CN114324661A - Method for detecting nitrosamine impurities in nitrogen-containing industrial wastewater - Google Patents
Method for detecting nitrosamine impurities in nitrogen-containing industrial wastewater Download PDFInfo
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Abstract
The invention provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which comprises the following steps: (1) enriching nitrosamine impurities in the wastewater by using an active carbon solid phase extraction column; (2) and (2) detecting the enrichment liquid obtained in the step (1) by adopting a high performance liquid chromatography-triple quadrupole tandem mass spectrometer, and calculating the content of nitrosamine impurities in the enrichment liquid by adopting an external standard method. The detection method for the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater provided by the invention has the advantages of excellent performance, good precision and high accuracy; the quantitative limit is low, the sensitivity is high, and the trace detection of the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater can be realized.
Description
Technical Field
The invention belongs to the technical field of chemical detection, and particularly relates to a method for detecting nitrosamine impurities in nitrogen-containing industrial wastewater.
Background
Nitrosamines are one of the N-nitroso compounds having the general structure R2(R1)N-N＝O。R1And R2Can be the sameMay also be different when R1Is equal to R2When referred to as symmetric nitrosamines, such as N-nitrosodimethylamine and N-nitrosodiethylamine; when R is1Is not equal to R2And are referred to as asymmetric nitrosamines, such as N-nitrosomethylethylamine, N-nitrosohexylamine, and the like.
N-nitrosodimethylamine has a carcinogenic action, and the mechanism is as follows: under the action of enzyme, the carbon atom of alkyl is hydroxylated to form hydroxyl nitrosamine, then dealdehydization is carried out to form monoalkylnitrosamine, and then denitrification is carried out to form electrophilic alkyl free radical, and the electrophilic alkyl free radical can alkylate nucleic acid in liver or cell to generate alkyl guanine, thereby causing cytogenetic mutation and displaying carcinogenicity.
CN101596447A discloses a filter medium for removing NDMA in water and a preparation method thereof, the method is that various raw materials such as zeolite powder, diatomite, molecular sieve, active carbon and the like are mixed according to a certain proportion, and then the mixture is pressed, sintered and cooled. The method is adopted to remove the NDMA in the water, and the removal rate reaches more than 92 percent; however, the method has complex filler proportion and needs means such as pressing and the like during preparation; if the method is applied to the solid phase extraction technology, the problems of complicated manufacturing process and high cost are exposed.
CN112946107A discloses an analysis method of N-nitrosodimethylamine and N-nitrosodiethylamine in argatroban bulk drugs or preparations, which adopts a method of combining ultra-performance liquid chromatography and mass spectrometry, and comprises the following steps: preparing a test solution and a reference solution: dissolving a sample to be tested by using methanol as a solvent, and preparing to obtain a test solution; dissolving a standard substance of N-nitrosodimethylamine and N-nitrosodiethylamine by using methanol as a solvent to prepare a reference substance solution; respectively injecting sample into the test solution and the reference solution, detecting by using an ultra-high performance liquid chromatography-mass spectrometer, and recording a chromatogram; calculating the contents of N-nitrosodimethylamine and N-nitrosodiethylamine according to an external standard method; however, this method requires the use of gradient elution and is only suitable for the analysis of N-nitrosodimethylamine, N-nitrosodiethylamine in argatroban drug substances or preparations.
The method can not accurately quantify the content of the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater. Therefore, how to provide a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater becomes a problem to be solved urgently at present.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater. The method for detecting the nitrosamine impurities in the nitrogenous drug industrial wastewater can be used for accurately and quantitatively detecting N-nitrosodimethylamine and N-nitrosodiethylamine through one-time sample injection, so that the detection efficiency is improved.
The invention provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which comprises the following steps:
(1) enriching nitrosamine impurities in the wastewater by using an active carbon solid phase extraction column;
(2) and (2) detecting the enrichment liquid obtained in the step (1) by adopting a high performance liquid chromatography-triple quadrupole tandem mass spectrometer, and calculating the content of nitrosamine impurities in the enrichment liquid by adopting an external standard method.
The detection method for the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater provided by the invention has the advantages of excellent performance, good precision and high accuracy; the quantitative limit is low, the sensitivity is high, and the trace detection of the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater can be realized.
In the present invention, in the step (1) and the step (2), the nitrosamine-based impurities include N-Nitrosodimethylamine (NDMA) and/or N-Nitrosodiethylamine (NDEA).
In the invention, in the step (1), the activated carbon used in the activated carbon solid-phase extraction column comprises any one or a combination of at least two of coal-based activated carbon, wood activated carbon, fruit shell and kernel activated carbon or resin activated carbon, and is preferably fruit shell and kernel activated carbon.
