CN115656369A - Method for simultaneously determining organic acid and nicotine content in electronic cigarette liquid by liquid chromatography-tandem mass spectrometry - Google Patents
Method for simultaneously determining organic acid and nicotine content in electronic cigarette liquid by liquid chromatography-tandem mass spectrometry Download PDFInfo
- Publication number
- CN115656369A CN115656369A CN202211295316.9A CN202211295316A CN115656369A CN 115656369 A CN115656369 A CN 115656369A CN 202211295316 A CN202211295316 A CN 202211295316A CN 115656369 A CN115656369 A CN 115656369A
- Authority
- CN
- China
- Prior art keywords
- acid
- mass spectrometry
- flow rate
- nicotine
- solution
- 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.)
- Pending
Links
Images
Landscapes
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The invention relates to a method for simultaneously determining the content of organic acid and nicotine in electronic cigarette liquid by liquid chromatography-tandem mass spectrometry, which comprises the following steps: (1) Weighing electronic cigarette liquid into a centrifuge tube, adding an internal standard solution and methanol, and performing vortex oscillation for later use; (2) Performing liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis on the extract liquid obtained in the step (1); (3) And (4) quantitatively calculating the content of the target object in the sample by adopting an internal standard curve method. The method disclosed by the invention has the advantages of high flux, good separation degree, simplicity and convenience in operation, good recovery rate and repeatability and the like, and is suitable for analyzing non-volatile organic acids and nicotine in the electronic cigarette liquid.
Description
Technical Field
The invention belongs to the technical field of chemical analysis, and particularly relates to a method for simultaneously determining the contents of organic acid and nicotine in electronic cigarette liquid by liquid chromatography-tandem mass spectrometry.
Background
The electronic cigarette liquid is an important component of an electronic cigarette product, and the electronic cigarette liquid is atomized by an electronic cigarette device to generate inhalable aerosol containing nicotine and other substances. The types of nicotine salts in the electronic cigarette mainly include nicotine benzoate, lactate, levulinate, salicylate, tartrate and the like, but no literature report is available on an analysis method for simultaneously measuring nicotine and various types of organic acids in the cigarette liquid of the electronic cigarette. Harvanko et al (Nicotine Tob Res,2019,22 (7): 1239-1243) use gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) to qualitatively determine the type of organic acid in the electronic cigarette smoke solution, however, GC-MS sample pretreatment requires a complex derivatization process, the analysis process is time-consuming and labor-consuming, HPLC-MS only carries out qualitative analysis on the organic acid, no method verification is carried out, quantitative analysis on a target object is not realized, and the method does not realize simultaneous determination of Nicotine (Nicotine is quantitatively analyzed by using gas chromatography). Moreover, because electronic cigarettes have various tastes and various matrixes, hundreds of additives can exist (Sci Rep,2019,9, 1-9.), and HPLC-MS uses a single quadrupole for qualitative analysis, so that matrix interference can exist, and the qualitative accuracy of a target object is insufficient. The liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has the advantages of high speed, sensitivity, good reproducibility, high flux and the like, and has wide application prospect in the aspect of simultaneously measuring the contents of organic acid and nicotine in the cigarette liquid of the electronic cigarette, but no relevant report is found at present.
Disclosure of Invention
The invention aims to establish a method for simultaneously determining the contents of organic acid and nicotine in the cigarette liquid of the electronic cigarette by using a liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) based on the technical defects, and the method realizes the simultaneous determination of the contents of nicotine, benzoic acid, salicylic acid, pyruvic acid, citric acid, malic acid, tartaric acid, levulinic acid, lactic acid, sorbic acid and succinic acid in the cigarette liquid of the electronic cigarette.
The purpose of the invention is realized by the following technical scheme:
a method for simultaneously determining the content of organic acid and nicotine in cigarette liquid of an electronic cigarette by liquid chromatography-tandem mass spectrometry comprises the following steps:
(1) Weighing electronic cigarette liquid into a centrifuge tube, adding an internal standard solution and methanol, and performing vortex oscillation for later use;
(2) Performing liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis on the extract liquor obtained in the step (1);
(3) And (3) quantitatively calculating the content of the target in the sample by adopting an internal standard curve method.
Preferably, in step (1), the internal standard solution is a methanol solution of benzoic acid-d 5 and nicotine-d 3 at concentrations of 500. Mu.g/mL and 400. Mu.g/mL, respectively.
