LACTONE FORMATION FROM GLUCONIC ACIDS AND THE STRUCTURE OF GLUCOSE.

BY P. A. LEVENE AND H. S. SIMMS.

~(P’rem &e Laboratories of The Rockefeller Institute for Medical Research, New York.)

(Received for publication, April 21, 1926.)

INTRODUCTION.

In a previous paper’ it was shown that when gluconic (A), galactonic (IS), glucoheptonic (C), and mannonic (D) acids are allowed to stand in aqueous solution, each forms two types of lactones. The lactone with a 6-membered ring forms rapidly but to a small extent, while the “5-ring” lactone forms slowly and to a larger extent. Each acid gives a curve for rate of lactone formation which is the resultant of the two reactions. The la&one formation was observed polarimetrically.

Furthermore, 4-methyl glucoheptonic acid (E) and 2,3,4,6- tetramethyl mannonic acid (8’) form only the 5-ring lactones, while 2,3,5,6-tetramethyl mannonic acid (G) forms only the 5-ring la&one, as predicted from chemical evidence for the structure of these substances. This method was shown to be useful in disclosing the ring structures of methylated sugars, particularly when the corresponding acids permit the formation of only one lactone.

1 Levene, P. A., and Simms, H. S., J. Biol. Chem., 1925, Ixv, 31. In that article the following corrections should be made:

The letter “t” is omitted from equation (5), footnote 10. Equations (6) and (8) should read:

K=fH+ (6) --cr

- log j = 0.30 d zivz (8)

“where Z iv2is the sum of all the ion concentrations i, each multiplied by the square of its valence Y. In this experiment 0.30 d=” etc.

737

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738 Lactone Formation from Gluconic Acids

Subsequent to the publication of the above paper, Charlton, Haworth, and Peat have applied the same principle2 to the tetra- methyl derivatives of glucose and galactose and the trimethyl derivatives of arabinose and xylose. They found that the “normal” derivatives obtained by methylation of the glucosides give 6-ring lactones; while the derivatives obtained by methyl- ating the lactones of the 2,3,6-trimethyl acids give 5-ring lac- tones. The latter observation is to be expected, while in the case of gluconic acid the former observation is contrary to the prevailing conception of the structure of glucose. Glucose has been supposed to contain a 5-membered ring, whereas they inter- preted these observations to indicate a 6-membered ring for glucose.

DISCUSSION.

In this paper we will give data on both the rates of formation and of splitting of the lactones of several derivatives of gluconic acid. Our observations are as follows (see Tables I and II and Figs. 1 to 4):

1. Both gluconic acid (L) and 2,3-dimethyl gluconic acid (M) form 6-ring and 5-ring lactones simultaneously (as would be expected).

2. The 3,5,6-trimethyl gluconic acid (N), obtained by methyla- tion of diacetone glucose, gives only a 5-ring lactone.

3. The trimethyl (0) and tetramethyl CP) derivatives, obtained by methylating the methyl glucoside, give 6-ring lactones. We must conclude that the methyl groups are in positions 2,3,4 and 2,3,4,6 respectively (rather than 2,3,5 and 2,3,5,6 as previously supposed).

4. From 2 it follows that both mono- and diacetone glucose have (i-membered rings;3 whereas (from 3) the tri- and tetramethyl glucoses derived from methyl glucoside possess 6-membered rings,

2 Charlton, W., Haworth W. N., and Peat, S., J. Chem. Sot., 1926, cxxviii, 89. They measured rates of splitting rather than rates of formation. Both methods lead to the same conclusions when only one lactone can form, but the rate of formation is of more significance when there is a possibility of two or more types.

3 This serves as proof that diacetone glucose has the structure 2, 3-5, 6- diacetone glucose.

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TABLE I.

Approximate Initial Reaction Rates in Formation (kf) and Xplitting (k8) of La&ones in Aqueous Solutions.

A, Data from Lactone Formation.

,a a &

(A) (E)

Acid.

I I

=4 8 2 -

UT

-- Gulonic. * 1.5 25 4-Methyl

glucoheptonic. 2.5 21 2,3,4,6-tetramethyl

mannonic. 6.0 30 2,3,5,6-tetramethyl

mannonic. --

Average.. . . . . . . . . . . . . . . . . . 3.3 25

I --

('3

T I B-ring. T

(L) Gluconic. 0.40 10

(M) 2,3-dimethyl gluconic. 0.33 15

(N) 3,5,6-trimethyl gluconic.

