CN111189949A - Method for simultaneously detecting linagliptin and saxagliptin in health food - Google Patents
Method for simultaneously detecting linagliptin and saxagliptin in health food Download PDFInfo
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Abstract
The invention belongs to the field of foods, and relates to a method for simultaneously detecting linagliptin and saxagliptin in health-care foods. The detection steps are as follows: preparing a solution required by an experiment; preparing a standard solution for drawing a standard curve; preparing a sample solution; liquid chromatography-tandem mass spectrometry; performing qualitative analysis; and (4) carrying out quantitative determination. The method has the advantages of simplicity, convenience, high efficiency, rapidness and accuracy, and can simultaneously and accurately determine the contents of linagliptin and saxagliptin in the health-care food.
Description
Technical Field
The invention belongs to the field of foods, relates to a detection method, and particularly relates to a method for simultaneously detecting linagliptin and saxagliptin in health-care foods.
Background
In recent years, the concept of regulating body functions and protecting body health with foods has been widely recognized by people. Diabetes is a disease seriously harming human health, and the number of attack people is on the rising trend. Since diabetics need to take medicines for life, many patients are concerned about toxic side effects of the medicines, and it is desired to improve and control blood sugar by foods having a function of regulating blood sugar. However, these products can only condition physiological functions, play a certain role in auxiliary health care, and do not have obvious effect of reducing blood sugar. Some illegal molecules have obvious blood sugar reducing effect for economic benefit by illegally adding blood sugar reducing chemical drugs, so as to entice diabetics to purchase. Such random addition often results in toxic reactions and serious or even life-threatening reactions to patients who take them for long periods of time and in overdose. In order to attack the act of illegally adding chemical drugs for reducing blood sugar, 13 drugs for reducing blood sugar such as tolbutamide, glibenclamide and the like have been listed as national health food safety supervision spot check items by the national market supervision and administration. Lawbreakers are continuously renovated for avoiding supervision and fake manufacturing, the added components are continuously changed, and blood sugar reducing medicines such as acarbose, vildagliptin and the like outside supervision and spot check projects are found to be illegally added.
The national standards of detection methods related to the determination of blood sugar-reducing drugs in health food include a national food and drug administration drug inspection supplement inspection method and inspection item approval part 2006005, 2007003, 2009029, 2011008, 2013001 and a national food and drug administration food supplement inspection method and detection of 75 illegally added chemical drugs in health food (BJS 201710), and the like, and the detection methods include 13 blood sugar-reducing drugs such as tolbutamide, glibenclamide, gliclazide, gliquidone, glimepiride, rosiglitazone maleate, repaglinide, pioglitazone hydrochloride, metformin hydrochloride, phenformin hydrochloride, buformin hydrochloride, glibornuride and the like.
The hypoglycemic drugs related in the literature include sulphonylurea insulin secretagogues (tolbutamide, chlorpropamide, tolazamide, acetohexamide, glibenclamide, glimepiride, gliborpizide, gliquidone, gliclazide), non-sulphonylurea insulin secretagogues (repaglinide, nateglinide, mitiglinide), biguanides (metformin, phenformin, buformin), α -glucosidase inhibitors (acarbose, voglibose, miglitol), insulin sensitizers (rosiglitazone, pioglitazone, troglitazone, ciglitazone), dipeptidyl peptidase-4 (DPP-4) inhibitors (vildagliptin, sitagliptin, alogliptin) and sodium-glucose cotransporter 2 (SGDAglicol-2) inhibitors (SGDAgliclan, GLIN, GLUTRAL) and the like.
Linagliptin and saxagliptin belong to dipeptidyl peptidase-4 (DPP-4) inhibitors, are used for treating type 2 diabetes, are on the market at home, and have the risk of being illegally added into health food by lawbreakers.
In domestic and foreign detection standards and documents, no detection method for linagliptin and saxagliptin in health food is available. Therefore, in order to prevent the behavior of illegally adding hypoglycemic drugs into health food and ensure the physical health of consumers, a detection method of linagliptin and saxagliptin in the health food is needed to be established.
Disclosure of Invention
The invention provides a method for simultaneously detecting linagliptin and saxagliptin in health-care food, which has the advantages of simplicity, convenience, high efficiency, rapidness and accuracy.
