Академический Документы
Профессиональный Документы
Культура Документы
Albany 6, Calif.
When sodium chlorite, glucose, and fructose are brought together in acidic aqueous solution, a t least three simultaneous
reactions occur which consume chlorite and produce chlorine
1
f
V O L U M E 2 6 , NO. 9, S E P T E M B E R 1 9 5 4
of glucose a t pH of 3.7, 4.3, and 4.7, showed good agreement with
first-order dependence on total chlorite, 0.006 t o 0.25M, and on
glucose concentrations, the values of the latter being 0.5, 1, and
2 mg. per ml. On the other hand, the dependence on [H+]
appeared to be definitely less than first order, the numerical values
ranging from slightly less than 0.7 to nearly 0.8. The effect of
changing buffer strength, adding salts to the buffer, or changing
from acetate to citrate buffer were qualitatively the same as
for the disproportionation reaction. An apparent activation
energy of 16,500 calories per mole was found.
Oxidation of Fructose. The stoichiometry of the oxidation of
fructose by chlorous acid was not investigated, but approximately
3000 times as much fructose was required in a reaction mixture
a t 25' C. to produce chlorine dioxide a t the same rate as a given
concentration of glucose. The comparatively little kinetic data
which were obtained on the fructose reaction showed approximately first-order dependence on fructose and total chlorite
concentrations, the same dependence on p H as the oxidation of
glucose, and an apparent activation energy of the order of 23,000
calories per mole.
325
315
495
WAVE LENGTH, p
C.
1479
tion in the 18-hour reaction period of about 500 times as much
chlorine dioxide due to oxidation of glucose as to oxidation of
fructose for the same initial concentrations of the two sugars.
ANALYTICAL METHOD
0.06M NaClOz
No. 42 filter
Spectrophotometric Measurement of Chlorine Dioxide Concentration. A Beckman Model DU spectrophotometer was used
in this investigation. Aqueous solutions of chlorine dioxide show
an absorption maximum at 358 mp ( A , Figure I ) , but because of
the absorption of sodium chlorite ( B , Figure 1) in this region, a
higher wave length should be used for spectrophotometric measurement of chlorine dioxide in reaction mixtures In order to
eliminate errors in wave length calibration and shift in wave length
a t a fixed wave length dial position due to changes in temperature of the the monochromator, much of the development work
was done with the 435 8 or 404.7 mp lines of a mercury arc source
such as is available for wave length calibration of the instrument.
However, the convenience of the tungsten source, resulting from
its greater stability, led to its use at 436 mp in later work. When
the tungsten source is used, the calibration should be checked
against the mercury arc, and the wave length dial should always
be set precisely and from the same direction. Use of either the
435.8 mp mercury line or the tungsten source of 436.0 mp is
recommended. A t these wave lengths, slit widths up to 1.0 mm.
for the mercury line or up to 0.5 uith the tungsten source can
be used without introducing errors in the absorbance measurements.
The absorption coefficients of chlorine dioxide in water solution
at 25 O C. were determined a t various wave length. and concentrations and were in accord with Beer's law. Values of 113.5,
115.3, and 495 liter per mole-cm. were found for the molar absorption coefficients a t 436.0, 435.8, and 404.7 mp, respectively.
ANALYTICAL CHEMISTRY
1480
Figure 2.
The second of these values agrees reasonably well with the value
found hy Launer et al. (18) who prepared chlorine dioxide by
acidifying sodium chlorite solution.
The authors chlorine dioxide solutions were prepared by the
method of B r w ( 4 )in which 150 Erama of oxalic acid, 40 mama
Ihlorine, throigh glass wool to remove spray, and into ice water
containing a small amount of acetic acid. The apparatus should
be all glass. The chlorine dioxide content is determined iodometrically.
In order t o permit measurement of chlorine dioxide roncentrations in the reaction tubes, a vertical light-tight extension of the
1-cm. cell compartment was constructed for the spectrophotometer. This is fastened by screws to the regular compartment of the instrument and accommodates the
standard cover (Figure 2). The reaction tubes fit
into the four positions of the I-em. square cell carrier
so that test and control reitation tubes can be
pared readily either with each other or with
erenee tube containing water.
