CN112034074B - Method for determining seven nonselective cyclooxygenase inhibiting drug residues in livestock meat - Google Patents
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Abstract
The invention discloses a method for determining seven nonselective cyclooxygenase inhibiting drug residues in livestock meat, which utilizes a novel dispersed solid phase extraction purification (dSPE) to extract and purify 7-seven nonselective COX inhibiting drug residues in the livestock meat, and adopts an ultra-high performance liquid chromatography-triple quadrupole mass spectrometry method which has the advantages of high sensitivity and strong anti-interference performance to qualitatively and quantitatively analyze the seven nonselective COX inhibiting drug residues in the livestock meat. Seven non-selective cyclooxygenase inhibiting drugs: salicylic acid, ibuprofen, diclofenac sodium, indometacin, piroxicam, naproxen and phenylbutazone. The invention well solves the problem in the process of detecting the residues of seven non-selective COX inhibitory drugs in poultry meat.
Description
Technical Field
The invention relates to a dispersed solid phase extraction purification-ultra-high performance liquid chromatography-tandem mass spectrometry detection method for inhibiting drug residues by seven non-selective cyclooxygenase enzymes in livestock meat.
Background
Non-steroidal anti-inflammatory drugs (NSAIDs) are drugs with a large class of non-cortical hormone structures and anti-inflammatory, analgesic, antipyretic and antiplatelet effects, are mainly used for symptomatic treatment of inflammatory and immune diseases, and can effectively relieve local pain, swelling and other symptoms of muscle, joint and inflammatory and immune diseases. Traditional NSAIDs mostly use carboxylic acid groups as active groups, and although the NSAIDs can be classified into phenylpropionic acids, phenylacetic acids, indoles, fenamic acids and the like according to the chemical structures of the NSAIDs, the carboxylic acid groups are mainly combined with 120-arginine of cyclooxygenase (COX, including COX-1 and COX-2), so that the curative effect and the side effect of the NSAIDs are not very different, and the NSAIDs are mostly nonselective Cyclooxygenase (COX) inhibitors.
The non-selective COX inhibitory drugs mainly comprise aspirin, phenylbutazone, indomethacin, pirfeixin and the like, have anti-inflammatory and analgesic effects, but have equivalent inhibitory effects on COX-1 and COX-2 and severe and high-incidence adverse reactions, such as hearing impairment, bone marrow suppression, gastrointestinal liver and kidney injury. Some drugs on the market in 70-80 th century, such as diclofenac, ibuprofen, meloxicam and the like, have the same curative effect as non-selective COX inhibitory drugs, and the adverse reaction is remarkably reduced compared with the former, so that the drugs with high curative effect and low adverse reaction are favored in clinic for a long time. The curative effect of the new coxib does not exceed that of the traditional medicine, the short-course ulcer has the advantage of low incidence, but the cardiovascular risk caused by long-course treatment and large dose is increased.
In addition to the wide clinical application, the non-selective COX inhibitory drugs have wide antimicrobial spectrum, low price and stable properties, so that the non-selective COX inhibitory drugs are widely used for preventing and treating the inflammatory infectious diseases of food-borne animals. Due to the lack of awareness of the risk of toxicity of non-selective COX inhibiting drugs, the purchase of such drugs by livestock breeders is not associated with limitations, leading to abuse of such drugs. Residues caused by abuse of nonsteroidal anti-inflammatory drugs in animal-derived foods can cause great harm to the health of human beings. The european union has specified maximum residual amounts of some non-selective COX inhibiting drugs.
Due to the complexity of the matrix of livestock meat, no more sensitive detection method has been reported to detect low levels of non-selective COX inhibitory drug residues in livestock meat. The general pretreatment method of the non-selective COX inhibitory drugs of complex matrixes mainly comprises the steps of solvent direct extraction, microwave-assisted extraction (MAE), ultrasonic extraction (USE), pressurized solvent extraction (PLE), QuEChERS method and the like to extract the non-selective COX inhibitory drug residues. After the sample is extracted, solid phase extraction SPE with certain stuffing is used for purification treatment. The detection method comprises a gas chromatography-mass spectrometry (GC-MS), a high performance liquid chromatography (HPLC-DAD/FLD) and a liquid chromatography-mass spectrometry (HPLC-MS/MS). Because the sample pretreatment of gas chromatography-mass spectrometry needs derivatization, the pretreatment is complex, and the sensitivity and specificity of the high performance liquid chromatography are lower.
Disclosure of Invention
The invention aims to provide a novel dispersive solid-phase extraction combined purification-ultra-high performance liquid chromatography-tandem mass spectrometry method which is simple, qualitative and quantitative and high in sensitivity and is suitable for detecting residues of seven nonselective Cyclooxygenase (COX) inhibitory drugs in livestock meat.
