OXIDATION OF FATS, NITROGENOUS SUBSTANCES, AND THEIR MIXTURES WITH CARBOHYDRATES BY AIR, AND METABOLISM I N NORMAL HEALTH AND DIABETES
BY C. C. PALlT AND N. R. DHAR
I n previous papers1 published from these laboratories, we have investigated the oxidation of carbohydra.tes by passing air under different conditions. From our researches on the oxidation of carbohydrates, we have been able to explain the metabolism in different forms of diabetes, glycosuria, etc. We have also proved experimentally that uric acid can be oxidised by passing air in presence of dilute solutions of caustic soda. Moreover, uric acid has been oxidised in neutral solutions by passing air in presence of ferrous hydroxide, manganous hydroxide, cerous hydroxide, cobaltous hydroxide, nickelous hydroxide, cupric hydroxide and sodium sulphite.
In this paper, we shall present the experimental results obtained in the slow oxidation of other nitrogenous products, fats, and Carbohydrates, either occurring singly or as a mixture of two or three of these substances. The experimental arrangement and the method of estimation of various substances were the same as given in previous papers. The experimental results given in Tables I and I1 show that, in presence of caustic soda or sodium bicarbonate, potassium stearate and potassium oleate are appreciably oxidised by passing air a t the ordinary temperature in absence of any catalyst. Moreover, our experimental results show that in presence of caustic soda, there is more oxidation than in presence of sodium bicarbonate. I n presence of the hydroxides of the different metals, the amount of oxidation of potassium stearate and potassium oleate is increased and the order of the accelerating effect is the following:Cerous hydroxide ) manganous hydroxide ) uranous hydroxide, cupric hydroxide, ferrous hydroxide ) nickelous hydroxide ) ferric hydroxide ) mercuric hydroxide, cobaltous hydroxide ) lead hydroxide ) chromic hydroxide with potassium stearate and uranous hydroxide ) cerous hydroxide ) mercuric hydroxide ) ferrous hydroxide ) manganous hydroxide ) ferric hydroxide ) cupric hydroxide ) chromic hydroxide ) lead hydroxide ) nickelous hydroxide ) cobaltous hydroxide with potassium oleate. With hippuric acid and glycine, we observe from Tables VI-XIII, that these substances can be oxidised by passing air at the ordinary temperature in presence of alkali and in presence of different reducing agents, the amount of oxidation is increased. J. Phys. Chem., 29, 376, 799 (1925);30, 939 (1926).
1664
C. C. PALIT AND N. R. DHAR
TABLEI Potassium Stearate Estimation of potassium stearate in presence of freshly precipitated hydroxides of different metals, caustic soda, sodium bicarbonate or sodium sulphite] in neutral and alkaline solutions. The volume of fat taken was I O C.C. and the volumes of different salt solutions, each of 1.0% concentration were 2 0 C.C. in each case. The hydroxides were precipitated from the salt solutions by the addition of the exact equivalent amount of caustic soda. The volume of air passed was 60 litres in 9 hours.
- .$& 2
Volume of N/IO sodium thiosulphate required sodium bicarbonate, or I O C.C. of or sodium sulphite) iodine trichloride used as catdyst in the taken with IO C.C. experiment of fat after 2 bra. (Blank) Substance (freshly precipitated hydrox-
a~ ides, caustic soda,
Volume of N/IO sodium thiosul!ha@ fequired or iodine trichloride left after the experiment in C.C.
A.-Neutral Solutions Ferrous hydroxide 14. z C.C. 17.9 I? 2 Ferric hydroxide I1 17.6 3 Cerous hydroxide 18.2 I1 4 Chromic hydroxide 17.2 11 5 Cobaltous hydroxide 17.4 11 6 Nickelous hydroxide 17.7 11 7 Lead hydroxide 17.3 11 8 Mercuric hydroxide 17.4 (wet HgO) 11 9 Manganous hydroxide 18.0 11 I O Uranous hydroxide 17.9 If 11 Cupric hydroxide 17.9 11 12 Sodium sulphite 15.0 (=o.1513 grm.) 11 13 Sodium bicarbonate 17.4 ( = I O C.C. of 1.0% sol’n.) 11 14 Caustic soda 17.7 ( = I O C.C. of N / 8 sol’n.)
3.7 3.4 4.0 3.0 3.2 3.5 3.1 3.2
I
B.-Alkaline
Solutions ( = IO
C.C.
Ferrous hydroxide 1 4 . ,, 2 C.C. 18. I Ferric hydroxide 11 17.9 3 Cerous hydroxide 18.5 11 77.5 4 Chromic hydroxide ,I 17.6 5 Cobaltous hydroxide )I 6 Nickelous hydroxide 18.0 11 17.5 7 Lead hydroxide ,1 1 7.8 8 Mercuric hydroxide (wet HgO) 11 18.2 9 Manganous hydroxide ,, IO Uranous hydroxide 18.1 11 I I Cupric hydroxide 18.0 I1 12 Sodium sulphite 15.1 (=o.1513 grm.) I O c.c of ICI3 KI 2 0 . 0 C.C. of X / I O NazSz03.) I O C.C. of potassium stearate IO C.C. IC13 = 14.2 I
+
Percentage amount of fat oxidised
63.8 58.6 68.9 51.7 55.1 60.3 53.4
55.1
3.8 3.7 3.7 0.8
65.5 63.8 63.8 13.7
3.2
55.1
3.5
60.3
of N / 8 NaOH)
2
+
Amount of fat oxidized in terms of N/IO sodium thiosulphate in C.C.
