Bobbitt 1956

PERIODATE OXIDATION OF CARBOHYDRATES
BY J. M. BOBBIW
Department o j C~emistry,The Ohio State Universitg, Columbus, Ohio
I. Introduction .............................................. 1
............................................. 2
111. Definition of the Reaction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
IV. Application.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
V. Reaction Conditions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
1. Oxidant . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
2. Solvent. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
3. Hydrogen-ion Concentration.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
4. Temperature. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .... 31
5. Light.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .... 31
6. Concentration of Reactants.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
VI. Methods of Analysis.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
1. Oxidant Consumption.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
2. Acidity Measurement. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
3. Aldehyde Analysis. . . . . . ......................................... 37
4. Carbon Dioxide Analysis.. . . . . . . . . . . . . . . . . . . . . . . . . ................ 39
5. Ammonia Analysis.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
VII. Course of the Reaction . . . . . . . . . . . . . . . . . .......................... 39
VIII. Conclusion.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
I. INTRODUCTION
Oxidation with periodate ion, resulting in 112-glycol scission, is one of
the most widely used reactions in carbohydrate chemistry. Since its dis-
covery by Malaprade in 1928, the periodate reaction has been applied to a
large variety of problems in all fields of organic chemistry. This reaction has
been used most spectacularly, however, in the study of the carbohydrates,
because of their polyhydroxylic nature. The mild conditions of the reaction
are especially well adapted for application to the sensitive carbohydrate
structures. The development and wide application of the reaction have been
occasioned by a number of factors, the principal one being the high degree
of selectivity shown by the periodate reaction when applied under the
proper conditions. Thus, the oxidation can be limited to 1,2-glycols,
2-amino alcohols, a-hydroxy ketones and aldehydes, a-amino aldehydes,
1 2-diketones, and certain activated methylene groups.
Another feature contributing to the wide usage of the periodate reaction
is its simplicity of application, The aqueous solvent conditions called for in
1
2 J. M. BOBBI'B!
periodate oxidation lend themselves very nicely t o use with the water-
soluble carbohydrates. The quantitative nature of the reaction, together
with the precise analytical procedures available for following its course,
make possible the obtaining of a great deal of information from experi-
ments utilizing minimal quantities of material. The knowledge that specific
groupings produce certain stable end-products not only makes periodate
useful as a n analytical tool, but permits the establishing of unknown struc-
tures. The insoluble nature of many inorganic salts of the periodate and
iodate ions simplifies their removal from reaction mixtures, thus facilitat-
ing isolation of the organic products. The pronounced effects (on the se-
lectivity) exhibited by such factors as temperature, pH, light, and the
nature of the solvent have been recently elaborated.
11. HISTORY
I n the course of work on the potentiometric titration of some oxidizing
acids, Malaprade' found the existing method for determination of periodic
acid in periodic-iodic acid mixtures too inaccurate for estimation of small
proportions of one component. He discovered that mannitol will reduce
only the periodic acid and, furthermore, that it only reduces the acid to
iodic acid.2 The following general reaction appeared to take place.
CHzOH HzC=O
I
CHzOH
+
HzC=O
Malaprade explored the application of this reaction to compounds in which

n was varied from zero to four. He carried out analyses for the periodic
acid consumed and for the formaldehyde and formic acid formed. Using
these methods, he worked out the details of the desired analysis for periodic
acid and, as a corollary, an analysis for the polyhydric alcohols employed.
Fleury and Lange3first pointed out that the reaction could be considered
selective for hydroxyl groups attached to adjacent carbon atoms; and they ex-
plored the application of this new reaction in the carbohydrate field. They
used analytical procedures for oxidant consumption4 somewhat different
from those employed by Malaprade ; and they applied new procedures for
(1) L. Malaprade, Bull. soc. chim. (France), [4] 39, 325 (1926).
(2) L. Malaprade, Compt. rend., 188. 382 (1928); Bull. soc. chim. (France), [4] 43,
683 (1928).
(3) P.F. Fleury and J. Lange, Compt. rend., 196, 1395 (1932).
(4) P. F. Fleury and J . Lange, J. phnrm. chim., [8! 17, 107 (1933).
PERIODATE OXIDATION OF CARBOHYDRATES 3
the assay of formaldehyde and formic acid.6 The procedures of Fleury and
Lange, as well as those of Malaprade, proved so satisfactory that they are
still extensively employed.
The scope of the reaction was considerably enlarged in 1935, when
Clutterbuck and Reuter6 observed that the compound tetrahydroterrein,
derived from the mold metabolite terrein, consumes more than the cal-
d a t e d amount of periodate per mole. They found that l ,2-diketones and
a-hydroxy ketones are also oxidized under the conditions used by Mala-
prade. Although this type of oxidation had been earlier noted [in a study7
of the action of periodate on 1,3-dihydroxy-2-propanone (dihydroxyace-
tone)], Clutterbuck and Reuter made a more thorough exploration of the
reaction.6 In a series of model compounds, R1 and RB were varied from
R , OH
I/ RI
C
I
RI
\ COzH
I + 2 H20 S OH 1049
+ + 2 HzO
c=o OH
COzH
I / I
c=o C' I
Rz
I I\
R2 R2 OH
R I OH
R,
RI C I/ I
COSH
I
C=O
I
+ Hz0 f + H20
I I HC=O
HCOH HCOH
I I
Rz
I
Rz
R?
hydrogen through a series of alkyl and aryl substituents. They did not
investigate the simplest members of the series, glyoxal and glycolic alde-
hyde.
The next step in the development of periodate oxidation was made by
Nicolet and Shinn.8They applied periodate oxidation t o a series of a-amino
acids, and found that those containing the 2-hydroxyamine structure are
almost instantaneously oxidized. For such an oxidation to proceed rapidly,
the amine could not be tertiary. An a-hydroxy N-acylamine was attacked
very slowly, if at all. Only a small number of compounds were investigated.
(5)P. F. Fleury and J . Lange, J . pharm. chim., [8] 17. 196 (1933).
(6) P.W.Clutterbuck and F. Reuter, J . Chem. SOC.,1467 (1935).
(7) P.F.Fleury and J. Lange, J . pharm. chim., IS] 17. 409 (1933).
(8) B. H. Nicolet and L. A. Shinn, J . Am. Chem. SOC.,61, 1615 (1939).
4 J. M. BOBBITT
It has since been learned that tertiary amines are slowly al-
R1
R1 I
I HC=O
HCNHRn
I
11040 ~
+ + NHpRz
HCOH HC=O
I I
R, Rs
(where R1 , RI , and Rt = H or alkyl)
though conditions have now been defined" which will obviate this oxida-
tion. Successful oxidations have also been carried out on 1 ,2-diamine~.~
Periodate oxidation has been successfully applied t o amino sugars and
their derivatives by Neuberger,12although difficulty was encountered with
D-glucosaminic acid.
The oxidation of some anhydroaldopento-benzimida~oles~~ was found
to entail uptake of more than the calculated amount of oxidant per mole.
This apparent anomaly was further explored by Huebner, Ames and Bubl,I4
and their work culminated in the discovery that periodate, under the usual
conditions, oxidizes certain methylene carbon atoms, namely, those ac-
tivated by two flanking carbonyl groups. This type of oxidation (a-hydrogen
oxidation) was simultaneously observed (by Fleury and Courtois16) to
occur on the periodate oxidation of malonic acid. A satisfactory reaction
was obtained under the following conditions.
Ri
Ri I
I
-
Ri Ri C OzH
I I COnH
+
c=o 109 1o.e
I AI +
1040
A RaCOzH
RaCH
I
RaCOH
I
RaC=O
I +
c=o c=o c=o COzH
I
R2
I
Rz
I
Rz
I
R2
(9) P. F. Fleury, J. E. Courtois and M. Grandchamp, Bull. SOC.chim. France,
88 (1949).
(10) E. H.Flynn, M. V. Sigal, Jr., P. F. Wiley aod K. Gerzon, J . Am. Chem. SOC.,
76, 3121 (1954).
(11) F.A. Hochstein and P. P. Regna, J . Am. Chem. SOC.,77, 3353 (1955).
(12) A. Neuberger, J . Chem. SOC.,47 (1941).
(13) C. F. Huebner, R. L. Lohmar, R. J. Dimler, S. Moore and K. P. Link, J .
Biol. Chem., 169, 503 (1945).
(14) C. F. Huebner, S. R. Ames and E. C. Bubl, J . Am. Chem, SOC.,68, 1621
(1946).
(15) P. F.Fleury and J. E. Courtois, Compt. rend., 228, 633 (1946).
1. Rl must be a hydrogen atom or a hydroxyl group.

2. Rz and R3 may be any permutation of hydrogen atom with alkyl,
hydroxyl, or 0-alkyl group.
3. If RBis an alkyl group and Rz is an 0-alkyl group, the reaction is
slow.
4. If R1or Rz is a hydroxyl group, one of the acids formed is carbonic
acid (which decomposes in acid solution to give carbon dioxide).
5. If Rzis an 0-alkyl group, the third reaction shown does not occur;
instead, the reaction ceases at the intermediate formed in the preceding
reaction.
6. If the grouping RCO- is replaced by the nitrile group, oxidation
does not take place.
It has now been shown that 1,3-diketones are oxidized if they constitute
part of a five- or six-membered carbon ring.lSa
7 c = o
+ coz
(where n = 2 or 3)
Still unexplained are the reactions, with periodate, of hydroxy phenols
and certain oxygenated aromatic compounds,16as well as of some a-amino
acids not containing the 2-hydroxyamine structure.8
111. DEFINITION
OF THE REACTION
A clear definition of selective oxidation by periodate, considered apart

from general oxidation (over-oxidation), is necessary before intelligent
use of this oxidant can be made. To draw a clear distinction between these
two reactions is difficult, as the governing factor appears to be the precise
experimental conditions employed. If compounds are oxidized at a high
temperature, or in very basic media, most of those containing oxygen are
oxidizable to a greater or lesser Choice of the optimal conditions
for promoting an ideal reaction will be discussed in later Sections.
(15a) M. L. Wolfrom and J . M. Bobbitt, J . Am. Chem. SOC., 78, 2489 (1956).
(16) D. E. Pennington and D. M. Ritter, J . Am. Chem. SOC.,69, 187 (1947).
(17) P. F. Fleury and Suzanne Uoisson, Compt. rend., 204, 1264 (1937).
(18) P . F. Fleury and Renee Roisson, Compt. rend., 208, 1509 (1939).
(19) P. F. Fleury and Ren6e Boisson, J. pharm. chim., [8]SO, 145 (1939).
(20) P. F. Fleury and Renee Boisson, J . pharm. chim., [8] SO, 307 (1939).
(21) J. E. Courtois, Ann. phatm. .franc.,2, 148 (1944).
(22) G. Lindstedt, Arkiv Kenti, Mineral. Geol., 2OA, No. 13 (1945).
(23) G. Lindstedt, Nature, 168, 448 (1945).
(24) F. S. H. Head, J. Textile Znsl.,38. T389 (1947).
(25) R . Delaby and P. Chabrier, Bull. S O C . chim. France, 113 (1946).
(26) L. Hartman, J . Chem. Soc., 4024 (1954).
6 J. M. BOBBITT
A practical distinction between selective oxidation and so-called

over-oxidation requires, first of all, the elaboration of a reliable, labora-
tory method. At present, this distinction can best be made by means of a
brief rate-study on the particular reaction. Most selective periodate
oxidations proceed quickly and quantitatively in a homogeneous reaction-
mixture. Aliquots of the reaction mixture are periodically removed and
analyzed for periodate consumption, or for the formation of oxidation
products, and the results are plotted graphically against time. If the rate
curve obtained displays a constant maximum after the elapse of a reasonable
reaction-time (for example, forty-eight hours, in a homogeneous reaction),
it can safely be assumed that the reaction involved is one of those already
described [in the historical Section (see p. 2)]. On the other hand, if the
rate curve exhibits no leveling off a t a maximum, during a period of days
or weeks, the reaction should be investigated more thoroughly. Often, a
simple change of conditions or reagents will appreciably decrease the un-
desirable side-reactions. If attempts to depress the side-reactions are com-
pletely unsuccessful, the curve can be back-extrapolated to provide a value
for the extent of selective oxidation. From the preparative standpoint, a
suitably limited amount of oxidant may then be used, in order to provide
the optimal yield of the main product.
A number of generalizations, other than those already discussed, may be
made concerning the periodate reaction. cis-Hydroxyl groups in ring
structures are more readily oxidized than are those disposed in the trans
form? Relative rates of oxidation have been determined for cis- and
trans-l,2-cyclohexanediol,28 the rate for the cis form being found the higher
of the two. The oxidation of those methyl aldohexopyranosides having cis
vicinal hydroxyl groups (for example, methyl a-D-mannopyranoside and
methyl a-D-galactopyranoside) has been shown to proceed faster than that
of those (such as methyl a-~-glucopyranoside~~) containing only a trans
vicinal hydroxyl system. This stereochemical effect has been considered
more recently (in the carbohydrate field) by Pratt, Richtmyer and Hudson30
and by Fleury, Courtois and Bieder.31I n the course of the investigation of
the structure of sedoheptulosari (2 ,7-anhydro-/3-~-altro-heptulopyranose),
Pratt, Richtmyer and Hudson30 proposed three general rules operative in
the periodate oxidation of cyclic sugars. These rules were based on those
(27) R. Criegee, Sitzber. Gcs. Bejordei. gcs. Natuiw. Mnrbrtrq, 69.25 (1034); Chcir~.
Abstracts, 29, 6820 (1935).
(28) C. C . Price and M. Knell, J . Am. Chem. Soc., 64, 552 (1942).
(29) T. 0. Halsall, E. L. IIirst and J . K . N . Jones, J . Cliem. Soc., 1425 (1947).
(30) J. W. Pratt, N . K . Richtmyer and C. S. Hudson, J . Am. Chem. Soc., 74,
2200 (1952).
(31) P. F. Fleury, J . E. Courtois and A. Bieder, Bull. SOC. chim. France, 118 (1952);
Compt. rend., 233, 1042 (1951).
propounded by Hockett, Dienes and R a m ~ d e nin~ order~ to explain the