Preferably, in step (1), the volume of methanol is 5 to 20mL, preferably 10mL.
Preferably, in step (1), the vortex is oscillated for 5 to 30 minutes, preferably for 5 minutes.
Preferably, in step (2), the column used in the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis is a Phenomenex Kinetex HILIC (4.6X 150mm,2.6 μm), phenomenex Luna C18 (2) (4.6X 150mm,3 μm), waters Xbridge BEH Amide (2.1X 100mm,2.5 μm) or Thermo SCIENTIENTIFIC accession Trinity P1 (3X 100mm,3 μm) column, preferably a Waters Xbridge BEH Amide (2.1X 100mm,2.5 μm) column.
Preferably, in step (2), the mobile phases used in the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis are water and acetonitrile, a 5mmol/L ammonium acetate solution and a solution of acetonitrile/water (V: V = 95/5) containing ammonium acetate at a concentration of 5mmol/L or a 5mmol/L ammonium acetate solution (pH 9.0) and a solution of acetonitrile/water (V: V =95/5, pH 9.0) containing ammonium acetate at a concentration of 5mmol/L, preferably a 5mmol/L ammonium acetate solution (pH 9.0) and acetonitrile/water (V: V =95/5, pH 9.0) containing ammonium acetate at a concentration of 5 mmol/L.
Preferably, in step (2), the chromatographic conditions of the liquid chromatography-tandem mass spectrometry further include: the column temperature is 40 ℃; the gradient elution procedure was: the initial flow rate is 0.6mL/min, the flow rate is reduced to 0.4mL/min at 0.5min, the flow rate is increased to 0.6mL/min at 7.2min, the flow rate is maintained for 1.8min, and the total operation time is 9min; the injection volume was 1. Mu.L.
Preferably, in step (2), the mass spectrometry conditions used in the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis are: an ion source: an electrospray ion source; an ionization mode: ESI (+); detection mode: an MRM mode; cone Temperature (Cone Temperature): 130 ℃; electrospray Voltage (Spray Voltage): 4000V (+), 3500V (-); cone orifice airflow (Cone Gas Flow): 20psi; heating Probe Temperature (Heated Probe Temperature): 350 ℃; probe Gas Flow rate (Probe Gas Flow): 50psi; atomizer air Flow rate (Nebulizer Gas Flow): 55psi.
Preferably, in step (2), the MRM mass spectrometric detection parameters of the target compound and the internal standard are as follows:
preferably, in step (3), the internal standard curve method is: preparing a series of standard working solutions containing a target object, adding internal standards (benzoic acid-d 5 and nicotine-d 3), taking the quantitative ion peak area ratio of the target object to the internal standard object in each standard working solution as a vertical coordinate, and taking the content of the target object in each standard working solution as a horizontal coordinate to prepare a standard working curve; and (3) substituting the analysis result obtained in the step (2) into the standard curve to obtain the content of the target object in the solution to be detected, and further calculating to obtain the content of each target object in the sample.
Preferably, in step (3), the concentration range of each target in the series of standard working solutions is: nicotine 0.10-100 mug/mL, citric acid 10-100 mug/mL, tartaric acid 5-100 mug/mL, pyruvic acid and lactic acid 1-100 mug/mL, levulinic acid 0.5-100 mug/mL, other organic acids 0.1-100 mug/mL.
Preferably, the organic acid includes benzoic acid, salicylic acid, pyruvic acid, lactic acid, succinic acid, levulinic acid, sorbic acid, malic acid, tartaric acid, citric acid, and the like.
Preferably, in a particular embodiment, the method comprises the steps of:
(1) Weighing the electronic cigarette liquid into a centrifuge tube, adding an internal standard solution and methanol, and performing vortex oscillation for later use;
(2) Performing liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis on the extract liquid obtained in the step (1);
wherein, the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis uses the chromatographic conditions: the chromatographic column is a Waters Xbridge BEH Amide chromatographic column (2.1 × 100mm,2.5 μm), and the column temperature is 40 ℃; the mobile phase was a 5mmol/L ammonium acetate solution (pH 9.0) and a 5mmol/L ammonium acetate in acetonitrile/water (V: V =95/5, pH 9.0) solution, and the gradient elution procedure was: the initial flow rate is 0.6mL/min, the flow rate is reduced to 0.4mL/min at 0.5min, the flow rate is increased to 0.6mL/min at 7.2min, the flow rate is maintained for 1.8min, and the total operation time is 9min; the injection volume was 1. Mu.L.
