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hplc chromatography in determination of Aldehydes and Ketones

The principle of hplc chromatography in stationary pollution source waste gas - Determination of aldehydes and ketones

The aldehydes and ketones in the waste gas from the fixed pollution sources react with the 2,4-dinitrophenylhydrazine (DNPH) in the absorption liquid in an acidic medium to generate stable 2,4-dinitrophenylhydrazone derivatives thing. After extracting and concentrating with dichloromethane, it is separated by high performance liquid chromatography and detected by ultraviolet or diode array detector. Qualitative by retention time, quantitative by external standard method.The chemical reaction is as follows

Instruments and Equipment

Instruments nameModel
High performance liquid chromatography(HPLC)LC3100
Quaternary low pressure gradient pumpP3200Q
Column ovenCT3200
Diode array detectorDAD3200

Chromatographic conditions

  • Column:C18 column, 4.6×250mm, 5μm, pH range: 2-11, double-terminated reversed-phase column or equivalent column with octadecylsilane-bonded silica gel (ODS) as packing material
  • Mobile phase:A: Acetonitrile B: Water C: Methanol
  • Flow rate:1.0 mL/min
  • Injection volume:10 μl
  • Column  temperature:35℃
  • Detector:DAD
  • Detection wavelength:360 nm

Gradient elution procedure


Chromatogram of 12 kinds of aldehydes and ketones-DNPH derivatives mixed standard

1-formaldehyde-DNPH; 2-acetaldehyde-DNPH; 3-acrolein-DNPH; 4-acetone-DNPH; 5-propionaldehyde-DNPH; 6-butenal-DNPH; 7-2-butanone-DNPH; 8-butyraldehyde-DNPH; 9-benzaldehyde-DNPH; 10-isovaleraldehyde-DNPH; 11-n-pentanal-DNPH; 12-n-hexanal-DNPH.

12 kinds of aldehyde and ketone compounds DNPH peak situation table

Peak No.Compound nameRetention timePeak heightPeak areaResolution
1Formaldehyde DNPH7.41718018919.029
2acetaldehyde DNPH10.000107124810.512
3Acrolein DNPH13.3848911871.249
4Acetone DNPH13.817709632.490
5propionaldehyde DNPH14.684729799.478
6crotonaldehyde DNPH18.167558574.158
72-Butanone DNPH19.967528220.664
8n-Butyraldehyde DNPH20.284487646.990
9Benzaldehyde DNPH23.350406433.658
10Isovaleraldehyde DNPH24.800456973.052
11n-valeraldehyde DNPH26.0174368013.588
12n-hexanal DNPH31.50037576--

Chromatograms of 12 aldehydes and ketones after derivatization

1-Formaldehyde DNPH, 2-acetaldehyde DNPH, 3-propanal DNPH, 4-propanal DNPH, 5-propanal DNPH, 3'-propanal DNPH polymer, 6-butenal DNPH, 7-2-butanone DNPH, 8-n-butyraldehyde DNPH, 9-benzaldehyde DNPH, 10-isovaleraldehyde DNPH, 11-n-valeraldehyde DNPH, 12-n-hexanal DNPH


  1. Wayeal instrument hplc chromatography can realize the separation and detection of 12 kinds of aldehydes and ketones;
  2. In order to ensure that the blank is not disturbed by the environment, the brown reagent bottle storing the DNPH saturated absorption solution should be washed with hot water and ultrapure water in advance, and then dried at 130 °C for 3 hours;
  3. The derivatization reaction produces water. When drying with anhydrous sodium sulfate, the particles should flow freely. At this time, the system reaches an anhydrous state, thus entering the concentration step.
hplc chromatography



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