In order to solve the technical problems, the invention provides a dispersive solid phase extraction purification-ultra high performance liquid chromatography-tandem mass spectrometry method for detecting the residue of seven nonselective Cyclooxygenase (COX) inhibitory drugs in livestock meat, which comprises the following steps:
1) preparing a sample solution to be detected:
1.1, crushing a sample to be detected by a crusher, and then uniformly mixing to obtain a sample;
1.2, weighing 1 to 10g (preferably 5g) of a sample, accurately weighing the sample to 0.01g, placing the sample in a centrifuge tube with a plug, adding 5 to 20ml (preferably 5ml) of 0.1 to 0.5mol/L (preferably 0.2mol/L) hydrochloric acid solution, adding 1 to 10ml (preferably 5ml) of 0.01 to 0.05mol/L (preferably 0.02mol/L) ascorbic acid solution, and carrying out vortex oscillation for 1 to 5min (preferably 2 min);
1.3, adding 50-200 ul (preferably 100ul) of 0.1-1 mg/ml (preferably 0.5mg/ml) of pepsin into the sample solution obtained in the step 1.2, and oscillating for 6-24 h (preferably 12h) in a constant-temperature water bath at 37 ℃;
1.4, precisely adding 10-100 mL (preferably 50mL) of acetonitrile solution containing 1-10% (v/v) (preferably 5%) formic acid into the sample solution obtained in the step 1.3, performing vortex oscillation extraction for 10-30 min (preferably 20min), adding 60-500 mg (preferably 250mg) of N-vinylpyrrolidone-divinylbenzene copolymer, performing vortex oscillation for 1-10 min (preferably 5min), adding 2-10 g (preferably 5g) of sodium chloride and 5-10 g (preferably 8g) of anhydrous sodium sulfate, performing vortex for 1-5 min (preferably 2min), centrifuging for 2-10 min (preferably 5min) at the speed of 4000-8000 r/min (preferably 5000r/min), precisely sucking 30mL of supernatant, transferring all the supernatant into a 50mL polypropylene centrifuge tube, and further purifying;
1.5, adding a purifying agent (containing 1000-5000 mg (preferably 3000mg) of anhydrous sodium sulfate, 100-500 mg (preferably 200mg) of C18-N and 100-500 mg (preferably 300mg) of NH) into the sample solution to be purified obtained in step 1.4 2 PSA), mixing uniformly by swirling for 1-5 min (preferably 2min), and centrifuging at 4000-8000 r/min (preferably 5000r/min) for 2-10 min (preferably 5 min);
1.6, precisely taking 25ml of the supernatant obtained in the step 1.5, and blowing nitrogen to be nearly dry in a water bath at 40 ℃. Adding 1.0mL of 0.1% formic acid-acetonitrile solution (50:50, v/v), mixing for 1min, dissolving the residue, and filtering with 0.22 μm microporous organic filter membrane; the purpose is to provide the ultra-high performance liquid chromatography-tandem mass spectrometer for qualitative and quantitative analysis;
2) preparing a standard solution
2.1, dissolving seven non-selective cyclooxygenase inhibiting drug standard substances (shown in table 1) by using a chromatographic pure organic solvent (methanol or acetonitrile) respectively to prepare seven non-selective cyclooxygenase inhibiting drug stock solutions with the concentration of 1000ug/mL, and storing at-20 ℃;
2.2, preparing a proper amount of the standard stock solutions into mixed standard stock solutions with the concentration of 100mg/L by using organic solvents (methanol or acetonitrile);
2.3, precisely absorbing the appropriate volume of the mixed standard stock solution respectively, diluting the mixed standard stock solution into a mixed standard intermediate solution of 1mg/L by using an organic solvent (methanol or acetonitrile), precisely absorbing the appropriate volume of the mixed standard intermediate solution into a 10mL volumetric flask respectively, diluting the mixed standard intermediate solution into a constant volume by using the organic solvent (methanol or acetonitrile), and preparing a mixed standard use solution of 5, 10, 20, 50, 100 and 200 mu g/L (wherein the linear range of naproxen, diclofenac sodium and ibuprofen does not include 5 ug/L);
3) injecting the mixed standard series working solution into a liquid chromatograph-mass spectrometer, determining peak positions of seven non-selective cyclooxygenase inhibiting drugs and qualitative and quantitative ion pairs thereof, and making a standard curve equation by taking the abundance of the quantitative ion pairs as a vertical coordinate and the concentration as a horizontal coordinate;
4) and taking the supernatant obtained in the step 1) to determine the peak area and the quantitative and qualitative ion pair of each non-selective cyclooxygenase inhibiting drug in the supernatant according to the method in the step 3), carrying out qualitative analysis according to each peak-appearing time and the abundance ratio of the quantitative and qualitative ion pairs, and calculating according to the standard curve equation obtained in the step 3) to obtain the content of each non-selective cyclooxygenase inhibiting drug in the sample to be detected.
The invention relates to a dispersive solid phase extraction purification-ultra high performance liquid chromatography-tandem mass spectrometry method for detecting seven non-selective cyclooxygenase inhibiting drug residues in livestock meat, which comprises the following steps: the seven non-selective cyclooxygenase inhibiting drugs: salicylic acid, ibuprofen, diclofenac sodium, indomethacin, piroxicam, naproxen, phenylbutazone;
the invention relates to an improvement of dispersed solid phase extraction purification-ultra-high performance liquid chromatography-tandem mass spectrometry for detecting seven non-selective cyclooxygenase inhibiting drug residues in livestock meat, which comprises the following steps:
the chromatographic conditions (namely, the liquid chromatographic conditions for Ultra Performance Liquid Chromatography (UPLC) detection) in the step 3) are as follows: flow rate: 0.3-0.8 mL/min (preferably 0.4 mL/min); column temperature: 20-50 deg.C (preferably 40 deg.C); the sample amount is 0.5-5 μ L (preferably 1 μ L);
the mobile phase consists of a mobile phase A and a mobile phase B: the mobile phase A is 0.1% formic acid aqueous solution; the mobile phase B is acetonitrile;
the gradient elution procedure was: 0-8 min, 90-45% of A; 8-9 min, 45% A; 9.1-12 min, 90% A.
The above% is volume%.