phenomena observed on oxidation of cyclic sugars by lead tetraacetate.
1. If two of three vicinal hydroxyl groups are themselves vicinal and
cis, the structure is oxidized more rapidly than if all hydroxyl groups are
vicinally trans.
2. An a-hydroxy aldehyde is attacked but the rate of oxidation is
often low.
3. An a-hydroxy aldehyde will be oxidized relatively rapidly if another
hydroxyl group in a position y or 6 to the carbonyl permits formation of a
pseudoglycol structure by cyclic hemiacetalization.
The above three generalizations were used in explaining the following
oxidation sequence.
FIH
II
-C-CHzOH -C-CHzOH
I I
HOCH HOCH
I I
HCOH HC=O
I
HCOH HC=O
I I
HCO- HCO-
I I
-0CH2 -0CHz
+;cHzo] I II
HOCH
104e
___f
HC=O
I
HCO
I
-0CHz
The production of formate esters in periodate work will be considered

subsequently (see p. 34).
Fleury, Courtois and Bieder31found that the relative rates of oxidation
of some simple carbohydrates lie in the following sequence, the highest
being given first : D-mannitol, D-glucitol (sorbitol), D-mannose, D-galactose,
and D-glucose. Explanation of these results from the steric viewpoint is
complicated and difficult.
(32) R . C. Hockett, Margaret T. Dienes and H. E. Ramsden, J. A m . Chem. SOC.,
66, 1474 (1943).
8 J. M. BOBBIlT
The oxidation of cyclic 2-amino alcohols has been ~tudied.3~ In the

2-amino-cyclopentanol series, the cis form was oxidized four times as fast
as the trans; but, in the 2-amino-cyclohexanol series, the cis form reacted
sIightly more slowly than did the trans.
The appearance of free iodine in periodate reaction-mixtures obviously
indicates that the iodate formed is being further reduced,34,3 6 sometimes
because of the addition of insufficient 37 This breakdown of
f
iodate to free iodine has been reported as accompanying the a-hydrogen type
of oxidation,29 a t least as regards the oxidation of those methyl aldohexo-
furanosides of which methyl a-D-mannofuranoside is a representative. The
CH,OH
- -
I
HCOH
2 I0,0 10'0
I
H H
ro4e
C=O + HzO L0.H + HzO
+
_ _ _ f
OH
~~
(33) G . E. McCasland and D. A. Smith, J . Am. Chem. SOC.,7 3 , 5164 (1951).

(34) R. J. Williams and M. A. Woods, J . A m . Chem. SOC.,69, 1408 (1937).
(35) W. A. Bonner and Ann Robinson, J . Am. Chem. SOC.,72,354 (1950).
(36) F. S. H. Head, Nature, 166, 236 (1950).
(37) P. B. Sarkar, Nature, 168, 122 (1951).
(38) H. G. Fletcher, Jr., H. W. Diehl and R. K. Ness, J . Am. Chem. SOC.,7 6 ,
3029 (1954).
(39) D. B. Sprinson and E. Chargaff, J . Biol. Chem., 164, 433 (1946).
PERIODATE OXIDATION O F CARBOHYDRATES 9
a-keto acids are slowly oxidized, to give carbon dioxide under the usual
periodate conditions. The rate of this reaction could be effectively de-
creased by lowering the temperature and the pH. Similar results had been
obtained by Fleury and B ~ n - B e r n a t e t s ,who
~ ~ recognized two distinct
phases in the oxidation of L-tartaric acid. The first was that in which
L-tartaric acid was rapidly oxidized to glyoxylic acid, and the second and
slower phase was that wherein the glyoxylic acid was further oxidized to
carbon dioxide and formic acid.
The oxidation of a-hydroxy acids is it has been comprehensively
studied, using glycolic acid as a m0de1.4~Three reactions have been pro-
posed as occurring.
CHzOH HzC=O + Hz0
I
I - +
IOke
COiH coz
CHzOH HCOzH + HzO
I
a 104e
COzH coz
- +
CHzOH
I
IOke
A 2 HCOzH
Oxalic acid is reported to be resistant to periodate,4. ae as are formic acid

and formaldehyde under normal condition^.^^
The oxidation of the a-amino acids has not been completely studied, but
it appears to be slow in most cases.*# The organic sulfide linkage is oxidized
by periodate t o a sulfone, with the concomitant liberation of iodine, but
the oxidation of a 1-deoxy-1-thioglycoside,without occurrence of this
side reaction, has been carried out at least once (in very dilute solution).M
An apparent anomaly is encountered when a reductone structure,
-C=C-C=O, is oxidized. The first equivalent of oxidant produces a
OHOH 1
diketone, as illustrated by the oxidation of dihydroxyfumaric acid.4BA
similar reaction, which, however, stopped at the diketone stage, was ob-
(40) P. F. Fleury and G. Bon-Bernatets, J . pharm. chim., [8] 23, 85 (1936).
(41) P. F. Fleury, G. Poirot and Yvonne Fibvet, Compt. rend., 220, 664 (1945).
(42) P. F. Fleury, J. E. Courtois, R . Perles and LBa Le Diset, Compt. rend., 237,
1019 (1953);Bull. S O C . chim. France, 347 (1954).
(43) W. A. Bonner and R. W.Drisko, J . Am. Chem. Soc., 73, 3699 (1951).
(44) M. L. Wolfrom, S. W. Waisbrot, D. I. Weisblat and A. Thompson, J . A m .
Chem. SOC.,66.2063 (1944).
(45) G . Lundblad, Arkiv Kemi, Mineral. Geol., 26A, No.5 (1918);the substance
is termed dihydroxymaleic acid in this publication.
10 J. M. BOBBITT
served in the oxidation of a 1,2-dihydroxyanthraquinone-3-sulfonate(and

of similar dyes) .46 The oxidation of 1,3-cyclohexanedione by periodate is
0
II
HO COH
\ /
C C0zH
7 II + 4 H I 0 4 + 2 Iz + 14 I + 2HzO
C C O,H
/ \
HOC OH
II
0
c=o
thought t o proceed through an intermediate reductone systern.lba The

intermediate 2 ,3-dihydroxy-2-cyclohexenonewas subsequently prepared
[/Jo] -0
and shown to give glutaric acid and carbon dioxide on periodate oxidation,
in accordance with this proposed reaction-sequence. These oxidations af-
ford free iodine unless performed in very dilute solution.'6a
When considering the course of a periodate reaction, it is of the utmost
importance to bear in mind that different bonds are broken at different rates
and that new structures are continuously being formed as a result. Conse-
quently, performance of a stepwise reaction often provides results permit-
ting the formulation of a feasible explanation for an apparently erroneous,
(46) L. K. Ramachandran and P. S. Sarma, J. Sci. Ind. Research (India), 10B,
147 (1951); Chem. Abstracts, 47, 2160 (1953).
fractional consumption of periodate. This type of reasoning is well illus-

trated in discussions of the results obtained on periodate oxidation of the
ketoses.3 * 70 47-49
(a) CHzOH CHtOH
I I
c=o 3 IO@ , c=o
I I
(CHOW8 HC=O +2 HCOtH + HiC=O
I
CHzOH
(b) HzC=O
CHaOH
I Io4e +
c=o __3
COzH
I I
HC=O HC=O
(c) CH40H CHnOH

II II
C=O xo4e ~ COzH
I
I
HC=O +
EICOeH
Reaction (a) takes place first, but two reactions, (b) and (c), can then
occur simultaneously, to give different products. Reaction (b) seems to
predominate, although (c) does take place. A stepwise reaction has also
been observed in the periodate oxidation of rnyo-in~sitol.~~*
The failure of a compound to undergo periodate oxidation can not be
regarded as positive proof that it contains no 1,2-glycol structures. If
the glycol is securely locked in a trans position, oxidation has been shown
not to occur. This resistance has been encountered in the bicyclic anhydro
hexoses, for example, 1,6-anhydro-P-~-glucofuranose~~ and 1,6-anhydro-a-
~-galactofuranose.~~ This work has already been summarized in this
(47) L. Malaprade, Bull. sac. chim. (France), [5] 1, 833 (1934).
(48) F. Rappaport and I. Reifer, Mzkrochim. Acta, 2, 273 (1937).
(49) Yvonne Khouvine and G. Arragon, Bull. S O C . chim. (France), [5] 8,676 (1941);
Compt. rend., 212, 167 (1941).
(50) H. G. Fletcher, Jr., Advances in Carbohydrate C'hem., 3 , 51 (1948).
(51) P. F. Fleury and M. Joly, J . pharm. chim., [S] 26, 341 (1937).
(51a) P. F. Fleury and M. Joly, J . pharm. chim., [ S ] 26,397 (1937).
(52) P. F. Fleury, G. Poirot and Yvonne FiBvet, Ann. pharm. franc., 6,209 (1947).
(53) R. J. Dimler, H. A. Davis and G. E. Hilbert, J . Am. Chem. Soc., 68, 1377
(1946).
(54) B. H. Alexander, R. J. Dimler and C. L. Mehltretter, J . Am. Chem. Sac.,
73, 4658 (1951).
12 J. M. BOBBITT
Series.66The monocyclic structures L-threitan (I) and erythritan (11) re-
? B
H H
I I1
act with periodate,KB the cis compound, 11, being oxidized the faster.
Trouble has often been encountered in reconciling the results obtained
on periodate oxidation of partially methylated sugars with those expected
on the basis of the then-current t h e ~ r y . ~This ? - ~accumulation
~ of apparently
inexplicable data has culminated in the hypothesis, proposed by Greville
and Northcote,B1that a hitherto-unrecognized kind of periodate oxidation,
which is virtually independent of hydroxyl groups, is here operative. The
rate of this type of oxidation reaches a maximum at a pH of 7.5, and varies
directly with the number of methyl ether groupings. It does not apply to
all partially methylated sugars and is not yet well understood. However, it
was found to proceed only very slowly at the pH values (3 to 5 ) customarily
employed. This work suffers greatly from failure to isolate products.
More successful applications of periodate oxidation to substituted sugars
were made employing the sugar phosphates and phosphoric acid^,^^-^^ the
c h l o r a l ~ s e s , ?*~the
~ ~benzyl ethers of sugars,I1sI2a dimethyleneoxy
(55) R. J. Dimler, Advances i n Carbohydrate Chem., 7, 46 (1952).
(56) H.Klosterman and F. Smith, J. Am. Chem. SOC.,74, 5336 (1952).
(57) R . W.Jeanloz, Helv. Chim. Acta, 27, 1509 (1944).
(58) D . J. Bell, J. Chem. SOC.,992 (1948).
(59) E. L.Hirst and J. K. N. Jones, J . Chem. SOC.,1659 (1949).
(59a) D.J. Bell, Anne Palmer and A. T. Johns, J. Chem. SOC.,1536 (1949).
(60) D.J. Bell and G. D. Greville, J. Chem. SOC.,1902 (1950).
(61) G. D. Greville and D . H. Northcote, J. Chem. SOC.,1945 (1952).
(62) W.E. A. Mitchell and Elizabeth E. Percival, J. Chem. SOC.,1423 (1954).
(63) P. F. Fleury and R . Paris, Compt. rend., 196, 1416 (1933);.I. pharm. chim.,
[8] 18, 470 (1933).
(64) H. von Euler, P. Karrer and B. Becker, Helv. Ciiim. A d a , 19. 1060 (1936).
(65) P. Karrer, P. Frei and H. Meerwein, Helv. Chim. Arta, 20, 79 (1937).
(66) J. E. Courtoie, Bull. SOC.chim. (France), [5]9, 136 (1942).
(67) J. E. Courtois and M. Ramet, Bull. soe. chim. (France), [5]11, 539 (1944).
(68) J. X.Khym, D. G . Doherty and W. E. Cohn, J . A m . Chem. SOC.,76, 5523
(1954).
(69) P. F. Fleury and Jacqueline Jolly, Compt. rend., 226, 688 (1947).
(70) H.Griffon, Ann. pharm. franc., 6 , 165 (1948).
(71) C. E. Ballou and H. 0. L. Fischer, J . Am. Chem. SOC.,76, 3188 (1954).
(72) C. E.Ballou and H. 0. L. Fischer, J . A m . Chem. SOC.,77, 3329 (1955).
(73) P. Andrews, L.Hough and J. K . N. Jones, J . Am. Chem. SOC.,77,125 (1955).
and 0-isopropylidene sugar^.'^-^^ The isopropylidene groups were slowly

