Improved anti-gout drug detection method
Through the detection method of liquid-mass synthesis and combined with liquid chromatography and mass spectrometry, the problem of long detection time in the existing technology is solved, and the rapid detection of 15 anti-gout drugs in functional foods is achieved, which improves the detection efficiency.
Patent Information
- Application Number
- CN202311030239.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-08-16
- Publication Date
- 2025-02-21
AI Technical Summary
The prior art is difficult to quickly and effectively detect anti-gout drugs illegally added in functional foods, and the detection time is long, making it difficult to meet the market's demand for rapid testing.
The liquid-mass synthesis detection method is used, and the gradient elution procedure and specific mass spectrometry conditions are set to detect 15 common anti-gout drugs simultaneously through the combination of liquid chromatography and mass spectrometry technology.
It has achieved rapid detection of 15 common anti-gout drugs in functional foods, shortened detection time and improved detection efficiency, and can easily identify conventional gout drugs.
Smart Images
Figure FT_1
Abstract
Description
Technical Field
[0001] The invention belongs to the field of detection technology, and specifically relates to a method for detecting anti-gout drugs illegally added into food or health food. Background Art
[0002] At present, the main drugs for the treatment of gout in clinical practice include xanthine oxidase inhibitors febuxostat, allopurinol, benzbromarone, a drug that promotes uric acid excretion, and colchicine, nonsteroidal anti-inflammatory drugs, adrenocorticotropic hormone, glucocorticoids, etc., which are often used during gout attacks. Since the quality of illegally added drugs is not guaranteed and the dosage is arbitrary, it is easy to cause serious adverse reactions, such as liver and kidney damage.
[0003] It is very urgent to establish an efficient and rapid method for detecting illegal additions to anti-gout drugs. The anti-gout drug detection method CN2022110237450 established by this project team in the early stage has a liquid phase detection time of 53 minutes and detects 11 common gout treatment drugs. Therefore, it is urgent to establish a detection method that shortens the detection time, improves the detection efficiency, and detects more gout treatment drugs. Summary of the invention
[0004] In view of the situation that anti-gout drugs are illegally added to functional foods that emerge in the market, the present invention provides a liquid chromatography-mass spectrometry detection method for illegally added anti-gout drugs in functional foods, covering 15 common anti-gout drugs.
[0005] The technical solution of the present invention is as follows: A liquid chromatography-mass spectrometry (LC-MS) detection method for illegally adding anti-gout drugs to functional foods is provided. The method is as follows: liquid phase elution conditions are as follows: mobile phase: A is 0.1% ammonium acetate aqueous solution, B is acetonitrile, gradient elution program: 0.0 min, 90% A, 10% B; 15 min, 65% A, 35% B; 30 min, 28% A, 72% B; 33 min, 5% A, 95% B; 35 min, 5% A, 95% B; 35.1 min, 90% A, 10% B; mass spectrometry conditions are as follows: ion source: electrospray ion source (ESI source); capillary voltage: 2.5 KV; sampling cone voltage: 40 V; ion source temperature: 100 ° C; desolvation temperature: 450 ° C; full scan primary mass spectrometry (MS), mass collection range 50 ~ 1200 m / z; full scan secondary mass spectrometry (MS / MS), mass collection range 50 ~ 1 200 m / z; cone gas flow: 50 L / h, desolvation gas flow 800 L / h, collision energy: low energy: 6 V, high energy 10-30 V.
[0006] Furthermore, the liquid chromatography column used was waters C18, 150*3, with a particle size of 3 um.
[0007] The optimized liquid phase spectrum of the reference substance is as follows Figure 1 As shown in the figure, 1 allopurinol, 2 acetaminophen, 3 phenacetin, 4 rofecoxib, 5 norloxicam, 6 meloxicam, 7 colchicine, 8 probenecid, 9 sulfinpyrazone, 10 lesinade, 11 diclofenac, 12 ibuprofen, 13 febuxostat, 14 benzbromarone, 15 dotinurad.
[0008] Furthermore, the mass spectrometry detection related parameters are as follows: Compound Detection Mode Precursor ion Product ions Allopurinol Negative ions 135.03 92.02 / 107.03 Acetaminophen Positive ions 152.07 110.06 / 92.05 / 65.04 Phenacetin Positive ions 180.09 138.09 Rofecoxib Positive ions 315.06 269.06 / 218.06 / 192.09 Norcloxicam Positive ions 371.99 164.08 / 121.04 / 95.06 Meloxicam Positive ions 352.03 141.01 / 115.03 Colchicine Positive ions 400.16 358.15 / 310.10 / 282.11 Probenecid Positive ions 286.11 244.06 / 202.01 / 184.99 Sulfinpyrazone Positive ions 427.10 301.09 / 186.05 Resinade Positive ions 405.42 220.86 Diclofenac Positive ions 295.03 278.01 / 250.01 / 214.04 Ibuprofen Negative ions 205.12 159.12 Febuxostat Positive ions 317.09 261.03 / 217.04 Benzbromarone Negative ions 420.90 248.85 Dotinurad Negative ions 356.93 160.96
[0009] In the actual sample detection process, the sample is crushed and ground, and then a monohydric or dihydric alcohol aqueous solution is used to extract the sample, and then the extract is tested. The detection method of the present invention is used to compare the peak time with the reference sample spectrum and compare the mass spectrometry data, so as to quickly identify the conventional gout drugs.