Preferably, the shell and kernel type activated carbon comprises any one or a combination of at least two of coconut shell carbon, apricot kernel carbon or walnut carbon, and is preferably coconut shell carbon.
In the present invention, in the step (1),the particle size of the activated carbon adopted by the activated carbon solid phase extraction column is 80-120 meshes, for example, 80 meshes, 85 meshes, 90 meshes, 95 meshes, 100 meshes, 105 meshes, 110 meshes, 115 meshes, 120 meshes and the like; the specific surface area is 4000-2Per g, may be, for example, 4000m2/g、4200m2/g、4400m2/g、4600m2/g、4800m2/g、5000m2/g、5200m2/g、5400m2/g、5600m2/g、5800m2/g、6000m2And/g, etc.
Preferably, the filling amount of the activated carbon solid phase extraction column is 1500-2500mg, for example 1500mg, 1600mg, 1700mg, 1800mg, 1900mg, 2000mg, 2100mg, 2200mg, 2300mg, 2400mg, 2500mg, etc.
In the present invention, in the step (1), the pressure of the enrichment is normal pressure; the atmospheric pressure refers to an atmospheric pressure, namely the gas pressure generated by the atmosphere in which people usually live; one standard atmospheric pressure is 101325 Pa.
Preferably, in step (1), the enrichment comprises activation, loading and elution performed sequentially.
Preferably, the solvent used for the activation comprises dichloromethane, methanol and water.
Preferably, the volume ratio of the dichloromethane, the methanol and the water is 1 (1.5-2.5) to (5.5-6.5), and can be 1:1.5:5.5, 1:1.7:6.2, 1:1.9:5.7, 1:2.1:6.2, 1:2.5:6.3, 1:2.5:6.5 and the like.
Preferably, the water comprises ultrapure water.
Preferably, the flow rate of the sample is 1-2mL/min, and may be, for example, 1mL/min, 1.1mL/min, 1.2mL/min, 1.3mL/min, 1.4mL/min, 1.5mL/min, 1.6mL/min, 1.7mL/min, 1.8mL/min, 1.9mL/min, 2mL/min, or the like.
Preferably, the solvent used for elution comprises any one of ethanol, n-hexanol or methanol or a combination of at least two thereof, preferably methanol.
Preferably, the flow rate of the elution is 1-2mL/min, and may be, for example, 1mL/min, 1.1mL/min, 1.2mL/min, 1.3mL/min, 1.4mL/min, 1.5mL/min, 1.6mL/min, 1.7mL/min, 1.8mL/min, 1.9mL/min, 2mL/min, or the like.
In the present invention, in the step (2), the mobile phase A comprises 0.1 to 1 vol% of glacial acetic acid aqueous solution, which may be, for example, 0.1 vol%, 0.2 vol%, 0.3 vol%, 0.4 vol%, 0.5 vol%, 0.6 vol%, 0.7 vol%, 0.8 vol%, 0.9 vol%, 1 vol% or the like; the mobile phase B comprises 0.1 to 1 vol% of a methanol glacial acetic acid solution, and may be, for example, 0.1 vol%, 0.2 vol%, 0.3 vol%, 0.4 vol%, 0.5 vol%, 0.6 vol%, 0.7 vol%, 0.8 vol%, 0.9 vol%, 1 vol%, or the like.
Preferably, the volume ratio of the mobile phase A and the mobile phase B is 1 (0.5-3), and can be 1:0.5, 1:0.7, 1:1, 1:1.2, 1:1.5, 1:1.7, 1:2, 1:2.2, 1:2.5, 1:2.8, 1:3, etc.
Preferably, isocratic elution is adopted in the detection of the high performance liquid chromatography.
In the present invention, in the step (2), the chromatographic column in the detection of the high performance liquid chromatography comprises Agilent Poroshell 120SB-AQ or Waters Xbridge C18, preferably Waters Xbridge C18.
Preferably, the inner diameter of the chromatography column is 4-5mm, for example, 4mm, 4.2mm, 4.4mm, 4.6mm, 4.8mm, 5mm, etc.; the column length is 80-120mm, such as 80mm, 90mm, 100mm, 110mm, 120 mm; the particle diameter is 4.5 to 5.5. mu.m, and may be, for example, 4.5. mu.m, 4.7. mu.m, 4.9. mu.m, 5.1. mu.m, 5.3. mu.m, 5.5. mu.m, or the like.
In the present invention, in the step (2), the flow rate in the detection by the high performance liquid chromatography is 0.45 to 0.55mL/min, and may be, for example, 0.45mL/min, 0.47mL/min, 0.49mL/min, 0.51mL/min, 0.53mL/min, 0.55mL/min, or the like.
Preferably, in step (2), the sample injection volume in the detection by high performance liquid chromatography is 15-25 μ L, such as 15 μ L, 16 μ L, 17 μ L, 18 μ L, 19 μ L, 20 μ L, 21 μ L, 22 μ L, 23 μ L, 24 μ L, 25 μ L, etc.