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis the mass spectrometry conditions used: an ion source: an electrospray ion source; an ionization mode: ESI (+); detection mode: an MRM mode; cone Temperature (Cone Temperature): 130 ℃; electrospray Voltage (Spray Voltage): 4000V (+), 3500V (-); cone orifice airflow (Cone Gas Flow): 20psi; heating Probe Temperature (Heated Probe Temperature): 350 ℃; probe Gas Flow rate (Probe Gas Flow): 50psi; atomizer airflow rate (Nebulizer Gas Flow): 55psi;
the MRM mass spectrometric detection parameters of the target compound and the internal standard are as follows:
note: "+" indicates the quantitative ion.
(3) And (4) quantitatively calculating the content of the target object in the sample by adopting an internal standard curve method.
Wherein the standard curve method of the internal standard is as follows: preparing a series of standard working solutions containing targets, adding internal standards (benzoic acid-d 5 and nicotine-d 3), taking the quantitative ion peak area ratio of the targets and the internal standards in each standard working solution as a vertical coordinate, and taking the content of the targets in each standard working solution as a horizontal coordinate to prepare a standard working curve; substituting the analysis result of the step (2) into a standard curve to obtain the content of the target object in the solution to be detected, and further calculating to obtain the content of each target object in the sample;
the concentration range of each target in the series of standard working solutions is as follows: nicotine 0.10-100 mug/mL, citric acid 10-100 mug/mL, tartaric acid 5-100 mug/mL, pyruvic acid and lactic acid 1-100 mug/mL, levulinic acid 0.5-100 mug/mL, other organic acids 0.1-100 mug/mL.
Compared with the prior art, the invention has at least the following beneficial technical effects:
based on LC-MS/MS technology, the invention inspects the selection of chromatographic column, the composition of mobile phase, the flow rate of mobile phase, the mass spectrum condition and the like, establishes an analysis method of non-volatile organic acid and nicotine in the electronic cigarette liquid, realizes the simultaneous analysis method of benzoic acid, salicylic acid, pyruvic acid, lactic acid, succinic acid, levulinic acid, sorbic acid, malic acid, tartaric acid, citric acid and nicotine, has the advantages of high flux, good separation degree, simple and convenient operation, good recovery rate and repeatability and the like, and is suitable for the analysis of non-volatile organic acid and nicotine in the electronic cigarette liquid.
Drawings
FIG. 1 is a Multiple Reaction Monitoring (MRM) chromatogram of a target and its internal standard in a standard working solution in the HPLC-MS/MS method in example 1.
Detailed Description
The technical solution of the present invention will be further described below with reference to specific embodiments.
Example 1: method for simultaneously determining content of 10 nicotine salts in cigarette liquid of electronic cigarette by liquid chromatography-tandem mass spectrometry
(1) Instruments and reagents
The instrument comprises: the liquid chromatography-tandem mass spectrometer (HPLC-MS/MS) system consists of an American Agilent 1290 ultra-high performance liquid chromatograph (comprising a G7129B autosampler, a G7120A quaternary solvent pump and a G1316B column incubator) and a Bruker EVOQ triple quadrupole mass spectrometer (matched with an ESI ion source), and data acquisition and processing Software is HyStar Software; ultrasonic apparatus (YM-100S, union); an electronic balance (AE 163, mettler, switzerland, feeling: 0.0001 g); talboys digital display multitubular vortex mixer.
Reagent consumables: benzoic acid, lactic acid, levulinic acid, salicylic acid, succinic acid, sorbic acid, citric acid, tartaric acid, malic acid, pyruvic acid and nicotine are purchased from Alfa Aesar, and the purity is more than 88%; benzoic acid-d 5 and nicotine-d 3 were purchased from Macklin with a purity of more than 98%, methanol (chromatographically pure) from MERCK and ammonium formate (purity of more than 99%) from Macklin, the ultrapure water used being made by the Milli-Q system (Milford, MA, USA).
