50?

V O L U M E 25, NO. 3, M A R C H 1 9 5 3
Table 111. Lactic Acid Content of Heavy Steep Liquor as
Determined by Ether Extraction for 58 Hours
% Lactic Acid (Dry Basis)
Conditions
22.8
Acidified and evaporatedbefore extraction
24.0
Acidified but not evaporated before extraction
12.4
S e i t h e r acidified nor evaporatedbefore extraction

traction time. Similar extractions have been conducted on steep

liquor which was acidified but not evaporated, and also on samples which were neither acidified nor evaporated (Figure 1, Table
111).
CONCLUSIONS

Lactic acid present in heavy steep liquor must exist partially

as a salt which may or may not be extracted with ether; in either
caqe it would not be included in the final alkali titration.
Direct extraction of heavy steep liquor after acidification with
sulfuric acid yields results which agree well with values obtained
by the oxidation-distillation technique on pretreated samples.
On the other hand, values obtained by the standard extraction
twhnique were about 1% lower. It does not seem likely that
thii. is due to loss of volatile acids during evaporation, as heavy
stwp liquor is produced by a concentration operation which should
rcmove such volatiles. Friedemann and Kendall(4) have shown
that determination of lactic acid in urine by ether extraction
alv ays results in a loss of lactic acid due to oxidation by organic

peroxides formed during evaporation and extraction. There

also exists the possibility of lactic acid condensation during evaporation, which would result in lowering the apparent lactic acid
content determined by extraction.
Although the extended ether extraction of acidified heavy steep
liquor (not evaporated) yields reliable results which are in agreement with the values obtained by the modified Friedemann technique, the oxidation-distillation procedure is recommended because of the short time required for analysis.
LITERATURE CITED

(1)

Elgart, S., and Harris, J. S I IND. ENG.CHEV.,ANAL. ED.,

12
758 (1940).

Friedemann, T. E., J . Biol. Chem., 76,75 (1928).

Friedemann, T. E., and Graeser, J. B., Ibid., 100, 291 (1933).
Frjedemann, T. E., and Kendall, A. I., Ibid., 82, 23 (1929).
Fries, H., Biochem. Z., 35,368 (1911).
Kerr, R. W., Chemistry and Industry of Ptarch, 2nd ed.,
Chap. 11,New York, Academic Press, 1960.
( 7 ) Kondo, K., Biochem. Z., 45,88 (1912).
(8) Leach, A. E., and Winton, A. L., Food Inspection and Analysis,4th ed., New York, John Wiley & Sons, 1936.
(9) Mendel, B., and Goldscheider, I., Biochem. Z., 164, 163 (1926).
(10) Pigman, 11. Ti., and Wolfrom, M. L., Advances in Carbohydrate
(2)
13)
(4)
(5)
(6)

Chem., 1, 257 (1944).

(11) Somogyi, M., J . B i d . Chem., 90,725 (1931).
112) Trov. H. C.. and Sharp. P. F., Cornell Cnic., Agr. Ezpt. Sta..
iGemoirs, 179 (1935):
(13) Van Slyke, D. D., J . B i d . Chem., 32, 455 (1917).
(14) Wolf, C. G. L.,J . Physiol., 48, 341 (1914).

RECEIVED
for review

Accepted October

January 17, 1952.

27, 1952.

Calibration of the Rolling Ball Viscometer

H. W. LEWIS
Bell Telephone Laboratories, Murray Hill, N. J .
N 1943, Hubbard and

Brown ( 1 ) carried out a systematic

1 experimental calibration and dimensional analysis of a rolling

ball viscometer. They determined a dimensionless calibration

curve, which enables one to design a viscometer of this type to
measure any given range of viscosities. It is the purpose of this
note to show that one can, to good approximation, derive this
calibration curve from a simple approximate treatment of the
problem in terms of the hydrodynamics of viscous fluids ( 2 ) .
IVe consider a cylindrical tube of diameter D, inclined a t an
angle 29 to the horizontal, and filled with a fluid of viscosity L,A,
and density p,. SVe suppose to be rolling down the tube with
velocity V, a spherical ball of diameter d, and density pa. We
will use a cylindrical coordinate system, with Z-axis parallel to
the axis of the tube, with polar angles referred to a vertical plane
through the 2-auk, and with the origin a t the center of the
sphere. We Tvill also make the approximation that the gap
between the sphere and the tube is small compared to the diameter of either one, so that the sphere nearly fills the tube-this
is the usual, practical situation, and makes the calculation possible. We will systematically neglect higher powers of ( D - d ) / D .
Now, if we call the distance betn-een the sphere and the wall
of the tube u (79, Z ) , a little consideration of the geometry will
show that

a narron- channel of this sort gives rise to a parabolic velocity

profile a t any given point. In particular, if L is the mean velocity of the fluid in the gap, then d 2 i l b r 2 = 12; /u2, where r is our
radial cylindrical coordinate. But the longitudinal gradient of
pressure is given by p LPu/dr* = 12 p u/u2, so that our problem may
now be stated as follows: rye have to determine the distribution
of G as a function of 9 and 2, so that the total difference of
pressure is enough to balance the reight of the sphere, and the
total flow across any plane Z = constant is equal to
D2V.
This will determine p as a function of V , n.hich is the problem
we have set ourselves.
K e will use the f o l l o ~ ~ i nprocedure:
g
for each value of 2, we
will distribute the flow velocity, as a function of 9, in such a way
that the longitudinal pi essure gradient is independent of $-this
implieb that we x-ill neglect all components of velocity perpendicular to the axis of the tube. Thus, we write

u (79,

(2)

