Method for Determining 16 Benzimidazole Drugs Using Anyeep TQ9100
This study established a method for the simultaneous detection of 16 benzimidazole drugs using Anyeep TQ9100, an ultra-high-performance liquid chromatography-tandem mass spectrometry system from Anyeep. The method demonstrated excellent linearity, with detection and quantification limits meeting relevant regulatory standards. This approach enables reliable detection and quantification of benzimidazole drugs.
Benzimidazole is an organic compound with the chemical formula C7H6N2. It serves as a key intermediate for producing fungicides such as imazodane and propiconazole. Benzimidazole-based drugs are systemic fungicides featuring a benzimidazole ring with fungicidal activity. They exhibit broad-spectrum bioactivity and are widely used in both pharmaceutical and agricultural sectors.
Routine testing involves animal-derived foods and feeds. Applicable standards include GB/T 22955-2008, GB/T 21324-2007, SN/T 2559-2010, SN/T 1753-2016, and Ministry of Agriculture Announcement No. 1730-1-2012. This method is based on GB/T 22955-2008 and GB/T 21324-2007, using the external standard method for quantification.
Instruments and Reagents
Anyeep TQ9100 Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry System
Acetonitrile (Mass Spectrometry Grade, Thermo Fisher Scientific, USA)
Formic acid (mass spectrometry grade, CNW Technologies, Germany)
Liquid Chromatography Conditions
Mobile Phase A: 0.1% Formic Acid/Water
Mobile Phase B: Acetonitrile
Column: Waters ACQUITY UPLC BEH C18 1.7μm, 2.1*50 mm
Column Temperature: 40 °C
Flow rate: 0.3 mL/min
Injection volume: 5 μL
Mass Spectrometry Conditions
Ionization Source: Electrospray Ionization Source (ESI+)
Source Temperature: 550 ℃
Spray Voltage: 5000 V
Curtain Airbag: 20 psi
Mist 60 psi
Auxiliary Air: 60 psi
Collision Gas: 6
Scan Mode: MRM
Gradient elution program
Time min | A% | B% | Flow Rate mL/min |
0 | 80 | 20 | 0.3 |
4 | 20 | 80 | 0.3 |
4.5 | 20 | 80 | 0.3 |
4.6 | 80 | 20 | 0.3 |
6 | 80 | 20 | 0.3 |
Analyte MRM Parameters
No. | Test Item | Q1 Ion | Q3 Ion | Residency Time (ms) | CE (V) | DP (V) |
1 | Albendazole-2-sulfone | 240.0 | 133.0* | 20 | 28 | 150 |
|
| 240.0 | 198.0 | 20 | 15 | 150 |
2 | 5-Hydroxy thiabendazole | 218.0 | 147.0* | 20 | 32 | 150 |
|
| 218.0 | 191.0 | 20 | 25 | 150 |
3 | Thiophanate | 202.0 | 131.0 | 20 | 35 | 150 |
|
| 202.0 | 175.0* | 20 | 25 | 150 |
4 | Aminotrimazole | 238.0 | 105.0* | 20 | 22 | 180 |
|
| 238.0 | 133.0 | 20 | 35 | 180 |
5 | Albendazole sulfoxide | 282.0 | 191.1 | 20 | 40 | 150 |
|
| 282.0 | 208.0* | 20 | 22 | 150 |
6 | 5-hydroxy mebendazole | 298.0 | 160.0 | 20 | 35 | 150 |
|
| 298.0 | 266.1* | 20 | 22 | 150 |
7 | 2-Aminofluorobenzimidazole | 256.0 | 95.0 | 20 | 48 | 180 |