(0) 2,3,4-trimethyl gluconic. 0.38 12

(P) 2,3,4,6-tetramethyl gluconic. 0.35 7.5

-- Average.. . . . . . . . . . . . 0.36 11

- \ d s -

11.1

5.1

3.t

- 7

-

17

$0

!2

.5 -- !l -

4 s a - 35

22

8

- 22

-

156

165

160

185 - 167

5-ring.

1 8 3 3

--

38 80

\ 02 d d 5: 2

--

1.4 0.36

117 75 --

78 --

16 5) mols

71 100 51-15 lO(t3(

I.44 0.15 -- I.9 0.25 --

I.8 (3.4)

1.5 0.5

50 44 j.6 0.8

-- -- 60 48 1.6 0.7

B. Data from Lactone Hydrolysis.

6-ring. 5-ring.

Symbol. I

lOOk, 1OOkf to.5 1OOA --

to.6 1OOA . - ~ -

CM) 1.27 15 (0.19)

43 11

WI (0) 1.33 78 41 12 (P) 1.2 91.5 53 5

--___--- Average.. . . . 1.3 85 46 9 108 53

look, lOOk,

--

(87) j 40-‘5 ‘oolGiZO 0.4 0.4

130 1 56 0.3 0.2

-- 0.35 0.3

* Galactonic (B), glucoheptonic (C), and mannonic (D) acids have essentially the same rates as gluconic acid (A).

to.6 = time to reach one-half equilibrium A

0 . 2

A = fraction changed at equilibrium (look = per cent changed). 739

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740 Lactone Formation from Gluconic Acids

which would indicate that the common form of methyl glucoside also has a 6-membered ring.

The question of the ring structure of the common form of glucose cannot, therefore, be decided solely from the structure of a derivative of glucose. Other data and observations are required to determine the ring structure of free sugars. This will be dis- cussed in a future paper.

There are certain anomalies in the data which require consideration.

1. The reactions in these acids have a higher velocity than in acids (A) to (G) (see Table I, A). In the formation of the 6-ring lactone kf is 3 times as large, and k, is 7.6 times as large.

FIG. 1. Lactone formation (f) of acids (L), (M), and (IV); and splitting (s) of lactones (ill) and (21’).

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P. A. Levene and H. S. Simms

IL 40 I 10 20 30 40 50 06 Lqu ilibrium I

FIG. 2. Lactone formation (f) and splitting (s) of (0) and (P).

2 - L L d 4 5 6 7 9 10

I I I I I I I I I I -

FIG. 3. Lactone formation of (L), (M), (N), (0), and (P) in the first 10 hours (see lower left-hand corner of Figs. 1 and 2).

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742 Lactone Formation from Gluconic Acids

FIG. 4. Rotations in the lactone formation and splitting of (F’) compared with the data of Charlton, Haworth, and Peat (Curve C., H., and P.) for the splitting of the same substance.

For the 5-ring la&one kj is about the same, while k, is about 3 times as great as in (A) to (G).

2. The resulting equilibria are different. The 6ring la&one reaches an equilibrium of 11 per cent rather than 25 per cent and the 5-ring gives 48 instead of 78 per cent. In other words, the tendency to hydrolyze (relative to the tendency to form la&one) is 3 times as great as in acids (A) to (G).

However, these rates are only approximate and are in the same order of magnitude as the corresponding values previously measured. It seems safe to assume that the lactones are those indicated in Table I (i.e., 6-membered and 5-membered respec- tively) .

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P. A. Levene and H. S. Simms

The only known difference between these experiments and the previous ones lies in the slightly lower temperature (22’C. com- pared with 25°C.) which would tend to lower the equilibria. The faster reaction rate must be attributed to chemical differences in the acids.

3. The b-lactone of Charlton, Haworth, and Peat is supposed to be identical with our acid (P). From Fig. 4 it will be seen that their rotations are considerably lower than ours. It would appear that their material was less pure.

EXPERIMENTAL AND MATHEMATICAL.

The lactone formation was observed by optical rotation. The method was the same as previously described, except that 2 dm. open polariscope tubes (holding 4 cc.) were used for the lactone formation. The solutions were 0.250 molar aqueous solutions, as before.

The lactone splitting was observed in 0.125 molar aqueous solutions which were first placed in 4 dm. tubes. Owing to cloudiness which developed, the solutions (M), (N), and (0) were filtered into 1 dm. tubes and (P) into a 2 dm. tube.

The substances (M), (N), (0), and (P) were lactones which were dissolved directly in water for studying lactone hydrolysis. These lactones were hydrolyzed with NaOH which was then neutralized with HCl for studying lactone formation.