C.C.
3.8 3.7 4.3 3.3 3.4 3.8 3.3 3.6
65.5 63.8 74. I 56.8 58.6 65.5 56.8 62 .o
4.0 3.9 3.8 0.9
68.9 67.2 65.5 15.5
of N / r o NakL03.)
OXIDATION O F FATS WITH CARBOHYDRATES
1665
TABLE I1 Potassium Oleate Estimation of potassium oleate in presence of (i) freshly precipitated hydroxoxides of different metals in neutral solutions, (ii) caustic soda, sodium bicarbonate and sodium sulphite. The volume of fat used was I O C.C. and the volumes of different salt solutions, each of 1.0% concentration, were 2 0 C.C. in each case. The hydroxides were precipitated from the salt solutions by the addition of the exact equivalent amount of caustic soda. The volume 17.4 C.C. of of air passed was 60 liters in 9 hours. (IO C.C. Ic13 K I N/IONa&Oa.)
+
-E .z 2
0
Substance (freshly precipitated hydroxoxides, caustic soda, sodium bicarbonate or sodium sulphite) used in the experiment aa catalyst
Volume of N/IO Volume of N/IO sodium thiosul- sodium thiosulpha,h required phate required or I O C.C. of or iodine triiodine trichloride chloride left after taken with I O C.C.the experiment of fat after 2 hrs. in C.C. (Blank)
Ferrous hydroxide Ferric hydroxide 3 Cerous hydroxide 4 Chromic hydroxide 5 Cobaltous hydroxide 6 Nickelous hydroxide 7 Lead hydroxide 8 Mercuric hydroxide (wet HgO) 9 Manganous hydroxide I O Uranous hydroxide I I Cupric hydroxide 1 2 Sodium sulphite I
I O .8 C.C. 71
2
(=
0.1513
11
I1
I, If 1,
Jl
11
11
I1 11
13.9 12.6 14.5 11.2
10.8 10.9
Amount of fat oxidised in terms of N/IO sodium thiosulphate in C.C.
Percentage amount of fat oxidised
3.1 1.8 3.7 0.4 0.9
47.0
0. I
1.5 4.5
27.3
56.I 6.1 0.0
14.I
0.3 3.3
13.8
3.0
15.0 11.5
4.2 0.7
45.4 63.6 10.6
12.3
1.5
22.7
11.9
1.1
16.6
13.8
3.0
45.4
11.1
50.0
m.)
13 Sodium bicarbonate (= IO C.C. of 1.0% sol’n.) 14 Caustic soda (= IO C.C. of N/8 sol’n.) I O C.C. potassium oleate f
1,
,, IO C.C.
Ic13= 10.8C.C. of N/IO Naz S203.
From our experimental results on the oxidation of the mixture of carbohydrates and fats as given in Tables 111-V, XIV, XV, we find that both the carbohydrates and fats are oxidised by passing air a t the ordinary temperature and the greater the amount of alkali, the greater is the amaunt of oxidation. Moreover, we have observed that a mixture of potassium stearate and
1666
C. C. PALIT AND N. R.
DHAR
different carbohydrates or a mixture of potassium oleate and different carbohydrates can be oxidised by passing air a t the ordinary temperature, though we have observed that solutions of carbohydrates cannot be oxidised in absence of alkali by passing air. The oxidation of carbohydrates in presence of potassium stearate or potassium oleate must be due to the existence of a small quantity of alkali obtained by the hydrolysis of potassium stearate or potassium oleate. It has also been observed that in presence of ferrous hydroxide or cerous hydroxide, the amount of oxidation of potassium stearate or potassium oleate and carbohydrates from a mixture of these substances is greatly increased. From our experimental results given in Tables XIV B and XIV C, we find that by passing air for 9 hours, the amount of oxidation of starch, cane ugBr, and glucose and other sugars from a mixture of each of these substances nd potassium stearate is the following:Glucose 9.5% Cane sugar 9 . I Starch 21 7G
Starch 41.37, Maltose 2 I . 5% Lactose 14.4Yo Laevulose I I .5yo Arabinose 3.5% Cane sugar I O . 5% Galactose I O %
TABLE I11 Estimation of potassium oleate in presence of freshly precipitated hydroxides of different metals and glucose occurring together in neutral solutions. The 17.4 C.C. volume of air passed was 60 litres in 9 hours. (IO C.C. of IC13 KI of N / I O Na2S203;I O C.C. of potassium oleate 10.8 C.C. of N / I O Na2S203 and I O C.C. of glucose = 15.6 C.C. of &'/IO Na2S203)
+
2
2
;. K I
2
3
4 5 6 7
8 9
Substance (freshly precipitated hydroxides) used in the experiment as catalyst
Volume of N / I O sodium thiosulghate required or I O C.C. of iodine trichloride taken with I O C.C. of fat after 2 hrs. (Blank)
Ferrous hydroxide Ferric hydroxide Cerous hydroxide Chromic hydroxide Colbaltous hydroxide Nckelous hydroxide Mercuric hydroxide (wet HgO) Manganous hydroxide Cranous hydroxide I O C.C. potassium oleate
Volume of K/ro sodium thiosulphate required or IO C.C. of iodine trichloride taken with I O C.C. of glucose after 2 hrs. (Blank)
1 5 . 6 C.C.