Preferably, in the step (2), the column temperature in the detection by high performance liquid chromatography is 35 to 45 ℃, and may be, for example, 35 ℃, 36 ℃, 37 ℃, 38 ℃, 39 ℃, 40 ℃, 41 ℃, 42 ℃, 43 ℃, 44 ℃, 45 ℃ or the like.
Preferably, in step (2), the sample chamber temperature in the detection by high performance liquid chromatography is 5-15 ℃, for example, 5 ℃, 6 ℃, 7 ℃, 8 ℃, 9 ℃, 10 ℃, 11 ℃, 12 ℃, 13 ℃, 14 ℃, 15 ℃ and the like.
Preferably, in the step (2), the running time in the detection of the high performance liquid chromatography is 5-7min, such as 5min, 6min, 7min and the like.
In the invention, in the step (2), the ion source in the detection of the triple quadrupole tandem mass spectrometer is APCI, and the polarity is positive.
Preferably, in the step (2), the drying gas temperature in the detection of the triple quadrupole tandem mass spectrometer is 240-260 ℃, and may be, for example, 240 ℃, 242 ℃, 246 ℃, 248 ℃, 250 ℃, 252 ℃, 254 ℃, 256 ℃, 258 ℃, 260 ℃ and the like; the dry gas flow rate is 3-6L/min, and may be, for example, 3L/min, 3.5L/min, 4L/min, 4.5L/min, 5L/min, 5.5L/min, 6L/min, etc.
Preferably, in the step (2), in the detection of the triple quadrupole tandem mass spectrometer, the temperature of the evaporation chamber is 340-; the atomization gas pressure is 30-40psi, and may be, for example, 30psi, 31psi, 32psi, 33psi, 34psi, 35psi, 36psi, 37psi, 38psi, 39psi, 40psi, etc.
Preferably, in the step (2), the capillary voltage in the detection of the triple quadrupole tandem mass spectrometer is 1400-1600V, which may be 1400V, 1420V, 1440V, 1460V, 1480V, 1500V, 1520V, 1540V, 1560V, 1580V, 1600V, etc.; the corona needle current is 4 to 7. mu.A, and may be, for example, 4. mu.A, 5. mu.A, 6. mu.A, 7. mu.A, or the like.
As a preferred technical scheme, the method for detecting the nitrosamine impurities in the nitrogen-containing medicament industrial wastewater comprises the following steps:
(A) and (3) activation: activated carbon solid phase extraction columns are activated by dichloromethane, methanol and water according to the volume ratio of 1 (1.5-2.5) to (5.5-6.5); the active carbon adopted in the active carbon solid phase extraction column is coconut shell carbon;
(B) loading: loading at a speed of 1-2 mL/min;
(C) and (3) elution: eluting with methanol solvent at a speed of 1-2mL/min to obtain a sample solution;
(D) and (3) detection: detecting the test solution obtained in the step (C) by adopting a high performance liquid chromatography-triple quadrupole tandem mass spectrometer, and calculating the content of nitrosamine impurities in the eluted solution by adopting an external standard method;
wherein, the testing conditions of the high performance liquid chromatography comprise: the mobile phase A is 0.1-1 vol% glacial acetic acid aqueous solution, the mobile phase B is 0.1-1 vol% glacial acetic acid methanol solution, and the volume ratio of the mobile phase A to the mobile phase B is 1 (0.5-3); the chromatographic column is Waters Xbridge C18, the inner diameter is 4-5mm, the column length is 80-120mm, the particle size is 4.5-5.5 μm, the flow rate is 0.45-0.55mL/min, the sample injection volume is 15-25 μ L, the column temperature is 35-45 ℃, the sample chamber temperature is 5-15 ℃, and the running time is 5-7 min;
wherein, the test conditions of the triple quadrupole tandem mass spectrometry comprise: the ion source is APCI, the polarity is positive, the temperature of the drying gas is 240-260 ℃, the flow rate of the drying gas is 3-6L/min, the temperature of the evaporation chamber is 340-360 ℃, the pressure of the atomizing gas is 30-40psi, the voltage of the capillary tube is 1400-1600V, and the current of the corona needle is 5-7 muA.
Compared with the prior art, the invention has the following beneficial effects:
(1) the method for detecting the nitrosamine impurities in the nitrogenous drug industrial wastewater can accurately quantify N-nitrosodimethylamine and N-nitrosodiethylamine simultaneously by one-time sample injection, thereby improving the detection efficiency;
(2) the detection method for the nitrosamine impurities in the nitrogen-containing drug industrial wastewater provided by the invention has excellent performance, the relative standard deviation range of the components of the nitrosamine impurities is 1.3-7.3%, and the standard recovery rate is 82.8-99.9%, which shows that the detection method provided by the invention has good precision and high accuracy;
(3) the method for detecting the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater provided by the invention has the advantages of low quantitative limit and high sensitivity, and can realize trace detection of the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater.
