Application range
This standard specifies HPLC methods for content determination of nadolol in feed.
This standard applies for the detection of nadolol in formula feed (The sample for this experiment is a kind of formula feed)
Reference Standard: No. 1063-4-2008 Announcement of Ministry of Agriculture Detection of nadolol in formula feed HPLC
Extraction steps
Accurately weigh 5g formula feed (accurate to 0.001g) in 50 mL centrifuge tube, then add 25mL methanol and vortex for 1 min and through ultrasound for 10 min, centrifuge at 8000r/min for 5 min. Take the supernatant.
Purification steps
SPE Column: Welchrom® P-SCX column, 150mg/6mL
Activation: 5mL methanol, 3mL water, discard
Sample loading: 5mL supernatant + 5mL water, mix well before loading sample, discard
Elution: 3mL methanol, drain and discard
Concentration: blow to dry with nitrogen at 50 ℃, then add the dilution to 1mL. Filter through 0.22 μm NY filter membrane for detection.
Chromatographic conditions
Column: Xtimate® C8, 4.6×150mm, 5μm
Flow rate: 1.0mL/min
Column temperature: 25 ℃
Injection volume: 10 μL
Detector: fluorescence detector, excitation wavelength: 230nm, emission wavelength: 300nm.
Gradient program:
| Time/min | 0.5% Methanic acid solution/% | Acetonitrile |
| 0 | 95 | 5 |
| 8 | 55 | 45 |
| 12 | 95 | 5 |
| 15 | 95 | 5 |
Chromatogram or spike recovery rate
Fig 1. Blank chromatogram of nadolol 1.0mg/L
Fig 2. Blank chromatogram of formula feed
Fig 3. Chromatogram of formula feed at 1.0mg/kg
Tab 1. Spike Recovery Rate
| Substance | Spike level | Average recovery rate | RSD（n=2） |
| Nadolol | 1.0mg/kg | 99.32% | 0.98% |
Ordering information
| P/N | Product | Specification |
| 00523-20043 | SPE column | Welchrom® P-SCX,150mg/6mL, 30pk |
| 00837-05006 | 50mL centrifuge tube with screw mouth and apical bottom | Disposable centrifuge tube, flat cap, conical bottom, RCF12000xg, 15 mL, 50/ pk without sterilization |
| 00824-31001 | Welch SPE instrument | 12 ports square cylinder |
| 00821-32291 | Caps and septa | Pre-slit red PTFE/white Silicone septa, 9mm blue short screw-thread polypropylene cap, 6mm centre hole, 100/pk |
| 00821-40927 | Vials | 2ml wide opening short screw-thread vial with write-on spot, clear, 11.6×32mm, 100/pk |
| 00102-21041 | HPLC column | Xtimate® C8,4.6×150mm, 5μm |
| 00802-02201 | Syringe filter | NY, 13mm×0.22μm,100pk |
If you have any problem or require further information, please contact info@welchmat.com.
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