The results are plotted in the figures and summarized in Table I. The per cent lactone is calculated from the rotations of the pure lactones given in Table II. Except for (L), these were obtained by extrapolation of our curves (drawn on large scale) to zero time and agree well with the values of Levene and Meyer.4 The lactone (M) is not a pure substance but a mixture of 5-ring and 6-ring lactones. The values for (L) were taken from the

4 Levene, P. A., and Meyer, G. M., J. Biol. Chem., 1925, lxv, 535. In that article the rotation of the free 2,3,5-trimethyl gluconic acid (now known to be 2,3,4) on p. 542, should read:

ral” = + 0.75” x 100 D 2 X 0.976

= + 38.4” + 47.2”

and these values should appear in Table I.

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744 La&one Formation from Gluconic Acids

literature. The material (L) was the calcium salt to which 1 equivalent of HCl was added for studying lactone formation. Since a trace of lactone may have been present at the start, 15 minutes were added to the observed time.

In Table I are the velocity constants for the various reactions. These are only approximate. They are calculated on the assump- tion of a monomole&lar reaction with uniform catalysis. If t0.5 is the time to reach one-half equilibrium, we may write:

CL) Of) WI (0) (P)

For p

-

-_

181 t B (Table I) lc, and k, are interchanged in these equations. When the 6-ring lactone is present during the formation or splitting of the 5-ring lactone it is necessary to make proper correction of (A).

A A 2.3 A kf=t,.,lnrx=----

to.5 log 2 = 0.69 6

1 k, = Ic, - - 1

( > A

TABLE II.

Molecular Rotations of Gluconic Acid Dmivatives.

2,s

3,5,6 2,3,4 2,3,4,6

___~ ~- 196.1 178.1 6and5 25 224.2 206.2 6and5 51 238.2 220.2 5 44 238.2 ‘220.2 6 12 252.2 234.2 6 8.5

-13.5’ +86”

$102O

-

.-

-

f122O +125” +129” +69” +loo” $113.5” +190° $104” +254” $152”

The reaction is not necessarily monomolecular and the catalysis (with H ion) is obviously not constant. However, these values give a relative idea of the reaction velocity.

Due to change in H ion activity, the experiments (Table I, A) in which the la&one is formed from free acid, should give a higher initial velocity than those (Table I, B) in which the lactone is allowed to hydrolyze. Thus the constants in Table I, A are 2 to 4 times greater than the corresponding ones in Table I, B.

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P. A. Levene and H. S. Simms 745

TABLE III.

Lactone Formation of Free Gluconic Acids at 22°C.

Experiment 1.

All concentrations were 0.250 molar.

Acid . . . .

Length of tube

2,3-dimethyl 3,5,6-trimethyl CM).* WI.

2 dm. 2 dm.

Time. WI,

hrs.

0 083 0.167 0.33 0.43 0.50 0.60 0.67 0.75

$62.2”

+66.8”

3-67.4’

1.0 1.5 3.0 7.0

I- (66.8”: +68.7” +70.9” +71.2O

23 31 48 72

120 -

+74.5” j-75.8” +78.8” +s3.0° +91.40

-

1

) ( 1 1 1

2 2 2 3 4

-

LW- LX- tone. Ml, tone. __~~ Per cent per cm

(-5.0”) (7.5, 3.2

(-S.OO) (4.81

(-8.0”) (4.8: 9.9

0.7 --- 9.9) -8.8” 4.1 2.6 -9.2” 3.8 5.8 -8.4” 4.5 6.5 -7.0” 5.7 ---

11 .o -1.6” 10.5 ;2.9 +O.S” 12.4 :7.2 +5.4” 16.7 83.3 +11.2” 21.7 .5.5

- “2,3,4-trimethyl

co).* 2,3,4,6-tetramethyl

cm.*

2 dm. 2 dm.

[Ml, LX- tone.

~-

per ten.

+88.4” 2.3

t

WI,

f102” f103”

+107.1

LSC- tone.

per cent

0 0.7

+92.0° 5.8 +94.2” 7.9 3.4

+96-O” 9.6

$97.2” 10.8 $98.0” 11.5 +98.8” 12.3 +99.6” 13.1

~-

f102.8” 16.2 t103.4” 16.7 t106.4’ 19.6 t109.4” 22.5 j-114.4” 27.3

+109.9 5.2 +110.0 5.3 f110.8 5.8 +112.7 7.0

$115.0 f115.8 f117.6 +118.8 f112 6 --

8.6 9.1

10.3 11.1 13.7

-

* These data are plotted in the figures (circles).