Drawings
FIG. 1 is a graph showing a standard curve of N-nitrosodimethylamine.
FIG. 2 is a graph showing a standard N-nitrosodiethylamine.
Detailed Description
The technical solution of the present invention is further explained by the following embodiments. It should be understood by those skilled in the art that the examples are only for the understanding of the present invention and should not be construed as the specific limitations of the present invention.
The nitrogen-containing industrial wastewater used in the following examples and comparative examples is the same sample, and the sources of the components in the following examples are as follows:
| components | Manufacturer of the product | Number plate |
| EU52112M6 | UCT | EU52112M6 |
| N-nitrosodimethylamine | TCI | D0761 |
| N-nitrosodiethylamine | Sigma-Aldrich | 442687 |
Preparation example
Preparing a standard sample: respectively adding 400uL, 600uL and 800uL of mixed standard working solution into 100mL of wastewater sample to be detected, wherein the concentrations of N-nitrosodimethylamine and N-nitrosodiethylamine in the mixed standard working solution are both 1 microgram/mL; three spiked samples of different concentrations were obtained.
Unlabeled sample: 100mL of wastewater sample to be tested.
Example 1
The embodiment provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which comprises the following steps:
testing a sample: preparation examples provided labeled and unlabeled samples
(1) And (3) activation: taking an EU52112M6 activated carbon solid phase extraction column (coconut shell carbon) (2000mg, 6mL), sequentially activating with 3mL of dichloromethane, 6mL of methanol and 18mL of ultrapure water, and ensuring that a solution in the column does not run dry in the activation process;
(2) loading: then, sampling at the speed of 1.5mL/min, and discarding the effluent liquid;
(3) and (3) elution: eluting with 20mL of methanol solvent at a speed of 2mL/min to obtain a test solution;
(4) and (3) detection: detecting the test solution obtained in the step (3) by adopting a high performance liquid chromatography-triple quadrupole tandem mass spectrometer, and calculating the content of nitrosamine impurities in the eluted solution by adopting an external standard method;
wherein, the testing conditions of the high performance liquid chromatography comprise: the mobile phase A is 0.5 vol% glacial acetic acid aqueous solution, the mobile phase B is 0.5 vol% glacial acetic acid methanol solution, the volume ratio of the mobile phase A to the mobile phase B is 4:6, and the elution procedure is isocratic elution; the chromatographic column is Waters Xbridge C18, the inner diameter is 4.6mm, the column length is 100mm, the particle size is 5 μm, the flow rate is 0.5mL/min, the sample injection volume is 20 μ L, the column temperature is 40 ℃, the sample chamber temperature is 10 ℃, and the running time is 5 min;
wherein, the test conditions of the triple quadrupole tandem mass spectrometry comprise: the ion source is APCI, the polarity is positive, the temperature of the drying gas is 250 ℃, the flow rate of the drying gas is 4L/min, the temperature of the evaporation chamber is 350 ℃, the pressure of the atomizing gas is 35psi, the voltage of the capillary tube is 1500V, and the current of the corona needle is 6 muA.
Example 2
The embodiment provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which comprises the following steps:
testing a sample: preparation examples provided labeled and unlabeled samples
(1) And (3) activation: taking an EU52112M6 activated carbon solid phase extraction column (coconut shell carbon) (2500mg, 7.5mL), sequentially activating with 3mL of dichloromethane, 5.8mL of methanol and 18mL of ultrapure water, and ensuring that a solution in the column does not flow dry in the activation process;
(2) loading: then, sampling at the speed of 1.4mL/min, and discarding the effluent liquid;
(3) and (3) elution: eluting with 25mL of methanol solvent at a speed of 2mL/min to obtain a test solution;
(4) and (3) detection: detecting the test solution obtained in the step (3) by adopting a high performance liquid chromatography-triple quadrupole tandem mass spectrometer, and calculating the content of nitrosamine impurities in the eluted solution by adopting an external standard method;
wherein, the testing conditions of the high performance liquid chromatography comprise: the mobile phase A is 0.6 vol% glacial acetic acid aqueous solution, the mobile phase B is 0.5 vol% glacial acetic acid methanol solution, the volume ratio of the mobile phase A to the mobile phase B is 4:5.8, and the elution procedure is isocratic elution; the chromatographic column is Waters Xbridge C18, the inner diameter is 4.6mm, the column length is 100mm, the particle size is 5 μm, the flow rate is 0.48mL/min, the sample injection volume is 18 μ L, the column temperature is 38 ℃, the sample chamber temperature is 8 ℃, and the running time is 5 min;
wherein, the test conditions of the triple quadrupole tandem mass spectrometry comprise: the ion source is APCI with positive polarity, the temperature of the drying gas is 255 ℃, the flow rate of the drying gas is 4.2L/min, the temperature of the evaporation chamber is 355 ℃, the pressure of the atomization gas is 36psi, the voltage of the capillary tube is 1550V, and the current of the corona needle is 6.2 muA.