1 0 . 8 C.C.
,,
19
,l
,!
J )
))
19
,I
,
+
,,
I,
9,
,,
1,
??
,,
I O C.C.
Volume of N / I OPercentage sodium thiosul- amount phate required of fat for iodine tri- oxidised chloride left after the experiment in C.C.
IC13
5
22.7
10.5
22.7
12.3
50.0
9.3 9.0
0.0
IO.
9.0
3.0 0.0
11.8
42 ' 4
11.6
39.3 56.0
12.7
10.8 C.C. of N / I O Na&O$.
OXIDATION OF FATS WITH CARBOHYDRATES
1667
TABLE IV Potassium Stearate Estimation of potassium stearate in presence of (i) freshly precipitated ferrous hydroxide ( = 0.0674 grrn.), glucose or urea and (ii) freshly precipitated cerous hydroxide ( = 0.0169 grm.) occurring singly or as a mixture of two or three of these substances. The volume of air passed was 60 litres in 9 hours. The volume of each of the solutions taken was I O C.C. in each case.
44.8 Glucose 1 4 . 2 C.C. I j . 6 c . c . 14.5 39.6 11 11 36.2 36.2 Urea 14.3 3 Ferroushy7) 11 63.8 65.5 droxide 17.9 4 Ferrous hydroxide and 7, 1, jj.1 glucose 154 j Ferrous hydroxide and 77 ,1 50.0 and urea 15.1 53.4 6 Ferrous hydroxide and glucose and lf 19 urea 12.6 41.3 77 68.9 7 Cerous hydroxide” 18.2 74.1 8 Cerous hydroxide and f1 J1 58.6 glucose 1j.6 9 Cerous hydroxide 17 11 60.3 56.8 and urea 15.5 I O Cerous hydroxide and ,, glucose and urea ” 12.8 44.8 I O C.C. of IC13 KI 2 0 . 0 C.C. of N / I O Na2S203. I O C.C.of potassium stearate f I O C.C. of IC13 3 14.2 C.C. of S/IO ?;azS201. I O C.C. of glucose I O C.C. IC13 I 5.6 C.C. of 3 / 1 0 ?ja2S203. IO C.C.of urea I O C.C. IC1, =- I 5.6 C.C. of X / I O ru’a&Os. I O C.C. IC& K I = 17.4 C.C. of X / I O Naz S203. I
2
+
+
+
+
1668
C. C. PALIT AKD N. R . DHAR
Whilst the amount of carbohydrates oxidised in absence of fat under otherwise identical conditions in 5+ hours is the following:-(Vide Table IX A) Glucose I I ,470 Cane sugar I 2 . 9 % Starch 30 %
Starch Maltose Lactose Laewlose Arabinose Cane sugar Galactose
49 31
% %
16.5% 1 7 (r, 4 70 21 12
70 Yo
Consequently these results prove conclusively that the amount of oxidation of carbohydrates is greatly decreased bythe presence of fats. Moreover from our experimental results given in Tables 111-V, we note that the amount of oxidation of fats in a definite time is greatly reduced by the presence of carbohydrates. Hence we are convinced that the presence of fat or carbohydrate which is itself undergoing oxidation retards the oxidation of the other. Moreover from our experimental results it will be seen that in presence of carbo-
TABLE V Potassium Oleate Estimation of potassium oleate in presence of freshly precipitated ferrous hydroxide ( = 0.0674 grm.) glucose or urea occurring singly or 89 a mixture of two or three of these substances. The volume of air passed was 60 litres in 9 hours. The volume of each of the solutions taken was I O C.C. in each case.
- .$
Substance or mixture of substances used in the experiment + f as catalyst
2
w I 2
3 4
5
6
Volume of N / I O Volume of X / I O Volume of N/Io sodium thiosul- sodium thiosul- sodium thiosulphate required hate required pha!e required for I O C.C. of IO C.C. of or iodine triiodine trichloride iodine trichloride chloride left taken with IO C.C. taken with 10c.c. after the exof fat after 2 hrs. of glucose or urea periment in C.C. (Blank) after 2 hrs. (Blank)
Glucose Urea Ferrous hydroxide Ferrous hydroxide and glucose Ferrous hydroxide and urea Ferrous hydroxide and glucose and urea
I O C.C.
IO C.C. I O C.C.
IO C.C. I O C.C.
+
fk
10.8C.C. 17
”
15.6 C.C. 9) ,I
Percentage amount of fat oxidised
11.5 11 5
37.8 37.8
13.9
47.0
ll
lj
IO.
5
22.7
1)
9)
10.5
22.7
99
1,
10.0
15.2
of IC13 KI E 17.6C.C. of N / I O IL’a2Sz03. potassium oleate I O C.C. IC13 I 10.8C.C. of Y / I O SatS203. 15.6 C.C. of N / I ONa&O3. of glucose I O C.C.IC1, 15.6C.C.of N / I O NasSzOa. of urea I O C.C. Ic13 of IC13 KI E 17.4 C.C. of N / I ONa2S203.