(2) Working conditions of the apparatus
The experiment adopts an Agilent 1290 ultra performance liquid chromatograph, and a chromatographic column is Waters Xbridge BEH Amide (2.1X 100mm,2.5 μm); mobile phase A:5mmol/L ammonium acetate solution (pH 9.0), mobile phase B: acetonitrile/water at a concentration of 5mmol/L ammonium acetate (V: V =95/5, ph 9.0); the sample volume was 1. Mu.L, and the column temperature was 40 ℃. The gradient elution procedure is shown in the following table:
| time (min) | A(％) | B(％) | Flow rate (μ L/min) |
| 0.00 | 0 | 100 | 600 |
| 0.50 | 0 | 100 | 400 |
| 7.00 | 50 | 50 | 400 |
| 7.20 | 0 | 100 | 600 |
| 9.00 | 0 | 100 | 600 |
Mass spectrum conditions: cone Temperature (Cone Temperature): 130 ℃; electrospray Voltage (Spray Voltage): 4000V (+), 3500V (-); cone orifice airflow (Cone Gas Flow): 20psi; heating Probe Temperature (Heated Probe Temperature): 350 ℃; probe Gas Flow rate (Probe Gas Flow): 50psi; atomizer air Flow rate (Nebulizer Gas Flow): 55psi; the MRM mass spectrometric detection parameters of the target compound and the internal standard are as follows:
the chromatogram of target substance and its internal standard Multiple Reaction Monitoring (MRM) in standard working solution in liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method is shown in FIG. 1.
(3) Sample pretreatment
Weighing 40mg of electronic cigarette liquid sample into a 15mL centrifuge tube, adding 100 μ L of mixed internal standard solution (the concentration of benzoic acid-d 5 is 500 μ g/mL, the concentration of nicotine-d 3 is 400 μ g/mL) and 10mL of methanol, shaking and extracting for 5min, filtering with a 0.22 μm organic phase filter membrane, and placing in a chromatographic bottle for detection.
(4) Standard working solution preparation
(1) And preparing an internal standard solution: 50mg of benzoic acid-d 5 and nicotine-d 3 are accurately weighed respectively and placed in 10mL brown volumetric flasks respectively, the volume is determined by methanol, and internal standard stock solutions with the concentration of about 5mg/mL are prepared. Taking 1mL of benzoic acid-d 5 internal standard stock solution and 0.8mL of nicotine-d 3 internal standard stock solution, and fixing the volume to 10mL by using methanol to obtain a benzoic acid-d 5 internal standard solution (internal standard) with the concentration of 500 mu g/mL and the concentration of 400 mu g/mL. Wherein nicotine-d 3 is used for measuring nicotine, and benzoic acid-d 5 is used for measuring benzoic acid, lactic acid, levulinic acid, salicylic acid, succinic acid, sorbic acid, citric acid, tartaric acid, malic acid and pyruvic acid.
(2) First-class standard stock solution preparation
Accurately weighing 2.0g of each of benzoic acid, lactic acid, levulinic acid, salicylic acid, succinic acid, sorbic acid, citric acid, tartaric acid, malic acid, pyruvic acid and nicotine, respectively placing the weighed materials in a 25mL brown volumetric flask, and metering the volume of methanol to a scale mark to prepare a standard substance stock solution with the concentration of about 80 mg/mL. The solution should be stored at 4-8 deg.C in the dark.
(3) Preparation of second-level standard stock solution
Precisely absorbing a certain volume of benzoic acid, lactic acid, levulinic acid, salicylic acid, succinic acid, sorbic acid, citric acid, tartaric acid, pyruvic acid, malic acid and nicotine stock solution, placing the stock solution in a 50mL brown volumetric flask, and preparing a mixed secondary standard stock solution with the concentration of 5mg/mL by metering the volume of methanol to a scale mark.
(4) Preparation of standard working solution
Accurately transferring the mixed secondary standard stock solution and the internal standard solution, and preparing standard working solutions with final concentrations of 0.1, 0.5, 1, 5, 10, 20, 40, 50, 80 and 100 mu g/mL by using methanol, wherein the final concentration of benzoic acid-d 5 in the internal standard is 5 mu g/mL, and the final concentration of nicotine-d 3 in the internal standard is 4 mu g/mL.