R:
where
= gARz
- 8,)
(4)
Bt
Ae)
- Be = (At CP
5
b
9 POO
(3)
...,,.--
~~~~~
E composition as in A hut
3
= 1.341
le composition a8
V O L U M E 2 6 , NO. 9, S E P T E M B E R 1 9 5 4
1481
BI
Concentrations Present in
Test Samples
Fructose
Glucose
mg./ml.
lIg,/ml.
%b
100.0
0 050
0.050
0.100
0.100
0,330
0.329
1,000
0.991
10.0
0,050
0,100
0.330
1.000
0.300
0,050
0.99
3.2
9.1
4.8
Glucose Found
(Mean Value),
.\Ig./Ml.
18 hr.C
19 hr.d
0 049
0.049
0 , l O O ~ 0.100;
0,330
0,331
0.966
0.969
0,0492
0.1000
0.330
0.981
0 300h
0.301h
0 299h
0.301
0.0502
0.1016
0.328
0.979
Error in Mean
Value&, Mg./Ml.
Glucose
18 hr,c
19 hr.d
-0.0002
-0.0001
0.0000 +0.0003
0,000
-0,034
f0.001
-0.0008
0.0000
0.000
-0,019
0.000h
t0.001h
-0.001h
+0.0002
+0.0016
-0.002
-0.021
-0.002h
Standard Deviation
from Mean
(Mg./Ml. Glucose)
18 hr C
19 hr.d
0.001
0 002
0.003
0,003
0 010
0.003
0.003
0.001
0,005
-0.031
0.005
0.005
0.015
0.005
0.003
0.005
0.009
0.004
0,003
0.001
6.0
0.298h
+O.O03h 0.001
3.0
0,300
Q.1
0.303h
-0.001h
0.003
0.003
0.299h
1.0
0.300
23.1
0.002
0.002
+0.002
0,302
fO.OO1
0.0
0,300
100.0
a Glucose found minus glucose present.
b Percentage of sugar content which was glucose.
C Spectrophotometer.
d Colorimeter.
e Controls containing 100 mg./ml. fructose shoved 426 X 10-534 ClOz a t 18 hours, 442 X 10-5.11 ClOz a t 19
hours.
f Controls containing 10 nig./ml. fructose shoned 239 X lo-5.M ClOz a t 18 hours 250 X 10-6.M C1Oz a t 19 hours.
0 Controls containing no fructose showed 226 X 10-634 ClOz a t 18 hours, 237 X IO-5.M ClOz a t 19 hours.
h Corrected for C102 evolred by fructose in sample.
C3
C,
CO
1.5 qBc
where C, is the initial total chlorite concentration, B, is the chlorine dioxide concentration in the control solution, and q is a factor
which reduces to unity for no fructose present and which is
somewhat less than unity for appreciable amounts of fructose.
Values calculated for expression (6) corresponding t o observed
values of B, of Table I11 for an 18-hour reaction period for controls containing 0, 10, and 100 mg. per ml. of fructose are 1.017,
1.017, and 1.057, respectively. Combining these figures with a
4.5% correction for loss of chlorine dioxide formed gives corresponding over-all correction factors, d, to the observed net chlorine
dioxide concentration of 1.063, 1.063, and 1.105. These are in
good agreement with the experimentally determined values 1.064,
1.073, and 1.108 based on stoichiometry of Equation 2.
The concentration of glucose in the test solution is then 90.1 d
( B , - B c )mg. per ml., where ( B t B,) is the observed net chlorine dioxide concentration and d is the appropriate theoretical correction factor just discussed. iin additional correction is made
for impurity of the fructose used in the control solution if the
latter is not glucose-free.
(11)
(111)
ANALYTICAL CHEMISTRY
1482
Table 11. Response of Various Sugars and Other
Substances to Chlorous .4cid Oxidation Procedure
Substance
Concentration,
M g ./ M 1.
