Issue date: 17/08/2020

Created By Sam
Formaldehyde
Chemicals:
 Buffer Solution (pH 5.0): 2.64g of sodium acetate in 500mL V-flask transfer 1 mL of con. Acetic
acid make up with D.I. water
 2,4-Dinitrophenylhydrazine (DNPH) solution: 120mg of DNPH in 200mL beaker make up with
ACN (light sensitive, FDG-01288, drybox-3, C)
 Extracting solvent: 100mL of DNPH solution and 100mL of Buffer solution in 250mL beaker (1:1)
 Saturated hexane in ACN: 100mL of ACN and 100ml of hexane in separating funnel discard
bottom layer.
 Mobile Phase: 70% methanol
 100ppm standard solution: ~30.0mg of 36.5% formaldehyde in 100mL V-flask make up with D.I.
Water
 10ppm intermediate solution: 1ml of standard solution in 10ml V-flask make up with water.

Calibration Curve & Program:

Vol. of Int. vol.Buffer solution 2,4-Dinitro-phenylhydrazine Make up vol. (mL) Expected
vol.(mL) sol. vol. (mL) conc.(ppm)
0.02 4.98 5 10 0.02
0.05 4.95 5 10 0.05
0.1 4.9 5 10 0.1
0.2 4.8 5 10 0.2
0.5 4.5 5 10 0.5
Follow procedure from step 2

Column: C18 column, 250 x 4.6mm, 5 um

Temperature: 40 oC UV wavelength: 365 nm
Injection volume: 20 uL Expected RT: 6 mins
Run time: 15 mins Flow: 1 mL/min
Test Procedure:
1. 2g solid sample (1ml of liquid sample) in 50ml Centrifuge tube and transfer 20mL of Extracting
solution for solid sample (10mL for liquid sample)
2. Vortex the tube and put the sample at 60 oC for 60 mins (shake the tube every 20 mins)
3. Cool down in room temperature and Centrifuge the tube at 4000 rpm in 5mins
4. Filtrate 0.45 um PDFE filter

QC parameter:
BLK, Duplicate, duplicate Spike (solid sample spike 0. 5 ppm by sample, spike 0.1ml of 10ppm
intermediate solution; liquid sample spike 2ppm by sample)

BLK, A: 2g, B:1g, sp1: 2g, sp2: 1g

 Sample high fat used by 5ml of saturated ACN: hexane solution Discard the top layer and follow
step 1 procedure