(5) Sample assay
And (4) respectively carrying out HPLC-MS/MS analysis on the standard curve solution obtained in the step (4) and the sample solution obtained in the step (3), wherein the chromatograms of the target substance and the internal standard Multiple Reaction Monitoring (MRM) thereof in the standard working solution are shown in the attached figure. Taking the quantitative ion peak area ratio of the target substance to the internal standard substance in each standard working solution as a vertical coordinate, and taking the content of the target substance in each standard working solution as the vertical coordinate to prepare a standard working curve; and (4) substituting the analysis result of the step (3) into the standard curve to obtain the content of the target object in the solution to be detected, and further calculating to obtain the content of each target object in the sample.
(6) Method verification
According to the lowest-level standard working solution, the detection limit of the method is calculated according to 3 times of signal-to-noise ratio, the standard addition recovery rate is calculated according to 3 addition levels of low, medium and high, 5 parallel samples are added at each level, and the test precision is calculated according to the parallel test result. The linear range, linear coefficient, detection limit, average recovery rate by adding standard, and average precision of the method are shown in table 1.
TABLE 1 Linear Range, linear coefficient, detection Limit, average spiked recovery and average precision for each target
(7) Analysis of actual samples
According to the measurement method, 5 electronic cigarette liquid samples are selected, and the content of the target substances is measured as shown in table 2:
TABLE 2 nicotine and organic acid content results (mg/g) in a typical e-cigarette smoke sample
Note: "/" indicates no detection
Example 2: selection experiment of chromatographic column
In view of the strongly polar chemical nature of organic acids, four column comparisons were chosen, kinetex HILIC (4.6X 150mm,2.6 μm) and Luna C18 (2) (4.6X 150mm,3 μm) from Phenomenex, xbridge BEH Amide (2.1X 100mm,2.5 μm) from Waters, and SCIENTIFIC Acclaim Trinity P1 (3X 100mm,3 μm) from Thermo. The specific experimental procedure is shown in example 1.
The research finds that:
(1) recommending a mobile phase (methanol-ammonium formate solution) according to the chromatographic column, wherein the Kinetex HILIC chromatographic column has an unsatisfactory organic acid separation effect, malic acid, tartaric acid, citric acid and nicotine are not retained on the chromatographic column, and the condition of the mobile phase is optimized to improve the unreserved compounds; the SCIENTIFIC Acclaim Trinity P1 chromatographic column has obvious organic acid separating effect, but no salicylic acid, malic acid, citric acid and nicotine remain on the chromatographic column.
(2) The Luna C18 (2) chromatographic column can retain all organic acids and nicotine, but the baseline noise of levulinic acid and lactic acid is large, the retention time of most organic acids is concentrated, and the separation effect is poor. Further optimizing the elution proportion and time of the mobile phase, and having no obvious improvement effect.
(3) And recommending a mobile phase (acetonitrile-ammonium acetate solution) according to the chromatographic column, wherein all the target substances can be retained on the Xbridge BEH Amide chromatographic column, and a more ideal separation effect is obtained by further optimizing the mobile phase and the gradient elution condition.
The column finally determined was therefore an Xbridge BEH Amide (2.1X 100mm,2.5 μm) column.
Example 3: selection experiment of mobile phase
In order to examine the influence of the flow rate on the detection method, a mobile phase of water and acetonitrile, a 5mmol/L ammonium acetate solution and acetonitrile/water at a concentration of 5mmol/L ammonium acetate (V: V = 95/5), a 5mmol/L ammonium acetate solution (pH 9.0) and acetonitrile/water at a concentration of 5mmol/L ammonium acetate (V: V =95/5, pH 9.0) were selected and studied. The rest of the process, except for the mobile phase, is referred to example 1.
As a result, it was found that:
(1) and solvents such as water or acetonitrile are used in the mobile phase, so that citric acid, tartaric acid, lactic acid and succinic acid in the target substance are not reserved, and the compound retention effect is influenced.
(2) Ammonium acetate (5 mmol/L ammonium acetate solution and acetonitrile/water (V: V = 95/5) with a concentration of 5mmol/L ammonium acetate) was added to the aqueous phase and the organic phase, respectively, and the target substance was retained, and the responses of citric acid and tartaric acid were low.
(3) The pH of the mobile phase was adjusted so that the pH of both the aqueous phase and the organic phase was 9.0 (5 mmol/L ammonium acetate solution (pH 9.0) and acetonitrile/water (V: V =95/5, pH 9.0) at a concentration of 5mmol/L ammonium acetate), and a preferable effect was obtained.
The final mobile phase composition thus determined becomes scheme (3).