The observations of lactone formation and splitting are given in Tables III to VIII.

The curves in Figs. 1 and 2 indicate that the acids (0) and (P) do not remain at the equilibrium reached in a few hours but rise slowly. This is because the solution evaporated at the rate of about 0.07 cc. per day in spite of the fact that the tubes were covered with small inverted dishes. The effect is obvious only when 1 dm. tubes were used (holding 2 cc.). It was less when 2 dm. tubes (4 cc.) were used, and was negligible with (25 cc.) 4 dm. tubes. This has been shown by repeating the lactone split-

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746 La&one Formation from Gluconic Acids

TABLE IV.

Lactone Formation of Free Gluconic Acids at 2S”C.

Experiment 9.

All concentrations were 0.250 molar. -

- _ Acid.. .

Length of tube

(W (MY

1 dm. 2 dm.

W

1 dm.

(0) (P)

2 dm. 2 dm.

Time. [Ml, LF%C- WI, t”,z- [Ml, LrtO- MID kni- Ml, LX- tone. tone. tone.

-- -- ----

hrs. Per Pm cent cent 87 celz Per Pm

cent cent 0.06 +61.2” 1.7 +87.0” 1.0+104o 1.3 0.10 $6.0” 7.2 0.20 -8.0 5.5 +95.6” 9.2 0.50 -8.0 5.5 +106.4” 2.9 0.60 $8 .O” 8.8 $67.0” 10.2

----- --~-

1.0 $9.6” 10.1+68.0” 11.6 -7.4 6.1 +96.4” 10.0 +llO” 5.3 2.5 +-12.0° 12.0’+70.6” 15.4 -6.5 7.0 $97.6” 11.2 +llO” 5.3 4.5 $72.0” 17.4-5.7 7.8 +98.0”11.5 +lll” 5.9 7.0 +72.4” 18.0 -4.1 9.4 $98.6” 12.1 f112’ 6.6

-- -- --~-

24 +76.0” 23.2 f2.1 13.8 t102.6”16.0+113.6” 7.6 30 +77.0” 24.7 $4.0 15.4 t103.0” 16.3 f113.4” 7.5 48 f79.2” 27.8+6.4 17.5 t-104.8” 18.1+114.8” 8.4 54.5 $80.6” 29.9 $9.6 20.3 +106.1”19.2 +115.6” 9.0

2 per cent concentrated HCl added (values corrected for volume).

72 +88.0” 40.6 +37” 44.5 +104.4”17.7+113° 7.2 96 +90.8” 44.7 +37” 44.5 f107” 20.2+115.4” 8.8

120 +90.8” 44.7 +37” 44.5 1 +lOS” I 21.2 +115.4” 8.8

* Some of the values for (L) and (M) in this table are plotted in the figures (triangles and circles with accent mark).

ting of (M) and (P) in 4 dm. tubes sealed nearly air-tight. The results (Table VII) show that (M) constantly approaches its equilibrium (compare with Table VI) while (P) remains at the equilibrium attained in a few hours (compare with Table VI).

SUMMARY.

The previously described method of studying lactone formation of sugar acids has been applied to the formation and the splitting

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P. A. Levene and H. S. Simms 747

TABLE V.

Lactone Formation of Free Gluconic Acids at 28°C.

Experiment 5.

All concentrations were 0.250 molar.

Acid.

-

- (LY (A’)*

Length of tube. 4 dm. 2 dm.

Time. Laotone.

h-8.

0.005 0.018 0.15 0.17 0.33 0.50

.-

. -

.-

.-

-

Ml, Laotone.

per cent

+3 3”

+9.20

5.4

9.8

-13.0” -13.2” -13.2’ -13.0” -12.74” -12.35”

-

.-

.- 1.0 2 4 7.25

10.7 -12,09O 1.2 11.4 -11.44” 1.8 12.2 -10.14” 3.0 14.7 -7.67” 5.1

24 72 80 96

120 144 168

+22.90 20.7 +25.6” 22.9 +25.5” 22.8 +25.8” 23.0 +26.2” 23.4 $25.8” 23.0 $26.2” 23.4

$1.69” 13.4 +15.6” 25.6 +16.3” 26.2 +18.3” 28.0 +20.8” 30.2

-

-

per cent

(0.4) (0.3) 0.3 0.4 0.7 1.0

1 per cent concentrated HCI added.

9-13 days. +27.2” 24.2 I

* The data in this table are plotted in the figures.

of lactones of gluconic acid and four of its methylated derivatives (see Table I and Figs. 1 to 3).