[0010] Compared with the prior art, the detection method of the present invention is convenient and rapid, and can simultaneously identify 15 common gout drugs. BRIEF DESCRIPTION OF THE DRAWINGS
[0011] Figure 1 Liquid chromatogram of common anti-inflammatory and uric acid-lowering drugs (i.e., liquid chromatogram of the mixed reference sample of Example 1 of the present invention) DETAILED DESCRIPTION Example 1
[0012] Instruments and reagents: Waters Arc HPLC liquid chromatograph, Waters Q-TOF-MS mass spectrometer, Milli-Q deionized water generator (Millipore); acetonitrile (chromatographic grade, Fisher); ammonium acetate (chromatographic grade, Merck); other reagents (analytical grade, Sinopharm Chemical Reagent Company), instrument conditions: Chromatographic conditions: injection volume, 1 μL, flow rate: 0.4 mL / min; column temperature: 35 °C, mobile phase: A is 0.1% ammonium acetate aqueous solution, B is acetonitrile, gradient elution program: 0.0 min, 90% A, 10% B; 15 min, 65% A, 35% B; 30 min, 28% A, 72% B; 33 min, 5% A, 95% B; 35 min, 5% A, 95% B; 35.1 min, 90%A, 10%B; mass spectrometry conditions are: ion source: electrospray ion source (ESI source); capillary voltage: 2.5KV; sampling cone voltage: 40V; ion source temperature: 100 ℃; desolvation gas temperature: 450℃; full scan primary mass spectrometry (MS), mass collection range 50 ~ 1200m / z; full scan secondary mass spectrometry (MS / MS), mass collection range 50 ~ 1200m / z; cone gas flow rate: 50L / h, desolvation gas flow rate 800L / h, collision energy: low energy: 6V, high energy 10-30V.
[0013] Preparation of mixed standard: prepare the reference substances (commercially available): allopurinol, acetaminophen, phenacetin, rofecoxib, norloxicam, meloxicam, colchicine, probenecid, sulfinpyrazone, lesinade, diclofenac, ibuprofen, febuxostat, benzbromarone, and dotinurad into a 200 ng / mL mixed reference standard. Pass the mixed reference standard through a filter membrane and place it in a liquid sample bottle.
[0014] The optimized chromatogram is as follows Figure 1 As shown, the peak order and corresponding names are as follows: 1 Allopurinol, 2 Acetaminophen, 3 Phenacetin, 4 Rofecoxib, 5 Norloxicam, 6 Meloxicam, 7 Colchicine, 8 Probenecid, 9 Sulfinpyrazone, 10 Lecithin, 11 Diclofenac, 12 Ibuprofen, 13 Febuxostat, 14 Benzbromarone, 15 Dotinurad.
[0015] The parent ions and product ions selected after optimization are shown in the following table: Compound Detection Mode Precursor ion Product ions Allopurinol Negative ions 135.03 92.02 / 107.03 Acetaminophen Positive ions 152.07 110.06 / 92.05 / 65.04 Phenacetin Positive ions 180.09 138.09 Rofecoxib Positive ions 315.06 269.06 / 218.06 / 192.09 Norcloxicam Positive ions 371.99 164.08 / 121.04 / 95.06 Meloxicam Positive ions 352.03 141.01 / 115.03 Colchicine Positive ions 400.16 358.15 / 310.10 / 282.11 Probenecid Positive ions 286.11 244.06 / 202.01 / 184.99 Sulfinpyrazone Positive ions 427.10 301.09 / 186.05 Resinade Positive ions 405.42 220.86 Diclofenac Positive ions 295.03 278.01 / 250.01 / 214.04 Ibuprofen Negative ions 205.12 159.12 Febuxostat Positive ions 317.09 261.03 / 217.04 Benzbromarone Negative ions 420.90 248.85 Dotinurad Negative ions 356.93 160.96
Claims
1. A liquid chromatography-mass spectrometry method for detecting illegal addition of anti-gout drugs in functional foods, characterized in that: Liquid phase elution conditions were as follows: mobile phase: A was 0.1% ammonium acetate aqueous solution, B was acetonitrile, gradient elution program: 0.0 min, 90% A, 10% B; 15 min, 65% A, 35% B; 30 min, 28% A, 72% B; 33 min, 5% A, 95% B; 35 min, 5% A, 95% B; 35.1 min, 90% A, 10% B; mass spectrometry conditions were as follows: ion source: electrospray ion source (ESI source); capillary voltage: 2.5 KV; sampling cone voltage: 40 V; ion source temperature: 100 °C; desolvation temperature: 450 °C; full scan primary mass spectrometry (MS), mass collection range 50 ~ 1200 m / z; full scan secondary mass spectrometry (MS / MS), mass collection range 50 ~ 1 200 m / z; cone gas flow: 50 L / h, desolvation gas flow 800 L / h, collision energy: low energy: 6 V, high energy 10-30 V.
2. The liquid chromatography-mass spectrometry detection method according to claim 1, characterized in that: The liquid chromatography column used was waters C18, 150*3, with a particle size of 3 um.
3. The liquid chromatography-mass spectrometry detection method according to claim 1, characterized in that: The relevant parameters of mass spectrometry detection are as follows: 。