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ORIGINAL ARTICLE
Center for Undergraduate Studies, University of the Punjab, New Campus, Lahore-54590, Pakistan
Institute of Chemistry, University of the Punjab, New Campus, Lahore-54590, Pakistan
KEYWORDS
Chromotropic acid;
Densitometry;
Formaldehyde;
Micro-analysis;
Quantication of spot test;
Spot test analysis
Abstract A simple, accurate, and economical method has been proposed to measure formaldehyde in
aqueous samples. The method is based on quantication of classical chromotropic acid formaldehyde violet spots developed on TLC. Different parameters such as concentration of chromotropic
and sulfuric acid, time of heating and order of application of reagents have been studied to nd out
the optimum working procedure. Spots have been quantied by scanning the spotted TLC and analyzing the image in computer with Visual Basic 6.0 based graphic application. The study consisted of
developing an appropriate calibration line, analyzing articial and real samples, and comparing the
new method with a standard spectrophotometric method (NIOSH Method-3500, 1994). It has been
concluded that the present method had the capability for measuring formaldehyde in aqueous samples
at trace level with high precision and accuracy, particularly when dealing with turbid and small volume
samples where the standard method failed.
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1. Introduction
Formaldehyde or methanal (HCHO) is the simplest aldehyde
that is an important precursor to many chemical compounds.
Yearly production of formaldehyde around the globe has
exceeded 23 million tons during last few years (Lide, 2004).
* Corresponding author. Tel.: +92 4299232068; fax: +92 4299232057.
E-mail address: research.group.pu@gmail.com (A. Dar).
Peer review under responsibility of King Saud University.
Formaldehyde is a gas at room temperature, but readily converts into a variety of other gaseous derivatives like trioxane.
Formaldehyde occurs in the environment up to 0.03 ppm parts
of air. Materials having formaldehyde, such as ureaformaldehyde foam insulation (UFFI) can release it in the form of gas or
vapor. Pressed-wood products are the major source of formaldehyde indoor pollution. Cigarette smoke, gas stoves, woodburning stoves, and kerosene heaters can also release formaldehyde. It is highly toxic to both plants and animals, and is particularly dangerous for the human eyes (Swenberg et al., 1980).
When present in the air at levels exceeding 0.1 ppm, it causes
severe irritation in the eyes, nose, and throat and usually results
in watery eyes with burning sensations, coughing, wheezing,
nausea, and skin irritation (Sakai et al., 2002). There is some
http://dx.doi.org/10.1016/j.jscs.2012.12.002
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This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
S353
proportional to the concentration of formaldehyde, a simple
analytical method has been developed for precise quantication
of formaldehyde in aqueous samples at trace levels. The color
densities of the spots were measured with simple software by
taking the image of spotty TLC in computer subsequent to
scanning. There are other techniques to quantify chromotropic
acid formaldehyde spots (Boyd and Logan, 1942; Boos, 1948);
the present method is only one of its kind in terms of measuring,
quantity of sample, simplicity and accuracy. The most appropriate application of the method is on-eld analysis of trace
amounts of formaldehyde in aqueous samples.
2. Materials and methods
2.1. Stock solution and standards
Stock solution of formaldehyde (1000 mg l 1) was prepared by
diluting analytical grade 10% formalin solution (SigmaAldrich, Inc.) with proper quantity of double distilled water.
Standard solutions of desired concentrations (210 mg l 1)
were prepared by appropriate dilution of the stock solution.
2.2. Chemicals
Technical grade chromotropic acid disodium salt dihydrate,
(HO)2C10H4(SO3Na)22H2O and reagent grade concentrated
sulfuric acid (SigmaAldrich, Inc.), 9598%, 1.840 g ml 1 at
25 C (lit.) were the chemicals that were used as it is or as doubled distilled water diluted solutions in order to evaluate the
effect of these on color intensity of spots. Desired solutions
of chromotropic acid were prepared in 50% sulfuric acid.
2.3. Procedure
TLC plate (Merck, Aluminum sheet, Silica gel 60F254,
3 5 cm) was used to develop spots since paper could not be
employed (burnt because of concentrated sulfuric acid). One
micro-liter drop of each, chromotropic acid, sulfuric acid
and aldehyde solution was employed with micropipette (Pipettman) one over the other. The order of application of reagents and aldehyde solution was altered to check the effect
on color density of the spot. Later, the TLC was placed in
an oven for 220 min at 60 C to notice the effect of time. Violet color spots intensifying on cooling were obtained.
2.4. Quantication of spots and analysis of samples
The spotty plate was scanned on a atbed scanner (HP 3670
CCD Reective Flatbed Scanner) and the image was imported
in Visual Basic 6.0 based graphic application to measure the
color density of spots. Detail of graphic application to read
color densities of the spots is given in (Anwar et al., 2010).
A calibration line was plotted taking concentration of formaldehydes standards as abscissa against the corresponding color
density of spots (ordinate). Slope, intercept, standard error of
estimates and correlation coefcient were measured. Finally,
synthetic as well as true aqueous samples of formaldehyde
were analyzed using the same set of conditions by measuring
their color densities and interpolating the corresponding concentrations from standards calibration line.