+ +
+
+
1669
OXIDATION O F FATS WITH CARBOHYDRATES
hydrates or fats, the oxidation of urea is appreciably decreased. Also in presence of urea the oxidation of fat or carbohydrate is also retarded. (Vide Tables 111-V, XIV-XVI.) From our experimental results given in Table XVI, it will be seen that in presence of a mixture of fat and carbohydrate, urea is not oxidised a t all. This is a very important point in understanding the metabolism in the animal body. I n normal health, the heat and energy of the body are supplied to the system from the combustion of carbohydrates, fate, and proteins. Our experimental results prove conclusively that the oxidation of fats is retarded by carbohydrates and less powerfully by nitrogenous products. It seems fairly certain now that the presence of either one or two of the above substances which are undergoing oxidation, retards the oxidation of the third. I n presence of an excess of fat, little protein should be burnt. It is evident that fat in the diet way avoid consumption of tissue fat and prevent too large catabolism of tissue proteins. The protein-sparing qualities of fats and carbohydrates were discovered by some of the earliest students of metabolism from feeding experiments and i t is believed that carbohydrate is the most efficient of the two in sparing the proteins. It is apparent, therefore, that our experimental results as recorded in this paper
TABLE VI Hippuric Acid Estimation of hippuric acid in presence of (i) carbohydrates, (ii) urea, (iii) fats, (iv) freshly precipitated ferrous hydroxide, and (v) sodium sulphite, in neutral solutions. The volume of each of the solutions taken was IO c.c . and the amount of ferrous hydroxide was 0.0674 gram and of sodium sulphite was 0.1513 grm. The volume of air passed was 36.5 litres in 53 hours. The strength of each of the solutions of Carbohydrates, urea and fats was practically 1.0%. ( I O C.C. of potassium stearate = 14.2 C.C. of N/IO XazS2O3:I O C.C. of potassium oleate = 10.8C.C. of N/IO NazSz03.)
-X .$ 2
Substance used in the experiment aa catalyst
w
q I 2
3 4 5 6 7 8 9 IO
Volume of N/IO caustic soda in C.C. required for IO C.C. of hippuric acid taken (Blank)
Glucose Laevulose Lactose Cane sugar Urea Potassium stearate Potassium oleate Ferrous hydroxide Sodium sulphite (=o.15 13 gm.)
Volume of N/IO caustic soda in C.C. required for hi puric acid leg after the experiment
Amount of hippuric acid oxidised in terms of N/IO caustic soda in C.C.
Nil
5.7
5.7
0.0
JJ
JJ
JJ
JJ
JJ
JJ
JJ
JJ
f,
3,
,, ,, 11 I,
J7 Jl
, JJ
3.9 3.0 3.8 2.9
JJ
1.8 2.7
1.9 2.8
cc. of hippuric acid = 5.7 C.C. of N/IO NaOH. of N/IO NaOH sol’n 0.0179 grm. of hippuric acid.
I C.C.
Percentage amount of hippuric acid oxidised
JJ
31.6 47.3 33.3 49.1
1670
C. C. PALIT A S D S . R. DHAR
have adduced for the first time evidence in support of the foregoing empirical observations of the physiologists and chemists. Moreover, our results show that the oxidation of hippuric acid and glycine is considerably retarded by the presence of glucose, lactose and laevulose (Vide Tables Xos. VI and XII). In$uence of dkdi:-In all our experiments, it will be seen that the amount of oxidation of fats, carbohydrates, and nitrogenous products is greater, the greater the amount of alkali. In previous papers,’ we have emphasized that alkali is beneficial for diseases like diabetes, gout, beriberi, rickets and other types of metabolism diseases, because we have experimentally proved that carbohydrates, fats and nitrogenous products are appreciably oxidised by passing air in presence of sodium bicarbonate. There is difference of opinion among the medical men about the efficiency of alkali treatment. Thus Fiirth2 is definitely against alkali treatment in gout as will be seen from the following lines:“The constant effort to favourably influence gout by flooding the body with curative alkaline waters is without the least theoretical foundation.” On the other hand Cushny3is in favour of alkaline treatment as will be evident
TABLE VI1 Hippuric Acid urea, (ii) fat glucose Estimation of hippuric acid in presence of (i) fat glucose urea. The volume of air passed was 36.5 liters in and (iii) fat 53 hours.
+
-X ,$ Y
5
Zk
w
+
+
Substances used in Volume of N / I O the experiment as caustic soda in C.C. required for catalysts I O C.C. of hippuric acid taken (Blank)
Volume of N / I O caustic soda in required for hippuric acid left after the experiment C.C.
+
Amount of hippuric acid oxidised in terms of IO/N caustic soda in C.C.
Potassium stearate and urea j.7 4.9 0.8 2 Potassium oleate 9) 4.7 1.0 and urea 3 Potassium stearate I, and glucose 4.8 0.9 4 Potassium oleate 1) 4.6 1.1 and glucose 5 Potassium stearate and glucose and urea ” 6 Potassium oleate and glucose and urea ” I O C.C. of potassium stearate = 14.2 C.C. of X / I O Na2S203. I O C.C. of potassium oleate = 10.8 C . C . of N / I O KazS203. I O C.C. of urea solution = 0.1grm. of urea. I O C.C. of glucose solution = 0.0962 gram. of glucose.
Percentage amount of hippuric acid oxidised
I
J. Phys. Chem., 29, 799 (1925);30, 277 (1926). of Metabolism”, p., 184. “Pharmacology and Therapeutlcs”, 7th Edition, p. 545.