The invention relates to a further improvement of dispersed solid phase extraction purification-ultra-high performance liquid chromatography-tandem mass spectrometry method for detecting seven non-selective cyclooxygenase inhibiting drug residues in livestock meat:
the liquid chromatographic column comprises: waters CORTECS C 18 (100×2.1mm，1.7μm)。
The mass spectrum conditions of the step 3) (i.e., the mass spectrum conditions for detecting triple quadrupole tandem mass spectrometry (MS/MS) are as follows: electrospray ionization (ESI), positive ion detection mode (ESI +); multiple reaction detection (MRM mode); temperature of drying gas: 200 ℃; flow rate of drying gas: 14L/min; the pressure of the sprayer is as follows: 200 KPa; capillary voltage: 3.5 KV; the calibration method comprises the following steps: automatic tuning correction of a mass axis; the MRM performs a segmented scan: : salicylic acid for 0-4.5 min; 4.5-6.0 min, piroxicam; naproxen for 6.0-7.5 min; 7.5-8.85 min, indometacin, diclofenac sodium and ibuprofen; 8.85-12.0 min, phenylbutazone; other mass spectrometry parameters are detailed in table 2.
TABLE 1 Mass spectrometric analysis parameters of seven nonselective cyclooxygenase inhibiting drugs
Note: is a quantitative ion pair
The invention is a further improvement of the dispersive solid phase extraction-ultra-high performance liquid chromatography-tandem mass spectrometry method for detecting the residue of seven non-selective cyclooxygenase inhibiting drugs in livestock meat: the organic solvent is chromatographic pure methanol and acetonitrile.
TABLE 2 chemical information table of seven non-selective cyclooxygenase inhibiting drugs
Compared with the prior art, the invention has the following remarkable effects:
(1) the method utilizes pepsin to carry out enzymolysis on high-content protein in the livestock meat, utilizes an enzymolysis technology to enable non-selective cyclooxygenase inhibiting drugs to be completely dissociated, more fully extracts the non-selective cyclooxygenase inhibiting drugs in high-protein substances such as the livestock meat, and simultaneously adds 0.02mol/L ascorbic acid solution into an enzymolysis medium to avoid oxidation of the non-selective cyclooxygenase in the enzymolysis process, thereby ensuring the stability of a target compound in the extraction process.
(2) The method utilizes the dispersive solid-phase extraction technology to deeply purify livestock and poultry meat with more complex matrix, and adopts the common dispersive solid-phase extraction technology (generally using C) 18 -N and NH 2 Based on the purification of animal-derived food substrates by PSA), by using N-vinylpyrrolidone-divinylbenzene copolymers, C 18 -N and NH 2 And the optimized combined use of three purifiers such as PSA and the like can fully remove interfering substances such as protein, lipid and the like, and meanwhile, the protein in the livestock meat is subjected to enzymolysis through pepsin.
(3) The method utilizes the special ultrahigh pressure advantage of an ultrahigh performance liquid chromatography system and implements a nuclear particle chromatography technology (Cortecs chromatographic column), optimizes the operating back pressure of the high performance liquid chromatograph, realizes the maximization of the separation efficiency of the liquid chromatograph by improving the separation degree and the peak capacity, obtains faster analysis speed while improving the separation degree of a target compound, and obtains higher sensitivity by combining the two chromatographic technologies, greatly shortens the analysis time and improves the analysis efficiency.
(4) The determination method disclosed by the invention has a good linear relation between 5-200 mug/L (the linear range of naproxen, diclofenac sodium and ibuprofen is 10-200 mug/L), the linear correlation coefficient is 0.9957, and the lowest detection limit of the method is 0.5-2 mug/g.
In conclusion, the novel dispersed solid phase extraction purification method for impurities in livestock meat has the advantages of simple pretreatment, less impurity interference and matrix effect, better recovery rate and higher reproducibility, and the ultra-high performance liquid chromatography-tandem mass spectrometry method can be used for quickly and accurately separating and qualitatively and quantitatively analyzing seven non-selective cyclooxygenase inhibiting drugs within 12 minutes, and is simple to operate, and better in sensitivity and accuracy.
The technical innovation points of the invention are mainly as follows:
1. the existing food safety standard related to detection of partial non-selective cyclooxygenase inhibiting drug residues in livestock meat products is SN/T2190-.
2. In the prior art, a target substance is separated from a complex matrix in the process of testing non-selective cyclooxygenase inhibiting drug residues, and the prior art discloses that the target substance is extracted by using the technologies of solvent direct extraction, microwave-assisted extraction (MAE), ultrasonic extraction (USE), pressurized solvent extraction (PLE) and the like. Meanwhile, in order to avoid part of non-selective cyclooxygenase inhibiting drugs from being oxidized in the enzymolysis process, a certain amount of ascorbic acid solution is added in the enzymolysis process, so that the stability of some non-selective cyclooxygenase inhibiting drugs is ensured.
3. In the prior art, impurities (including protein, fat, pigment and the like) are removed in the process of testing the non-selective cyclooxygenase inhibiting drug residues by removing the impurities through an organic solvent (such as normal hexane and the like) or purifying a sample through a solid phase extraction cartridge, but the prior art has the defects of complex operation, low testing efficiency and poor reproducibility, and C is adopted 18 -N and NH 2 And (4) removing impurities by a dispersed solid phase extraction technology consisting of PSA and the like, and finding that the matrix effect is obvious. By using N-vinylpyrrolidone-divinylThe optimized combination of three purificant of phenyl copolymer, C18-N and NH2-PSA, etc. can reduce the impurity content of the finally obtained sample solution, obviously reduce the matrix effect, and improve the efficiency, accuracy and precision of the detection.