hydr~lyzed?~ Whereas a tertiary amine group adjacent to a hydroxyl group
does not completely prohibit ~ x i d a t i o n , the
~ - ~N-acetyl
~ structure appears
fairly resistantls-7 except in 2-acetamido-2-deoxy-~-glucose?*~ 78a
IV. APPLICATION
The application of periodate oxidation to a problem can best be con-
sidered in terms of an approach from two separate standpoints. The first
generally involves the application of analytical techniques to semimicro
quantities of material, and sometimes suffices to yield the desired informa-
tion. After performing this preliminary, analytical investigation, the second
or preparative phase may be undertaken. This entails use of much larger
amounts of material, with the intention of isolating the end products of
oxidation. If these are obtained, their properties should serve to confirm
the results of the preliminary, analytical work; and, if this is proved correct,
the method is then available as a route to the desired synthetic compound.
The analytical phase generally involves the use of very dilute solutions
and a relatively high ratio of oxidant to substrate. Solutions of a concen-
tration of 0.01 M to 0.001 M (in periodate ion) should be employed in an
excess of two t o three hundred percent (of oxidant) over the expected
consumption, in order to elicit a valid value for the selective oxidation.
This value can best be determined by timed measurements of the oxidant
consumption, followed by the construction of a rate curve as previously
described. If extensive overoxidation occurs, measures should be taken
t o minimize it, in order that the break in the curve may be recognized,
and, thence, the true consumption of oxidant. After the reaction has, as
far as possible, been brought under control, the analytical determination
of certain simple reaction-products (such as total acid, formaldehyde,
carbon dioxide, and ammonia) often aids in revealing what the reacting
structures actually were. When possiblc, these values should be determined
a t timed intervals and be plotted as a rate curve. A very useful tool in this
type of investigation, particularly when applied to carbohydrates, has
been the polarimeter. With such preliminary information a t hand, a struc-
ture can often be proposed, or the best conditions for a synthetic operation
can be outlined.
Oftentimes, this information solves the problem, with no further work
(74) A. T. Ness, R . M. Hann and C. S. Hudson, J . A m . Chem. SOC.,64,982 (1942).
(75) E. J. Bourne, G. P. McSweeney, M. Stacey and L. F. Wiggins, J . Chem. SOC.,
1408 (1952).
(76) E. J. Bourne, G. P. McSweeney and L. F. Wiggins, J . Chem. SOC.,2642 (1952).
(77) C. Niemann and J. T. Hays, J . Am. Chem. Soc., 67, 1302 (1945).
(78) R . W. Jeanloz, Ezperientia, 6, 52 (1950).
(7%) R. W. Jeanloz and E. Forchielli, J. B i d . Chem., 188,361 (1951).
14 J. M. BOBBITT
necessary. Thus, the simple fact that DL-glyceritol 1-phosphate reacts with
periodate is sufficient63 to indicate its structure and to distinguish it from
DL-glyceritol 2-phosphate1 which possesses no vicinal glycol structure.
Similar differentiations between two carbohydrate structures can sometimes
be revealed by analytical methods. Thus, by this means, it was shown that
the disaccharidic alditol isomaltitol (111) contains a (1 -+ 6)-glycosidic
linkageP9 If, for example, the glycosidic linkage had been (1 4), an --f
CH,OH
:G\
H q F
6 IOde * HC=O
& O x HC=O OCH,
H I
H OH H ,,H
/OH ; H. ,CH,OH
+ 4HC0,H + H,C=O
Hh=O
HO,
H OH
111
activated methylene structure would have been formed, periodate con-

sumption would have been much higher, and different products would
have resulted.
Similar instances, in which analytical data were of prime importance
and where no products were isolated, are the oxidations of various di-D-
fructose dianhydridesIs0inositols,*l the trisaccharide moiety of the alkaloid
so1amargine,822-O-cu-~-fucopyranosyl-~-fucitol derived from fucoidins3 (a
polysaccharide sulfate ester) , trehalose and n e ~ t r e h a l o s emaltotetraose,86
,~~
D-galactopyranosyl-glyceritol,86and a 3 -p en tu lo ~ e .~ ~
As a corollary to this work, oxidation with periodate constitutes the
basis of analytical procedures for the determination of 6-deoxyhexoses
(methylpentoses) 188 calcium gluconatesg and gluconic acidIg0h e x i t o l ~ , ~ ~ - ~ ~
(79) M. L. Wolfrom, A. Thompson, A . N . ONeill and T . T. Galkowuki, J . Am.
Chem. SOC.,74, 1062 (1952).
(80) Emma J. McDonald and R. F . Jackson, J . Research Natl. Bur. Standards,
36,497 (1945).
(81) A . M. Stephen, J . Chem. SOC.,738 (1952).
(82) L. H. Briggs and E. G. Brooker, J . Chem. Soc., 2833 (1953).
(83) A. N . ONeill, J. Am. Chem. Soc., 76, 5074 (1954).
(84) R . U. Lemieux and H. F . Bauer, Can. J. Chem., 32,340 (1954).
(85) R . L. Whistler and J. L. Hickson, J . Am. Chem. SOC.,76, 1671 (1954).
(86) E. W. Putman and W. Z. Hassid, J. Am. Chem. SOC.,76, 2221 (1954).
(87) G . Ashwell and Jean Hickman, J. Am. Chem. SOC.,77, 1062 (1955).
(88) B. H. Nicolet and L. A. Shinn, J. Am. Chem. Soc., 63, 1456 (1941).
(89) P . F. Fleury, 6. E. Courtois and A. Wickstrglm, Ann. pharm. franc., 6, 338
(1948).
(90) J. E. Courtois and A. Wickstrglm, Ann. pharm. franc., 7, 288 (1949).
glycol mixtures,96glycerit01,~~~ 97 ephedrine,gs r i b o f l a ~ i n e threonine,10'J-*02

,~~
~ ~ ~ the extent of hydroxylation of a
serine,lo3,lo4 5 - h y d r o x y l y ~ i n e ,and
double bond.lo6
An excellent example of the use of polarimetric data in conjunction with
periodate studies is afforded in the work of Hudson, Jackson and Ma-
clay107-110on the glycosides. Their investigation involved an examination
of the configurational difference between the a and i3 series of the methyl
D-aldohexopyranosides, and can best be illustrated by the following series
of reactions. Periodate oxidation permits the quantitative destruction of
the asymmetry of C2, C3, and C4 while preserving C1 and C5 in optical
purity. Since, on oxidation, all of the (investigated) isomeric members of
the a - series
~ showed the same final rotation, it was established that they
possessed the same configuration on C1 and C5. I n the p-D series, solutions
with the same final rotation were also obtained from each member, but
this rotation was different from that of the end product given by members
of the CY-D group. Since both series possess the D configuration on C5, the
difference between the a-Dand /3-D isomers must lie in the opposite con-
figurations of C1. A similar series of experiments was carried out on the
a and p forms of the methyl D-aldopentopyranosides, with analogous re-
sults. Periodate oxidation in this series destroyed the asymmetry of all of
the carbon atoms except C1.
Analytical work, alone, has educed much helpful information on the
structure of polysaccharides.llOaResults in this field have come primarily
(91) F. Rappaport, I. Reifer and H. Weinmann, Mikrochim. Aeta, 1, 290 (1937).
(92) F. Silberstein, F. Rappaport and I. Reifer, Klin. Wochschr., 16, 1506 (1937).
(93) W. W. Smith, Norma Finkelstein and H. W. Smith, J . Biol. Chem., 136, 231
(1940).
(94) A. C. Corcoran and I. H. Page, J . Biol. Chem., 170, 165 (1947).
(95) G. Hoepe and W. D. Treadwell, Helv. Chim. Acta, 26, 353 (1942).
(96) P. F. Fleury and M. Fatome, J . pharm. chim., [8]21, 247 (1935).
(97) Marguerite Lambert and A. C. Neish, Can. J . Research, B28, 83 (1950).
(98) A. Wickstr@m,Ann. pharm. franc., 8, 86 (1950).
(99) A. Wickstr@m,Ann. pharm. franc., 10, 95 (1952).
(100) L. A. Shinn and B. H. Nicolet, J . Biol. Chem., 138, 91 (1941).
(101) T. Winnick, J. Biol. Chem., 142, 461 (1942).
(102) M. W. Rees, Biochem. J . (London), 40, 632 (1946).
(103) B. H. Nicolet and L. A. Shinn, J . Biol. Chem., 139, 687 (1941).
(104) M. J. Boyd and M. A. Logan, J . Biol. Chem., 146,279 (1942).
(105) D. D. Van Slyke, Alma Hiller and D. A. MacFadyen, J . B i d . Chem., 141,
681 (1941).
(106) G. Marinetti and E. Stotz, J . Am. Chem. Sac., 76, 1345 (1954).
(107) E. L. Jackson and C. S. Hudson, J . Am. Chem. Sac., 69,994 (1937).
(108) W. D. Maclay and C. S. Hudson, J. Am. Chem. Sac., 60,2059 (1938).
(109) E. L. Jackson and C. S. Hudson, J . Am. Chem. Sac., 61, 959 (1939).
(110) E. L. Jackson and C. S. Hudson, J . Am. Chem. Sac., 63, 1229 (1941).
(110a) See C. T. Greenwood, this volume, p. 335.
16 J. M. BOBBITT
HOCH
2 IOie
HCOH
HCO
I
CHzOIl
IV
Methyl a-n-mannopyranoside
[al% +
79"
I
HCOCH3
I
HC=O
HC=O
I
HCO-
I
CHzOH
VI
7
HCOCHI
I
HCOH
I
HOCH
I
HCO-
I
CH20H
V
Methyl a-n-glucopyranoside
+
[ a l z " ~ 159"
CHaOCH
HOCH 2 10.3
HCOII
HCO
I
CHaOH
VII
Methyl 8-D-mannopyranoside
[alaoD -69"
I
CHsOCH
1
HC=O
HC=O
I
HCO-
I
CH2OH
IX
[CY:~OD -161"
HOCH
I 2 IOP
HCOH
HC
I
I
CHiOH
VIII
Methyl 8-D-glucopyranoside
[CY]~OD-34"
from the measurement of the amount of formic acid evolved in the oxida-
tion of (1 4 4)-linked polymers with or without (1 -+ 6)-chilin branching;
examples are s t a r ~ h , l land
~ - ~cellul0se.~4~
~~ 117 119 lZ1 140-146
8 9
(111) T. G . Halsall, E. L. Hirst andJ. K. N. Jones, J. Chem. SOC.,1399 (1947).

