Nadia Citta

12/329521/TP/10309
Muktiadin Latifah

12/333186/TP/10447
Imasni Satriani
13/346395/TP/10548
Norma Dwi Septian
13/346454/TP/10561
Duanti Oktarani
13/346483/TP/10577
Wea Aanisah
IN 13/348620/TP/10717
HOUSE VALIDATION FROM DIRECT

DETERMINATION OF 5-HYDROXYMETHYL2-FURFURAL (HMF) IN BRAZILIAN CORN

AND CANE SYRUPS SAMPLES BY HPLC-UV

Introduction
Corn

syrups
are
applied
as
sweeteners and also as a humectant.

Cane

syrup has been applied as an

ingredient in confectionery products,
soft drinks, candies, and even as a
syrup substitute in certain types of
fruit preserves.

Syrup

HMF

Diseas
es

New methods have been developed for

HMF determination in many different
foods.

The analytical methodology more usually

adopted is the liquid chromatography with
ultraviolet detection (HPLCUV).

Method validation can be realized at two

different levels: full validation and in
house validation.

Material
HMF

99%
Formic acid 95%
Acetonitrile (HPLC grade)
Ultrapure water (0.055 S/cm)
Six corn syrups samples coded (S1S6)
Six cane syrups coded (CS1CS6

Sample and Standard

Solutions Preparation
Standard : 1.05.0 mg/L
S1-S6
: 0.5 g in 250 mL
CS1CS6: 0.5 and 3.0 g in 100 mL

Chromatographic
conditions
Breeze

HPLC system
Phenomenex column (Luna C18, 5.0
m, 4.6 x 150 mm) at 30 oC
Mobile phase : water (with 0.5%
formic acid) and acetonitrile in the
ratio 90:10 (v/v)
Detected at 285 nm
Total run time : 8 min

Validation Study
Selectivity
Calibration and linearity
Detection (LOD) and quantification

(LOQ) limits
Precision
Accuracy

Results and discussion

Method development

Initially the chromatographic conditions reported by Lemos et al. (2010)

were tested for the cane syrup sample (CS1). It showed that the peaks
were not well separated (resolution < 1.5).

Therefore, different aqueous mobile phases with acetonitrile consisting

of ratios between (80:20, v/v) until (90:10, v/v) were tested. Better
results were observed with the ratio of water:acetonitrile (90:10, v/v).

It was observed by other authors using similar ratios of water and

acetonitrile (92:8 or 90:10, v/v). The flow rate varied from 1.0 to 0.6
mL/min using the same mobile phase composition. Flow rate of 0.8
mL/mi1 with separation time around 4.6 min gave higher sensitivity and
was therefore selected for further optimization.

CS1 were encountered values for k of 2.84 0.01, tailing factor of HMF
peak of 1.23 0.02, and resolution of HMF with an adjacent peak (Rs) of
3.13 0.04.

S1 were encountered values for k of 1.57 0.01, Rs of 3.31 0.05, and

Selectivity

Linearity

LOD and LOQ

Detection (LOD) and quantitation (LOQ) limits were 0.09 and
0.26 mg/L

Precision and accuracy

Method application to real

samples

Conclusions
A

quick and direct method of

determination of HMF in corn and
cane syrup samples has been
developed and validated.
Results showed that the HMF
concentration in Brazilian corn and
cane syrup samples are very high.