The technical scheme adopted by the invention is as follows:
a method for simultaneously detecting linagliptin and saxagliptin in health food comprises the following detection steps:
(1) preparation of the solution
① 0.1.1% aqueous formic acid 1mL of formic acid was measured and diluted with water to 1000 mL.
② 0.1.1% formic acid acetonitrile solution formic acid 1mL is measured and diluted to 1000mL with acetonitrile.
(2) Preparation of Standard solutions
① Standard stock solutions, accurately weighing 10.0mg (accurate to 0.0001g) of each of saxagliptin and linagliptin standard substances, respectively placing in 10mL volumetric flasks, dissolving with methanol, metering to a certain volume, shaking up to obtain standard stock solutions with concentration of 1mg/mL, storing at-18 deg.C, and prolonging effective period of 3 months.
② mixing standard intermediate solution, accurately sucking 100 μ L of saxagliptin and 1000 μ L of linagliptin standard stock solution into a 100mL volumetric flask, adding methanol to constant volume to scale, and shaking to obtain mixed standard intermediate solution, wherein the concentration of linagliptin is 10 μ g/mL, and the concentration of saxagliptin is 1 μ g/mL.
③ matrix matching mixed standard working solution, accurately sucking mixed standard intermediate solution 0.02mL, 0.04mL, 0.08mL, 0.1mL and 0.5mL respectively, diluting with blank sample solution to scale in a 10mL volumetric flask, shaking up to obtain series matrix matching standard working solution, wherein the concentrations of saxagliptin are 2ng/mL, 4ng/mL, 8ng/mL, 10ng/mL and 50ng/mL in sequence, and the concentrations of linagliptin are 20ng/mL, 40ng/mL, 80ng/mL, 100ng/mL and 500ng/mL in sequence.
The blank sample solution is a sample solution obtained after pretreatment of a sample which does not contain saxagliptin and linagliptin.
(3) Method of pretreating sample (preparation of sample solution)
Accurately weighing 1g (accurate to 0.0001g) of sample, placing in a 50mL centrifuge tube with a plug, adding 45mL of methanol, mixing uniformly by vortex, carrying out ultrasonic extraction for 30min, centrifuging for 3min at 12000r/min, transferring supernatant into a 50mL volumetric flask, cooling, adding methanol to a constant volume to scale, shaking uniformly, and filtering with a 0.22 μm filter membrane. Taking the filtrate, diluting the filtrate to a linear range according to the actual concentration, and analyzing the filtrate by using a liquid chromatography-tandem mass spectrometer.
(4) Reference condition for liquid chromatography-tandem mass spectrometry
① reference conditions for chromatography
a) A chromatographic column: agilent SB-Aq (2.1 mm. times.100 mm, 1.8 μm);
b) mobile phase: a: 0.1% aqueous formic acid solution, B: 0.1 percent formic acid acetonitrile solution, and gradient elution is carried out;
c) flow rate: 0.2 mL/min;
d) column temperature: 25 ℃;
e) sample introduction amount: 1 mu L of the solution;
the gradient elution procedure is shown in table 1.
② reference conditions for mass spectrometry
An ion source: electrospray ion source (ESI);
the detection mode is as follows: multiple Reaction Monitoring (MRM);
the scanning mode is as follows: a positive ion mode;
capillary voltage: 4000V;
ion source temperature: 300 ℃;
flow rate of drying gas: 15L/min;
atomizing gas pressure: 35 psi;
temperature of sheath gas: 350 ℃; sheath gas (N2) flow rate: 11L/min;
other mass spectral parameters are shown in table 2.
TABLE 1 gradient elution procedure
| Time/min | Mobile phase A/%) | Mobile phase B/%) |
| 0 | 98 | 2 |
| 6.5 | 40 | 60 |
| 6.6 | 10 | 90 |
| 7.6 | 10 | 90 |
| 7.7 | 98 | 2 |
| 10 | 98 | 2 |
TABLE 2 qualitative, quantitative ion and Mass Spectrometry parameters for the Compounds
*And (4) quantifying ion pairs.