(112) M. Schlamowite, J. B i d . Chem., 188, 145 (1951).
18 J. M. BOBBITT
If a representative formula (X) for this type of structure is examined, it

may readily be seen that, if the formate ester on the (originally reducing)
end of the oxidation product (XI) remains intact, each end will give rise to
one molecule of formic acid per mole. But, if the formate ester is broken,
(113) M. Abdel-Akher and F. Smith, J. Am. Chem. SOC.,73, 994 (1951).
(114) D. J. Bell and D . J. Manners, J . Chem. Soc., 3641 (1952).
(115) D. J. Bell and D. J. Manners, J . Chem. SOC.,1891 (1954).
(116) D. J. Manners and K. Maung, J . Chem. Soc., 867 (1955).
(117) E. L. Jackson and C. S. Hudson, J . Am. Chem. SOC.,69, 2049 (1937).
(118) C. G. Caldwell and R. M. Hixon, 1.BioE. Chem., 123, 595 (1938).
(119) E. L. Jackson and C. S. Hudson, J . Am. Chem. SOC.,60, 989 (1938).
(120) D. Gottlieb, C. G. Caldwell and R. M. Hixon, J. Am. Chem. SOC.,62, 3342
(1940).
(121) D. H. Grangaard, E. K. Gladding and C. B. Purves, Zaper Trade J., 116,
No. 7, 41 (1942).
(122) F. Brown, Sonia Dunstan, T. G. Halsall, E. L. Hirst and J. K . N. Jones,
Nature, 166, 785 (1945).
(123) T. G. Halsall, E. L. Hirst, J. K . N. Jones and A. J. Roudier, Nature, 160,
899 (1947).
(124) E . L. Hirst, J. K. N. Jones and A. J. Roudier, J . Chem. SOC.,1779 (1948).
(125) F. Brown, T. G. Halsall, E. L. Hirst and J. K. N. Jones, J. Chem. SOC.,27
(1948).
(126) A. L. Potter and W. Z . Hassid, J . Am. Chem. SOC.,70, 3488 (1948).
(127) T. G. Halsall, E. L. Hirst, J. K. N. Jones and F. W. Sansome, Biochem. J .
(London), 43, 70 (1948).
(128) K . H. Meyer and P. Rathgeb, Helv. Chim. Acta, 31, 1545 (1948).
(129) A. L. Potter, W. 2. Hassid and M. A. Joslyn, J . Am. Chem. SOC.,71, 4075
(1949).
(130) G. C. Gibbons and R. A. Boissonnas, Helv. Chim. Acta, 33, 1477 (1950).
(131) I. C. MacWilliam and E. G. V . Percival, J. Chem. SOC.,2259 (1951).
(132) 0. A. Moe, S. E. Miller and Marjorie I. Buckley, J . Am. Chem. SOC.,73,
4185 (1951).
(133) R. W. Kerr and F. C. Cleveland, J . Am. Chem. SOC.,74, 4036 (1952).
(134) K. H. Meyer and W. Settele, Helv. Chim. Acta, 36, 197 (1953).
(135) -4. L. Potter, V. Silveira, R. M. McCready and H. S. Owens, J . Am. Chem.
SOC.,76, 1335 (1953).
(136) R. L. Lohmar, J . Am. Chem. SOC.,76, 4608 (1954).
(137) C. T. Greenwood and J. S. M. Robertson, 6.Chem. SOC.,3769 (1954).
(138) I. A. Wolff, B. T . Hofreiter, P. R . Watson, W. L. Detherage and M. M.
MacMasters, J . Am. Chem. SOC.,77, 1654 (1955).
(139) D. M. W. Anderson, C. T. Greenwood and E. L. Hirst, J . Chem. Sac., 225
(1955).
(140) G. F. Davidson, J . Textile Znst., 31, TS1 (1940).
(141) G . F. Davidson, J . Textile Znst., 32, T109 (1941).
(142) H. A. Rutherford, F. W. Minor, A. R. Martin and M. Harris, J . Research
Natl. Bur. Standards, 29, 131 (1942).
(143) C. Goldfinger, H. Mark and S. Seggia, Ind. Eng. Chem., 36, 1083 (1943).
(144) G. Jayme and S. Maris, Ber., 77, 383 (1944).
(145) E. Pacsu, Textile Research J . , 16, 364 (1945).
the originally reducing end gives two molecules of formic acid per mole.
I n both cases, no formic acid would be obtained from the center units.
CH,OH CH,OH CH,OH
'- H OH H j OH
x
HC=O HC=O + 2HCOzH

0
n
XI
In such a non-branched polymer as1l0aamylose,126slZ9, l33 13* measurement
8
of the evolved formic acid yields direct information concerning the degree
of polymerization. An ambiguity exists in interpreting the results of such
work unless it is first definitely ascertained that the formate ester is saponi-
fied. In one instance,138the formate ester was carefully guarded against
hydrolysis during the initial, acid titration. A subsequent saponification
and titration then yielded a more reliable measure of the degree of poly-
merization.
In the more highly branched polysaccharidesllOa(for example, glyco-
gen111-ll6 and amylopectin126n129-131 9, the type of branching present is
such that the reducing terminal unit (A in XII) is a negligible fraction of
the total polymer. Thus, measurement of the yield of formic acid here
gives the ratio of nontermind D-glucose residues (B in XII) to nonreducing,
terminal portions (B to C in XII), which, in turn, is a measure of the num-
ber of polymerizing units in the branched chains.
The isolation124sl3O' lB1,l37 of, or failure to isolate,ll6' u9 D-glucose from
oxidized, hydrolyzed starches and glycogens has been regarded as an indi-
cation of the presence or absence of (1 + 2) or (1 --+ 3) branching.
If branching is (1 + 3 ) , no glycol grouping will be present in any central
unit and it may be seen that reaction will only occur on the terminal units.
Such is the case with the polysaccharide laminarin, with the glucan of
yeast membrane, and with a galactose-containing polysaccharide isolated
from agar.146-148
With such polysaccharidesllOaas the d e x t r a n ~ , ~ ~ which
~ - ~ 6contain
~
primarily CX-D-(~ -+ 6) linkages, periodate oxidation has been most useful.
Each (1 ---f 6)-linked D-glucose unit contains three contiguous hydroxyl
20 J. M. BOBBITT
/ B
xIr
[In formula XII,A = reducing terminal unit (negligible). B = non-reduc-
ing terminal unit; B consumes two molecules of oxidant to give one mole-
cule of formic acid. C = point of branching. Each non-terminal unit (in
the chains BC and elsewhere in the interior of the molecule) consumes one
molecule of oxidant but produces no formic acid.]
groups, and therefore yields one equivalent of formic acid per mole on
periodate oxidation. Thus, a measurement of the formic acid formed yields
a value which represents the sum of the terminal residues and those which
are (1 -+ 6)-linked. These figures, in conjunction with total periodate
consumption, can be employed for calculating values for (1-+ 4)-like
and (1 -+ S)-like linkages.
Other than in these examples, periodate oxidation has been employed in
(146)V. C. Barry, T. Dillon and Winifred McGettrick, J . Chem. SOC.,183 (1942).
(147)V. C. Barry, J . Chem. SOC.,578 (1942).
(148)V. C. Barry, Nature, 162, 537 (1943).
(149)Allene Jeanes and C. A. Wilham, J . Am. Chem. SOC.,72,2655 (1950).
(150)R. L. Lohmar, J . Am. Chem. Soc., 74, 4974 (1952).
(151)J. W. Sloan, 3. H. Alexander, R. L. Lohmar, I. A. Wolff and C. E. Rist,
J . Am. Chem. SOC.,76, 4429 (1954).
(152) J. C. Rankin and Allene Jeanes, J . A m . Chem. SOC.,76, 4435 (1954).
(153) Allene Jeanes, W. C. Haynes, C. A. Wilhrtm, J. C. Rankin, E. H . Melvin,
Marjorie J. Austin, J. E. Cluskey, B. E. Fisher, H. M. Tsuchiya and C . E. Rist, J.
Am. Chem. Soc., 70, 5041 (1954).
the determination of the structure of heparin,lS4 yeast mannan,22 the

Schardinger dextrins,166'66 x ~ l a n s , ~ 6 ~a - pento~an,'6~
'~~ blood group-A
substance,l64' 166 a glucose-containing protozoal polysaccharide,16a 167 chon-
8
droitinsulfuric acid,lasalginic acid,laS and a polyuronidic hemicellulose from

wheat Periodate oxidation has also been employed in the elucida-
tion of the structure of a g l u c ~ m a n n a n ,a~ polyglucan,
~~ "nigeran,"'7* a
fructan,l73 a galactan,'74 a galactomannan,lTb an algal polysa~charide,'7~
certain gum^,'^^-'*^ a polygalactosiduronic acid methyl ester,'*' a jelly-coat
substance from Echinocardium cordatum,xs2and a seed m~cilage.'~3
Periodate oxidation in conjunction with a color-developing reagent has
been employed as an indicator in the paper chromatography of carbohydrate
(154) J. E. Jorpes, B. Werner and B. Aberg, J . B i d . Chem., 176, 277 (1948).See
also, A. B. Foster and A. J. Huggard, Advances i n Carbohydrate Chem., 10,335 (1955).
(155) K.Myrback and T. Jarnestrom, Arkiv Kemi, 1, 129 (1949).
(156) D.French and R. L. McIntire, J . A m . Chem. Soc., 72, 5148 (1950).
(157) G.Jayme, M. Satre and S. Maris, NatzLrwissenschaften, 29, 768 (1941).
(158) G.Jayme and M. Satre, Ber., 76, 1840 (1942).
(1.59)G.Jayme and M. Satre, Ber., 77,242 (1944).
(159a) G.Jayme andM. Satre, Ber., 77,248 (1944).
(160) S.K. Chanda, E. L. Hirst, J. K. N. Jones and E. G. V. Percival, J . Cheni.
SOC.,1289 (1950).
(161) C. T. Bishop, Can. J . Chem., 31, 793 (1953).
(162) I. Ehrenthal, R. Montgomery and F. Smith, J . Am. Chem. SOC.,76,5509
(1954).
(163) A. S.Perlin, Cereal Chem., 28, 382 (1951).
(164) D.Aminoff and W. T. J. Morgan, Biochem. J . (London), 44, xxi (1949).
(165) D. Aminoff and W. T. J. Morgan, Biochem. J . (London), 48,74 (1951).
(166) G.Forsyth, E.L. Hirst and A. E. Oxford, J . Chem. SOC.,2030 (1953).
(167) G.Forsyth and E. L. Hirst, J . Chem. SOC.,2132 (1953).
(168) K. H. Meyer, M. E. Odier and A. E. Siegrist, HeZv. Chim. Acta, 31, 1400
(1948).
(169) H. J. Lucas and W. T. Stewart, J . A m . Chem. SOC., 62,1792 (1940).
(170) G.A. Adams, Can. J . Chem., 30,698 (1952).
(171) P. Andrews, L. Hough and J. K. N. Jones, J . Chem. Soc., 1186 (1953).
(172) 5. A. Barker, E. J. Bourne and M. Stacey, J . Chem. SOC., 3084 (1953).
(173)V. D. Harwood, R. A. Laidlaw and R . G. J. Telfer, J . Chem. SOC.,2364 (1954).
(174) P. Andrews, L.Hough and J. K. N. Jones, J . Chem. Soc., 806 (1954).
(175) E. 3. Larson and F. Smith, J . Am. Chem. SOC.,77, 429 (1955).
(176) Joyce W. E.Brading, M. M. T. Georg-Plant and Doreen M. Hardy, J . Chem.
SOC.,319 (1954).
(177) B.W.Lew and R. A. Gortner, Arch. Biochem., 1, 325 (1943).
(178) E.L. Hirst and A. S. Perlin, J . Chem. SOC.,2622 (1954).
(179) S.Multherjee and H. C. Srivastava, J . Am. Chcm. SOC.,77, 422 (1955).
(180) G.0.Aspinall, E. L. Hirst and A. WickstrGm, J . Chem. Soc., 1160 (1955).
(181) P.A. Levene and L. C. Kreider, J . B i d . Chem., 120,591 (1937).
(182) E.Vasseur, Acta Chem. Scand., 6,376 (1952).
(183) E.L.Hirst, E. G. V. Percival and Clare B. Wylam, J . Chem. SOC.,189 (1954).
22 J. M. BOBBITT
materials.1s3aColor-developing reagents which have been used include a

benzidine solution,184an alkaline permanganate solution,186Schiff reagent,186
and a borate-buffered, potassium iodide and starch mixture.lS7The benzidine
technique was more successful with glycosides than with free sugars.
Periodate followed by Schiff reagent has been successfully used for
staining carbohydrate structures on histological slides,ls8~ IE9 but the possi-
bility of interference b y amino acids has been pointed out.lgO

Although the analytical treatment of periodate oxidation is sometimes
sufficient, and certainly always desirable, confirmation of the results by
actual isolation of the oxidation products is often necessary. This can be
considered as a preparative phase and necessitates somewhat different
operating conditions. As compared with analytical conditions, only a slight
excess of oxidant is permitted to react with a larger proportion of substrate,
in a more concentrated solution. The separation and isolation of the oxida-
tion products brings to the fore several new problems. It is always ad-
vantageous to separate the organic products from the periodate ions, and
it is often necessary that all of the inorganic salts be removed from the
reaction medium. This poses no problem when the organic products are
soluble in organic solvents and so can be extracted, but such a situation is
but seldom encountered in carbohydrate studies.
The removal of inorganic salts from reaction mixtures afforded by poly-
meric materials may be simply and effectively accomplished by dialy-
sis,1*5, after decomposition of remaining periodate with ethylene gly-
~01130- or butylene glycol.161* Alternatively, the iodate and periodate
ions may be removed as such, or after reduction to free iodine. The iodate
and periodate ions have been effectively precipitated by means of sodium
carbonate plus manganous sulfate,6 or by lead dithionate,lgl barium chlo-
ride124,I g 2 . lg3 strontium h y d r o ~ i d e ~ or
~ 4 barium
- ~ ~ ~ hydroxide,203 204 lead
8
(183a) G . N . Kowkabany, Advances i n Carbohydrate Chem., 9,303 (1954).