* “Chemistry 3
14.0 17.5
15.8
19.3
OXIDATION O F FATS WITH CARBOHYDRATES
1671
from the following quotations :- “When diabetes induces an increased acid formation in the tissues as is almost invariably the case in its later stages, the alkalis are of undoubted benefit in neutralising the oxybutyric acid formed and thus economising the alkalis of the blood. I n diabetic coma, temporary improvement may be attained by the uses of large doses of alkalies.” Yon Soorden was also a great advocate of alkaline treatment in diabetic coma. From our experimental results, it will be seen that uric acid and other substances can be readily oxidised by passing air in presence of alkali at the ordinary temperature. Hence we are definitely of opinion that alkaline treatment should be efficacious in gout, diabetes, beri-beri, and other metabolism diseases, because in presence of alkali, the amount of metabolism is increased. In previous papers,’ we have emphasised that when glucose is not oxidised in the body, fats are likely to be oxidised rapidly not to carbon dioxide and
TABLEVI11 Urea Estimation of urea in presence of freshly precipitated hydroxides of different metals in neutral solutions. The volume of urea used was IO C.C. and the concentration, were 20 C.O. volumes of different salt solutions, each of 1.07~ in each case. The hydroxides were precipitated from the salt solutions by the addition of the exact equivalent amount of alkali. The volume of air passed was 36.jlitres in j+ hours.
- .s
Substance (freshly precipitated hy0 droxides or sul0 phite) used as catalyst in the W experiment 13
8
& I
2
3 4
5 6 7 8 9
IO 11 12
+mount, of urea in grm. in I O C.C. of the solution taken. (Blank)
Cerous hydroxide 0.20 Manganous hydroxide ” Ferrous hydroxide Ferric hydroxide 1, Uranous hydroxide Chromic hydroxide !I Cobaltous hydroxide ” Sickelous hydroxide ” Mercuric hydroxide ” (wet HgO) Cupric hydroxide 1, ,, Lead hydroxide Sodium sulphite ( = IO C.C. of 1 . 0 7 ~sol’n.)
Amount of urea Amount of urea Percentage left in grm. oxidised in amount after the exgrm. of urea periment oxidised
0.17
0.03
Ij.0
0.1625
0.0375
18.75
f,
0.I 8
0.02
10.0
,f
0.18 0.16
0.02
10.0
0.04
20.0
0.1725
0,0275
13.75
0.18 0.I 8
0.02
10.0
0.02
10.0
0.162 j
0.0375
18.75
0.165
0.035 0.03
,)
0.17 0.176
0.024
17.5
15.0 12.0
1 J. Phys. Chem., 29, 376 (1925); “Chemie der Zelle und Gewebe,” 12, 217 ( 1 9 2 j ) ;13, 1x9 (1926).
1672
C. C . PALIT AND N. R. DHAR
water but to the stage of acetone bodies and that the oxidation of the glucose leads to the complete oxidation of the fats. When glucose is not burnt appreciably in the body as happens in the case of severe diabetes, the fats and proteins have to be oxidised for the supply of heat to the body and hence in diabetes, fats and proteins have to be burnt more readily than in normal health. From our experimental results, it will be seen that in the presence of carbohydrates which are undergoing oxidation, the velocity of oxidation of fats is greatly decreased. Hence in normal health and with a mixed diet, the easily oxidisable carbohydrates which have been proved to act as negative catalysts in the oxidation of fats, are necessary for the complete oxidation of fatty food materials, and the oxidation of both the substances can go on simultaneously and slowly. When the negative catalysts, the carbohydrates, are not present, the oxidation of fats proceeds rapidly and the end products TABLEIX Carbohydrate Estimation of glucose, cane sugar and starch in presence of (i) I O C.C. of caustic soda containing 0.05 grm. and (ii) I O C.C. of urea of 2.0% solution I O C.C. of caustic soda containing 0.05 grm. Volume of air passed was 36.5 litre-s in s$ hours.
+
e
Substance Actual amount Amount of
Amount of Percentage substance oxi- amount of dised in grm. substance oxidised
Remarks
mat (Blank
A I
Glucose
*
z Canesugar
3 Starch
i1
0.0962
0.0853
0.01og
11.4
0.0964
0.0839
0.0125
12.9
0.1027
0.0717
0.0310
30.0
Experiment performed in alkaline solution ( = I o C.C. of NaOH containing 0.05 grm.) and in absence of urea.
B I
Glucose
0.0962
0.0897
0.0065
6.7
2
Cane sugar 0.0964
0.0935
0.0029
3.0
0.0792
0.0235
22.8
3 Starch
0.1027
Experiment performed in presence of urea under identical conditions as above.
*These results have been taken from our previous publication:-Palit and Dhar: J. Phys. Chem., 30, 941 (1926).
OXIDATION O F FATS WITH CARBOHYDRATES
I673
TABLE X--Urea Estimation of urea in presence of freshly precipitated ferrous hydroxide (=0.0674 grm. of Fe (OH)*) and excess of alkali. The volume of air passed was 36.5 litres in s i hours and the volume of urea solution used was IO C.C. of 2.0% strength. A Fj.
3
Volume of caustic
Amount of caustic soda
Amount of urea in grm.
for the expt.
of the solution experiment (Blank)
Amount of urea left in
Amount of Percenturea oxidised age amount of urea oxidised
3 ,g soda added added in grm. takenin I O C.C. grm. after the in grm.
i
In
I 2
3 4 5
c’c’ 0 C.C. IO C.C.