(5) Qualitative analysis
And (3) determining the sample and the matrix matching standard working solution according to the high performance liquid chromatography-tandem mass spectrometry conditions, recording the chromatographic retention time of each compound in the sample and the standard solution, taking the percentage of the abundance of the strongest ions as the relative abundance of the qualitative ion pair, and recording the relative ion abundance of corresponding components in the sample and the matrix matching standard working solution with the same concentration. When a chromatographic peak corresponding to the retention time of a chromatographic peak of a standard of 2 compounds is detected in a sample (within. + -. 2.5%), and the allowable deviation of the relative ion abundance does not exceed the range specified in Table 3, the detection of the corresponding compound in the sample can be determined.
TABLE 3 maximum permissible deviation of relative ion abundance in qualitative confirmation
| Relative abundance (%) | Maximum allowable deviation (%) |
| k>50％ | ±20 |
| 50％≥k>20％ | ±25 |
| 20％≥k>10％ | ±30 |
| K≤10％ | ±50 |
(6) Quantitative determination
And (3) respectively measuring the matrix matching mixed standard working solution according to the reference conditions of the instrument to obtain the chromatographic peak area of the corresponding standard solution. And drawing a standard curve by taking the concentration of the matrix matching mixed standard working solution as an abscissa and taking the peak area of a chromatographic peak as an ordinate.
The content of the target in the sample solution is determined by a standard curve external standard method. The content of the target in the sample is calculated according to the formula (1).
In the formula:
x represents the content of each target object in the sample, mg/kg;
c-concentration of each target in the sample solution, ng/mL, read from the standard curve;
v, the final constant volume of the sample solution, namely mL;
m-mass represented by the sample solution, g;
k is the dilution multiple.
The calculation results retain three significant digits.
Compared with the prior art, the invention has the beneficial effects that: the method has the advantages of simplicity, convenience, high efficiency, rapidness and accuracy, and can simultaneously and accurately determine the contents of linagliptin and saxagliptin in the health-care food.
Drawings
FIG. 1 is an MRM chromatogram of a blank sample of oral liquid without saxagliptin and linagliptin; wherein, (a) is saxagliptin qualitative ions, (b) is saxagliptin quantitative ions, (c) is linagliptin qualitative ions, and (d) is linagliptin quantitative ions;
FIG. 2 is an MRM chromatogram of an oral liquid matrix standard solution with saxagliptin of 2ng/mL and linagliptin of 20 ng/mL; wherein, (a) is saxagliptin qualitative ions, (b) is saxagliptin quantitative ions, (c) is linagliptin qualitative ions, and (d) is linagliptin quantitative ions;
FIG. 3 is a schematic diagram of a matrix calibration curve of saxagliptin oral liquid;
FIG. 4 is a schematic diagram of a matrix standard curve of an oral liquid of linagliptin;
FIG. 5 is an MRM chromatogram of a blank sample of a tablet without saxagliptin and linagliptin; wherein, (a) is saxagliptin qualitative ion, (b) is saxagliptin quantitative ion, (c) is linagliptin qualitative ion, and (d) is linagliptin quantitative ion;
FIG. 6 is an MRM chromatogram of a tablet matrix standard solution with saxagliptin at 2ng/mL and linagliptin at 20 ng/mL; wherein, (a) is saxagliptin qualitative ions, (b) is saxagliptin quantitative ions, (c) is linagliptin qualitative ions, and (d) is linagliptin quantitative ions;
FIG. 7 is a schematic diagram of a tablet matrix calibration curve for saxagliptin;
FIG. 8 is a schematic diagram of a tablet matrix calibration curve for linagliptin;
FIG. 9 is an MRM chromatogram of a blank sample of a capsule without saxagliptin and linagliptin; wherein, (a) is saxagliptin qualitative ion, (b) is saxagliptin quantitative ion, (c) is linagliptin qualitative ion, and (d) is linagliptin quantitative ion;
FIG. 10 is an MRM chromatogram of a standard solution of capsule matrix with saxagliptin of 2ng/mL and linagliptin of 20 ng/mL; wherein, (a) is saxagliptin qualitative ions, (b) is saxagliptin quantitative ions, (c) is linagliptin qualitative ions, and (d) is linagliptin quantitative ions;
FIG. 11 is a schematic diagram of a standard curve for the capsule matrix of saxagliptin;
fig. 12 is a schematic diagram of a standard curve of the capsule matrix of linagliptin.