(184) d . A. Cifonelli and F. Smith, Anal. Chem., 26, 1132 (1954).
(185) R. U. Lemieux and H. F. Bauer, Anal. Chem., 26,920 (1954).
(186) J . G. Buchanan, C. A. Dekker and A. G. Long, J . Chem. Sac., 3162 (1950).
(187) R . L. Metzenherg and H. K . Mitchell, J. A m . Chem. Sac., 76,4187 (1954).
(188) J. F. A. McManus, Nature, 168,202 (1946).
(189) R . D . Hotchkiss, Arch, Biochem., 16, 131 (1948).
(190) J. F. Lhotka, Nature, 171, 1123 (1953).
(191) J. F. ODea and R. A. Gibbons, Biochem. J. (London), 66,580 (1953).
(192) J. W. Pratt and N. K . Richtmyer, J . Am. Chem. Soc., 77. 1906 (1955).
(193) Laura C. Stewart and N. K . Richtmyer, J . A m . Chem. Sac., 7 7 , 1021 (1955).
(194) E. L. Jackson and C. S. Hudson, J. A m . Chem. Sac., 68, 378 (1936).
(195) E. L. Jackson and C. S.Hudson, J. A m . Chem. Soc., 61,1530 (1939).
(196) W. D. Maclay, R . M. Hann and C . S.Hudson, J. A m . Chem. SOC.,61, 1660
(1939).
(197) E. L. Jackson and C. 5. Hudson, J. A m . Chem. Sac., 62,958 (1940).
PERJODATE OXIDATION O F CARBOHYDRATES 23
a ~ e t a t e , 2 ~ or
~-~lead
~ nitrate.22 Periodate ion, alone, may be effectively
precipitated by potassium ion in cold, acid solution.208
Periodate and iodate ions may both be removed by reaction with iodide
ion in acid solution; this yields free iodine which can be removed by extrac-
tion209 or by centrifugation2l0-or the iodine can be converted to iodide ion
by titration with sodium thiosulfate.211Periodate and iodate ions are also
readily removed from solution by use of a n anion-exchange resin.2118
The dialdehydes, such as VI and XI, produced by periodate oxidation of
sugar glycosides or polymers, very probably do not exist in the aldehydo
f0rm~2~2-2~4 but in a hydrated212and cyclized structure.213 2149
Since aldehydes are notoriously polymerizable and difficult to manipu-

late, the products of periodate oxidation are oftentimes further oxidized,
with hypohalite, to carboxylic acids, or are reduced to the corresponding
alcohols. Oxidation has been more usually employed than reduction, since
acids frequently form crystalline salts and other conveniently prepared
derivatives. A process of oxidation of these aldehydic products by hypo-
bromite, in the presence of barium carbonate or strontium carbonate, was
developed and used extensively by Hudson and his coworkers.107-110 1
194-199, 203 Their method can best be illustrated by an example: the further
oxidation of the dialdehyde, VI, shown previously (see p. 16) to be obtained
by the oxidation of the methyl a-D-aldohexopyranosides.The isolation of
(198) N. K. Richtmyer and C. S. Hudson, J . Am. Chem. SOC.,62,961 (1940).
(199) A. E. Knauf, R. M. Hann and C. S. Hudson, J . Am. Chem. SOC.,63, 1447
(1941).
(200) J. E.Courtois and Angela Valentino, Bull. sac. chim. biol., 26,469 (1944).
(201) J. E.Courtois, B d . S O C . chim. biol., 29, 248 (1947).
(202) J. E.Courtois and A. WickstrZm, B d . S O C . chim. biol., 32,759 (1950).
(203) N. K.Richtmyer and C. S. Hudson, J . Org.Chem., 11.610 (1946).
(204) P. F. Fleury, J. E. Courtois and R. Perlks, Jaikrochemie ver. Mikrochim.
Acta, 36/37, 863 (1951).
(205) S.Akiya and S. Okui, J . Pharm. SOC.Japan, 71,865 (1951);Chem. Abstracts,
46, 8018 (1952).
(206) S . Akiya, S. Okui and S. Suauki, J . Pharm. Sac. Japan, 72, 1554 (1952);
Chem. Abstracts, 47, 8657 (1953).
(207) S.Akiya, S.Okui and S. Suzuki, J . Pharm. SOC. Japan, 72,785 (1952);Chem.
Abstracts, 47, 7446 (1953).
(208) P.Desnuelle and M. Naudet, Butt. SOC. chim. (France), [5]12,871 (1045).
(209) 0.Th. Schmidt and P. Gunthert, Ber., 71,493 (1938).
(210) F.Eisenberg, Jr., and S. Gurin, J . Am. Chem. Soc., 73, 4440 (1951).
(211) G. Arragon, Compt. rend., 211, 558 (1940).
(211a) M.A. Smith and B. R. Willeford, Jr., Anal. Chem., 26,751 (1954).
(212) J. W. Rowen, Florence H. Forziati and R. E. Reeves, J. A m . Chem. SOC.,
73, 4484 (1951).
(213) C. D.Hurd, P. J. Baker, Jr., R. P. Holysa and W. H. Saunders, Jr., J . Org.
Chem., 18, 186 (1953).
(214) V. C. Barry and P. W. D. Mitchell, ,I.Chem. SOC., 3631 (1953).
24 J. M. BOBBITT
these oxidation products established the relative configuration of C1 in the

a!and B series of the methyl D-aldohexopyranosides and confirmed the
I
SrCOa
HC=O
I
I
CHzOH
HCO
I
CHzOI-I
I
CHzOH
VI
results indicated by the changes in optical rotation. In addition, the isola-
tion verified the presence of the pyranoside ring and the D-configuration
for C5. This technique has since been used extensively in oxidations of the
mono- and oligo-saccharides,1s2~ lg3 201 * 2 0 2 * 216, 216 and their derivatives.200*
+
,O6p2oD Applications have been made to cellulose,144 to an oxidized poly-

galactosiduronic acid,lB1to an oxidized ~ y l a n , ~l69* ~ ' ,and to alginic acid.lG9
The reduction of the aldehydic structures to the corresponding alcohols
-"Ao
has been applied, in the main, to polymeric materials.l44- lK1, 167 169a- lE2, 217
3
CH,OH
HC=O
0 4 HC=O
O=CH
XI
CH,OH CH,OH SH,OH
HOCH,
0
HOCH, CH,OH
n
CH,OH
CH,OH I
I HC=O CHOH
CHOH + ( n + 1) I + ( n + 1) I + HCO,H
I CH,OH CHOH
CH,OH I
CH,OH
(215) P. F. Fleury and J. E. Courtois, Compt. rend., 214, 366 (1942); Bull. soc.
chim. (France), [5] 10,245 (1943).
(216) L. H. Briggs and L. C. Vining, J . Chem. SOC.,2809 (1953).
(217) M. Abdel-Akher, J. K . Hamilton, R. Montgomery and F. Smith, J . Am.
Chem. SOC.,74, 4970 (1952).
This sequence is exemplified by the further reaction of the aldehydic

substance XI, obtained by the periodate oxidation of a representative
carbohydrate polymer.
The reduction, to the corresponding alcohols, of the aldehydic compounds
derived from the periodate oxidation of the glycosides has provided an
excellent confirmation of the work of Hudson and his coworkers in the
a- and P-D-aldohexopyranoside series. Smith and Van Cleve218noted that
such reduction destroys the asymmetry of C5 (those of C2, C3 and C4 had
already been destroyed by periodate) and leaves only C1 in its original
asymmetric state, thus providing a more direct route to effecting a dis-
tinction between the a-D and p-D series of the aldohexopyranosides. This
may be illustrated by inspecting the reduction of the aldehydic fragment
derived from the periodate oxidation of a methyl a-D-aldohexopyranoside
(VI, having asymmetric carbon atoms marked with an asterisk). The re-
I
I I C ! HC*OCH3
'
HC *O
I
A I
HCO-
I
CHiOH CHzOH
VI
agents so far used in the reduction of these aldehydes have been sodium
borohydridez17# and hydrogen catalyzed by Raney n i ~ k e 1 . l161
~ ~ 159s, 8
162, 217. 218
The reaction of these aldehydes, derived from periodate oxidation, with

carbonyl reagents has also been investigated. Studies 14', 148 were made on
oxidized laminarin, a (1 + 3)-linked polysaccharide, in which only the
terminal residues had been oxidized. The addition of phenylhydrazine
acetate detached the remainder of the terminal residue as glyoxal phenyl-
osazone. When the aldehydic compounds derived from the periodate oxida-
tion of raffinose and trehalose2188
were reacted with p-nitrophenylhydrazine,
the authors were surprised to find that one molecule of oxidized raffinose,
containing six aldehydic functions, reacts with only three molecules of
the reagent, and that the four aldehydic functions of the oxidized trehalose
molecule react with only two. The reactions of periodate-oxidized carbo-
(218) F. Smith and J. W. Van Cleve, J . A m . Chem. Sac., 77,3091 (1955).
(218a) S. Akiya, S. Okui andS. Suzuki, J. Pharm. Sac. Japan, 72,891 (1952); Chem.
Abstracts, 47, 7447 (1953).
26 J. M. BOBBITT
hydrates with nitrogenous bases have since been extensively

219-222 and a reaction sequence was proposedZ14
which entailed (tentatively)
the formation of a new hydrated, aldehydic, cyclic structure (XIII) to ac-
count for the absence of a carbonyl group,212,213 and of a final structure
(XIV) capable of explaining the reaction of two aldehydic groups per
molecule with only one molecule of base.218aA cyclic structure of this type
had previously been established for such a dialdehyde.222a-222c Controlled
hydrolysis of these reaction products afforded the corresponding nitrogen
---&TI r-
CH
I
HCOH
I
HCOH
I I
R-HN-N
I
0
R-NH-NHa ~
I
HCOH HCOH
I
.--OCH
I
HCO-
I
CH,OH
I
CHaOH
XI11 XIV
derivatives of the simple aldehydic fragments; these are more readily

handled than the free comp0unds.~47~ 148* 167, 206-207* 214, 218a-222 The periodate
oxidation of an or-glycol grouping concerned in a ring structure yields a

dialdehyde which reacts with only one molecule of carbonyl reagent per
moleZ14921*a, 221; consequently, the percentage of nitrogen in the product
from a polymer so treated (by periodate oxidation and subsequent reaction
with the hydrazine derivative) has been used as a measure of the percentage
of or-glycol groups present in the original
Still another reaction of these aldehydic structures (formed by periodate
oxidation), cyanohydrin formation, has been inve~tigated.2~~
Care must be taken that the periodate-oxidation products are not
(219) T. Dillon, Nature, 166, 546 (1945).
(220) V. C. Barry and P. W. D. Mitchell, J. Chem. Soc., 3610 (1953).
(221) V. C. Barry, Joan E. McCormick and P. W. D. Mitchell, J. Chem. Soc.,
3692 (1954).
(222) V. C. Barry and P. W. D. Mitchell, .J. Chem. SOC.,4020 (1954).
(222a) H. E. Carter, R. K. Clark, Jr., S. R. Dickman, Y. H. Loo, P. S. Skell and
W. A. Strong, Science, 103, 540 (1946).
(222h) R. U. Lemieux and M. L. Wolfrom, Advances in Carbohydrate Chem., 3 ,
349 (1948).
(222c) E. Schreier, G. Stlihr and E. Hardegger, Helu. Chim. Acta, 37, 574 (1954).
(223) H. S. Isbell, Abstracts Papers Am. Chem. Soc., 122, 17A (1952).
modified in some undesirable or unknown way by these methods involving

isoIation of derivatives. The precipitation of free iodine can be deleterious,
since a reaction has been shown to take place between iodine and certain
periodate-oxidation products.43Treatment of the common dialdehydic type
of oxidation product with a strong base is performed a t the risk of inducing
~ ~Indeed,
an internal Cannizzaro r e a ~ t i 0 n . l 224 . this reaction has been found
to occur quantitatively with the dialdehyde (XVI) engendered by the
oxidation of methyl P-L-rhamnopyranoside (XV) . This Cannizzaro reaction
I 1
HCOCH, HCOCHI 1 1
- -
I I HCOCH, HCOCHj
HCOH HC=O I
CHzOH
I
I COzH
HCOH
2 10'0 NaOH
+
1 COzH CHnOH
HOCH HC=O I I
I I -0CH -0C H
-0CH -0CH I I
1 I CHI CHa
CH.7 CH3
XV XVI
is complete within five minutes a t loo", using N sodium hydroxide.
I n addition to the foregoing examples, periodate oxidation has been
applied to isotopically labeled sugars (for the determination of label dis-
tribution), 226, 226 to certain branched-chain sugars,227-229 and to some
nitrogenous derivatives of the simple s ~ g a r s . 2 " ~ - ~ ~ ~
V. REACTION CONDITIONS
1. Oxidant
Periodic acid, sodium metaperiodate, and potassium metaperiodate are
now commercially available. Trisodium paraperiodate, Na3H2106, may be
(224) E. M. Fry, E. J. Wilson, Jr., and C. S. Hudson, J. Am. Chem. SOC., 64, 872
(1942).
(225) A. P. Doerschuk, J . Am. Chem. SOC.,73, 5453 (1951).
(226) F. Eisenberg, Jr., J . Am. Chem. SOC.,76, 5152 (1954).
(227) N. G . Brink, F. A. Kuehl, Jr., E. H. Flynn and K . Folkers, J . Am. Chem.
Soc., 68,2405 (1946).
(228) M. L. Wolfrom and C. W. DeWalt, J . Am. Chem. Soc., 70, 3148 (1948).
(229) P. P. Regna, F. A. Hochstein, R . L. Wagner, Jr., and R. B. Woodward,
J . Am. Chem. SOC.,76,4625 (1953).
(230) P. Karrer and K. Pfaehler, Helo. Chim. A d a , 17,766 (1934).
(231) C. Niemann and J. T. Hays, J. Am. Chem. SOC.,62,2960 (1940).
(232) C. Niemann, A. A. Benson and J. F. Mead, J. Org. Chem., 8,397 (1943).
(233) E. Chargaff and B. Magasanik, J . Am. Chem. SOC.,69, 1459 (1947).
(234) J. F. Mead and E. A. Bartron, J. Am. Chem. Soc., 70, 1286 (1948).
(235) C. F. Huebner and K. P. Link, J. Am. Chem. Soc., 72, 4812 (1950).
28 J. M. BOBBITT
prepared in 90 % yield by the interaction of bromine and sodium iodide in