0.0
2 0 C.C.
0 .IO
40 C.C. 60 C.C.
0.20
0.I800
0.0200
10.0
17
0.1750
0.0250
12.5
11
0.1675 0.1625 0.1600
0.0325
16.2 18.7
0.20
0.05
’1
11
0.30
0.0375 0.0400
20.0
TABLE XI-Glycine Estimation of glycine in presence of (i) freshly precipitated hydroxides of different metals in neutral solutions and (ii) caustic soda, sodium bicarbonate and sodium sulphite. The volume of glycine used was I O C.C. and the volumes of different salt solutions, each of 1.0% concentration, were 2 0 C.C. in each case. The hydroxides were precipitated from the salt solutions by the addition of the exact equivalent amount of caustic soda. The volume of air passed was 36.5 litres in 53 hours.
-
Substance (freshly precipitated hy8 droxides, caustic 9 ,g soda, sodium bi0 b carbonate SUIphite) used aa H catalvst in the expeiiment 4
Volume of N/IO caustic soda in C.C. required for I O C.C. of glycine taken. (Blank)
Volume of N/xo caustic soda in C.C. required for glycine left after the experiment
Amount of glycine oxidised in terms of N/IOcaustic soda in C.C.
Percentage amount of glycine oxidised
Ferrous hydroxide 13.3C.C. 8.6 4.7 35.3 11 36.8 Cerous hydroxide 8.4 4.9 27.8 ” 9.6 3.7 3 Manganous hydroxide 11 30.8 4.1 4 Uranous hydroxide 9.2 11 11.6 12.7 1.7 5 Cobaltous hydroxide 11 Nickelous hydroxide 10.2 6 3.1 23.3 11 27.8 9.6 3.7 7 Lead hydroxide 11 8 Mercuric hydroxide 9.4 29.3 3.9 (wet HgO) 9.9 3.4 25.5 9 Chromic hydroxide 11 1 0 Ferric hydroxide 28.5 3.8 9.5 17 I 1 Cupric hydroxide 8.8 4.5 33.8 30.0 I 2 Causticsoda(=Ioc.c.ofN/8)” 4.0 9.3 11 42.1 7.6 5.7 I3 Sodium bicarbonate (= IO C.C. of 1.07~sol’n.) I4 Sodium sulphite 73.6 3.5 9.8 ( = IO C.C. of 1.07~ sol’n.) IO C.C. of glycine = 13.3 C.C. of N/IO NaOH = 0.09975 grm. of glycine. I C.C. of N/ro NaOH = o.ooj5 grm. of glycine. I
2
1,
1,
1674
C. C. PALIT AND N. R. DHAR
are not mainly carbon dioxide and water as in normal health but are P-hydroxybutyric acid, diacetic acid, acetone, etc. The rapid oxidation of fats in the absence of carbohydrates leads to the formation of acetone bodies as will be apparent from the following facts:( I) Ingestion of a large amount of fats without carbohydrates or proteins leads to the formation of acetone bodies. ( 2 ) I n starvation, after a few days, acetone bodies are generally found in the urine. It is well known that by injection of insulin, acetone bodies disappear from the urine of the diabetic. This is due to the fact that by the presence of insulin, the glucose which was passing unoxidised from the diabetic system, is oxidised and the oxidation of the glucose leads to the more complete though more slow oxidation of fats. Lusk, Landegren and others’ proved that the withdrawal of carbohydrates from food increases the protein metabolism. Moreover, it is well known that there is a considerable tissue waste in diabetes. These facts become clear from our experimental results already recorded which prove that carbohydrates markedly retard the oxidation of nitrogenous matter. According to J. Bar; simple withdrawal of carbohydrates is followed in normal human beings accustomed to a mixed diet and also in apes by an acidosis, i.e. by an execretion of acetone, diacetic acid and P-oxybutyric acids along with coincident increase in the ammonia elimination.
TABLE XI1 Glycine Estimation of glycine in presence of (i) carbohydrates, (ii) urea and (iii) fats in neutral solutions. The volume of each of the solutions taken was I O c.c and the volume of air passed was 36.5 litres in 54 hours. The strength of each of the solutions of carbohydrates, urea and fats was about 10% ( I O C.C. of glycine 3 13.3 C.C.of N / I O XaOH; 10c.c. potassium stearate + I O C.C. IC13 14.2 C.C. of K/IO Na2S203 and I O C.C. potassium oleate I O C.C. IC13 10.8 C.C. of N/IO Na2SzO3.)
+
Substance used in the experiment as catalyst I
2
3 4 5
6 7 8
Volume of N/IO caustic soda in C.C. required for IO C.C. of lycine taken. (Blank) 13.3
Glucose Galactose Lactose Laevulose Cane sugar Urea Potassium stearate Potassium oleate
,, J)
,)
,, ,I
” J l
Volume of N / I O caustic soda in C.C. required for glycine left after the experiment 13.0
Percentage amount of glycine oxidised
0.3
2.2
12.7
0.6
13. I
0.2
4.4 1.4 3.0 0.3
12.9
0.4
13.25 12.j
0.05
7.3 8.2
Compare Lusk: “Science of Nutrition”, p. 269 (1919). exp. Pathol., 54, 153 (1905).