Detailed Description
The present invention will be further described with reference to the following examples.
Example 1 determination of linagliptin and saxagliptin in oral liquid
(1) Sample pretreatment
Weighing 1g (accurate to 0.0001g) of oral liquid to be detected, placing the oral liquid into a 50mL centrifuge tube with a plug, adding 45mL of methanol, uniformly mixing by vortex, carrying out ultrasonic extraction for 30min, centrifuging for 3min at 12000r/min, transferring supernatant into a 50mL volumetric flask, cooling, adding methanol to a constant volume to scale, shaking up, and filtering by using a 0.22 mu m filter membrane. Taking the filtrate, diluting the filtrate to a linear range according to the actual concentration, and analyzing the filtrate by using a liquid chromatography-tandem mass spectrometer. (2) Preparation of standard solution and blank solution
① Standard stock solutions, accurately weighing 10.0mg (accurate to 0.0001g) of each of saxagliptin and linagliptin standard substances, respectively placing in 10mL volumetric flasks, dissolving with methanol, metering to a certain volume, shaking up to obtain a single standard stock solution with a concentration of 1mg/mL, storing at-18 deg.C, and prolonging the shelf life of 3 months.
② mixing standard intermediate solution, accurately sucking 100 μ L of saxagliptin and 1000 μ L of linagliptin standard stock solution into a 100mL volumetric flask, adding methanol to constant volume to scale, and shaking to obtain mixed standard intermediate solution, wherein the concentration of linagliptin is 10.0 μ g/mL, and the concentration of saxagliptin is 1.0 μ g/mL.
③ oral liquid matrix blank solution, which is to take oral liquid samples without saxagliptin and linagliptin, and carry out pretreatment according to the step (1) to obtain the oral liquid matrix blank solution, wherein the oral liquid sample blank matrix MRM chromatogram is shown in figure 1.
④ preparation of matrix matching standard solution, which is to accurately absorb 0.02mL, 0.04mL, 0.08mL, 0.1mL and 0.5mL of mixed standard intermediate solution respectively, dilute the mixed standard intermediate solution to scale with an oral liquid matrix blank solution in a 10mL volumetric flask, shake the solution evenly to serve as series matrix standard working solutions S (1) to S (5), wherein the concentrations of saxagliptin are sequentially 2ng/mL, 4ng/mL, 8ng/mL, 10ng/mL and 50ng/mL, and the concentrations of linagliptin are sequentially 20ng/mL, 40ng/mL, 80ng/mL, 100ng/mL, 500ng/mL, 2ng/mL of saxagliptin and 20ng/mL of linagliptin are shown in figure 2 of an MRM chromatogram of the oral liquid matrix standard solution.
(3) Liquid chromatography-Mass Spectrometry/Mass Spectrometry (HPLC-MS/MS) determination
And respectively injecting the series of oral liquid matrix standard working solutions S (1) -S (5), the oral liquid matrix blank solution and the sample solution into HPLC-MS/MS for determination, establishing a matrix matching correction curve, and quantifying by an external standard method.
① the chromatographic conditions are:
a chromatographic column: agilent SB-Aq (2.1 mm. times.100 mm, 1.8 μm), or equivalent;
mobile phase: a is 0.1% formic acid water, B is 0.1% formic acid acetonitrile solution, the procedure of gradient elution is shown in Table 4;
flow rate: 0.2 mL/min;
column temperature: 25 ℃;
sample introduction amount: 1 μ L.
TABLE 4 gradient elution schedule
| Gradient time/min | Mobile phase A/%) | Mobile phase B/%) |
| 0 | 98 | 2 |
| 6.5 | 40 | 60 |
| 6.6 | 10 | 90 |
| 7.6 | 10 | 90 |
| 7.7 | 98 | 2 |
| 10 | 98 | 2 |
② Mass Spectrometry conditions were:
an ion source: electrospray ion source (ESI);
the detection mode is as follows: multiple Reaction Monitoring (MRM);
the scanning mode is as follows: a positive ion mode;
capillary voltage: 4000V;
temperature of the drying gas: 300 ℃;
flow rate of drying gas: 15L/min;
atomizing gas pressure: 35 psi;
temperature of sheath gas: 350 ℃; sheath gas (N2) flow rate: 11L/min;
other mass spectral parameters are shown in table 5.