Example 3
The embodiment provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which comprises the following steps:
testing a sample: preparation examples provided labeled and unlabeled samples
(1) And (3) activation: taking an EU52112M6 activated carbon solid phase extraction column (coconut shell carbon) (1800mg, 5.5mL), sequentially activating with 3mL of dichloromethane, 6mL of methanol and 15mL of ultrapure water, and ensuring that a solution in the column does not flow dry in the activation process;
(2) loading: then, sampling at the speed of 1.6mL/min, and discarding the effluent liquid;
(3) and (3) elution: eluting with 30mL of methanol solvent at a speed of 2mL/min to obtain a test solution;
(4) and (3) detection: detecting the test solution obtained in the step (3) by adopting a high performance liquid chromatography-triple quadrupole tandem mass spectrometer, and calculating the content of nitrosamine impurities in the eluted solution by adopting an external standard method;
wherein, the testing conditions of the high performance liquid chromatography comprise: the mobile phase A is 0.4 vol% glacial acetic acid aqueous solution, the mobile phase B is 0.5 vol% glacial acetic acid methanol solution, the volume ratio of the mobile phase A to the mobile phase B is 4:6.1, and the elution procedure is isocratic elution; the chromatographic column is Waters Xbridge C18, the inner diameter is 4.6mm, the column length is 100mm, the particle size is 5 μm, the flow rate is 0.52mL/min, the sample injection volume is 22 μ L, the column temperature is 42 ℃, the sample chamber temperature is 12 ℃, and the running time is 5 min;
wherein, the test conditions of the triple quadrupole tandem mass spectrometry comprise: the ion source is APCI with positive polarity, the temperature of the drying gas is 248 ℃, the flow rate of the drying gas is 3.8L/min, the temperature of the evaporation chamber is 348 ℃, the pressure of the atomizing gas is 37psi, the voltage of the capillary tube is 1450V, and the current of the corona needle is 5.8 muA.
Example 4
The embodiment provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which is different from the embodiment 1 only in that an apricot kernel activated carbon solid-phase extraction column is adopted, and other steps are the same as the embodiment 1.
Experimental example 5
The embodiment provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which is different from the embodiment 1 only in that a wood activated carbon solid-phase extraction column is adopted, and other steps are the same as the embodiment 1.
Example 6
The embodiment provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which is different from the embodiment 1 only in that n-hexanol is adopted for elution in the step (3), and other steps are the same as the embodiment 1.
Example 7
The embodiment provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which is different from the embodiment 1 only in that ethanol is adopted for elution in the step (3), and other steps are the same as the embodiment 1.
Example 8
This example provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which is different from example 1 only in that mobile phase A is 0.5 vol% formic acid aqueous solution, mobile phase B is 0.5 vol% formic acid methanol solution in step (4), and the other steps are the same as example 1.
Example 9
The embodiment provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which is only different from the embodiment 1 in that an Agilent Poroshell 120SB-AQ chromatographic column is adopted in the step (4), and other steps are the same as the embodiment 1.
Example 10
This example provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, which differs from example 1 only in that ESI ion source is used in step (4), and the other steps are the same as example 1.
Comparative example 1
The comparative example provides a method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater, and the method is different from the method in the example 1 only in that a C18 solid phase extraction column is adopted, and other steps are the same as the steps in the example 1.
Drawing of standard curve
The test method comprises the following steps: respectively preparing N-nitrosodimethylamine standard working solutions and N-nitrosodiethylamine standard working solutions with 5 concentration levels, injecting samples according to the test conditions of the high performance liquid chromatography-triple quadrupole tandem mass spectrometer in the embodiment 1, determining peak areas, drawing a standard working curve and establishing a linear equation by taking the concentration ratio of the N-nitrosodimethylamine or the N-nitrosodiethylamine in the standard working solutions as a horizontal coordinate and the peak area ratio as a vertical coordinate; the experimental data are shown in Table 1, and the standard graphs are shown in FIGS. 1 and 2.
TABLE 1
As can be seen from the data in Table 1, the correlation coefficients of the linear equations of N-nitrosodimethylamine and N-nitrosodiethylamine both reach 1.00, which indicates that the detection method of N-nitrosodimethylamine and N-nitrosodiethylamine in the nitrogenous drug industrial wastewater provided by the invention has wide linear range and good linear relationship.