Moles of ClOn
per Mole of
Substances
2 00
1 90
1 98
1.92
1.98
0 0034
0 0035
0 0038
0 0047
0 00019
1 44
0 013
Glucose
0.1
Galactose
0.1
Mannose
0.1
Lactose monohydrate
0.1
Maltose monohydrate
0.1
Fructose
100
Sorbose
50
Sucrose
50
Raffinose pentahydrate
50
E t h y l alcohol
50
Isobutyraldehyde
0.158
Betaine hvdrochloride
5
Lysozymeb
2
9 5
Galacturonic acid monohydrate
0.2
2 01
a Corrected for hydrolytic loss of chlorine dioxide and difference in chlorite concentration in test and control. Except for glucose and fructose these
figures are the mean results of duplicate determinstiona.
b Molecular weight 14,600.
Fructose esssentially free of glucose can be prepared by crystallization as dihydrate. Material prepared in this manner can
be used as a reference standard in cont,rol solutions for determining the aldose (glucose-equivalent) content of a good grade of
commercial fructose. The latter can then be used in controls
for other analyses.
Preparation. A cold (0' C.) aqueous solution (about 65% by
weight,) of the best fructose commercially available is seeded with
fructose dihydrate crystals (16) which are crushed in the solution
and well dispersed. After crystallization a t 0" for 12 hours or
more, the crystals are freed of mother liquor so far as possible by
use of a Biichner funnel (without paper), a sintered glass filter, or
a centrifuge. The crystals arc then washed by intimate mixing
with eit,her undersaturated solution of previously purified fructose
or cold water, in the latter case using about one fift,h the volume
of the original solution. The n-ashed crystals are separated as
before. The product may be air dried a t 0" C. or stored below
10' C. as wet crystals (about 80% fructose), or as a sirup (diluted
to about, 70%). Fructose content can be found from refractive
index measurement (8) or by dichromate oxidation (11).
1/2,
= 1/2 and a =
1/2, #J = 2/3.
0.00
50
100
150
TIME, MINUTES
Figure 4.
I.
11.
D) added
V O L U M E 2 6 , NO. 9, S E P T E M B E R 1 9 5 4
1483
Preparationb
A
B
C
Df
E
F
a
Source of
Original
Fructose
Xd
Xd
Y
1
z
Z
Oxidizable Impuritiesa
Original fructose,
Purified Fructose
photometric
Photometric
Rate curve
method
method
intercept C
6
0 10
- 0 004
0
0
0
0
10
18
18
+o
70
0 70
005
0.008
0 021
0 031
0 039
0
0
0
0
009
024
016
019
fer solutions, and minor fluctuations of temperature of the solutions during the reaction period.
Minimum exposure of the reaction mixtures to light is recommended, because chlorine dioxide solutions are known to be photosensitive, but the brief exposure incident to transfer of tubes
from the bath and photometric measurement was found to produce no significant change. Errors due to light exposure or other
procedural details can be detected by check analyses using Sational Bureau of Standards dextrose as a test sample.
The sensitivity of the method for measuring glucose as an impurity in fructose is ultimately limited by the reproducibility of
replicate control and replicate test solutions run simultaneously.
It is seen from Table I that a standard deviation of about 0.003
mg. per ml. of glucose is to be expected if the chlorine dioxide is
measured spectrophotometrically, indicating that glucose impurity as low as 0.01% could probably be detected and estimated
with considerable uncertainty provided a suitable fructose standard were available. Suitahle aldose-free standard fructose can
be prepared by recrystallization of fructose dihydrate without
resort to chlorite treatment as shown by the fact that preparations
A and B of Table I11 did not differ significantly from chloritetreated fructose in their yields of chlorine dioxide produced in
the analytical procedure. Fructose, purified by recrystallization as the dihydrate to the point where successive crystallizations produce no change in response to the analytical procedure,
is accordingly thought to be equivalent to fructose freed from
aldose by treatment with chlorous acid. The results of Table
I11 further show that reference fructose containing less than
0.03% of glucose-equivalent can be prepared by a single recrystallization of commercially available material as dihydrate provided the product is adequately washed.