aqueous sodium hydroxide at 80:36 or in 80% yield by the oxidation of
sodium iodate with chlorine in the same s0lveiit.2~~ From this salt, sodium
metaperiodate is readily preparable by crystallization from dilute nitric
acid (used in the ratio of 150 ml. of water:45 ml. of concentrated nitric
acid:100 g. of salt.) Paraperiodic acid may be obtained in 93% yield
from trisodium paraperi0date,2~0or by the electrolytic oxidation of iodic
acid.238. 239
An aqueous solution of periodic acid, free from metal ions, may be pre-
pared through the conversion of potassium metaperiodate to the slightly
soluble barium dimesoperiodate, Ba2120,, which, through reaction with an
equivalent amount of sulfuric acid, yields pure periodic acid.lo7 Hudson
found that nitrate as an impurity in periodic acid gives rise t o erroneous
results in this field.239a
A novel method for the electrolytic regeneration of periodate ion in
oxidizing solutions has been rep0rted.~4OThis procedure shows industrial
promise, since i t circumvents the initial, high cost of periodates.
The preparation of periodate compounds has been well summarized by
Smith.24l Periodate solutions appear to be stable in the dark at room
t e m p e r a t ~ r e .242-244
~~
Another factor to be considered in the use of periodate ion as Oxidizing
agent is the desired pH. Potassium metaperiodate in solution givesz9a p H
value of about 4. For values of pH below 3 to 4,pure periodic acid or acid-
salt solutions must be used. For values of pH above 5 , solutions containing
potassium hydroxide and potassium metaperiodate must be employed.245
It was found29 111 114 122 124 125 127, 131 135 137 174 that, if an equimolar
8 I 1 v I I I
mixture of sodium metaperiodate and potassium chloride was used, partial

precipitation of potassium metaperiodate occurred, and the resulting, low,
constant concentration of periodate ion evoked less overoxidation.
(236) J. Lange and R. Paris, J . pharm. chim., [8]21,403 (1935).
(237) A . E. Hill, J . A m . Chem. Soc., 60, 2678 (1928).
(238) A. Hickling and S. H. Richards, J . Chem. SO C., 256 (1940).
(239) H. H. Willard and R. R. Ralston, Trans. Electrochem. SOC.,62, 239 (1932).
(239a) C. S. Hudson, personal communication.
(240) W. Dvonch and C. L. Mehltretter, J . Am. Chem. SOC.,74,5522 (1952). C. L.
Mehltretter, U. S. Pat. 2,713,553 (1955); Chem. Abstracts, 49, 13E06 (1955).
(241) G. F. Smith, Analytical Applications of Periodic Acid and Iodic Acid and
Their Salts, G . Frederick Smith Chemical Co., Columbus, Ohio, 5th Edition, 1950,
p. 7 (Manufacturers circular).
(242) F. S. H. Head and G. Hughes, J . Chem. SOC.,2046 (1952).
(243) F. S. H . Head and H. A. Standing, J . Chem. SOC.,1457 (1952).
(244) F. S. H. Head, J . Testile Inst., 44, T209 (1953).
(245) K. H. Meyer and P . Rathgeb, H e l v . Chim. Acta, 31, 1540 (1948).
2. Solvent
The use of water or of an aqueous solvent as the reaction medium is a
characteristic of periodate oxidation, as performed in this field. This situ-
ation is ideal for most work on carbohydrates, but imposes on applications
to other fields of organic chemistry a limitation which, nevertheless, has
been partially circumvented in a number of ways.
Palfray and S a b e t a added
~ ~ ~ ~an emulsifying agent, Gardinal, to aid in
the oxidation of the water-insoluble l-O-benzylglyceritol. Aqueous solu-
tions of rnethan01,~4~'248 ethanol,13 249-261 di0xane,1~*
262-266 acetic 266
and acetic acid buffered with lithium acetate16*have been used. The use
of lithium periodate or triethylammonium periodate in aqueous alcohol
solution has been suggested,267because of the solubility of these salts in
this medium.
The rate of oxidation in aqueous ethanoP1, 268 and in aqueous acetic acid31
is lower than that in water; and this means of decreasing the rate has been
suggested31as an improvement on lowering the temperature for this purpose.
3. Hydrogen-ion Concentration
The pH of the periodate reaction medium is of paramount importance
in such oxidation of carbohydrates, principally because of the sensitive
acetal and ester linkages already present or formed in the course of the
reaction. In many cases, any hydrolysis of these linkages will free more
than the original theoretical number of reactive groups. The pH essential
for preserving the blocking groups intact must be compromised with the
optimal pH for the periodate oxidation.
The rate of oxidation of ethylene glycol was found268-269 to attain a
broad maximum between pH 2.5 and 6. I n a very complete study of the
effect of pH on the periodate oxidation of carbohydrates, Neumuller and
Vasseur2B0showed that the oxidation of maltose, melibiose, methyl LY-D-
(246) L. Palfray and S. Sabetay, Bull. SOC. chim. (France), [5] 4,950 (1937).
(247) T. Reichstein, C. Meystre and J. von Euw, Helv. Chim. Acta, 22,1107 (1939).
(248) L. F. Fieser, M. Fields and S. Lieberman, J . Biol. Chem., 166, 191 (1944).
(249) H. L. Mason, C. S. Myers and E. C. Kendall, J . Biol. Chem., 116,267 (1936).
(250) G. King, J. Chem. SOC.,1788 (1936).
(251) G. King, J . Chem. SOC.1826 (1938).
(252) J. von Euw and T. Reichstein, Helv. Chim. Acta, 23, 1114 (1940).
(253) H. G. Fuchs and T. Reichstein, H e l v . Chim. Acta, 24, 804 (1941).
(256) 1'. Karrer and R . Hirohata, Helv. Chim. Acta, 16,959 (1933).
(257) R. A. Gibbons, unpublished results.
(258) J. E. Taylor, B. Soldano and G. A. Hall, J . Am. Chem. SOC.,77,2656 (1955).
(259) J. E. Taylor, J . Am. Chem. Soc., 76, 3912 (1953).
(260) G. Neumuller and E. Vasseur, Arlciv K e m i , 6,235 (1953).
30 J. M. BOBBIW
glucopyranoside, yeast glucan, and dextran proceeded with a minimum of

overoxidation in the p H range 3 to 3.5. The most nearly theoretical
oxidation of starch occurred in the p H range 2 to 5, although overoxida-
tion could not be completely suppressed.lZ1A solution buffered a t p H
3.7 to 5 was recommended for the oxidation of starch and c e l l ~ l o s e lS7 .~~~~
LindstedtZ3found that the results obtained in neutral and acidic media
were similar when carbon-carbon bonds were broken and no hydroxy
aldehydes were formed. Otherwise, results higher than the theoretical were
obtained in neutral solution. He showed that the yeast mannan molecule
is oxidized almost completely to carbon dioxide and water if the p H is
above 5. However, he worked in a temperature range near 50, which is
high for periodate work. Meyer and Rathgeb246 successfully used a p H
of 4.2 t o 4.5 in the oxidation of polyhydric alcohols. I n the oxidation of
methyl 8-cellobioside and methyl 6-D-glucopyranoside, Head24 found that
a p H of 9 to 10 promotes hydrolysis of the acetal linkages of the intermedi-
ate dialdehydes and consequently enhances overoxidation. The degrada-
tion of isotopically labeled D-glucose, by periodate oxidation of the derived
D-gluconic acid:26 was found to proceed more smoothly a t p E 5.8 than at
pH 1; and the oxidation of glyceritol was erratic at p H 13.z6
In a study of the oxidation of 2-amino-2-deoxy-~-g~ucose (D-glucosamine)
derivatives, NeubergeP obtained better results using sodium metaperiodate
(at its accompanying p H of 3 to 4) than with periodic acid a t lower p H
values; and a p H of 4.5 was found by Jeanloz and F ~ r c h i e l l i t~o~be
optimal
for these derivatives.
Contrary to these results, a p H of 7.5 gave the most rapid oxidation of
the simple amino alcohols,g ephedrine,g8serine, and threonine.l00. lo1 l o 3 4
8
A pH of 7.5 was found to be necessary for the successful oxidation of

partially methylated D-glucose derivatives.67~68, s g a # 6o It is conceivable
that these apparently strange results may be related to the oxidation of
the sterically hindered glycol, pinacol, which also has a rate maximum at
pH 7.5.261I n a study of the oxidation of a series of progressively more
hindered glycols (from ethylene glycol to pinacol), a change in reaction
course accompanied by a change in maximum of p H rate was observed for
2-methyl-2,3-butanediol to pinacol, Since partially methylated
D-glucose is certainly more hindered than D-glucose, a different reaction
may occur with the more hindered compounds.
If these results are considered in the light of the excellent work of Crouth-
amel and associates263 (wherein it was found that, in aqueous periodate
(261) C. C. Price and H. Kroll, J. Am. Chem. SOC.,
60,2726 (1938).
(262) F. R. Duke and V. C. Bulgrin, J . Am. Chem. SOC.,7 6 , 3803 (1954).
(263) C. E. Crouthamel, A. M. Hayes and D. S. Martin, J . Am. Chem. SOC.,7 3 ,
82 (1951).
solutions, the univalent, non-hydrated periodate ion, IO$, prevails between

approximately pH 3 and 8 a t 25), the major glycol-cleaving ability of
periodate solutions must be ascribed to that ion. This was established, by
kinetic means, for ethylene
Certain solubility factors are to be considered in work with periodate
at different pH values. Above pH 5 , disodium paraperiodate (Na2HoIOa)
tends to crystallize from aqueous Below p H 5 , the sodium
salt of periodic acid is more soluble, whereas above p H 5 , the potassium
salt is best employed, Phosphate-buffer solutions cause erroneous re-
s u l t ~ ,6 9~a ,~6 0, , and should be avoided in oxidations of carbohydrates.
I n the carbohydrate field, p H values outside the range 3 to 5 should
be avoided whenever possible. When such factors as the presence of blocking
groups render this infeasible, some reactions can still be performed a t
pHs of 7.5264and 5.7.265The partially methylated sugars,67*6 8 , 69a, 6o the
highly hindered glycols,261and certain amino alcohol^,^^ 98, l o o *lol- are
exceptions and tend to be most readily oxidized a t a p H of 7 . 5 .
4. Temperature
Most periodate oxidations have been performed a t room temperature,
but there is some reason to believe that a t least some of them might have
been more successfully accomplished a t lower temperatures. Evidence has
been accumulated indicating that a more random type of oxidation occurs
at higher than at lower temperatures.-23
Lowering the temperature of the reaction would certainly decrease the
rate of acetal hydrolysis and thereby partially remove one of the causes
of overoxidation. This would simultaneously decrease the rate of oxida-
tion by periodate. Although no comprehensive study of the effect of tem-
perature on oxidation rates has been made, the number of reactions suc-
cessfully dealt with in the temperature range of 0 to 6O3l# 7 8 -
-
39s 126, 1303
133 164, 1769 2O3 2 1 0 2 6 6 , 267 indicates that lowered temperatures do not affect
the rates unfavorably. In order to obtain the maximum of selective oxida-
tion and the minimum of overoxidation, periodate oxidations should be
performed a t as low a temperature as is practicable in relation to the solvent
system used and the solubility of the reactants therein.
5 . Light
The study of the effect of light has been one of the more recent contri-
butions to the literature on periodate oxidations. It was shown4. 36, 242-244
(264) R . E. Reeves, J. A m . Ghem. Soc., 63, 1476 (1941).
(265) A. S. Perlin, J. A m . Chem. SOC.,76, 4101 (1954).
(266) R . U. Lemieux and H . F. Bauer, Can. J . Chem., 31, 814 (1953).
(267) K . H. Meyer and P. Rathgeb, Helv. Chim. Acta, 32,1102 (1949).
32 J. M. B O B B I a
that aqueous sodium metaperiodate slowly decomposes spontaneously in

