44.4.

15
AOAC Official Method 980.23
Hydroxymethylfurfural in Honey
Spectrophotometric Method
First Action 1980
Final Action 1983

(Do not heat honey before this determination.)

A. Apparatus and Reagents
(a) Spectrophotometer.—UV, to measure A at 284 and 336 nm.
(b) Carrez solution I.—Dissolve 15 g K4Fe(CN)6⋅3H2O and di-
lute to 100 mL with H2O.
(c) Carrez solution II.—Dissolve 30 g Zn(CH3COO)2⋅2H2O and
dilute to 100 mL with H2O.
(d) Sodium bisulfite solution.—0.20%. Dissolve 0.20 g NaHSO3
(technical grade is satisfactory) and dilute to 100 mL with H2O. Di-
lute 1 + 1 for dilution of reference solution if necessary. Prepare
fresh daily.
B. Determination
Accurately weigh ca 5 g honey in small beaker and transfer with
total of ca 25 mL H2O to 50 mL volumetric flask. Add 0.50 mL
Carrez solution I, mix, add 0.50 mL Carrez solution II, mix, and di-
lute to volume with H2O. Drop of alcohol may be added to suppress
foam. Filter through paper, discarding first 10 mL filtrate.
Pipet 5 mL filtrate into each of two 18 × 150 mm test tubes. Add
5.0 mL H2O to 1 tube (test solution) and 5.0 mL NaHSO3 solution to
other (reference). Mix well (Vortex mixer is useful) and determine A
of test solution against reference at 284 and 336 nm in 1 cm cells. If A
is >0.6, dilute test solution with H2O and reference solution with
0.1% NaHSO3 solution to same extent and correct A for dilution.
mg Hydroxymethylfurfural (HMF) (A284 − A336 ) × 14.97 × 5
=
100 g honey g test sample

Factor = 14.97 = (126/16 830)(1000/10)(100/5)

where 126 = molecular weight HMF; 16 830 = molar a of HMF at

284 nm; 1000 = mg/g; 10 = centiliters/L; 100 = g honey reported; 5 =
nominal test portion weight.
Reference: JAOAC 62, 509, 515, 921(1979).
CAS-67-47-0 (hydroxymethylfurfural)

© 2000 AOAC INTERNATIONAL