Example 4: selection experiment of mobile phase flow rate
In order to examine the influence of the flow rate of the mobile phase on the detection method, the flow rate of the mobile phase is set to be 0.4mL/min; and the initial flow rate is 0.6mL/min, the flow rate is reduced to 0.4mL/min at 0.5min, the flow rate is increased to 0.6mL/min at 7.2min, the flow rate is maintained for 1.8min, and the total operation time is 9min. The rest of the process is referred to example 1, except for the flow rate of the mobile phase.
The research finds that:
(1) under the experimental condition of 0.4mL/min, the separation effect of nicotine and salicylic acid is poor, and the analysis time is too long.
(2) 0.6mL/min, the flow rate is reduced to 0.4mL/min at 0.5min, the flow rate is increased to 0.6mL/min at 7.2min, a gradient flow rate mode of 1.8min is maintained, and the separation degree and the analysis time are considered.
Therefore, the finally determined flow rate conditions of the mobile phase are that the initial flow rate is 0.6mL/min, the flow rate is reduced to 0.4mL/min at 0.5min, the flow rate is increased to 0.6mL/min at 7.2min, and the flow rate is kept for 1.8min.
Example 5: selection experiment of mass spectrometric temperature
In order to examine the influence of the taper hole temperature on the detection method, several taper hole temperatures of 100 ℃, 120 ℃, 130 ℃, 150 ℃, 200 ℃ and 300 ℃ were selected and studied, and the method other than the taper hole temperature was referred to example 1.
As a result, it was found that:
(1) the temperature of the taper hole has great influence on the citric acid, the lactic acid, the tartaric acid and the succinic acid, when the temperature of the taper hole is 100-130 ℃, the citric acid has the maximum response when the temperature of the taper hole is 120 ℃, and the tartaric acid, the lactic acid and the succinic acid have the maximum response when the temperature of the taper hole is 130 ℃.
(2) And when the temperature of the taper hole is 130-300 ℃, the response of citric acid, lactic acid, tartaric acid and succinic acid is decreased progressively.
The final cone temperature was determined to be 130 ℃.
Claims (10)
1. A method for simultaneously determining the content of organic acid and nicotine in cigarette liquid of an electronic cigarette by liquid chromatography-tandem mass spectrometry comprises the following steps:
(1) Weighing the electronic cigarette liquid into a centrifuge tube, adding an internal standard solution and methanol, and performing vortex oscillation for later use;
(2) Performing liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis on the extract liquor obtained in the step (1);
(3) And (4) quantitatively calculating the content of the target object in the sample by adopting an internal standard curve method.
2. The method according to claim 1, wherein in step (1), the internal standard solution is a methanol solution of benzoic acid-d 5 and nicotine-d 3 at concentrations of 500 μ g/mL and 400 μ g/mL;
preferably, in step (1), the volume of methanol is 5 to 20mL, preferably 10mL.
3. The method according to claim 1 or 2, wherein in step (1), the vortex is oscillated for a period of 5 to 30 minutes, preferably for 5 minutes.
4. The method according to any one of claims 1 to 3, wherein in step (2), the chromatography column is a Phenomenex Kinetex HILIC (4.6 x 150mm,2.6 μm), phenomenex Luna C18 (2) (4.6 x 150mm,3 μm), waters Xbridge BEH Amide (2.1 x 100mm,2.5 μm) or Thermo SCIENTIFIC accession Trinity P1 (3 x 100mm,3 μm) chromatography column, preferably a Waters Xbridge BEH Amide (2.1 x 100mm,2.5 μm) chromatography column.
5. The process according to any one of claims 1 to 4, characterized in that in step (2), the mobile phase used in the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis is water and acetonitrile, a 5mmol/L ammonium acetate solution and acetonitrile/water at a concentration of 5mmol/L ammonium acetate (V: V = 95/5) or a 5mmol/L ammonium acetate solution (pH 9.0) and acetonitrile/water at a concentration of 5mmol/L ammonium acetate (V: V =95/5, pH 9.0), preferably a 5mmol/L ammonium acetate solution (pH 9.0) and acetonitrile/water at a concentration of 5mmol/L ammonium acetate (V: V =95/5, pH 9.0).
