Evaluation of artificially contaminated fish
with formaldehyde under laboratory
conditions and exposure assessment in
freshwater fish in Southern Bangladesh
Md. Hoque, S., Jacxsens, L., Md. Rahman,
B., Alam A.K.M. Nowsad, S.M. Azad, O.,
Meulenar, B., Lachat, C., Rahman, L. (2018)
Chemosphere 195 (2018) 702-712.
https://www.sciencedirect.com/science/articl
e/pii/S0045653517320817
Formalin can be added as preservative to fresh
foods to prevent spoilage and extend shelf life.
Formalin contains 37–40% formaldehyde, which
is classified as carcinogenic to humans. To assess
the public health risk associated with
formaldehyde exposure in freshwater fish in
Southern Bangladesh, formaldehyde
concentrations (mg/kg) were determined in
tilapia, Indian major carp rui, Chinese carp and a
minor carp from local market and in laboratory
simulations with spectrophotometric and high
performance liquid chromatography (HPLC)
methods.
I. Determination of formaldehyde concentration
using spectrophotometry
The spectrophotometric method with some
modification (Benjakul et al. 2003) using Nash’s
reagent was applied to determine the
formaldehyde content (mg/kg) in fish. Nash’s
Reagent was used as an indicator which helps to
detect the absorbance of formaldehyde (Nash,
1953). To prepare Nash’s reagent, 15 g
ammonium acetate was diluted in a 100 ml
Erlenmeyer flask with an addition of 0.3 ml of
acetyl acetone and 0.2 ml of acetic acid. About
30 g fish flesh was blended for 10 minutes, using
a Philips HR-2106 blender. Sixty (60) mL of 6%
w/w TCA was added for extraction of
formaldehyde from the fish flesh. The extracted
solution was then filtered by a Whatman No.1
filter paper. The pH of the solution was
determined by a pH meter (pH 211, Hanna
Instruments, Italy). The addition of TCA resulted
in a reduced pH value of the sample which was
then adjusted between 6.00-6.50 by using
potassium hydroxide (KOH) and hydrochloric
acid (HCl). Five (5) mL of sample solution was
taken in a 50 mL volumetric/conical flask and
kept in a freezer (Walton W2D-1H5,
Bangladesh) at -20 ° 159 C for 1 h. The sample
was taken out of the freezer and 2 mL of
previously prepared Nash’s reagent was added as
indicator. The fish sample was then heated in the
water bath (Fisherbrand FB60301, China) at 60 °
161 C for 30 minutes. The absorbance of the
sample in a cuvette was measured at 415 nm
immediately by UV/Vis spectrophotometer (T60
UV/Visible Spectrophotometer, PG Instruments,
U.K). The sample reading (triplicate) was placed
in the standard curve for the calculation of
formaldehyde concentration in the fish sample.

II. Determination of formaldehyde concentration

using HPLC

Analyzing Time Dynamic Concentration of
Formaldehyde in Fresh and Formalin
Treated Fish ‘Labeo rohita’
Md. Uddin, M., Amit, Sadat Kamal, S.M.
Islam, R., Rahman, R., Sameera, S., Khan,
Mohdius Samad. (2014). Proceedings of the
ICChE (2014), 29-30 December, Dhaka,
Bangladesh.
https://www.researchgate.net/profile/Mohidu
s_Samad_Khan/publication/279529832_Anal
yzing_Time_Dynamic_Concentration_of_Fo
rmaldehyde_in_Fresh_and_Formalin_Treate
d_Fish_'Labeo_rohita'/links/55954b6908ae2
1086d201e09.pdf
Food is a naturally perishable substance that
undergoes degradation because of the spoilage
actions of chemical, microorganisms or
enzymatic reactions. To retain the freshness of
food items different physical and chemical
preservative techniques are used. In Bangladesh,
it is reported that formalin, a 37-40% solution of
formaldehyde, is widely used as a chemical
preservative to increase the shelf-life of food
items, especially fishes
I. Formaldehyde Detection
Bones of the fishes were removed. Portions of
the fishes taken for the experiment were blended
with equal weight of water for approximately 1
min. Equal weight of Trichloroacetic acid (10
w/w %) was added with the blend and was
mixed properly with the help of glass rod. The
fish blend was filtered by Whatman 42 filter
paper. 5ml of filtrate was taken and 10ml of
distilled water was added with it. pH was
adjusted between 6 to 6.5. Volume of the extract
was diluted to 25ml by adding water. 5ml of this
extract was taken and 5ml of Nash was added.
Mixture was kept at 60°C water bath for 5min.
Then mixture was cooled to room temperature
and optical density (absorbance) was measured
at 415nm by using UV mini-1240
spectrophotometer (Smith 1973). Calibration
curves (Fig. 1 and 2) for formaldehyde solutions
were prepared to quantify formaldehyde in fresh
and treated fish samples.
II. Time dynamic behavior of formaldehyde in
formalin treated fish: fish sample dipped in
formalin solution
III. Time Dynamic Behavior of Formaldehyde in
Formalin Treated Fish: Formalin Sprayed on
Fish Sample
OUTLINE

Evaluation of artificially contaminated fish with formaldehyde under laboratory conditions and
exposure assessment in freshwater fish in Southern Bangladesh

I. Determination of formaldehyde concentration using spectrophotometry

II. Determination of formaldehyde concentration using HPLC

Analyzing Time Dynamic Concentration of Formaldehyde in Fresh and Formalin Treated Fish
‘Labeo rohita’

I. Formaldehyde Detection
II. Time dynamic behavior of formaldehyde in formalin treated fish: fish sample dipped
in formalin solution
III. Time Dynamic Behavior of Formaldehyde in Formalin Treated Fish: Formalin
Sprayed on Fish Sample