* Archiv.
Amount of elycine oxidised in terms of N / I O caustic soda in C.C.
0.8 6.0
44.3
6.0
j.1
37.5
I675
OXIDATION OF FATS WITH CARBOHYDRATES
TABLE XI11 Glycine
+
+
+
Estimation of glycine in presence of (i) fat glucose (ii) fat urea, (iii) fat glucose urea and (iv) excess of fats. The volume of air passed was 36.5 litres in 5; hours.
+
potassium stearate potassium oleate
I O C.C.
+
c Substances used E in the experiment 6 'i: as catalyst
0 0
2 %
4 I
2
3 4 j
6
7 8 9 IO
II
+
+
I O C.C.
IC13 14.2 C.C.of N / I O Na2S203. IC13 3 10.8 C.C.of K/IO N&03.
I O C.C.
I O C.C.
Volume of X / I O caustic soda in C.C. required for I O C.C. of glycine taken . (Blank)
Potassium stearate 13.3 and glucose Potassium stearate JJ and urea Potassium oleate JJ and glucose Potassium oleate JJ and urea Potassium stearate 1J and glucose and urea Potassium oleate and JJ glucose and urea JJ Glucose and urea Excessof potassiumstearate (20 c.c.) and glucose Excess of potassium oleate ( 2 0 c.c.) and glucose Excess of potassium stearate 11 (20 c.c.) and urea Excess of potassium oleate JJ ( 2 0 c.c.) and urea J
J
J
Amount of Percentage glycine oxiamount disedin terms of glycine of N / I Ocaustic oxidised soda in C.C.
9.8
3.5
26.3
10.2
3.1
23.3
11.1
2.2
16.5
12.0
1.3
9.7
11.5
1.8
13.5
12.8
0.j
3.9
13.3
0.0
0.0
6.5
6.8
51.1
6.3
7.0
52.6
9.7
3.6
27.0
9.4
3.9
29.3
of glycine 13.3 C.C. of N / I O NaOH = 0.09975 grm. of glycine. of N/IO KaOH 0.007 j grm. glycine.
I O C.C. I C.C.
J
Volume of N / I O caustic soda in C.C. required for glycine left after the experiment
1676
C. C. PALIT AND N. R. DHAR
TABLE XIV Carbohydrates Estimation of carbohydrates in presence of fats. The volume of air passed was 60 litres in 9 hours. The volume of fat taken was I O C.C. in each case. IO C.C. of uotassium stearate = 14.2 C.C. of N/IO NazS2O3; IO C.C. of pot. oleate 5 10.8 C.C. of N/IO Na&03. Substance Actual amount used in of substance the experi- in grm. in IO C.C.of the solution taken merit (Blank)
.L
8
9 .$
wB A.
I 2
3 4 5 6 7 8
B.
I
2
3 4 5 6 7 8
C.
I 2
3 4
5 6 7 8
Arabinose Galactose Glucose Laevulose Lactose Canesugar Maltose Starch
Amount of of substance left in grm. after the experiment
Amount of of substance
oxidised in ,grm.
Percentage amount of substance oxidised
0.1027
0.0974 0.0788 0.0906 0.0904 0.0859 0.0896 0.0889 0.0823
0.0026 0.0073 0.0056 0.00145 0.0138 0.0068 0.0208 0.0204
13.8 7.0 18.9 19.9
Arabinose Galactose Glucose Laevulose Lactose Canesugar Maltose Starch
0.1000 0.0861 0.0962 0.09185 0.0997 0.0964 0 . I097 0.1027
0.0899 0.0756 0.0866 0.0723 0.0786 0.0876 0.0829 0.0811
0.0101
IO. I
0.01og 0.0092 0.0195
12.2
21.2
0.0211
21.2
0.0088 0.0268 0.0216
9.1 24.4 21
Arabinose Galactose Glucose Laevulose Lactose Cane sugar Maltose Starch
0.1000 0.0861 0.0962 0.09185 0.0997 0.0964 0.1097
0.0965 0.0774 0,0880 0.08125 0.0853 0.0862 0.0861
0. I027
0.0602
0.0035 0.0087 0.0076 0.0106 0.0144 0.0102 0.0236 0.0425
3.5 10.0 7.9 11.5 14.4 10.5 21.5 41.3
0.1000
0.0861 0.0962 0.09185 0.0997 0.0964 0.1097
2.6 8.5 5.8
1.5
9.5
.o
Remarks
Experiment performed in presence of potassium stearate ( = 10c.c.) in neutral solutions. Experiment performed in presence of potassium stearate ( = I O c.c.) in alkaline solutions. The volume of alkali used = I O C.C. of N/8 NaOH. Experiment performed in presence of potassium stearate ( = IO c.c.) ferrous hydroxide
+
(=0.0674
grm. of Fe(0H)Z) in neutral solutions.
OXIDATION O F FATS WITH CARBOHYDRATES
-
I677
TABLE XIV (Continued)
Substance Actual amount $ used in of substance 0 .@the experi- in grm. in 2 3 ment I O C.C. of the c.n solution taken (Blank) e
D.
E.