TABLE 5 qualitative, quantitative ion and Mass Spectrometry parameters for the Compounds
Quantitation of ion pairs.
The obtained calibration curves are shown in table 6, fig. 3, and fig. 4.
TABLE 6 matrix matching calibration curves for saxagliptin and linagliptin in oral liquids
| Compound (I) | Regression equation | Linear range | Coefficient of correlation (r)2) | Detection limit |
| Saxagliptin | Y＝29996.1906X+10085.1120 | (2～50) | 0.9999 | (0.1) |
| Linagliptin | Y＝889.4713X–4397.3791 | 20～500 | 0.9995 | 1.0 |
(4) Recovery and repeatability of spiked samples
1.0mg/kg, 2.0mg/kg and 10.0mg/kg of linagliptin standard solutions and 0.1mg/kg, 0.2mg/kg and 1.0mg/kg of linagliptin standard solutions are respectively added into the oral liquid without the saxagliptin and linagliptin, and the content is determined according to the steps after standing for 30min at room temperature. Each addition level was assayed in parallel 6 times. The results of the measurements were compared to the theoretical additive concentrations to obtain the average recovery and relative standard deviation, the results of which are shown in Table 7.
Table 7 recovery and precision of saxagliptin and linagliptin in oral liquid (n ═ 6)
Table 7 shows that the average recovery ranges of the three spiked levels of saxagliptin are 96.5% to 103.6%, and the relative standard deviation ranges are 6.6% to 8.7%; the average recovery rate range of linagliptin is 87.6-92.5%, and the relative standard deviation range is 3.2-5.3%, which indicates that the recovery rate and the repeatability of the method in the embodiment both meet the requirements.
(5) Sensitivity of the probe
Taking the lowest detection limit of the actually added sample as a detection limit, in the embodiment, the detection limit of linagliptin in the oral liquid is 1.0 mg/kg; the detection limit of the saxagliptin in the oral liquid is 0.1 mg/kg.
Example 2 determination of linagliptin and saxagliptin in tablets
(1) The preparation method of the tablet sample to be measured, the pretreatment method, the preparation method of the standard solution and the blank solution, and the measurement method were the same as those of example 1. The MRM chromatogram of the blank matrix of the tablet sample is shown in FIG. 5; the MRM chromatogram of the tablet matrix standard solution with saxagliptin of 2ng/mL and linagliptin of 20ng/mL is shown in FIG. 6; the obtained standard curves are shown in table 8, fig. 7, and fig. 8.
TABLE 8 matrix matching calibration curves for saxagliptin, linagliptin in tablets
(2) Recovery and repeatability of spiked samples
1.0mg/kg, 2.0mg/kg and 10.0mg/kg of linagliptin standard solutions and 0.1mg/kg, 0.2mg/kg and 1.0mg/kg of linagliptin standard solutions are respectively added into the tablets without the saxagliptin and linagliptin, and the contents of the solutions are measured according to the steps after standing at room temperature for 30 min. Each addition level was assayed in parallel 6 times. The results of the measurements were compared to the theoretical additive concentrations to give average recovery and relative standard deviation, the results of which are shown in table 9.
Table 9 recovery and precision of saxagliptin and linagliptin in tablets (n ═ 6)
Table 9 shows that the average recovery ranges of the three spiked levels of saxagliptin are 75.6% to 83.9%, and the relative standard deviation ranges are 5.6% to 8.5%; the average recovery rate range of linagliptin is 73.6-82.3%, and the relative standard deviation range is 3.8-8.6%, which indicates that the recovery rate and the repeatability of the method of the embodiment both meet the requirements.
(3) Sensitivity of the probe
Taking the lowest detection limit of the actually added samples as a detection limit, in the embodiment, the detection limit of linagliptin in the tablet is 1.0 mg/kg; the detection limit of saxagliptin in the tablets was 0.1 mg/kg.