The standard graph of N-nitrosodimethylamine shown in FIG. 1; the N-nitrosodiethylamine standard curve as shown in FIG. 2; the method for detecting N-nitrosodimethylamine and N-nitrosodiethylamine in the industrial wastewater of nitrogenous drugs has good linear relation.
Experimental example 2
Recovery test
The test method comprises the following steps: the test results of examples 1 to 10 and comparative example 1 (5 parallel tests with an average value) were counted, and the average contents of N-nitrosodimethylamine and N-nitrosodiethylamine in the spiked solution and the untagged solution were calculated, respectively, and the recovery was calculated according to the following formula;
the calculation formula of the standard recovery rate is as follows:
in the formula: r is the recovery rate of the added standard; c0The concentration of the mixed standard solution is added, mg/mL; v0Volume to add mixed standard solution, uL; c1The detection concentration of the standard sample is mg/mL; v1The volume is the fixed volume (mL) of the sample before the sample is added; c2The detection concentration of the sample without the added standard is mg/mL; v2The volume is the fixed volume (mL) before the sample is injected;
the test results are shown in tables 2 and 3:
TABLE 2
As can be seen from the data in Table 2, the recovery of N-nitrosodimethylamine on the normalized scale was between 82.8 and 96.6% and the RSD value was between 2.1 and 7.3% using the detection method provided by the present invention (examples 1 to 9); by adopting the detection method provided by the preferred technical scheme (embodiment 1-3), the recovery rate of the N-nitrosodimethylamine is 92.8-96.6%, and the RSD value is 2.1-3.6%; the detection method for the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater provided by the invention is good in precision, high in recovery rate and small in RSD value.
TABLE 3
As can be seen from the data in Table 3, the recovery of N-nitrosodiethylamine by the detection method provided by the present invention (examples 1-9) was between 82.8-99.9% and the RSD value was between 1.3-7.3%; by adopting the detection method provided by the preferred technical scheme (embodiment 1-3) of the invention, the recovery rate of the N-nitrosodiethylamine is between 91.2 and 99.9 percent, and the RSD value is between 1.3 and 2.8 percent; the detection method for the nitrosamine impurities in the nitrogen-containing pharmaceutical industrial wastewater provided by the invention is good in precision, high in recovery rate and small in RSD value.
As can be seen from comparison of the examples 1 and 4-5 with the comparative example 1, the coconut shell carbon is more suitable for detecting nitrosamine impurities in nitrogen-containing industry, and the coconut shell carbon has high standard recovery rate and small RSD value.
As can be seen from the comparison between example 1 and examples 6 to 7, methanol was more suitable as the eluent, and methanol was used as the eluent, and the recovery rate of the spiked product was high.
As can be seen from the comparison between example 1 and example 8, the RSD value of the mobile phase is smaller by using the mixture of glacial acetic acid aqueous solution and glacial acetic acid methanol solution.
By comparing example 1 with example 9, the RSD value is smaller with a Waters Xbridge C18 column.
The comparison between the example 1 and the example 10 shows that the NDMA and NDEA detection limits are low, and NDMA and NDEA cannot be detected by adopting an ESI ion source, so that APCI serving as an ion source is more suitable for the detection method of the nitrosamine impurities in the nitrogen-containing industrial wastewater provided by the invention.
Test example 3
Method quantitative limit determination
The test method comprises the following steps: 10mL of standard solutions of known concentrations were added to each of a N-nitrosodimethylamine blank and a N-nitrosodiethylamine blank, and repeated measurement was performed for a plurality of times under the chromatographic conditions as provided in example 1, and the peak areas were measured to determine the limit of quantitation (LOQ) of the detection method of the present invention with a 10-fold signal-to-noise ratio, and the test results are shown in Table 4.
TABLE 4
| Sample name | LOQ(ng/mL) | Number of measurements | Average area | RSD(％) | Signal to noise ratio |
| N-nitrosodimethylamine | 3.2406 | 6 | 1946 | 9.8 | 10 |
| N-nitrosodiethylamine | 3.0471 | 6 | 921 | 7.2 | 10 |
As can be seen from the data in Table 4, the quantitative limit of the N-nitrosodimethylamine and the quantitative limit of the N-nitrosodiethylamine in the method for detecting the nitrosamine compounds in the nitrogen-containing industrial wastewater provided by the invention are 3.2406ng/mL and 3.0471ng/mL, which indicates that the method for detecting the nitrosamine compounds in the nitrogen-containing industrial wastewater provided by the invention has the characteristics of low quantitative limit and high sensitivity for detecting the N-nitrosodimethylamine and the N-nitrosodiethylamine.