In conclusion, the invention utilizes a novel dispersed solid phase extraction purification (dSPE) to extract and purify 7-seven non-selective COX inhibitory drug residues in livestock and poultry meat, and adopts an ultra-high performance liquid chromatography-triple quadrupole mass spectrometry method which has the advantages of high sensitivity and strong anti-interference performance to qualitatively and quantitatively analyze the seven non-selective COX inhibitory drug residues in the livestock and poultry meat. The method well solves the problem in the process of detecting the residue of seven nonselective COX inhibitory drugs in the poultry meat, and has important practical significance for guaranteeing human health, promoting the development level of the self technology of enterprises, promoting the development of the detection technology of China, promoting the international position of the quality safety detection technology of the poultry meat in China, increasing employment, driving the overall progress of the poultry meat industry, and having great economic and social benefits for the progress of the technology and the transformation and upgrade of the industry.
Drawings
The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.
FIG. 1 is a qualitative and quantitative ion pair plot of seven non-selective cyclooxygenase inhibiting drugs, i.e., 7 non-selective COX inhibitory components (20 μ g/L) MRM spectra; in the figure, 1 to 7 represent the seven non-selective cyclooxygenase inhibiting drugs, respectively.
In fig. 1, qualitative and quantitative ion graphs of salicylic acid, ibuprofen, naproxen, diclofenac sodium, phenylbutazone, piroxicam and indomethacin are sequentially arranged from top to bottom.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of the invention is not limited thereto.
1 reagents and materials
Unless otherwise indicated, all reagents used in the analysis were chromatographically pure grades, and all water used was primary water.
1.1 methanol
1.2 acetonitrile
1.3 formic acid
1.4 sodium chloride: analytical purity
1.5 anhydrous sodium sulfate: analytical purity
1.6 ascorbic acid: analytical purity
1.7 hydrochloric acid: analytical purity
1.8 pepsin: 250U/mg
1.9N-vinylpyrrolidone-divinylbenzene copolymer (Waters Co.)
1.10 octadecyl bonded silica gel adsorbent (C) 18 -N)
1.11NH 2 -propylethylenediamine adsorbent (NH) 2 PSA), shimadzu corporation.
1.12 nonselective cyclooxygenase inhibiting drug standard: salicylic acid, ibuprofen, diclofenac sodium, indometacin, piroxicam, naproxen and phenylbutazone, and the purity is more than or equal to 99.0 percent.
1.13 standard stock solutions: the seven non-selective cyclooxygenase inhibiting drug standard substances are respectively treated as follows: preparing seven non-selective cyclooxygenase inhibiting drug standard substances into standard stock solutions respectively by using acetonitrile (or other suitable organic solvents); precisely preparing a proper amount of the standard stock solution into a mixed standard stock solution with the concentration of 1mg/L by using acetonitrile (or other suitable organic solvents); namely, the concentrations of seven non-selective cyclooxygenase inhibiting drugs (salicylic acid, ibuprofen, naproxen, diclofenac sodium, phenylbutazone, piroxicam, indomethacin) in the mixed standard stock solution were all 1 mg/L.
1.14 standard working solution: and (3) preparing the mixed standard stock solution into a mixed standard working solution with the concentration distribution of 5-200ug/L by using acetonitrile (or other suitable organic solvents).
Note: the standard stock solution is stored at-20 ℃ in the dark, and the validity period is half a year. The standard working solution is stored at 4 ℃ in the dark, and the validity period is 1 month.
2 instruments and devices
2.1 Ultra Performance Liquid Chromatography (UPLC).
2.2 triple quadrupole tandem mass spectrometer (MS/MS).
2.3 analytical balance: the dose is 0.0001g and 0.01 g.
2.4 vortex oscillator.
2.5 high-speed centrifuge: the maximum can reach 10000 r/min.
2.6 constant temperature water bath oscillator.
2.7 extractor: polyethylene centrifuge tube, 50 mL.
2.8 organic phase filtration membrane: 0.22 um.
Example 1, a dispersive solid-phase extraction purification-ultra high performance liquid chromatography-tandem mass spectrometry method applicable to the residue of seven non-selective cyclooxygenase inhibiting drugs in livestock meat, sequentially comprising the following steps:
1) preparing a sample solution to be detected:
(1) taking livestock meat as a sample to be detected, crushing the livestock meat into meat powder by using a crusher, and uniformly mixing the meat powder and the meat powder to be detected to obtain a sample (matrix);
the livestock meat includes pork, beef, mutton, chicken, duck, etc.
