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INDUSTRIAL AND ENGINEERING CHEMISTRY

VOL. 11, NO. 10

I n several cases the method has been applied to milk products 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 because of its rapidity, and because biological assays may be complicated at times by the presence or absence of nutritional 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 standard. 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 calculating 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 decomposition of the hyposulfite solution.

Certain unstable colored impurities may be destroyed by including from 1 to 3 per cent of hydrogen peroxide in the solvent. 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 absorption 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 absorption 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 University 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). and Khaustov, N. V., Biochem. J., 29, 3 6 7 (3) Kharit, A. Yu., (1935). (4) Kosohara, W., phyaiol. Chem., 232, 101-16 (1935). 2. (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, 291328 (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 the Division of Agricultural and Food Chemistry at before
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.

Conclusions A photometer has been described which utilizes a zeropotential circuit in connection with a blocking-layer photocell. 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.

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 considerable 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 difficulties 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

N HUMID regions, boron often exists in soils largely as tourmaline, a boroaluminum silicate which is not readily

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 titrimetric 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 hydroxyanthraquinones 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 determination of boron. For this purpose, Feigl and Krumholz (6) tried purpurin, alizarin S, and quinalizarin. Quinaliz-