The applicant states that the present invention is illustrated by the above examples of the process of the present invention, but the present invention is not limited to the above process steps, i.e. it is not meant that the present invention must rely on the above process steps to be carried out. It will be apparent to those skilled in the art that any modification of the present invention, equivalent substitutions of selected materials and additions of auxiliary components, selection of specific modes and the like, which are within the scope and disclosure of the present invention, are contemplated by the present invention.
Claims (10)
1. A detection method of nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater is characterized by comprising the following steps:
(1) enriching nitrosamine impurities in the wastewater by using an active carbon solid phase extraction column;
(2) and (2) detecting the enrichment liquid obtained in the step (1) by adopting a high performance liquid chromatography-triple quadrupole tandem mass spectrometer, and calculating the content of nitrosamine impurities in the enrichment liquid by adopting an external standard method.
2. A method for detecting nitrosamine impurities in industrial wastewater containing nitrogen according to claim 1, wherein in steps (1) and (2), said nitrosamine impurities comprise N-nitrosodimethylamine and/or N-nitrosodiethylamine.
3. A method for detecting nitrosamine impurities in industrial wastewater containing nitrogen according to claim 1 or 2, wherein in step (1), the activated carbon used in said activated carbon solid phase extraction column comprises any one or a combination of at least two of coal-based activated carbon, wood activated carbon, nutshell and fruit pit activated carbon or resin activated carbon, preferably nutshell and fruit pit activated carbon;
preferably, the shell and kernel type activated carbon comprises any one or a combination of at least two of coconut shell carbon, apricot kernel carbon or walnut carbon, and is preferably coconut shell carbon.
4. The method for detecting nitrosamine impurities in industrial wastewater containing nitrogen drugs as claimed in any one of claims 1-3, wherein in step (1), the activated carbon used in said activated carbon solid phase extraction column has a particle size of 80-120 mesh and a specific surface area of 4000-6000m2/g；
Preferably, the filling amount of the activated carbon solid phase extraction column is 1500-2500 mg.
5. A method for detecting nitrosamine impurities in industrial wastewater containing nitrogen according to any one of claims 1 to 4, wherein in step (1), said enrichment pressure is normal pressure;
preferably, in step (1), the enrichment comprises activation, loading and elution which are sequentially performed;
preferably, the solvent used for the activation comprises dichloromethane, methanol and water;
preferably, the volume ratio of the dichloromethane to the methanol to the water is 1 (1.5-2.5) to (5.5-6.5);
preferably, the water comprises ultrapure water;
preferably, the flow rate of the sample loading is 1-2 mL/min;
preferably, the solvent used for elution comprises any one or a combination of at least two of ethanol, n-hexanol or methanol, preferably methanol;
preferably, the flow rate of the elution is 1-2 mL/min.
6. A method for detecting nitrosamine impurities in industrial wastewater containing nitrogen according to claim 1-5, wherein in step (2), in the detection of high performance liquid chromatography, mobile phase A comprises 0.1-1 vol% glacial acetic acid solution, and mobile phase B comprises 0.1-1 vol% glacial acetic acid solution in methanol;
preferably, the volume ratio of the mobile phase A to the mobile phase B is 1 (0.5-3);
preferably, isocratic elution is adopted in the detection of the high performance liquid chromatography.
7. A method for detecting nitrosamine impurities in industrial wastewater containing nitrogen according to claim 1-6, wherein in step (2), said chromatographic column for high performance liquid chromatography comprises Agilent Poroshell 120SB-AQ or Waters Xbridge C18, preferably Waters Xbridge C18;
preferably, the inner diameter of the chromatographic column is 4-5mm, the column length is 80-120mm, and the particle size is 4.5-5.5 μm.
8. A method for detecting nitrosamine impurities in industrial wastewater containing nitrogen according to any one of claims 1 to 7, wherein in step (2), the flow rate in the detection by high performance liquid chromatography is 0.45 to 0.55 mL/min;
preferably, in the step (2), the injection volume in the detection of the high performance liquid chromatography is 15-25 μ L;
preferably, in the step (2), the column temperature in the detection of the high performance liquid chromatography is 35-45 ℃;
preferably, in the step (2), the temperature of a sample chamber in the detection of the high performance liquid chromatography is 5-15 ℃;
preferably, in the step (2), the running time in the detection of the high performance liquid chromatography is 5-7 min.
9. The method for detecting nitrosamine impurities in industrial wastewater containing nitrogen according to any one of claims 1 to 8, wherein in step (2), the ion source in the detection of said triple quadrupole tandem mass spectrometer is APCI, and the polarity is positive;
preferably, in the step (2), the temperature of the drying gas in the detection of the triple quadrupole tandem mass spectrometer is 240-260 ℃, and the flow rate of the drying gas is 3-6L/min;
preferably, in the step (2), in the detection of the triple quadrupole tandem mass spectrometer, the temperature of the evaporation chamber is 340-;
preferably, in the step (2), the capillary voltage in the detection of the triple quadrupole tandem mass spectrometer is 1400-1600V, and the corona needle current is 4-7 muA.