(2) Weighing 5g of the sample, accurately weighing the sample to 0.01g, placing the sample in a polyethylene centrifugal tube, adding 5ml of 0.2mol/L hydrochloric acid solution, simultaneously adding 5ml of 0.02mol/L ascorbic acid solution, and carrying out vortex oscillation for 2 min;
(3) adding 100ul of 0.5mg/ml pepsin into the product obtained in the step (2), and oscillating in a thermostatic water bath at 37 ℃ for 12 h;
(4) precisely adding 50mL of acetonitrile solution containing 5% (v/v) formic acid into the product obtained in the step (3), carrying out vortex oscillation extraction for 20min, then adding 250mg of N-vinylpyrrolidone-divinylbenzene copolymer, carrying out vortex oscillation for 5min, then adding 15g of sodium chloride and 15g of anhydrous sodium sulfate, carrying out vortex oscillation extraction for 2min, carrying out centrifugation for 5min at the speed of 5000r/min, precisely absorbing 30mL of the supernatant, and transferring the whole supernatant into a 50mL polypropylene centrifuge tube to serve as a sample solution to be purified;
acetonitrile solution containing 5% (v/v) formic acid, namely formic acid: acetonitrile in a volume ratio of 5: 95;
(5) adding a purifying agent into the sample solution to be purified, wherein the purifying agent is prepared from 3000mg of anhydrous sodium sulfate and 200mgC 18 -N (octadecyl)Bonded silica gel adsorbent) and 300mgNH 2 -PSA(NH 2 -propyl ethylenediamine adsorbent), mixing uniformly by swirling for 2min, and centrifuging for 5min at the speed of 5000 r/min;
(6) precisely taking 25ml of the supernatant obtained in the step (5), and placing the supernatant in a water bath at 40 ℃ to blow nitrogen till the supernatant is nearly dry. Adding 0.1% (volume%) formic acid solution-acetonitrile (50:50, v/v) to constant volume to 1.0mL, vortex mixing for 1min to dissolve residue, and filtering with 0.22 μm microporous organic filter membrane; the obtained filtrate is used for qualitative and quantitative analysis of the ultra-high performance liquid chromatography-tandem mass spectrometer; the change in filtrate volume was very small (negligible), i.e. the filtrate was obtained for 1 min.
0.1% formic acid in water-acetonitrile (50:50, v/v), i.e., 0.1% formic acid in water: acetonitrile in a volume ratio of 1: 1;
2) preparing a standard solution
(1) Dissolving seven non-selective cyclooxygenase inhibiting drug standard substances with acetonitrile respectively to prepare seven non-selective cyclooxygenase inhibiting drug standard stock solutions with the concentration of 1 mg/mL;
(2) preparing a proper amount of the seven standard stock solutions into mixed standard stock solutions with the concentration of 100mg/L by using acetonitrile;
(3) precisely sucking the appropriate volume of the mixed standard stock solution respectively, diluting the mixed standard stock solution into a mixed standard substance intermediate solution of 1mg/L by using acetonitrile, precisely sucking the appropriate volume of the mixed standard substance intermediate solution into a 10mL volumetric flask respectively, diluting the mixed standard substance intermediate solution into a constant volume by using acetonitrile, and preparing a mixed standard substance use solution with the concentration of 5, 10, 20, 50, 100 and 200 mu g/L (wherein the linear range of naproxen, diclofenac sodium and ibuprofen does not comprise 5 ug/L);
3) respectively carrying out the following operations on the standard series working solution obtained in the step 2): injecting into an ultra-high performance liquid chromatography-triple quadrupole mass spectrometer, determining peak positions of the seven non-selective cyclooxygenase inhibiting drugs and qualitative and quantitative ion pairs thereof (shown in figure 1), taking the abundance of the quantitative ion pairs as a vertical coordinate, taking the concentration of the seven non-selective cyclooxygenase inhibiting drugs in the solution to be detected as a horizontal coordinate, and making a standard curve equation.
The method comprises the following specific steps:
a) and (3) chromatographic column: waters Cortecs C18, 50 mm. times.2.1 mm, 1.6um column or equivalent;
b) flow rate: 0.5 ml/min;
c) column temperature: 40 ℃;
d) sample introduction volume: 1 ul;
e) mobile phase:
the system consists of a mobile phase A and a mobile phase B;
the mobile phase A is 0.1 percent formic acid aqueous solution; the mobile phase B is acetonitrile;
f) the gradient elution procedure is shown in Table 3
TABLE 3 gradient elution schedule
The above% is volume%.
g) An ion source: electrospray ion source (ESI);
h) a monitoring mode: multiple reaction monitoring mode (MRM); and (3) performing segmented scanning: salicylic acid for 0-4.5 min; 4.5-6.0 min, piroxicam; naproxen for 6.0-7.5 min; 7.5-8.85 min, indometacin, diclofenac sodium and ibuprofen; 8.85-12.0 min, phenylbutazone;
i) dry gas (nitrogen): temperature: 200 ℃; flow rate: 14 ml/min;
j) capillary voltage: 3.5 KV;
k) capillary exit voltage (Fragmentor): 380V;
l) other mass spectrometry parameters are detailed in table 4.
TABLE 4 Mass spectrometric analysis parameters of seven nonselective cyclooxygenase inhibiting drugs
Note: is a quantitative ion pair
When the concentration range of the ultra-high performance liquid chromatography-triple quadrupole mass spectrometry is 5-200ug/L (the linear range of naproxen, diclofenac sodium and ibuprofen is 10-200 mug/L), the linear relationship of the seven non-selective cyclooxygenase inhibiting drugs is good, and the table 5 shows that the two drugs are combined together.
TABLE 5 Linear relationship, detection limits and quantitation limits for seven non-selective cyclooxygenase-inhibiting drugs
In Table 5, Y represents the quantitative ion pair abundance and X represents the concentration of the target (data unit ug/L)
4) And taking the supernatant obtained in the step 1) (namely, the filtrate obtained by filtering through a 0.22-micron microporous organic filter membrane) to perform quantitative analysis on the abundance of the quantitative ions of the metabolic products of the non-selective cyclooxygenase inhibiting drugs in the supernatant according to the method in the step 3), performing qualitative analysis according to peak-appearing time and the quantitative ion-to-abundance ratio, calculating according to the standard curve equation obtained in the step 3) to obtain the content c (ug/L) of the non-selective cyclooxygenase inhibiting drugs in the supernatant, and finally calculating according to the following conversion formula to obtain the content of the non-selective cyclooxygenase inhibiting drugs in the sample to be detected.