The total flux through any plane perpendicular to the axis of

the tube \vi11 then he
flux =

So we have to consider the flow of a viscous fluid through a narrow channel of width given by ( 1 ) . The total flow through this
channel will be D2V, since we will, everyn-here except in the
expression ( D - d ) , neglect the differencebetu-een D and d.
Now, it is well known that the flow of a viscous liquid through

Z) = B ( 2 ) uy9, Z )

a fT

L(79, Z )

flux

x
Fz

fx

D ( D - d ) 3B ( Z )
16

u(8,

Z ) X D d9
(3)

;B(2)

Substituting the expression for

ing the integral, we find

u3(9,

Z ) d8

from Equation 1, and perform-

[5

+ 18a + 24aa + 1 6 4

(4)

A N ALY TICAL CHEMISTRY

508
where

(Y

[D

4 D 2 - 422]/2(D

- d)

1 .o

But this must be independent of Z, and must be equal to

so that we find

B(Z) =

T H E O R E T I C A L . NO

10

4 DT.

(D

- d)3[5 + 18a + 24a2 + 16a3]

(5)

1 CY

48 p DT

(D

TI0
D

Consequently, the longitudinal pressure gradient is

grad p =

0 - E X P E R I M E N T A L , LIQUIDS
0 -EXPERIMENTAL, A IR

DV,

- d ) 3 [ 5 + 18a + 24a2 + 1601~1

(6)

and the total pressure drop along the tube, due to the ball, is
AP =

grad p dZ =
Jr
m n

[&d]62x
dE
5

(7)

where, if we call the integral 8 r I / 3 , thereby defining a number I ,

we have

I =

1 [.\/a
1/2 - + 21
45

0.398

(7)

In passing from Equation 6 to 7, we have defined a quantity

by the equation $2 = a,and have again neglected terms which
contribute only in higher orders of ( D - d ) / D . Thus, for example,
the limits on the integrals are not really infinite, but depend upon
D/(D - d).
This pressure drop must now support the sphere, so that
? r ~ a

(i

p l ) g sin 8 =

7r

- ~

a
10

u
IC

lo5

10-6

I04

10-3

Figure 1. Plot of Theoretical Equation 9with Experimental

Points
with the calibration constant K of Hubbard and Brown, in terms
of which Equation 9 is

K = -2o 7

[ DT ]- d

512

D-d 7
= 0.0891 [
]
(9)

I n Figure 1 has been plotted the expression for K given by

Equation 9, the points measured by Hubbard and Brown, and
the points calculated by them from other experimental data.
The fit, considering the simplifications in the calculation, is
satisfactory. I t can serve for the design of rolling ball viscometers, although, for precision work, the usual calibration in
terms of fluids of known viscosity should be carried out. Equation 9 should, of course, be used directly, without going through
the definition of K .

LITERATURE CITED

and

(1) Hubbard, R. M., and Brown, G. G., IND.ENG.CHEM.,ANAL.

ED.,15, 212 (1943).
(2) Lamb, H., Hydrodynamics, New York, Dover Publications,
1945.

which is our final result.

It will be convenient to compare this

RECEIVED
for review

July 7, 1952.

Accepted October 15, 1952.

Identification of Flavonoid Compounds by Filter Paper Chromatography

Additional Rf Values and Color Tests
HELEN WARREN CASTEEL AND SIMON H. WENDER
University of Oklahoma, Norman, Okla.
APER chromatographic techniques applicable to flavonoid
Pcompounds have been developed by Bate-Smith and Westall
(1, 9) and by Gage, Douglass, and Wender ( 3 ) . Because of the
interest indicated by many research workers in these paper chromatographic studies of flavonoids, the present investigation was
undertaken to extend the usefulness of this technique by the determination of R f values for a number of flavonoid compounds not
yet reported in the seven solvent systems listed.
The colors produced by chromogenic sprays when considered in
conjunction with the R, value often aid in the tentative classification of an unidentified flavonoid pigment into one of the major
subdivisions of flavonoid compounds. Therefore, the colors produced on paper by chromogenic sprays and certain of the newly
studied flavonoids were also determined.

EXPERIMENTAL

Experimental apparatus, materials. and procedures used correspond BB nearly as possible to those of Gage et al. (3). A newer

model Chromatocab chamber (Chromatography Division,

University Apparatus Co., Berkeley, Calif.) was used in the present study, however. This chamber was much better sealed and
better insulated than the previous model used. Thus, uniform
saturation, indicated by movement of solvent fronts through
equal distances for all strips within the chamber, was obtained if
sufficient time (usually 24 hours) was allowed for saturation.
Also, rate of movement on the paper was usually much more
rapid in the newer chamber.
A 250-ml., all-glass spraying flask (University Apparatus Co.)
operated by compressed air a t 5 pounds pressure delivered an even
mist of chromogenic reagent. The spray was controlled by an
air hole covered by the thumb during delivery.
RESULTS AND DISCUSSION

Table I lists the R / values obtained for twenty-one flavonoid

compounds in seven different solvent systems. These listed
values represent average R f values for each compound. Some
variation in Rt of a pigment occurred from time to time, but the
variation was usually less than 1 0 . 0 4 Rf value and, in most cases,
was less than 50.02 Rr value.
Some of the flavonoid samples used in this study were found by