S354
A. Dar et al.
SO 3
SO 3H
HO
- H2 O
HO3S
HO
OH
HO
OH
HCHO
HO
SO 3
SO 3H
HO3S
-H2O
+O
SO 3H
HO3S
HO
HO
OH
SO 3H
HO3S
Violet-red
Scheme 1
Seses.
OH
O3 S
HO
O
SO 3
SO 3
SO 3
Scheme 2
S355
Asasa.
the nature of this chromogen and its formation has never been
conclusively proven.
The effect of concentration of chromotropic acid on color
intensity of the spot was studied. It was found that the color
intensity had increased with an increase in the concentration
of chromotropic acid up to 2%. However, afterward there
was no prominent increase in color density with further increase in concentration. In the same way, time to heat TLC
at 60 C was changed from 2 to 20 min and it was found that
14 min were optimum to obtain maximum intensity of the spot.
To check the selectivity of the method, different compounds
were tested separately and with formaldehyde to study the
interference if any. It was noted that among acetaldehyde,
Figure 1
Table 1
propanal, butanal, isovaleric aldehyde, chloral hydrate, glyoxal, aromatic aldehydes, formic acid, phenoxyacetic acid
and isopropyl ester, only the last two produced violet spots
at optimum conditions (2% chromotropic acid, 50% sulfuric
acid, and heating at 60 C for 14 min), while furfural, fructose
and sucrose produced yellow spots. The order of application of
reagents and aldehyde solution was also changed and it was
noted that the correct order was to apply formaldehyde solution foremost, then sulfuric acid followed by chromotropic
acid. When chromotropic acid was applied foremost, the nal
color of spot was brownish rather than violet. It may be because of silica or some impurity on TLC that interfered with
chromotropic acid and did not allow it to produce violet chromogen (Scheme 2) on contact with formaldehyde.
TLC containing violet spots was imported to specially designed Visual Basic 6.0 based graphical application using atbed scanner at 300dpi, 24-bit RGB mode. Each spot was
selected inside a selection marquee and the color density was
measured. The software measured the red, green, and blue components of each pixel inside the margin and sums up the values
to give a nal color density. Calibration lines (Fig. 1) were plotted for formaldehyde standards in the range of 210 mg l 1 by
the present method (color density against concentration) and
the standard NIOSH method (absorbance against concentra-
Calibration lines of the present and standard method using Microsoft Excel 2007.
Comparison of calibration lines of the present and standard method using Microsoft Excel 2007.
Parameter
Standard method*
Present method
Slope
Y-intercept when X = 0.0
X-intercept when Y = 0.0
1/slope
0.049 0.002
0.086 0.015
1.778
20.580
18.700 0.627
38.000 4.160
2.032
0.053
0.041 to 0.056
0.038 to 0.135
3.219 to 0.692
16.700 to 20.700
24.760 to 51.240
3.043 to 1.206
Goodness of t
r2 **
Sy.x ***
Result for 5 mg l
Percentage error
0.993
0.014
4.745
5.100%
0.997
3.967
5.044
0.880%
solution
Spectrophotometric method (NIOSH Method-3500 (1994), NIOSH Manual of Analytical Methods (NMAM), ALS Columbia).
Coefcient of determination.
***
Standard error of estimates.
**
S356
tion, Method-3500 published in 1994). The calibration data are
compared in Table 1. Coefcient of determination for the present method (0.997) was better than the standard method
(0.993). The coefcient of determination is useful as it gives
the amount of the uctuation of one variable that is predictable
from the other variable. In addition to the standard statistical
analysis of calibration, an articial sample (5 mg l 1) was tested
by both methods. The output of the present method was
5.044 mg l 1 (0.88% ) whereas the standard method found it
to be 4.745 mg l 1 (5.10% ). The present method used only
1 ll of the sample while the standard method used 5 ml. Moreover, present method was applicable to turbid samples but the
standard method could only analyze clear solutions.
Actual samples (tap and canal water from university campus,
and textile efuent from local industry) were also tested by both
methods. Tap water was used as such and no formaldehyde was
detected by both methods. Canal water and efuent were allowed to settle for 30 min and then ltered ahead of analysis with
the standard method, whereas the sample was used without ltration in case of the present method. The formaldehyde content
of canal water was 2.546 and 2.429 mg l 1 while the concentrations of formaldehyde in textile efuent were 8.926 and
8.654 mg l 1 by the present and standard method, in that order.
The present method is not only economical but also accurate,
and it can be used for on-eld analysis with the help of portable
scanners and laptops/smart phones (having color density measuring software) since it does not involve painstaking sample
preparations and sophisticated instruments.
4. Conclusion
A trouble-free, precise, and inexpensive method of formaldehyde analysis for aqueous samples has been proposed. The
method was based on quantication of classical chromotropic
acid formaldehyde violet spots developed on TLC. Optimum
working conditions were 2% chromotropic acid, 50% sulfuric
acid, and 14 min heating at 60 C to get utmost color intensity
spots. The correct order of reagents application was sample
(formaldehyde), followed by sulfuric acid and then chromotropic acid (1 ll drop of each). Spots were quantied by scanning
the spotted TLC and analyzing the image in computer with
homemade Visual Basic 6.0 based graphical application. Calibration line of the present method was compared with that of
the standard spectrophotometric method. The present method
was found to have better capability for measuring formaldehyde in aqueous samples at trace level with high precision
and accuracy, especially when dealing with turbid and small
volume samples where the standard method was inapplicable.
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