Conversion formula
In the formula:
x is the residual amount of the component to be measured in the sample, and the unit is microgram per kilogram (ug/kg);
c-concentration of the measured component solution in nanograms per milliliter (ug/L) from the standard curve;
v-final volumetric volume of sample solution in milliliters (mL);
m-the mass of the final sample represented by the sample solution in grams (g);
f-dilution factor 2.
5) Qualitative analysis of the cells
Corresponding to step 4), there were 2 detection channels per component (except ibuprofen which had only one ion channel), one for each monitoring ion pair. When a sample is detected, if a plurality of (2) channels corresponding to a certain component have a spectrum peak consistent with the retention time of a reference substance, and the relative abundance of the several daughter ions is consistent with that of the reference substance, the component can be judged to be detected in the sample. The abundance data of each component conforms to the allowable deviation range of relative ion abundance in qualitative judgment according to European Union 2002/657/EC regulations, and the relative abundance of the daughter ions can be judged to be consistent. If the qualitative process meets the requirement of 4 minutes and more than 3 minutes according to the calculation of the value division method (the number of identification points of the mass spectrometry analysis method) of the regulation, the sample contains the non-selective cyclooxygenase inhibiting drug residue.
6) Quantitative analysis of the sample
Corresponding to the step 4), the method adopts an external standard method for quantification, selects standard working solution with similar concentration according to the content of the substance to be measured in the sample solution, inserts the same volume of the standard working solution and the sample solution into a sample for measurement, and the response values of seven non-selective cyclooxygenase inhibiting drugs in the standard working solution and the sample solution to be measured are all in a linear range.
Note 1: if the detection response value of the sample liquid exceeds the linear range, the standard series working liquid can be properly adjusted.
Note 2: the MRM ion pattern of the seven non-selective cyclooxygenase inhibiting drugs under the chromatographic and mass spectrometric conditions described above is shown in figure 1.
7) Detection lower limit
The lowest detectable concentration (LOD) is determined by 3 times of signal-to-noise ratio (S/N is 3), the lowest quantitative concentration (LOQ) is determined by 10 times of signal-to-noise ratio (S/N is 10), and the detection limit of the method for the dispersed solid phase extraction combined purification-ultra high performance liquid chromatography-tandem mass spectrometry detection method for seven nonselective cyclooxygenase inhibiting drugs in livestock meat is shown in table 5.
The method of SN/T2190-2008 < method for detecting residual quantity of non-steroidal anti-inflammatory drugs in import and export animal derived food by liquid chromatography-mass spectrometry/mass spectrometry > is adopted to detect and ensure that the pork which does not contain seven non-selective cyclooxygenase inhibiting drugs is used as a blank pork sample.
The ground substrate formed by the crushed blank pork sample is treated as follows:
the recovery rates of the seven non-selective cyclooxygenase-inhibiting drugs added to the meat samples of the poultry were determined by the pretreatment and analysis method of example 1 above, using a standard mixture of seven non-selective cyclooxygenase-inhibiting drugs added at concentrations of 2.5ug/kg, 5ug/kg, and 25ug/kg, in 6 replicates each, and the results are shown in table 6 (the linear equation is shown in table 1), which is an average of the 6 replicates.
TABLE 6 results of the non-selective cyclooxygenase-inhibited drug recovery and relative standard deviation (n ═ 6) for the samples added at different concentrations
From table 6, it can be seen that: the recovery rate of the seven non-selective cyclooxygenases added into the pork matrix at three levels is within the range of 85.1-95.2%, and the recovery rate is better.
25 batches of livestock meat (including 5 batches of pork, 5 batches of beef, 5 batches of mutton, 5 batches of chicken and 5 batches of duck) randomly extracted in a laboratory, wherein the No. 1 to 5 samples are pork, the No. 6 to 10 samples are beef, the No. 11 to 15 samples are mutton, the No. 16 to 20 samples are chicken and the No. 21 to 25 samples are duck, and a sample 26 (0.05 ml of mixed standard intermediate solution is added into 5g of pork confirmed to be blank), a sample 27 (0.05 ml of mixed standard intermediate solution is added into 5g of beef confirmed to be blank) and a sample 28 (0.05 ml of mixed standard intermediate solution is added into 5g of mutton confirmed to be blank) are also arranged, sample 29 (5 g of chicken confirmed as blank with 0.05ml of the mixed standard intermediate solution added) and sample 30 (5 g of duck confirmed as blank with 0.05ml of the mixed standard intermediate solution added).
The concentration of 7 non-selective cyclooxygenase inhibiting drugs in the mixed standard intermediate solution was 1 mg/L.
According to the above operation steps, extraction and purification were performed, each sample was performed 2 times in parallel, and the results of measurement by LC MS were shown in Table 7.
TABLE 7 actual sample measurement results (ug/kg)
ND: indicating no detection.
From table 7 we know that: ibuprofen is detected in sample 2, diclofenac sodium is detected in sample 4, and diclofenac sodium is detected in sample 7.
Comparative example 1-1, the procedure (2) "of step 1) of example 1 was changed to the procedure of adding 5ml of 0.2mol/L hydrochloric acid solution and simultaneously adding 5ml of 0.02mol/L ascorbic acid solution, vortex shaking for 2 min" to adding 5ml of 0.2mol/L hydrochloric acid solution, vortex shaking for 2min "; the rest is equal to example 1, and the recovery rate of phenylbutazone is lower than 60%, and the sensitivity and the accuracy are obviously reduced.