6. The method according to any one of claims 1 to 5, wherein in step (2), the chromatographic conditions of the liquid chromatography-tandem mass spectrometry further comprise: the column temperature is 40 ℃; the gradient elution procedure was: the initial flow rate is 0.6mL/min, the flow rate is reduced to 0.4mL/min at 0.5min, the flow rate is increased to 0.6mL/min at 7.2min, the flow rate is maintained for 1.8min, and the total operation time is 9min; the injection volume was 1. Mu.L.
7. The method according to any one of claims 1 to 6, wherein in step (2), the mass spectrometry conditions used in the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis are: an ion source: an electrospray ion source; an ionization mode: ESI (+); detection mode: an MRM mode; cone Temperature (Cone Temperature): 130 ℃; electrospray Voltage (Spray Voltage): 4000V (+), 3500V (-); cone orifice airflow (Cone Gas Flow): 20psi; heating Probe Temperature (Heated Probe Temperature): 350 ℃; probe Gas Flow rate (Probe Gas Flow): 50psi; atomizer air Flow rate (Nebulizer Gas Flow): 55psi;
preferably, in step (2), the MRM mass spectrometric detection parameters of the target compound and the internal standard are as follows:
8. the method according to any one of claims 1 to 7, wherein in step (3), the internal standard curve method is: preparing a series of standard working solutions containing a target object, adding internal standards (benzoic acid-d 5 and nicotine-d 3), taking the quantitative ion peak area ratio of the target object to the internal standard object in each standard working solution as a vertical coordinate, and taking the content of the target object in each standard working solution as a horizontal coordinate to prepare a standard working curve; substituting the analysis result of the step (2) into a standard curve to obtain the content of the target object in the solution to be detected, and further calculating to obtain the content of each target object in the sample;
preferably, in step (3), the concentration range of each target in the series of standard working solutions is: nicotine 0.10-100 mug/mL, citric acid 10-100 mug/mL, tartaric acid 5-100 mug/mL, pyruvic acid and lactic acid 1-100 mug/mL, levulinic acid 0.5-100 mug/mL, other organic acids 0.1-400 mug/mL.
9. The method of any one of claims 1 to 8, wherein the organic acid comprises a nicotine salt formed from benzoic acid, salicylic acid, pyruvic acid, lactic acid, succinic acid, levulinic acid, sorbic acid, malic acid, tartaric acid, citric acid and nicotine.
10. Method according to any one of claims 1 to 9, characterized in that it comprises the following steps:
(1) Weighing electronic cigarette liquid into a centrifuge tube, adding an internal standard solution and methanol, and performing vortex oscillation for later use;
(2) Performing liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis on the extract liquor obtained in the step (1);
wherein, the liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis uses the chromatographic conditions: the chromatographic column is a Waters Xbridge BEH Amide chromatographic column (2.1 × 100mm,2.5 μm), and the column temperature is 40 ℃; the mobile phase was a 5mmol/L ammonium acetate solution (pH 9.0) and acetonitrile/water at a concentration of 5mmol/L ammonium acetate (V: V =95/5, pH 9.0), and the gradient elution procedure was: the initial flow rate is 0.6mL/min, the flow rate is reduced to 0.4mL/min at 0.5min, the flow rate is increased to 0.6mL/min at 7.2min, the flow rate is maintained for 1.8min, and the total operation time is 9min; the sample injection volume is 1 mu L;
liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis the mass spectrometry conditions used: an ion source: an electrospray ion source; an ionization mode: ESI (+); detection mode: an MRM mode; cone Temperature (Cone Temperature): 130 ℃; electrospray Voltage (Spray Voltage): 4000V (+), 3500V (-); cone orifice Flow rate (Cone Gas Flow): 20psi; heating Probe Temperature (Heated Probe Temperature): 350 ℃; probe Gas Flow rate (Probe Gas Flow): 50psi; atomizer airflow rate (Nebulizer Gas Flow): 55psi;
the MRM mass spectrometric detection parameters of the target compound and internal standard are as follows:
note: "+" indicates quantitative ions
(3) Quantitatively calculating the content of the target object in the sample by adopting an internal standard curve method;
the internal standard curve method is as follows: preparing a series of standard working solutions containing a target object, adding internal standards (benzoic acid-d 5 and nicotine-d 3), taking the quantitative ion peak area ratio of the target object to the internal standard object in each standard working solution as a vertical coordinate, and taking the content of the target object in each standard working solution as a horizontal coordinate to prepare a standard working curve; substituting the analysis result obtained in the step (2) into the standard curve to obtain the content of the target object in the solution to be detected, and further calculating to obtain the content of each target object in the sample;
