
Get m1g, adding concentrated nitric acid and anhydrous ethanol (the volume ratio of the concentrated nitric acid to the ethanol is 3:1) into calcium stearate of a certain company, heating, stopping heating when an oil layer becomes clear, cooling, filtering, and drying a filter cake at 50 ℃ to obtain stearic acid m2g。
Respectively take m3Adding absolute ethyl alcohol into stearic acid g in a volumetric flask of 100ml, fully and uniformly mixing, fixing the volume, diluting the sample according to the concentration range of a standard curve by f times to obtain a solution to be detected, and carrying out liquid quality analysis. Determining the liquid to be detected under the chromatographic and mass spectrum conditions, and recording the peak area of the quantitative ions; substituting into the peak area-concentration standard curve or linear equation in step III to obtain stearic acid concentration c in the solution to be tested, wherein the stearic acid content is 35.44% by adopting the following formula.
Wherein:
x represents the content of stearic acid in a sample to be detected in mg/kg;
c, checking the concentration of stearic acid in the solution to be detected by a standard curve, wherein the unit is mg/L;
f is the dilution multiple of the sample;
m1-mass of stearate, g;
m2-mass of stearic acid after pretreatment, g;
m3-mass of sample to be tested stearic acid, g.
Control experiment
1. Determination of the precision of the method
Adding anhydrous ethanol into 7 sodium stearate bottles of different masses of a certain company in 100ml volumetric flasks, fully and uniformly mixing, fixing the volume, diluting the sample according to the concentration range of a standard curve by f times, and obtaining the solution to be detected. Determining the liquid to be detected under the chromatographic and mass spectrum conditions, and recording the peak area of the quantitative ions; substituting into the peak area-concentration standard curve or linear equation in step three to obtain stearic acid concentration c in the solution to be tested, unit mg/L, obtaining stearic acid content by adopting the following formula,
the calculation results are shown in the following table:
| |
1 | 2 | 3 | 4 | 5 | 6 | 7 | RSD |
| Concentration of | 12.23％ | 12.28％ | 12.95％ | 12.12％ | 12.32％ | 13.04％ | 12.60％ | 2.923％ |
As can be seen from the data in the table, the experimental results of this method are very reproducible.
2. Determination of the accuracy of the method
Adding anhydrous ethanol into 5 sodium stearate bottles of different masses of a company in 100ml volumetric flasks, fully and uniformly mixing, then respectively adding stearic acid standard solutions of 50uL, 100uL, 200uL, 300uL and 400uL into the volumetric flasks, and carrying out constant volume to obtain a solution to be detected. Determining the liquid to be detected under the chromatographic and mass spectrum conditions, and recording the peak area of the quantitative ions; and (5) substituting a peak area-concentration standard curve or a linear equation in the third step to obtain the concentration c of the stearic acid in the solution to be detected in unit mg/L, and obtaining the content of the stearic acid by adopting the following formula.
The calculation results are shown in the following table:
| |
1 | 2 | 3 | 4 | 5 |
| Recovery rate of added standard | 98.9％ | 99.1％ | 98.9％ | 102.0％ | 103.0％ |
3. Determination of detection limits of methods
The detection limit of the method is calculated according to the condition that the ratio of the response value of the instrument to the noise is 3, and the detection limit of stearic acid is 0.005 ug/L.
As mentioned above, the quantitative method for analyzing stearate by liquid chromatography-mass spectrometry provided by the invention only needs simple treatment and direct sample injection analysis on a sample, so that the content of stearic acid in stearate is simply, quickly and accurately analyzed, the accuracy is high, the precision is good, the recovery rate is high, and the detection limit is low.
Claims (1)
1. A method for analyzing the stearic acid content in stearate is characterized in that: analyzing a sample to be detected by adopting a liquid chromatography-mass spectrometer, and specifically comprising the following steps:
chromatographic and mass spectrum conditions:
(1) the used instrument is a high performance liquid chromatography-quadrupole-time-of-flight mass spectrometer, and the chromatographic column is an ODS series liquid chromatographic column;
(2) liquid phase conditions: the mobile phase A is methanol; the mobile phase B is water; flow rate: 0.1-1.0 mL/min, elution program: contains A: 80-95 percent of the total weight of the composition, and the balance of B; the column temperature is 15-25 ℃, and the sample injection amount is 1-20 mu L;
(3) mass spectrum conditions: ESI ionization source, anion detection mode, ion source temperature 300 ℃, dry gas flow 5 ~ 15L/min, atomizing gas pressure 30 ~ 40psi, capillary voltage 2500 ~ 3000V, capillary exit voltage 100 ~ 120V, taper hole voltage 55 ~ 65V, octupole rod radio frequency voltage 650 ~ 750V, ration ion: 283.2650; and (3) qualitative ion: 329.2697;
drawing of (II) standard curve
Weighing a stearic acid standard substance, preparing a series of standard solutions, determining by adopting the chromatographic and mass spectrum conditions, and drawing a peak area-concentration standard curve; carrying out regression analysis according to the ion peak area corresponding to the quantitative ions and the stearic acid concentration to obtain a linear equation of y-827095.7811 x-25950.0581, R20.9993; x is the concentration of stearic acid in the stearic acid standard solution, mg/L, and y is the ion peak area corresponding to the quantitative ions;
(III) detection of sample to be detected
Get m1g, adding stearic acid salt, adding concentrated nitric acid and absolute ethyl alcohol, heating, stopping heating when an oil layer becomes clear, cooling, filtering, drying a filter cake to obtain pretreated stearic acid m2g；
Taking the stearic acid m3g, adding absolute ethyl alcohol into a 100ml volumetric flask as a sample to be detected, fully and uniformly mixing, fixing the volume, diluting the sample according to the concentration range of a standard curve by a dilution multiple of f times to obtain a liquid to be detected, and performing liquid quality analysis; determining the liquid to be detected by adopting the chromatographic and mass spectrum conditions, and recording the peak area of the quantitative ions; substituting the peak area-concentration standard curve or linear equation in the step (II) to obtain the concentration c of stearic acid in the solution to be detected in unit mg/L, and obtaining the content of stearic acid by adopting the following formula;
wherein: x represents the content of stearic acid in a sample to be detected in mg/kg;
c, checking the concentration of stearic acid in the solution to be detected in unit mg/L by using a standard curve;
f is the dilution multiple of the sample;
m1-mass of stearate, g;
m2-mass of stearic acid after pretreatment, g;
m3-mass of stearic acid of the sample to be tested, g;
v-constant volume;
the volume ratio of the concentrated nitric acid to the ethanol is 3: 1;
the stearate is single stearate.
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