Comparative examples 1-2, the procedure (2) "of step 1) of example 1 was changed to the procedure of adding 5ml of 0.2mol/L hydrochloric acid solution and simultaneously adding 5ml of 0.02mol/L ascorbic acid solution and vortex shaking for 2 min" to the procedure of adding 5ml of 0.2mol/L hydrochloric acid solution and simultaneously adding 5ml of 0.03mol/L ascorbic acid solution and vortex shaking for 2min "; the rest is equal to example 1, and the result shows that the recovery rate of the piroxicam is lower than 80%, and the sensitivity and the accuracy are obviously reduced.
Comparative examples 1 to 3, 100uL of 0.5mg/mL pepsin was added to the dissolved sample solution (3) "obtained in step 1) of example 1, and the solution was changed from" 12 hours in 37 ℃ constant temperature bath shaking to "16 hours in 37 ℃ constant temperature bath shaking" after dissolving the sample solution; the rest is equal to example 1, and the results show that the recovery rate of the seven non-selective cyclooxygenase inhibiting medicaments is lower than 75%, the detection limit of the seven non-selective cyclooxygenase inhibiting medicaments is 5ug/kg, and the sensitivity and the accuracy are obviously reduced.
Comparative examples 1 to 4, 50ml of acetonitrile solution containing 5% (v/v) formic acid was added to the above sample solution of (4) "in step 1) of example 1, vortex-extracted for 20min, 250mg of N-vinylpyrrolidone-divinylbenzene copolymer was added, and vortex-extracted for 5 min" was changed to "50 ml of acetonitrile solution containing 5% (v/v) formic acid was added to the above sample solution, vortex-extracted for 20 min"; the rest is equal to example 1, and the results show that the recovery rate of the seven non-selective cyclooxygenase inhibiting medicaments is lower than 70 percent, the detection limit of the seven non-selective cyclooxygenase inhibiting medicaments is 6ug/kg, and the sensitivity and the accuracy are obviously reduced.
Comparative examples 1 to 5, 50ml of acetonitrile solution containing 5% (v/v) formic acid was added to the above sample solution in (4) "of step 1) of example 1, vortex extraction was performed for 20min, 250mg of N-vinylpyrrolidone-divinylbenzene copolymer was added, and vortex extraction was performed for 5 min" changed to "50 ml of acetonitrile solution containing 5% (v/v) formic acid was added to the above sample solution, vortex extraction was performed for 20min, 500mg of N-vinylpyrrolidone-divinylbenzene copolymer was added, and vortex extraction was performed for 5 min"; the rest is equal to example 1, and the results show that the recovery rates of the seven non-selective cyclooxygenase inhibiting medicaments are all lower than 75%, the detection limits of the seven non-selective cyclooxygenase inhibiting medicaments are all 8ug/kg, and the sensitivity and the accuracy are obviously reduced.
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is shown that the present invention is not limited to the above embodiments, but many variations are possible. All such modifications which may become apparent to those skilled in the art from this disclosure are deemed to be within the scope of the invention.
Claims (3)
1. The method for determining seven nonselective cyclooxygenase inhibiting drug residues in livestock meat is characterized by comprising the following steps:
1) preparing a sample solution to be detected:
1.1, crushing a sample to be detected by a crusher, and then uniformly mixing to obtain a sample;
1.2, weighing 1-10 g of a sample, placing the sample in a centrifuge tube with a plug, adding 5-20 ml of 0.1-0.5 mol/L hydrochloric acid solution, adding 1-10 ml of 0.02mol/L ascorbic acid solution, and carrying out vortex oscillation for 1-5 min;
1.3, adding 50-200 mu l of 0.1-1 mg/ml pepsin into the sample solution obtained in the step 1.2, and oscillating in a constant-temperature water bath at 37 ℃ for 12 hours;
1.4, precisely adding 10-100 ml of acetonitrile solution containing 1-10% v/v formic acid into the sample solution obtained in the step 1.3, performing vortex oscillation extraction for 10-30 min, adding 250mg of N-vinylpyrrolidone-divinylbenzene copolymer, performing vortex oscillation for 1-10 min, adding 2-10 g of sodium chloride and 5-10 g of anhydrous sodium sulfate, performing vortex extraction for 1-5 min, centrifuging at the speed of 4000-8000 r/min for 2-10 min, and absorbing supernate to serve as a sample solution to be purified;
1.5, adding a purifying agent into the sample liquid to be purified obtained in the step 1.4, uniformly mixing the mixture in a vortex mode for 1-5 min, and centrifuging the mixture at the speed of 4000-8000 r/min for 2-10 min;
the purificant contains 3000mg of anhydrous sodium sulfate, 200mg of C18-N and 300mg of NH 2 -PSA；
1.6, taking 25ml of supernatant obtained in the step 1.5, and blowing nitrogen to be nearly dry in water bath at 40 ℃; adding 1.0mL of an aqueous solution containing 0.1% formic acid-acetonitrile solution =50:50, v/v, vortex mixing uniformly for 1min to dissolve residues, and filtering with a 0.22 mu m microporous organic filter membrane; the purpose is to provide the ultra-high performance liquid chromatography-tandem mass spectrometer for qualitative and quantitative analysis;
2) preparing a standard solution: obtaining mixed standard series working solution;
3) injecting the mixed standard series working solution into a liquid chromatograph-mass spectrometer, determining peak positions of seven non-selective cyclooxygenase inhibiting drugs and qualitative and quantitative ion pairs thereof, and making a standard curve equation by taking the abundance of the quantitative ion pairs as a vertical coordinate and the concentration as a horizontal coordinate;
the chromatographic conditions are as follows: the liquid chromatography column is Waters CORTECS C 18 (ii) a Flow rate: 0.3-0.8 ml/min; column temperature: 20-50 ℃; the sample feeding amount is 0.5-5 mu L;
the mobile phase consists of a mobile phase A and a mobile phase B: the mobile phase A is 0.1 percent formic acid aqueous solution; the mobile phase B is acetonitrile;
the gradient elution procedure was: 0-8 min, 90% -45% A; 8-9 min, 45% of A; 9.1-12 min, 90% A;
the above% is volume%;
the mass spectrum conditions are as follows: electrospray ionization source ESI, multi-reaction detection mode; temperature of the drying gas: 200 ℃; flow rate of drying gas: 14L/min; the pressure of the sprayer is as follows: 200 KPa; capillary voltage: 3.5 KV; the calibration method comprises the following steps: automatic tuning correction of a mass axis; the mass spectrometry parameters of the seven nonselective cyclooxygenase inhibiting drugs are as follows:
4) and taking the filtrate obtained in the step 1) to determine the peak area and the quantitative and qualitative ion pair of each non-selective cyclooxygenase inhibiting drug in the supernatant according to the method in the step 3), carrying out qualitative analysis according to each peak-appearing time and the abundance ratio of the quantitative and qualitative ion pairs, and calculating according to the standard curve equation obtained in the step 3) to obtain the content of each non-selective cyclooxygenase inhibiting drug in the sample to be detected.