|
| 256.0 | 123.0* | 20 | 25 | 180 |
8 | Albendazole sulfone | 298.0 | 224.0 | 20 | 25 | 150 |
|
| 298.0 | 159.0* | 20 | 38 | 150 |
9 | albendazole | 250.0 | 176.0 | 20 | 25 | 150 |
|
| 250.0 | 218.0* | 20 | 15 | 150 |
10 | Cyanthidazole | 303.0 | 217.1* | 20 | 28 | 150 |
|
| 303.0 | 261.1 | 20 | 15 | 150 |
11 | Albendazole | 316.0 | 159.0* | 20 | 35 | 180 |
|
| 316.0 | 191.0 | 20 | 20 | 180 |
12 | Fenbendazole sulfone | 332.0 | 159.0 | 20 | 38 | 180 |
|
| 332.0 | 300.0* | 20 | 20 | 180 |
13 | Albendazole | 266.0 | 191.0 | 20 | 32 | 150 |
|
| 266.0 | 234.0* | 20 | 15 | 150 |
14 | Mebendazole | 296.0 | 105.0 | 20 | 35 | 150 |
|
| 296.0 | 264.0* | 20 | 18 | 150 |
15 | Fenbendazole | 314.0 | 123.0 | 20 | 35 | 150 |
|
| 314.0 | 282.0* | 20 | 20 | 150 |
16 | Fenbendazole | 300.0 | 159.0 | 20 | 35 | 150 |
|
| 300.0 | 268.0* | 20 | 20 | 150 |
Note: * indicates the quantification ion pair.
Linear Range
No. | Test Item | Retention Time min | Limit of Detection ng/mL | Linear Range ng/mL | Linear Equation 1/x2 weighted | correlation coefficient r |
1 | Albendazole-2-sulfone | 0.998 | 0.01 | 0.025-50 | Y = 60850.6 * X + 146.098 | 0.9969 |
2 | 5-hydroxy thiabendazole | 0.929 | 0.01 | 0.025-50 | Y = 92767.1 × X - 584.292 | 0.9988 |
3 | Thiabendazole | 1.012 | 0.025 | 0.05-50 | Y = 164176 × X - 957.247 | 0.9965 |
4 | Aminotrimazole | 1.579 | 0.025 | 0.05-50 | Y = 39139.8 × X - 151.423 | 0.9956 |
5 | Albendazole sulfoxide | 1.247 | 0.025 | 0.05-50 | Y = 55068.8 × X - 444.04 | 0.9975 |
6 | 5-hydroxy mebendazole | 1.537 | 0.01 | 0.025-50 | Y = 213003 × X - 565.917 | 0.9988 |
7 | 2-Aminofluorobenzimidazole | 1.717 | 0.01 | 0.025-50 | Y = 67638.3 * X + 822.987 | 0.9949 |
8 | Albendazole sulfone | 1.758 | 0.01 | 0.025-50 | Y = 84191.9 × X - 595.787 | 0.9972 |
9 | albendazole | 1.827 | 0.005 | 0.01-50 | Y = 314783 * X + 1055.93 | 0.9970 |
10 | Cipendazole | 1.648 | 0.005 | 0.01-50 | Y = 147755 * X + 223.444 | 0.9984 |
11 | Albendazole | 1.758 | 0.01 | 0.025-50 | Y = 58823.2 * X + 296.146 | 0.9960 |
12 | Fenbendazole sulfone | 2.242 | 0.005 | 0.01-50 | Y = 90512.6 × X - 0.721244 | 0.9962 |
13 | Albendazole | 2.352 | 0.005 | 0.01-50 | Y = 283031 * X + 552.487 | 0.9978 |
14 | Mebendazole | 2.291 | 0.005 | 0.01-50 | Y = 174624 * X + 401.96 | 0.9978 |
15 | Fenbendazole | 2.463 | 0.005 | 0.01-50 | Y = 165351 * X + 28.2079 | 0.9971 |
16 | Fenbendazole | 2.850 | 0.005 | 0.01-50 | Y = 274587 * X + 1388.05 | 0.9972 |
Summary
This study established an ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) system using the Anyeep TQ9100 for the analysis of 16 benzimidazole drugs. Results demonstrate that this method offers excellent sensitivity, stability, and linear range, making it suitable for routine analysis and detection of benzimidazole drugs.