the presence of light, t o yield sodium iodate and ozone.
These various workers found that formic acid, formaldehyde, methy1
alcohol, oxalic acid, glycolic acid, and glyoxal are oxidized in the presence
of periodic acid and light within ten to twenty days, but are unaffected
by the oxidant in the dark. Acetaldehyde was found to be oxidized only
very slowly in the presence of light, whereas acetic acid is completely un-
affected. Over a period of seventeen days in the light, cellulose was found
to take up 1.6 equivalents of oxidant per anhydro-D-glucose residue, but
only the theoretical one equivalent in the dark. Similar results were ob-
tained in the oxidation of methyl /3-cellobioside and methyl /3-D-gluco-
pyranoside. It was demonstrated244that, in such reactions permitted to
proceed over a very long time, a small proportion of overoxidation does
occur in the dark, but the odor of ozone, denoting the spontaneous de-
composition of sodium metaperiodate, was absent. A sample of methyl
8-D-glucopyranoside, treated with a 50 % excess of sodium metaperiodate,
reacted completely with the oxidant in forty-five days in the light, whereas
two and a half years were required in order to reach the same stage in the
dark.
The effect of light needs consideration only in long-term reactions, but
it can then be very important.
6. Concentration of Reactants
I n homogeneous reactions, the upper limits of concentration are de-
termined by the (limited) solubility of the salts of periodic acid and by the
low p H values produced by periodic acid itself. Apart from these considera-
tions, the concentration conditions to be selected are governed by the type
of information desired. A very dilute solution having a high 0xidant:sub-
strate ratio is used in the exploratory or preliminary phase defined earlier
(see p. 13), but a more concentrated solution, in which the oxidant is
only slightly in excess of the theoretical, is recommended for the preparative
phase.
VI. METHODS OF ANALYSIS
1. Oxidant Consumption
Calculations derived from the measurement of final periodate consump-
tion indicate the number of reactive groups and can often be interpreted
to reveal the extent of overoxidation. Chemically, this determination
involves the use of one of two general reactions. These are (a) the reduction
of periodate and iodate t o free iodine in acid solution, and (b) the reduction
of periodate to iodate in neutral solution.
The most common reagent used for reducing periodate and iodate to iodine
is the iodide ion. The free iodine is then titrated with standard thiosulfate
solution.
lode + 7 + 5 He + 4 12 + 4 HzO
Ie
10se + 5 Ie + 6 Hm + 3 Iz + 3 Ha0
I2 + 2 2 Ie +
S203ee-+ s40aee
This method was used by Malaprade2t 47 and is sometimes employed to-

days6,lz0, 2b9 because of its very fast action. A disadvantage of the method
is the appearance, in the reaction mixture, of free iodine which can react
with some oxidation products.
The most widely used of the methods involving a reduction of periodate,
only, to iodate employs the arsenite ion4 as reductant, in a solution main-
tained at neutrality with sodium bicarbonate, with iodide ion as catalyst.
IOde + hoee 5 ASOae -k IOae
The standard procedure is t o drop aliquots of the reaction mixture into a n
aqueous quenching solution consisting of a small proportion of potassium
iodide, an excess of sodium bicarbonate, and a known excess of standard
sodium arsenite. After fifteen minutes, the solution is back-titrated with
standard iodine having about the same normality as the arsenite solution.
This method obviates the appearance of free iodine for long periods of
time and has the additional advantage of incorporation of a primary
standard, sodium arsenite solution. A serious disadvantage of the pro-
cedure, when applied to studies of the rate of very fast reactions, is its
relatively low rate.269It has been recommended that the final quenching
reaction be carried out162at 4. Moreover, the quenching mixture must
notZB8 have a pH above 7.5, since, above this value, di- and tri-sodium
paraperiodates are precipitated and this may cause erroneous results. The
method has been used extensively14, 101-110 162 172, 173 203 * 209 268 269 * 270 and
I I I I 8
has been described in excellent detail b y Jacks0n.2~The suggestion has

been madeZ7lathat it gives more accurate results when compounds contain-
ing sulfide structures43 or intermediates containing active hydrogen atoms
are not present.
The technique of Willard and Greathouse2Y2uses iodide ion as the reduc-
ing agent, in a solution maintained near neutrality with borax-boric acid
(268) G. Lundblad, A r k i v K e m i , Mineral. Geol., 24A, No. 25 (1947).
(269) P. F. Fleury, Mikrochemie ver. Mikrochim. Acta, 26.263 (1938).
(270) M. A. Smith and B. R. Willeford, Jr., J . Am. Chem. SOC.,76,2502 (1954).
(271) E. L. Jackson, in Organic Reactions, R. Adams, ed., John Wiley and Sons,
Inc., New York, N. Y., 1944, Vol. 2, p. 341.
(271a) J . C. P. Schwara, Chemistry & Industry, 1000 (1954).
(272) H. H. Willard and L. H. Greathouse, J . Am. Chem. Soc., 60,2869 (1935).
34 J. M . BOBBITT
buffer. The free iodine liberated by the reduction of periodate to iodate is

IOae + 2 Ie + 2 H ---f Iz + IOae + HzO
then titrated as usual. This method has been used less often169- 231- 234
than either the Malaprade or the Fleury arsenite procedure.

A method has been for the simultaneous removal of iodate
and periodate by passing the aqueous solution over an anion-exchange
resin. The two ions can then be recovered (in 98 % yield), and determined
by one of the usual methods. This procedure has been applied to highly
furthermore, it is the method of choice if the reagents
colored solutions211a;
employed in determining oxidant consumption react with the organic
compounds present.
The analyses of periodate and iodate mixtures is readily accomplished
colorimetrically at 222.5-230 mp (in the ultraviolet region).263273 The 3
method utilizes very low concentrations of reactants, but is useless when

the reactants or products have structures which absorb light in this region
of the ultraviolet absorption spectrum. Also, certain undesirable side-
reactions are catalyzed by ultraviolet light.273
2. Acidity Measurement
Measurement of the acidity produced in the periodate oxidation of
carbohydrates is confined mainly to the determination of formic acid. Only
from unsubstituted ketoses is the formation of such acids as glyoxylic and
glycolic to be expected in carbohydrate chemistry.274
One of the more difficult problems encountered in obtaining a valid
assay of formic acid is that of formate ester formation. The formate is
derived from the cyclic, hemi-acetal structure which is an equilibrium
form of many free sugars in solution. For example, the oxidation of one
of the cyclic forms of D-glucose can readily be seen to give a formate ester
(as well as a C-formyl group) on the atom originally denoted as C5. It
HpF
CH,OH
1
3 10
L Q A-O
O=CHH
\C=O
I + 2HC02H
HO H
H OH (A) (B)
[(A) is the C-formyl group; (B) is the 0-formyl group]
then becomes necessary to know whether the ester is saponified in the

reaction mixture. If it is, D-glycerose will be formed and this will react
(273) J. S. Dixon and D. Lipkin, Anal. Chem., 26, 1092 (1954).
(274) P. F. Fleury and Yvonne Fibvet-Guinard, Ann. pharm. franc., 6, 404 (1947).
further with periodate. On the other hand, the oxidation of the acyclic
sugar will not give a formate. That these formates are formed was proved
by Barker and Smith.276They oxidized 3-O-methyl-~-glucose (XVII), and
obtained 4-0-formyl-2-0-methyl-~-arabinose (XVIII) as a sirup. When
this was treated with acetone in the presence of dry hydrogen chloride, a
crystalline derivative of D-arabinose, 3 ,4-0-isopropylidene-2-O-methyl-~-
arabinose (XIX), was obtained. Formation of formate ester was suggested,
FH,OH CH,OH
XVII XVIII
I
CH,OH
XIX
without actual isolation thereof, to explain low yields of formic acid in the
oxidation of lactose,267 3,4-di-O-methyl-~-glucose,~~ 3,4-di-O-methyl-~-
rhamn0se,2?~2,7-anhydro-/3-~-altro-heptulopyranose,~~ methylated D-glu-
cose derivatives,266and c e l l ~ b i o s e .Low
~ ~ ~yields of formic acid were also
encountered in the oxidation of ~ - m a n n i t o and . ~series
l ~ ~ of a d o d e ~ i t o lA ~~
of reactions was proposed29in order to explain these results with D-mannitol.
The D-mannitol, by stepwise oxidation, could be converted initially t o any
one of the following: a pentose, a tetrose, a triose, or a diose. The pentose
and tetrose, if formed, would be in equilibrium with a cyclic isomer which
would, in turn, give rise t o formate ester production. The experimental
conditions for retention or saponification of the formate ester have been
rather nicely defined by Meyer and Rathgeb.267The formate arising in
lactose oxidation could be retained if the oxidation was performed at O",
and in a pH range of 4.2 to 5.7. By this means, they were able to liberate
only two equivalents of formic acid from lactose and to preserve the third
(275) G . R. Barker and D. C. C. Smith, Chemistry & Industry, 1035 (1952).
(276) F. Brown, L. Hough and J. K. N. Jones, J . Chem. SOC.,1125 (1950).
(277) F. S. H. Head and G. Hughes, J. Chem. SOC.,603 (1954).
(278) M. L. Wolfrom, W. W. Binkley, C. C . Spencer and B. W. Lew, f.A m . Chewz.
SOC.,
73,3357 (1951).
36 J. M. BOBBIrP
as a formate ester. For titration of the formic acid, exclusive of the formate
ester, they used sodium hydroxide to an end-point with Bromocresol
Purple (pH 5.6 to 6.8). A slow titration with sodium hydroxide, to an end
point with Bromothymol Blue, both saponified and titrated the third
equivalent of formic acid. Very similar experiments have been carried out
on maltose and starch.138 Careless treatment with base should obviously
be avoided, as this can lead to Cannizzaro 224 (see p. 27).
The analysis for formic acid has been largely developed for dealing with
terminal-group determinations on polymers. The simplest and most widely
used method employs a salt of periodic acid as the oxidant. After destruc-
tion of the excess periodate, a direct titration with a strong base to the
proper end-point will provide valid values for formic acid. The excess
periodate is generally destroyed with ethylene glycol, since this substance
is readily available and is readily oxidized to formaldehyde, which does not
interfere with the titration. Two bases have experienced general usage:
sodium hydroxide,6gr8 6 . 114. 130, 1 3 6 , 1 3 9 , 174, 246 and barium hydroxide.111,122.
126, l33- There is some controversy concerning the choice of an indicator.
The majority of workers have used Methyl 85, l Z 6 ,160, but
thym~lphthalein,~~,66 Phenol
499 and Methyl Red-Methylene
l35. 174 have been used in addition to the indicators used by Meyer
and Rathgeb,267 Bromocresol Purple and Bromothymol Blue. Phenol-
phthalein2vl b 4 , 156v 278 affords results which are 1% high.164.166 A logical
treatment and a partial review of the indicator problem was undertaken
in a study of the potentiometric titration curves of pure formic acid solu-
tions and of formic acid present in periodate reactions.279After investigat-
ing a series of model compounds, these workers concluded that the titra-
tion curves are superimposable only to pH 6, and that, at this pH, about
81 % only of the formic acid had been titrated. Use of Bromocresol Purple,
with a correction factor of 81 % to 100 %, was proposed. A similar study
has shown that the potentiometric curves of free formic acid and of formic
acid in oxidized-dextran solutions are identical between pH values'62 of 6.0
and 8.5. Thus, a titration to a phenolphthalein end-point produces only a
1% error. Others have preferred pH values of 7.1133and 6.25.139 Formic acid
has also been determined iodimetri~a11y.l~~ 9 177
In the determination of formic acid in more complicated reaction-mix-
tures (for example, in the presence of buffers,22.59a in solutions containing
non-volatile 67 and in solutions containing ammonia234), it was
necessary to distil the formic acid from the reaction solution (after destruc-
tion of the excess periodate with ethylene glycol or arsenite) before it could
be titrated.
(279) M. Morrison, A. C . Kuyper and J. M. Orten, J . Am. Chem. SOC.,76, 1502
(1953).
A different technique utilizes the Warburg respirometer to measure the

carbon dioxide liberated by formic acid from a dilute, sodium bicarbonate
. ~Successful
s ~ l u t i o n26s ~~ application of the method necessitates a pH value
of 5.7, which is somewhat high for optimal periodate oxidations, but good
results were reported in experiments on a number of materials, including
polysaccharides. The procedure is applicable to very small amounts and
does not require prior destruction of periodate.
3. Aldehyde Analysis
Since aldehydes are very common amongst periodate-oxidation products,
analysis for them has been well developed and widely used. Most emphasis
has been placed on methods for the assay of formaldehyde, as this is
the most common product. Formaldehyde was first determined (in periodate
work) by an oxidation method? 47 66 9 1
HaC=O + HgK& + 3 OHe -+ 2 KI + HCOze + Hg + 2 Ie + 2 HzO