Although this work has not been extensive enough to evaluate
the method fully, the results of Table I indicate that it can be
used to determine glucose in mixtures of glucose and fructose
over the entire range of composition. For the spectrophotometric procedure, approximate standard deviations in the percentage glucose are 0.003, for samples containing less than 0.5%
glucose, 0.03 for samples containing from 0.5 to 5 % glucose, and
about 1% of the glucose content for samples containing more than
5% glucose. For the colorimeter procedure the corresponding
standard deviations are larger by a factor 1.5 t o 2. The accuracy
of the results for glucose contents below a few per cent is determined by the accuracy with which the glucose content of the
reference fructose employed in the method is known.
ACKNOWLEDGMENT
The analytical method presented has the advantages and disadvantages characteristic of most difference methods. Thus
about as much chlorine dioxide is formed in the control solution
containing no fructose as is produced by the oxidation of 0.2
mg. per ml. of glucose. Similarly, the chlorine dioxide produced
in control solutions containing 100 mg. per ml. of fructose is approximately equivalent to that resulting from 0.4% of glucose
impurity in a fructose sample. This emphasizes the importance
of obtaining maximum precision in the photometric measurements, especially if a colorimetric procedure is used where the
difference in chlorine dioxide concentrations of the test and control solutions cannot be measured directly in a single measurement.
The comparison of test and control solutions prepared from
the same reagents and subject to the same temperature history
practically eliminates, as sources of error, the slight differences
in absolute reaction rates observed for different buffer preparations, the slight differences in the small amount of chlorine
dioxide produced immediately upon mixing the chlorite and buf-
(1) Baldwin, R. W., Campbell, H. A., Thiessen, R., Jr., and Lorant,
ANALYTICAL CHEMISTRY
1484
Jeanes, Allene, and Isbell, H. S., J . Research Natl. Bur. Standurds, 27, 125 (1941) (RP 1408).
Keilin, D., and Hartree, E. F., Biochem. J . , 42, 221 (1948).
Launer, H. F., and Tomimatsu, Y.,ANAL. CHEM.,25, 1767
(16)
(17)
Young, F. E., Jones, F. T., and Lewis, H. J., J . Phys. Chem., 56,
1 1 , 1952).
738 (1952).
(1953).
(12)
Launer,.
(1952).
(15) White, J. F., Taylor, M . C., and Vincent, G. P., Ind. Eng.
Chem., 34,782 (1942).
RECEIVED
for review December 21, 1953. Accepted June 7, 1954. Presented
before the joint sessions of the Divisions of Analytical and Carbohydrate
Chemistry. Symposium on Analytical Methods and Instrumentation Applied
to Sugars and Other Carbohydrates at the 124th RIeeting of the AMERICAN
CHEMICAL
SOCIETY,
Chicago, Ill. Mention of products by specific manufacturers does not imply that they are endorsed or recommended by the
Department of Agriculture over others of a similar nature not mentioned.
W. Va.
An effort has been made to evaluate the use of conductometric methods for end-point determinations in the
titration of solutions of the disodium salt of ethylenediaminetetraacetic acid and divalent metallic ions.
Conductance methods may be used for accurate standardization of solutions of copper(II), zinc, lead, nicltel(11), cobalt, calcium, barium, strontium, magnesium,
manganese, cadmium, iron(IL), and mercury(I1) in the
concentration range from 0,001 to 0.5M, before dilution
in the titration vessel.
ECENTLY the disodium salt of ethylenediaminetetraacetic acid (Versenate, Sequestrene, Complexone 111)
has been proposed as a standard for establishing the concentrations of solutions of certain divalent cations ( 2 ) . The stability
constants of the complexes are great enough t o make precise
end-point determinations possible ( 1 7 , 18, 20). The stoichiometric relations for the reactions between the metallic ions and
the reagent have already been determined by several methods
with reported accuracies within 0.05 to 2.0%. Metal ion concentrations have been determined potentiometrically (1, 10, 11,
19),by use of indicators ( I ! 4 , 5 , 9 , 1 6 , 19), spectrophotometrically
(12, I S , 22, 23), polarographically (6, 14,1.5,21),and by a specialized high-frequency technique ( 3 ) .
The present work shows that conventional conductometric
methods may be used for the standardization of solutions of several common cations. The accuracy compares favorably with
the best previously described methods.
REAGEVTS