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The Coleman universal spectrophotometer, model 11, with wave band
set at 620 rnp, was used in this study. A resistance was placed in the
circuit of the exciter lamp so that the brilliance of the light could be
* Contribution No. 298, Department of Chemistry.
181
182 DETERMINATION OF VITAMIN A
following mixing of the reagent with vitamin A. The values shown for the
Coleman photometer were all obtained by using the calibration curve pre-
pared with the incident light at 13 per cent normal brilliance.
The data in Fig. 1 and Table I show clearly that light exerts a powerful
effect in the fading of the blue color. As the incident light of the Coleman
photometer is changed from 100 to 13 per cent of full normal brilliance, the
rate of loss of color during the first minute decreases from 39 to 18.2 per
cent. Exposure of the solution to light at 13 per cent of normal for only
long enough to read the galvanometer reduces this loss of color to 11.7 per
cent. The loss of color is 89.4 per cent if the light of a 200 watt bulb is
allowed to strike the solution. A smaller loss, 7.8 per cent, is observed if
light of low intensity such as that employed in the Evelyn photometer is
used. The extrapolation of all curves to zero time tends to bring all the
curves toward a common point. As would be expected from a study of the
curves, there is a tendency for the photometric reading to be lower when
184 DETERMINATION OF VITAMIN A
TABLE I
Rate of Fading of Carr-Price Color from Initial to 60 Second Reading
Photometer Light intensity Rate of color loss
through six pieces of glass and an aqueous solution before reaching the
absorption cells; and yet fading was very rapid.
Hock (6) studied the kinetics of the Carr-Price reaction, using a photo-
graphic device to record color density. Fig. 4, a of his paper shows that
vitamin A naphthoate fades so rapidly that if the same rate had continued
for 70 secondsthe value would have been practically 0 per cent of the maxi-
mum color developed. The results of this study indicate that a lessintense
light source would have causedlessrapid fading. In making kinetic studies
of the Carr-Price reaction, one might be led to assume either a zero or a
first order reaction, depending upon the intensity of light employed in the
photometer. Meunier and Raoul (9) studied the kinetics of the Carr-
Price reaction of vitamins A1 and A,. The intensity of the light source
may causethe two forms of the vitamin to fade at different rates than those
found in their study and merits further investigation. Since the deter-
mination of correction factors for the presence of carotenoids interfering
M. J. CALDWELL AND D. B. PARRISH 185
SUMMARY
1. The intensity of the incident light influences the rate of fading of the
blue color developed in the Carr-Price reaction for vitamin A.
2. Investigations of the kinetics of the Carr-Price reaction should take
into account the effects of the illumination in the photometer.
3. Photometers for determining vitamin A by the Carr-Price reaction
should employ low intensity of incident light to reduce fading of the blue
color to a minimum and make possible more precise determinations.
BIBLIOGRAPHY
1. Dann, W. J., and Evelyn, K. A., Biochem. J., 32, 1008 (1938).
2. Koehn, C. J., and Sherman, W. C., J. Biol. Chem., 132, 527 (1940).
186 DETERMINATION OF VITAMIN A
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