Measurement of the decrease in basicity gave valid values for formaldehyde.
The assay for formaldehyde is today mainly performed by either of two
general methods. The one most widely used depends on the reaction of
5,5-dimethyl-l,3-cyclohexanedione (Dimedon) with formaldehyde; when
applied after the destruction of excess periodate, it proceeds quantitatively,
&H2;bcH
0 0 0
II
CHs-i()=O + H&=O -+ CH3 + H20
CH3 CH, 0 0 CH3

and the weight of the precipitate is an accurate measure of formaldehyde
(if no other aldehydes are present). In earlier work, the formaldehyde was
distilled from the solution and determined in the di~tillate.'~. 57. 6 6 , 118,23O
The determination of formaldehyde in the reaction mixture was first per-
formed by Fleury6and the same general method was developed by Reeves2"
into a widely usable procedureK8'69a, 8 s . 164, 16s, 270; it uses arsenite
22Kn
for destroying the excess periodate. Another excellent reagent for periodate
destruction is butanedione (biacetyl) , although commercial lots of this
reagent can develop a foul and penetrating odor. Biacetyl is readily oxidized
by periodate to acetic acid, which does not interfere in the estimation.
The Dimedon reaction must be used with some degree of caution, since
other aldehydes react with Dimedon, although to a lesser extent,76*76n 280
and the melting points of the "methones" of the simple aldehydes are not
(280) D. Vorliinder, 2.anal. Chem., 77,241 (1929).
38 J. M. BOBBITT
sufficiently far apart for accurate differentiation for identification. Observa-

tion of mixed melting points is mandatory.
A method for the determination of formaldehyde in the presence of
acetaldehyde was developed by Nicolet and Shinn.88sl o o , lo3 After the
excess periodate had been destroyed, these workers swept the acetaldehyde
(from the neutral reaction mixture) into a sodium bisulfite solution by
means of a stream of carbon dioxide. The acetaldehyde was measured by
conventional, bisulfite methods, and the residual formaldehyde was pre-
cipitated with Dimedon. This procedure was applied to protein hydroly-
zates and to terminal deoxy structures of carbohydrates.88*mn.
The other common method for determination of formaldehyde is the
colorimetric. It depends on the color developed through the specific reaction
of formaldehyde with 4,5-dihydroxy-2,7-naphthalenedisulfonic acid (chro-
motropic acid). The technique was developed by MacFadyen,281and it,
too, requires the prior destruction of periodate. Stannous chl0ride,~4sodium
a r ~ e n i t e and
, ~ ~ lead dithionateLglare among the reagents used for this
purpose. Chromotropic acid is added to the reaction mixture, and the color
of the warmed solution is determined in a colorimeter. Like all colorimetric
procedures, this requires a calibration curve derived from the readings
given by known solutions; for this purpose, periodate-oxidized solutions
of ethylene glycol or D-glucose are re~ommended.'~~ Formaldehyde has
also been distilled and then determined (in the distillate) with chromotropic
acid.Io4
A micro-diffusion method has been devised by WinnickLolfor the deter-
mination of acetaldehyde. Procedures for the assay of glycerose were de-
veloped by Jayme, Satre and M a r i ~ . ' 158~ ~They
, dehydrated glycerose to
pyruvic aldehyde by distillation from mineral acid, and determined the
pyruvic aldehyde as its phenylosazone.
The early addition of Dimedon is reported234to depress the formation
of hexamethylenetetramine from formaldehyde in the presence of ammonia.
O'DeaZs2found that the activated methylene center, sometimes formed in
periodate oxidations of carbohydrates, reacts appreciably with the form-
aldehyde formed, thus giving low yields of apparent formaldehyde. He was
able to depress this side reaction by the use of lowered temperatures and
by the addition of benzaldehyde or of p-hydroxybenzaldehyde. The analy-
ses for formaldehyde have often been more successful at a pH of 7.5 than
a t lower pH values.57~58, 59a, 8O, 264
(280a) M. L. Wolfrom, F. B. Moody, M. Konigsberg and R. M. Goepp, Jr., J. Am.
Chem. SOC.,68, 578 (1946).
(281) D. A. MacFadyen, J. B i d . Chem., 168, 107 (1945).
(282) J. F. O'Dea, Chemistry & Industry, 1338 (1953).
4. Carbon Dioxide Analysis

Carboii dioxide is not a common oxidation product in periodate work,
but it does appear in the oxidation of ketoses,49 a-keto a ~ i d s , ~ 4and #3~
a-hydroxy 3 9 and it is often a product23,I41 of '(overoxidation."
Carbon dioxide analyses have been carried out using the Plantefol ap-
p a r a t ~ s the
, ~ ~ Warburg apparatu~,'~, 23 and the Van Slyke-Neil1 mano-
metric apparatus,39 and by absorption in standard sodium hydroxide141

followed by back-titration with acid. A most convenient method is the
very old, barium hydroxide absorption scheme.16aThe carbon dioxide is
swept from the reaction mixture into a saturated, filtered barium hydroxide
solution by means of a stream of pure nitrogen. The precipitated barium
carbonate is filtered, dried, and weighed. This method is essentially a
terminal assay. The manometric methods permit kinetic measurements,
but involve use of much more complicated apparatus.
5. Ammonia Analysis
Ammonia, like carbon dioxide, is not over-common in periodate oxidation
mixtures; but it is quite important when it does appear, as in the oxidation
of aminodeoxy sugars. The analysis for ammonia has been successfully
performed using a Van Slyke-Neil1 apparatus,lo2lo6 and by distillation
8
and titration?s
VII. COURSEOF THE REACTION
The exact course of the periodate reaction has not yet been established.
That an intermediate complex, compound, or ion is involved has been
determined 2 6 9 , 261 2 6 2 - 283-286 The exact structure of this inter-
1
mediate is still in doubt. The most universally accepted structure is a cyclic
ester intermediate propounded by Criegee,270 analogous to his cyclic
ester intermediate for another agent oxidizing 1,2-glycols,lead tetraacetate.
i [
2 HzO + MO, S HJOS
-C-OH -C-OIO~H,
+Hme $ + Hz0
-C-OH -C-OH
I
(283) F. R. Duke, J . Am. Chem. SOC.,69, 3054 (1947).
(284) G. J. Buist, C . A. Bunton and V. J. Shiner, Research (London), 6 , 5 S (1953).
(285) G.J . Buist and C. A. Bunton, J . Chem. SOC.,1406 (1954).
(285a) R . Criegee, L. Kraft and B. Rank, Ann., 607, 159 (1933).
40 J. M. BOBBIlT
I I
-c=o
+ HzO + HIOa
~ -C-~ > I O J L -+ ]-c=o
I
The interatomic distances for such an intermediate were calculated,286and,
using the resulting data in conjunction with certain st,eric and oxidizing-
potential considerations, it was successfully predicted that sodium bis-
muthate and trivalent silver ion would also specifically oxidize 1,2-glycols.
These reactions have since been 288
The intermediate has been reported to be more stable in base than in

and it is thought to be formed by a backside The inter-
mediate is also believed to be an ion (XX),284- 286 perhaps hydrated (XXI) .
XX xx I
It has not been clearly demonstrated, kinetically or otherwise, that one
of these structures for the intermediate is correct, or even that the inter-
mediate breaks down to form the known end-products.
VIII. CONCLUSION
This article has not been in any sense an attempt to enumerate all of
the known examples of periodate oxidation of carbohydrates. It has en-
deavored to explain and illustrate as many of the different types of periodate
(286) L. J. Heidt, E. K. Gladding and C. B. Purves, Paper Trade J . , 121, No.9,
35 (1945).
(287) W.Rigby, J. Chern. SOC.,1007 (1950).
(288) F. P. Greenspan and H. M. Woodburn, J . A m . Chern. Sac., 76,6345 (1954).
(289) G. Hughes and T. P. Nevell, Trans. Faraday SOC.,44,941 (1948).
oxidation as possible, a.nd to indicate the various kinds of application. An

extensive discussion of the effect of experimental conditions on the problem
of "overoxidation" has here been undertaken, and some of the advantages
and disadvantages of the common analytical procedures used in periodate
work have been pointed out.
Since this is not a complete review, it seems desirable t o cite the different
reviews or partial reviews that have been written; the author gratefully
acknowledges the help obtained from them. The most complete review is
that of E. L. Jacks0n.2~'Other general reviews have been writte11.2~~-~~~
The periodate oxidation of the glycogens294and starcheslloa have been re-
viewed. The assays for formic and f~rmaldehyde'~'have also been
given consideration.
(290) L. N. Owen, Ann. Repts. on Progr. Chem. (Chem. SOC. London), 40, 115
(1943).
(291) J. K. N. Jones, Ann. Repts. on Progr. Chem. (Chem. SOC. London), 43, 180
(1946).
(292) A. W. Johnson, Science Progr., 39.96 (1951).
(293) J. E. Courtois, Mededel. Vlaam. Chem. Ver., 13, 67 (1951); Chem. Abstracts,
48, 3004 (1952).
(294) D. J. Manners, Biochem. J. (London), 66, xx (1953).

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Bobbitt 1956

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PERIODATE OXIDATION OF CARBOHYDRATES

Malaprade explored the application of this reaction to compounds in which

1. Rl must be a hydrogen atom or a hydroxyl group.

A clear definition of selective oxidation by periodate, considered apart

A practical distinction between selective oxidation and so-called

propounded by Hockett, Dienes and R a m ~ d e nin~ order~ to explain the

The production of formate esters in periodate work will be considered

The oxidation of cyclic 2-amino alcohols has been ~tudied.3~ In the

(33) G . E. McCasland and D. A. Smith, J . Am. Chem. SOC.,7 3 , 5164 (1951).

Oxalic acid is reported to be resistant to periodate,4. ae as are formic acid

served in the oxidation of a 1,2-dihydroxyanthraquinone-3-sulfonate(and

thought t o proceed through an intermediate reductone systern.lba The

fractional consumption of periodate. This type of reasoning is well illus-

(c) CH40H CHnOH

Series.66The monocyclic structures L-threitan (I) and erythritan (11) re-

and 0-isopropylidene sugar^.'^-^^ The isopropylidene groups were slowly

activated methylene structure would have been formed, periodate con-

glycol mixtures,96glycerit01,~~~ 97 ephedrine,gs r i b o f l a ~ i n e threonine,10'J-*02

(111) T. G . Halsall, E. L. Hirst andJ. K. N. Jones, J. Chem. SOC.,1399 (1947).

If a representative formula (X) for this type of structure is examined, it

HC=O HC=O + 2HCOzH

the determination of the structure of heparin,lS4 yeast mannan,22 the

droitinsulfuric acid,lasalginic acid,laS and a polyuronidic hemicellulose from

materials.1s3aColor-developing reagents which have been used include a

bility of interference b y amino acids has been pointed out.lgO

(183a) G . N . Kowkabany, Advances i n Carbohydrate Chem., 9,303 (1954).

Since aldehydes are notoriously polymerizable and difficult to manipu-

these oxidation products established the relative configuration of C1 in the

,O6p2oD Applications have been made to cellulose,144 to an oxidized poly-

CH,OH CH,OH SH,OH

This sequence is exemplified by the further reaction of the aldehydic

162, 217. 218

The reaction of these aldehydes, derived from periodate oxidation, with

hydrates with nitrogenous bases have since been extensively

derivatives of the simple aldehydic fragments; these are more readily

oxidation of an or-glycol grouping concerned in a ring structure yields a

modified in some undesirable or unknown way by these methods involving

prepared in 90 % yield by the interaction of bromine and sodium iodide in

mixture of sodium metaperiodate and potassium chloride was used, partial

glucopyranoside, yeast glucan, and dextran proceeded with a minimum of

A pH of 7.5 was found to be necessary for the successful oxidation of

solutions, the univalent, non-hydrated periodate ion, IO$, prevails between

that aqueous sodium metaperiodate slowly decomposes spontaneously in

This method was used by Malaprade2t 47 and is sometimes employed to-

has been described in excellent detail b y Jacks0n.2~The suggestion has

buffer. The free iodine liberated by the reduction of periodate to iodate is

than either the Malaprade or the Fleury arsenite procedure.

method utilizes very low concentrations of reactants, but is useless when

then becomes necessary to know whether the ester is saponified in the

A different technique utilizes the Warburg respirometer to measure the

HaC=O + HgK& + 3 OHe -+ 2 KI + HCOze + Hg + 2 Ie + 2 HzO

CHs-i()=O + H&=O -+ CH3 + H20

CH3 CH, 0 0 CH3

sufficiently far apart for accurate differentiation for identification. Observa-

4. Carbon Dioxide Analysis

metric apparatus,39 and by absorption in standard sodium hydroxide141

The intermediate has been reported to be more stable in base than in

oxidation as possible, a.nd to indicate the various kinds of application. An

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