Example 3 determination of residual amounts of linagliptin and saxagliptin in capsules
(1) The pretreatment method, the preparation method of the standard solution and the blank solution, and the measurement method of the capsule sample to be measured were the same as those in example 1. The MRM chromatogram of the blank matrix of the capsule sample is shown in FIG. 9; the MRM chromatogram of the capsule matrix standard solution with saxagliptin of 2ng/mL and linagliptin of 20ng/mL is shown in FIG. 10; the obtained standard curves are shown in table 10, fig. 11, and fig. 12.
TABLE 10 matrix matching calibration curves for saxagliptin, linagliptin in capsules
(2) Recovery and repeatability of spiked samples
1.0mg/kg, 2.0mg/kg and 10.0mg/kg of linagliptin standard solutions and 0.1mg/kg, 0.2mg/kg and 1.0mg/kg of linagliptin standard solutions are respectively added into the capsules without the sagliptin and linagliptin, and the contents of the solutions are determined according to the steps after the solutions are kept stand at room temperature for 30 min. Each addition level was assayed in parallel 6 times. The results of the measurements were compared with the theoretical additive concentrations to obtain the average recovery and relative standard deviation, the results of which are shown in Table 11.
Table 11 recovery and precision of saxagliptin and linagliptin in capsules (n ═ 6)
Table 11 shows that the average recovery ranges of the three spiked levels of saxagliptin are 89.3% to 95.3%, and the relative standard deviation ranges are 6.3% to 9.2%; the average recovery rate range of linagliptin is 93.5-97.8%, and the relative standard deviation range is 4.8-8.6%, which indicates that the recovery rate and the repeatability of the method meet the requirements
(3) Sensitivity of the probe
Taking the lowest detection limit of the actually added sample as a detection limit, in the embodiment, the detection limit of linagliptin in the capsule is 1.0 mg/kg; the detection limit of saxagliptin in the capsule is 0.1 mg/kg.
Appendix A information on Compounds
TABLE A.1 Compound Chinese name, English name, CAS number, molecular formula, relative molecular mass
Appendix A illustrates: CAS number, molecular formula and relative molecular mass of saxagliptin and linagliptin.
The above-mentioned embodiments only express the embodiments of the present invention, but not should be understood as the limitation of the scope of the invention patent, it should be noted that those skilled in the art can make several variations and modifications without departing from the concept of the present invention, and these all fall into the protection scope of the present invention.
Claims (2)
1. A method for simultaneously determining linagliptin and saxagliptin in a health food is characterized by comprising the following steps of:
(1) preparing 0.1% formic acid water solution and 0.1% formic acid acetonitrile solution;
(2) preparation of Standard solutions
① Standard stock solutions, namely accurately weighing standard substances of saxagliptin and linagliptin, respectively dissolving the substances with methanol to prepare standard stock solutions with the concentrations of 1mg/mL, and storing at-18 ℃;
② mixing standard intermediate solution, namely placing the standard stock solutions of saxagliptin and linagliptin in volumetric flasks, and adding methanol to prepare the mixed standard intermediate solution, wherein the concentration of linagliptin is 10 μ g/mL, and the concentration of linagliptin is 1 μ g/mL;
③ matrix matching and mixing standard working solution, namely putting quantitative mixing standard intermediate solution into five volumetric flasks respectively, adding blank sample solution for dilution to serve as serial matrix matching standard working solutions, wherein the concentrations of saxagliptin are 2ng/mL, 4ng/mL, 8ng/mL, 10ng/mL and 50ng/mL in sequence, and the concentrations of linagliptin are 20ng/mL, 40ng/mL, 80ng/mL, 100ng/mL and 500ng/mL in sequence;
(3) method for pretreating sample, and sample solution prepared by the method
Placing the sample in a centrifuge tube with a plug, adding methanol, performing vortex mixing, performing ultrasonic extraction, performing centrifugal treatment, and filtering the supernatant with a 0.22 μm filter membrane; taking the filtrate, properly diluting the filtrate to a linear range according to the actual concentration, and analyzing the filtrate by using a liquid chromatography-tandem mass spectrometer;
(4) reference condition for liquid chromatography-tandem mass spectrometry
① reference conditions for chromatography