in step (3), the concentration range of each target in the series of standard working solutions is as follows: nicotine 0.10-100 mug/mL, citric acid 10-100 mug/mL, tartaric acid 5-100 mug/mL, pyruvic acid and lactic acid 1-100 mug/mL, levulinic acid 0.5-100 mug/mL, other organic acids 0.1-100 mug/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211295316.9A CN115656369A (en) | 2022-10-21 | 2022-10-21 | Method for simultaneously determining organic acid and nicotine content in electronic cigarette liquid by liquid chromatography-tandem mass spectrometry |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211295316.9A CN115656369A (en) | 2022-10-21 | 2022-10-21 | Method for simultaneously determining organic acid and nicotine content in electronic cigarette liquid by liquid chromatography-tandem mass spectrometry |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115656369A true CN115656369A (en) | 2023-01-31 |
Family
ID=84990394
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211295316.9A Pending CN115656369A (en) | 2022-10-21 | 2022-10-21 | Method for simultaneously determining organic acid and nicotine content in electronic cigarette liquid by liquid chromatography-tandem mass spectrometry |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115656369A (en) |
-
2022
- 2022-10-21 CN CN202211295316.9A patent/CN115656369A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109828044A (en) | A kind of method that ultra high efficiency closes 8 kinds of phenolic acids in phase chromatography concatenation QDa while quickly detection alcohol product | |
| CN109696514B (en) | Method for measuring 12 polyphenol compounds in fresh tobacco leaves | |
| Markelj et al. | Optimization of High Performance Liquid Chromatography Method for Simultaneous Determination of Some Purine and Pyrimidine Bases. | |
| CN105954402A (en) | Gas chromatography-flame ionization/tandem mass spectrometry detection method of alkaloids in cigarette main stream smoke | |
| CN109991318A (en) | A kind of tobacco juice for electronic smoke and fume component analysis method | |
| Xu et al. | Determination of nicotine and cotinine in human plasma by liquid chromatography-tandem mass spectrometry with atmospheric-pressure chemical ionization interface | |
| CN107688062A (en) | A kind of method of trace nicotine in measure tobacco juice for electronic smoke | |
| CN104655753A (en) | Method for determining 3-acetyl-2, 5-thioxene in food additive | |
| CN115541789A (en) | Method for measuring nicotine salt content in tobacco and tobacco products | |
| CN114019062A (en) | Method for detecting related substances in rifampicin | |
| CN105842377A (en) | High performance liquid chromatography detection method for pyrazine compounds in Baijiu | |
| CN112505227A (en) | Analysis method for simultaneously determining 11 components in electronic cigarette liquid | |
| CN107356689A (en) | One grow tobacco in 12 kinds of Amadori compounds assay method | |
| CN115656369A (en) | Method for simultaneously determining organic acid and nicotine content in electronic cigarette liquid by liquid chromatography-tandem mass spectrometry | |
| CN111983109B (en) | Chiral analysis method of alkaloid in tobacco or tobacco product | |
| CN115728408A (en) | Method for simultaneously determining seven artemisinin related compounds in artemisia annua | |
| CN113671086B (en) | Isolation and measurement of posaconazole Z 2 And method for impurity thereof | |
| CN111983108B (en) | Chiral analysis method for main alkaloids in electronic smoke sol | |
| CN111307992B (en) | Pre-column derivative liquid chromatography-mass spectrometry analysis method for quantitatively detecting organic acid in PM2.5 | |
| CN114577950A (en) | Method for determining anti-infective drugs in cosmetics | |
| CN113866305A (en) | Method for rapidly and accurately analyzing theanine in fresh tea leaves based on liquid chromatography-mass spectrometry technology | |
| CN110672735A (en) | Method for detecting alpha-dicarbonyl compound in simulated Maillard reaction system by high performance liquid chromatography-tandem mass spectrometry | |
| CN112461982A (en) | Detection method of L-camphorsulfonic acid methyl ester and L-camphorsulfonic acid ethyl ester | |
| CN111024872A (en) | Method for rapidly detecting 3-methylthio propanol in sesame-flavor liquor | |
| CN113686993B (en) | UPLC-MS detection method for nitrophenol substance and derivative thereof |
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 |