The Improvised Spectrophotometric M

The Improvised Spectrophotometric Method for Fumaric Acid Quantification
All the quantification of FA was performed spectrophotometrically following the method of
Marshall, Orten, and Smith. However, some technical improvisations were made in the
original protocol that did not cause any significant difference in the measured values of FA.
The technical comparison of the procedure mentioned in the original work and followed in the
present investigation is shown in Table 1. The outcome of the technical improvisation of the
existing technique could be evaluated in terms of cost (volumetric reduction of reagents used)
and time effectiveness (total estimation time). However, the chemical requirement was the
same in both cases. As can be summarized from Table 1, there was an almost fourfold decrease
in the total time of estimation in the improvised technique. Practically, there was no drastic
change in the FA concentration determined following both procedures.
Previously, there was no report on the spectrophotometric determination of FA in biological
sample. The method being followed in the present investigation could be a good alternative to
conventional methodological approach such as HPLC for concentration measurement of FA.
Literature on fermentation-based production of FA shows that the presence of (only qualitative)
fumaric acid was determined by the formation of insoluble mercurous fumarate in 5 %
nitric acid as no method for exact quantification of FA was available [20]. This might have
urged many researchers to develop a reliable colorimetric method for the determination of FA
concentration in biological samples. However, probably pertaining to the rapid development in
the modernization and sophistication of analytical instruments such as HPLC, this method of
FA estimation was not explored anymore. The present investigation has highlighted and
improvised a method important in both economical and analytical prospective.