a) A chromatographic column: agilent Agilent SB-Aq;
b) mobile phase: a: 0.1% aqueous formic acid solution, B: 0.1 percent formic acid acetonitrile solution, and gradient elution is carried out;
c) flow rate: 0.2 mL/min;
d) column temperature: 25 ℃;
e) sample introduction amount: 1 mu L of the solution;
the gradient elution procedure is shown in table 1;
TABLE 1 gradient elution procedure
② reference conditions for mass spectrometry
a) An ion source: electrospray ion source (ESI);
b) the detection mode is as follows: multiple Reaction Monitoring (MRM);
c) the scanning mode is as follows: a positive ion mode;
d) capillary voltage: 4000V;
e) ion source temperature: 300 ℃;
f) flow rate of drying gas: 15L/min;
g) atomizing gas pressure: 35 psi;
h) temperature of sheath gas: 350 ℃; sheath gas N2Flow rate: 11L/min;
i) other mass spectral parameters are shown in table 2;
TABLE 2 qualitative, quantitative ion and Mass Spectrometry parameters for the Compounds
*Quantifying ion pairs;
(5) qualitative analysis
Determining a sample and a matrix matching standard working solution according to a high performance liquid chromatography-tandem mass spectrometry condition, recording the chromatographic retention time of each compound in the sample and the standard solution, taking the percentage of the strongest ion abundance as the relative abundance of a qualitative ion pair, and recording the relative ion abundance of corresponding components in the sample and the matrix matching standard working solution with the same concentration; when a chromatographic peak corresponding to the retention time of a chromatographic peak of a standard substance of 2 compounds is detected in the sample, namely the variation range is within +/-2.5 percent, and the allowable deviation of the relative ion abundance does not exceed the range specified in the table 3, the corresponding compound can be determined to be detected in the sample;
TABLE 3 maximum permissible deviation of relative ion abundance in qualitative confirmation
(6) Quantitative determination
Respectively measuring the matrix matching mixed standard working solution according to the reference conditions of the instrument to obtain the chromatographic peak area of the corresponding standard solution; taking the concentration of the matrix matching mixed standard working solution as a horizontal coordinate, taking the peak area of a chromatographic peak as a vertical coordinate, and drawing a standard curve;
determining the content of the target object in the sample solution by using a standard curve external standard method;
the content of the target object in the sample is calculated according to the formula
Calculating; in the formula: x is the content of each target object in the sample, mg/kg; c is the concentration of each target in the sample solution read from the standard curve, ng/mL; v is the final constant volume of the sample solution, mL; m is the mass represented by the sample solution, g; k is the dilution factor.
2. The method for simultaneously determining linagliptin and saxagliptin in a health food according to claim 1, wherein in the step (3), the ultrasonic extraction time is 30 min; the centrifugal speed is 12000r/min, and the centrifugal time is 3 min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201911289056.2A CN111189949A (en) | 2019-12-13 | 2019-12-13 | Method for simultaneously detecting linagliptin and saxagliptin in health food |
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| CN113607846A (en) * | 2021-08-03 | 2021-11-05 | 苏州科技城医院 | Method for determining dapagliflozin concentration in blood plasma |
| CN114487200A (en) * | 2022-01-28 | 2022-05-13 | 大连市检验检测认证技术服务中心 | Mass spectrum detection method for screening and quantifying prometryn metabolite by adopting plant hydroponics technology |
| CN114577928A (en) * | 2022-01-27 | 2022-06-03 | 大连市检验检测认证技术服务中心 | Method for detecting Edwardsient in food by using deuterium internal standard substance |
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| CN112326842A (en) * | 2020-11-27 | 2021-02-05 | 合肥科颖医药科技有限公司 | Saxagliptin quantitative detection and analysis method |
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| CN114487200A (en) * | 2022-01-28 | 2022-05-13 | 大连市检验检测认证技术服务中心 | Mass spectrum detection method for screening and quantifying prometryn metabolite by adopting plant hydroponics technology |
| CN114487200B (en) * | 2022-01-28 | 2024-03-22 | 大连市检验检测认证技术服务中心 | Mass spectrum detection method for screening and quantifying prometryn metabolite by adopting plant hydroponic technology |
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