2. The method for determining residues of seven nonselective cyclooxygenase-inhibiting drugs in livestock meat according to claim 1, characterized in that: the step 2) comprises the following steps:
2.1, respectively dissolving seven non-selective cyclooxygenase inhibiting drug standard substances by using an organic solvent to prepare seven non-selective cyclooxygenase inhibiting drug stock solutions with the concentration of 1000 mu g/mL, and storing at the temperature of minus 20 ℃;
2.2, preparing the standard stock solutions into mixed standard stock solutions with the concentration of 100mg/L by using organic solvents;
and 2.3, respectively sucking appropriate volumes of the mixed standard storage solutions, diluting the mixed standard storage solutions into mixed standard intermediate solutions with 1mg/L by using an organic solvent, respectively sucking the mixed standard intermediate solutions, diluting the mixed standard intermediate solutions with the organic solvent to a constant volume, and preparing the mixed standard series working solutions with 5 mug/L, 10 mug/L, 20 mug/L, 50 mug/L, 100 mug/L and 200 mug/L.
3. The method for determining residues of seven nonselective cyclooxygenase-inhibiting drugs in livestock meat according to claim 2, characterized in that: the organic solvent is methanol or acetonitrile.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009109406A (en) * | 2007-10-31 | 2009-05-21 | Kowa Co | Simultaneous determination method of ibuprofen, tipepidine, and thiamine, method of manufacturing tablet preparation ensuring effective component content using the method |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2629367C1 (en) * | 2016-11-02 | 2017-08-29 | федеральное государственное автономное образовательное учреждение высшего образования "Казанский (Приволжский) федеральный университет" (ФГАОУ ВО КФУ) | Nonsteroid anti-inflammatory agent on basis of naproxen with low gastrotoxicity |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009109406A (en) * | 2007-10-31 | 2009-05-21 | Kowa Co | Simultaneous determination method of ibuprofen, tipepidine, and thiamine, method of manufacturing tablet preparation ensuring effective component content using the method |
Non-Patent Citations (9)
| Title |
|---|
| Determination of non-steroidal anti-inflammatory drugs residues in animal muscles by liquid chromatography–tandem mass spectrometry;Piotr Jedziniak 等;《Analytica Chimica Acta》;20100424;第672卷;第85-92页 * |
| HPLC-MS/MS法同时测定鸡肉中40种糖皮质激素和9种非甾体抗炎药物残留;陈晶燕 等;《质谱学报》;20180727;第40卷(第02期);第123-130页 * |
| Simultaneous determination of thirty non-steroidal anti-inflammatory drug residues in swine muscle by ultra-high-performance liquid chromatography with tandem mass spectrometry;Ting Hu 等;《Journal of Chromatography A》;20120106;第1219卷;第104-113页 * |
| 分散固相萃取-超高效液相色谱-串联质谱法测定奶粉中7种非选择性环氧化酶抑制药物;励炯 等;《色谱》;20190228;第37卷(第02期);第177-182页 * |
| 屠宰动物肌肉组织中的保泰松残留量的LC-UV测定方法研究;庞国芳 等;《检验检疫科学》;20071031;第17卷(第05期);第3-7页 * |
| 微波辅助提取-分散固相萃取净化-超高效液相色谱-串联质谱法测定畜禽肉中的7种非选择性环氧化酶抑制药物残留;励炯 等;《分析科学学报》;20190808;第35卷(第04期);第479-484页 * |
| 猪组织中氟尼辛残留量的HPLC和LC-MS/MS检测及消除研究;孙庆;《中国优秀博硕士学位论文全文数据库(硕士)农业科技辑》;20140415(第04期);第D050-246页 * |
| 超声波-微波辅助提取-高效液相色谱法同时检测羊肉组织中4种非甾体抗炎药物残留;康永锋 等;《色谱》;20101130;第28卷(第11期);第1056-1060页 * |
| 通过式固相萃取净化-超高效液相色谱-串联质谱法测定鱼肉中7种非选择性环氧化酶抑制药物残留;励炯 等;《中国食品卫生杂志》;20190331;第31卷(第02期);第116-121页 * |
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