540 INDUSTRIAL AND ENGINEERING CHEMISTRY
10
I n several cases the method has been applied to milk prod-
ucts which were also assayed for riboflavin by means of
chicks. The chick assays upon six samples gave an average
value of 21.2 micrograms per gram. An average value
of 21.5 micrograms per gram was obtained by the photometric
test. The method presented in this paper is preferred be-
cause of its rapidity, and because biological assays may be
complicated at times by the presence or absence of nutri-
tional factors which are not as yet clearly defined.
The absolute accuracy of this method depends upon the
purity of the sample of synthetic riboflavin used as a stand-
ard. Several samples of crystalline riboflavin were obtained
from different sources a t the time of standardizing the method
and none of these appeared to be quite so pure as the sample
used. This conclusion was based both upon the light absorp
tion and the completeness of reduction. Since that time an
occasional sample has been encountered which appeared to be
reduced to a greater extent than the standard. When a
satisfactory standard of reference has been obtained it may
be necessary to alter slightly the above correction factor for
calculating the concentration of riboflavin.
When other instruments are set up according to the above
directions the same correction factor may be used for cal-
culating riboflavin concentration. Some care must be taken
to assure that exactly 90 per cent reduction of the riboflavin is
obtained. Variation in the completeness of reduction may
occur through incomplete removal of the peroxide or decom-
position of the hyposulfite solution.
Conclusions
A photometer has been described which utilizes a zero-
potential circuit in connection with a blocking-layer photo-
cell. Such an instrument gives a linear response to varying
light intensities, and offers advantages in regard to proper use
of the photocell, reproducibility of results, and application to
certain spectrophotometric measurements.
Riboflavin may be extracted from dried milk products by
refluxing with a dilute solution of acid in 75 per cent acetone.
Boron Determination in Soils and Plants
Using the Quinalizarin Reaction
K. C. BERGER AND E. TRUOG, University of Wisconsin, Madison, Wis.
I N HUMID regions, boron often exists in soils largely as
tourmaline, a boroaluminum silicate which is not readily
available. The available boron, probably existing often largely
as the calcium salt, is usually present in humid regions in
amounts of less than one part per million. I n plant tissue,
boron is often present to the extent of 25 parts per million,
and since it is now recognized as an essential plant nutrient,
its determination in both soils and plants assumes consider-
able importance. Methods used in the past for making these
determinations have not been very satisfactory in some r e
spects, and the present paper reports the results of an attempt
at improvement.
I n the past, the turmeric paper test described by Bertrand
and Agulhon (I) has often been used in the determination of
small amounts of boron. As the turmeric paper is not very
sensitive to small differences in amoynts of boron, and difficul-
ties are encountered in making accurate comparisons with
the standard, the method leaves much to be desired. The
spectroscopic method has recently been used by a number of
VOL. 11, NO.
Certain unstable colored impurities may be destroyed by in-
cluding from 1 to 3 per cent of hydrogen peroxide in the solv-
ent. The resulting solution, after neutralizing and filtering,
may be used for quantitative measurements since riboflavin
is not preferentially adsorbed by the precipitated milk solids.
By reduction with sodium hyposulfite, 90 per cent of the
color of riboflavin is removed. Observation of the light ab-
sorption before and after reduction enables one to calculate
the concentration of the solution to 10.05 microgram per ml.
Acknowledgments
The sample of synthetic riboflavin was kindly supplied by
Richard Kuhn of the Kaiser Wilhelm Institut, Heidelberg,
Germany. The transmission curve of the filters and the ab-
sorption curve of riboflavin were measured by A. E. Sidwell,
Jr., of the University of Chicago. The work reported in this
paper was made possible by the establishment a t Cornel1 Uni-
versity of a special temporary fellowship, by the Kraft-Phenix
Cheese Corporation of Chicago, Ill.
Literature Cited
(1) Cohen, F. H., Arch. nderland. phyaiol., 20, 167-74 (1935).
(2) Karrer, P.,and Fritache, H., Helv. Chim. Acta, 18, 911-14
(1935).
(3) Kharit, A. Yu.,and Khaustov, N. V., Biochem. J., 29, 3 6 7
(1935).
(4) Kosohara, W.,2. phyaiol. Chem., 232, 101-16 (1935).
(6) Kuhn, R . , Rudy, Hermann, and Weygand, Friedrich, Ber., 68,
625-34 (1935).
(6) Wagner-Jaurreg, Th., Angew. Chem., 47, 318-20 (1934).
(7) Warburg, Otto, and Christian, Walter, Biochem. Z . , 287, 291-
328 (1936).
(8) Withrow, R: B.,Shrewsbury, C. L., and Kraybill, H. R., IND.
E N G .CHEM.,Anal. Ed., 8, 214-19 (1936).
(9) Wood, L. A., Rev. Sci. Znatrumenta, 7, 157 (1936).
PRE~ENTBD
before the Division of Agricultural and Food Chemistry at
the Mid-West Meeting of the American Chemical Society, Omaha, Nebr.
This paper was compiled largely from a thesis presented to the faoulty of
Cornell University in partial fulfillment of the requirement for the degree
of dootor of philosophy, February, 1937, and was part of an experiment
station project under the direction of L. C. Norris.
investigators (3, 6) to determine the boron content of plant
materials. It seems to give fairly good results, but requires
rather elaborate equipment and is time-consuming. The ti-
trimetric procedure, involving the use of polyhydroxy alcohols
and discussed in detail by several workers (8, II), is not well
adapted for the rapid determination of the small amounts
sometimes encountered in soil and plant analysis, because
the method involves distillation which is time-consuming,
and because traces of buffering substances which are difficult
to remove may seriously interfere with the results.
Quinalizarin Reaction
It has been known for a considerable number of years
that the addition of boric acid to many of the hydroxyanthra-
quinones in concentrated sulfuric acid will cause a marked
color change which may be used for the identification of these
quinones. Recently this color change was applied to the de-
termination of boron. For this purpose, Feigl and Krumholz
(6) tried purpurin, alizarin S, and quinalizarin. Quinaliz-