From the rates of reaction it was found that gluconic acid (L) and also 2,3-dimethyl gluconic acid (M) give mixtures of 5-ring lactone and 6-ring lactone (as would be expected).

The trimethyl (0) and tetramethyl (P) derivatives from methyl- ating the glucoside give 6-ring lactones in agreement with the

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748 Lactone Formation from Gluconic Acids

TABLE VI.

Hydrolysis of Gluconic La&ones at 22°C. Experiment I.

All concentrations were 0.125 molar. (Plotted in Figs. 1 and 2.)

Lactmm. CM)

Length of tube --

4 dm.

Time. mfl,

hrs.

0.005 0.167 0.25 0.42 0.50 0.67 0.75

+12s0 98.5

+127” 97.1

$127” 97.1

1.0 1.5 2.0 2.5 3 4 5 6

+125” 94.2 $124”’ 92.7 $123” 91.3

24 30 48 72

120 144

$118”

f115” $115” +112” +109” $111” f115”

LX- tone.

- (N) (0)

2 dm. 2 dm.

84.0

79.7 79.7 75.4 71.1 72.5 79.7

-

1

i

- I

. -

-c

-

t- looO* k100” f98”

+96” +98”

$96”

+91” +86.4 +82.4 +77.6 +64.0 +60.0

-

- -

P

11 11

--

0 0 0 0 0

-

--

+ 184” +180”

00 +178” 00 98.2 f167’ -I__

96.5 f154” 98.2 +148”’

+13s” +131o +124” f122O

96.5 $116” + (121”

--

92.1 -f-113” 88.0 +114o 84.5 +117o 79.4 +120° 68.3 +124’= 65.0 +128”

I,%- tone.

per cm

94.2 90.2 88.5

77.8

65.4 59.6 50.0 43.3 36.5 34.6 28.8

(33.6

-

. -

26.0 27 .O 29.8 32.7 36.6 40.4

) -

-

7 days. 8 “

10 “

.t

2 per cent concentrated HCl added.

(P)

2 dm. -

!Ml, LX- tone.

+252” +247o $242”

rxr cent 98.7 95.4 92.2

$225” 81 .o

S209” 70.4

+193” 60.0 + 170” 44.8

f140” 25.0 + 129” 17.8 +122” 13.5 +118” 10.5 $117” 9.9

$115” $115” $116” +119” +124” +127”

8.6 8.6 9.2

11.2 14.5 16 5

--

37 42 48

* Owing to cloudiness the solution was filtered into a 1 dm. tube at this point.

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P. A. Levene and H. S. Simms 749

TABLE VII.

Hydrolysis of Gluconic Lactones at 22°C.

Experiment 2.

The first reading was not taken until 19 hours, when the solutions were filtered and remained clear in 4 dm. tubes, nearly air-tight.

Lact,one.

Length of tube.

Time.

hrs.

19 23.5 42 68 90 96 (4 days)

120 (5 “ ) 168 (7 ‘( )

-

CM)

4 dm

WI, Lactone.

pet cent

+llS” 81 +114” 78 +llO” 72 $106” 66 $104” 64 +104” 64 $103” 62 +lOl” 60

- (P)

4 dm.

WI,

+123” +122” +122O $123” +123” $122” +122” +122o

Lactone.

per cent

4 13 13 14 14 13 13 13

TABLE VIII.

Lactone Hydrolysis with 2 to Q Per Cent HCI.

All concentrations 0.250 molar.

Lactone. 00 (N)

Time. [Ml, Lactone [Ml, Lactone

-- dlZY8 per cent per cent

1 +93.6” 48.7 +36.0” 43.6 3 +95.2” 51 .o f35.2” 42.9

-7

(0)

Ml, Lrtc-

[Ml, La+

tone. tone. -- ~-

per een, per cent j-107.2” 20.4 t111.20 7.9 t107.2” 20.4 +114.0” 6.1

observations of Charlton, Haworth, and Peat which lead them to postulate that glucose has a Bmembered ring.

However, the trimethyl gluconic acid (N), obtained by methyl- ating diacetone glucose, gives only a 5-ring lactone, which shows that a derivative may have a different composition from the parent substance. Hence, the type of ring in the common form of glucose cannot yet be decided.

The reaction rates and equilibria were somewhat different from those of the acids previously studied.

1 dm.

- (P)

1 dm.

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P. A. Levene and H. S. SimmsSTRUCTURE OF GLUCOSE

GLUCONIC ACIDS AND THE LACTONE FORMATION FROM

1926, 68:737-749.J. Biol. Chem. 

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