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/min0.5% Methanic acid solution/%Acetonitrile

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

SubstanceSpike levelAverage recovery rateRSD(n=2)

Ordering information

00523-20043SPE columnWelchrom® P-SCX,150mg/6mL, 30pk
00837-0500650mL centrifuge tube with screw mouth and apical bottomDisposable centrifuge tube, flat cap, conical bottom, RCF12000xg, 15 mL, 50/ pk without sterilization
00824-31001Welch SPE instrument12 ports square cylinder
00821-32291Caps and septaPre-slit red PTFE/white Silicone septa, 9mm blue short screw-thread polypropylene cap, 6mm centre hole, 100/pk
00821-40927Vials2ml wide opening short screw-thread vial with write-on spot, clear, 11.6×32mm, 100/pk
00102-21041HPLC columnXtimate® C8,4.6×150mm, 5μm
00802-02201Syringe filterNY, 13mm×0.22μm,100pk

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