10. The method for detecting nitrosamine impurities in nitrogen-containing pharmaceutical industrial wastewater according to any one of claims 1 to 9, wherein said method comprises the steps of:
(A) and (3) activation: activated carbon solid phase extraction columns are activated by dichloromethane, methanol and water according to the volume ratio of 1 (0.5-1.5) to (0.5-1.5); the active carbon adopted in the active carbon solid phase extraction column is coconut shell carbon;
(B) loading: loading at a speed of 1-2 mL/min;
(C) and (3) elution: eluting with methanol solvent at a speed of 1-2mL/min to obtain a sample solution;
(D) and (3) detection: detecting the test solution obtained in the step (C) by adopting a high performance liquid chromatography-triple quadrupole tandem mass spectrometer, and calculating the content of nitrosamine impurities in the eluted solution by adopting an external standard method;
wherein, the testing conditions of the high performance liquid chromatography comprise: the mobile phase A is 0.1-1 vol% glacial acetic acid aqueous solution, the mobile phase B is 0.1-1 vol% glacial acetic acid methanol solution, and the volume ratio of the mobile phase A to the mobile phase B is 1 (0.5-3); the chromatographic column is Waters Xbridge C18, the inner diameter is 4-5mm, the column length is 80-120mm, the particle size is 4.5-5.5 μm, the flow rate is 0.45-0.55mL/min, the sample injection volume is 15-25 μ L, the column temperature is 35-45 ℃, the sample chamber temperature is 5-15 ℃, and the running time is 5-7 min;
wherein, the test conditions of the triple quadrupole tandem mass spectrometry comprise: the ion source is APCI, the polarity is positive, the temperature of the drying gas is 240-260 ℃, the flow rate of the drying gas is 3-6L/min, the temperature of the evaporation chamber is 340-360 ℃, the pressure of the atomizing gas is 30-40psi, the voltage of the capillary tube is 1400-1600V, and the current of the corona needle is 5-7 muA.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104568562A (en) * | 2014-12-31 | 2015-04-29 | 中国地质大学（武汉） | Water sample and pretreatment method of nitrosoamine compound in suspended matter of water sample |
| CN108008040A (en) * | 2017-12-01 | 2018-05-08 | 暨南大学 | A kind of detection method of underwater trace N- nitrosamine |
| CN108508111A (en) * | 2018-04-08 | 2018-09-07 | 哈尔滨工业大学 | Analysis method that is a kind of while detecting 9 kinds of trace nitrosamines disinfection by-products in water |
-
2021
- 2021-12-30 CN CN202111648656.0A patent/CN114324661A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104568562A (en) * | 2014-12-31 | 2015-04-29 | 中国地质大学（武汉） | Water sample and pretreatment method of nitrosoamine compound in suspended matter of water sample |
| CN108008040A (en) * | 2017-12-01 | 2018-05-08 | 暨南大学 | A kind of detection method of underwater trace N- nitrosamine |
| CN108508111A (en) * | 2018-04-08 | 2018-09-07 | 哈尔滨工业大学 | Analysis method that is a kind of while detecting 9 kinds of trace nitrosamines disinfection by-products in water |
Non-Patent Citations (6)
| Title |
|---|
| 唐雪惠;吴艳芬;李朝晖;朱宏;黄媛媛;: "超高效液相色谱质谱法测定饮用水中亚硝胺", 化学研究与应用 * |
| 张力群 等: "超高效液相色谱-串联质谱法检测生活饮用水8种N-亚硝胺类化合物", 预防医学 * |
| 张小敏 等: "饮用水中 9 种痕量亚硝胺类化合物的分析方法研究", 分析试验室 * |
| 张蓓蓓;赵永刚;孙慧婧;史震宇;: "固相萃取-超高效液相色谱/三重四极杆质谱法同时测定地表水中8种亚硝胺类化合物", 环境监控与预警 * |
| 王帅;姚常浩;贾立明;杨阳;王妍;: "固相萃取-液相色谱/串联质谱测定水中亚硝胺化合物", 环境化学 * |
| 韩莹;王济禾;李军;倪永炯;田雪颐;李微微;姚杰;: "高效液相色谱同时测定水中四种亚硝胺类物质", 中国给水排水 * |
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Address after: 226009 No. 1, Guangxing Road, comprehensive free trade zone, economic and Technological Development Zone, Nantong City, Jiangsu Province Applicant after: Nantong Lianya Pharmaceutical Co.,Ltd. Address before: 226009 No. 1, Guangxing Road, comprehensive free trade zone, economic and Technological Development Zone, Nantong City, Jiangsu Province Applicant before: NOVAST LABORATORIES (CHINA) Ltd. |