Arabinose Galactose 3 Glucose 4 Laevulose 5 Lactose 6 Cane sugar 7 Maltose 8 Starch
0.0029 0.0041 0.0046 0.0065 0.0109 0.0008
0.0198 0.0861 0.0122 0.0962 0.0098 0.09185 0.01395 0.0813 0.0997 0.0964 0.0795 0.1097 0.0617
0,0802
0.0169 0.0480
0.1027
0.0757
0.1000
0.0861 0.0962 0.09185 0.0997 0.9964 0.I097
I
3 4 j
6 7 8
F.
0.0971 0.0820 0.0916 0.08725 0.0888 0.0956 0.0973 0.0642
I
2
2
I
2
3 4
5 6 7 8
Arabinose Galactose Glucose Laevulose Lactose Cane sugar Maltose Starch
Arabinose Galactose Glucose Laevulose Lactose Cane sugar Maltose Starch
Amount of Amount of substance left substance oxiin grm. after dised in grm. theexperiment
0.1027
0,1000
0.0270
0.0124
0.0385
0.0739 0.0864 0.0779 0.0184
0.1000
0.1000
0.0000
0.0861 0.0962 0,09185 0.0997 0.0964 0.1097
0.0861 0.0962 0.0918 0.0977 0.0963
0.0000
0 .I020
0.1027
0.0784
0.0077 0.0243
0.0000
0.0000
Percentage amount of substance oxidised
Remarks
2.9 Experiment performed in pres4.8 ence of potas7.1 sium stearate 10.9 ( = 10c.c) +fer0.8 rous hydroxide 11.3 ( = o . 0674grm. 37.5 of Fe (OH)*) urea ( = I O C.C. of 1.07~ solution) in neutral solutions. 4.7
+
Experiment performed in presence of potassium oleate ( = 10c.c.) +fer17.5 rous hydroxide 43 ’ 7 (=0.0674grm. 73.7 of Fe (OH)z)in neutral solutions 80.2 85.8 89.7 84.8 18.4
Nil Nil Kil Nil
0.0020
2.0
0.0001
Si1 7.0 23.7
Experiment performed in presence of potassium oleate ( = I o C.C.)
+
ferrous hydroxide (=0.0674 grm. of Fe (OHM urea ( = 1oc.c. of I .oyGsolution) in neutral solutions.
+
1678
C. C. PALIT AND N. R. DHAR
XI Glucose Estimation of glucose in presence of potassium oleate and freshly precipitated hydroxides of different metals. The volume of potassium oleate used was I O C.C. and the volumes of different salt solutions used were 2 0 C.C. in each case. The hydroxides were precipated from the salt solutions by the addition of the exact equivalent amount of caustic soda. The volume of air passed was 60 litres in 9 hours. ( I O C.C. of Pot. oleate 3 10.8 C.C. of N / I O Na2S203). TABLE
l
e
Substance * c (freshly )
E ppted. hy6 '2 droxides) 3
Z % usedin the
;ri:
experiment as catalyst
Cerous hydroxide 2 Cobaltous hydroxide 3 Ferrous hydroxide 4 Ferric hydroxide 5 Manganous hydroxide 6 Mercuric hydroxide (wet HgO) 7 Kickelous hydroxide 8 Uranous hydroxide
Amount of glucose in grm. in I O C.C. of the solution taken (Blank)
Amount of Amount of glucose left in glucose oxigrm. after the dised in grm. experiment
Percentage Remarks amount of glucose oxidised
I
0.0962
0.0764
0.0198
20.6
,,
0.0;19
0.0443
46.0
0.0098
0.0864
89.7
0.0344
0.0618
64.2
0.0604
0.0358
37.2
0.0362
0.0600
62.4
0.0461
0.OjOI
52.0
0.0503
0,0459
47 7
,I
,It
,, f,
,,
'
Experiment performed in presence of freshly precipitated hydroxides of different metals, potassium oleate and glucose in neutral solutions. The strength of salt solutions used was 1.0% in each case.
OXIDATION OF FATS WITH CARBOHYDRATES
1679
TABLE XVI Urea Estimation of urea in presence of mixtures of freshly precipitated ferrous hydroxide ( = 0.0674 grm.) and carbohydrates and potassium oleate in neutral solutions. The volume of air passed was 60 litres in 9 hours. The volume of each solution taken was I O C.C. in each case. ( I O C.C. of potassium oleate I O C.C. of Ic13 IO. 8 C.C. of K/IO Na&o3.)
+
%
8 E
6
6 2” I 2
3 4
5 6 7 8
Substance used as catalyst in the experiment
Arabinose Galactose Glucose Laevulose Lactose Cane sugar Maltose Starch
Amount of urea in grm. in I O C.C. of the solution taken (Blank) 0.20 I,
JJ JJ
Amount of urea left in grm. after the experiment
Amount of urea oxidised in grm.
0.19
0.01
0.19 0.19
0.01 0.01
Percentage amount of urea oxidised 5.0 JJ JJ
0.0
Nil
11
1,
19
1,
11
Jt
,,
,,
1J
11
JJ
JJ
,I
,I
71
1,
0.20
Strength of carbohydrate solutions was about
1.0%.
It is generally believed by physiologists that carbohydrates are oxidised in the animal economy most readily but our experimental results show that fats are also readily oxidised. I n a previous paper,’ we have explained that glycosuria induced by the intake of phosphorus, arsenic, or phlorhizin or ether, is really due to the decrease in the velocity of oxidation of carbohydrates by the presence of the above reducing agents. As soon as the oxidation of carbohydrates is decreased by the presence of reducing agents like phosphorus, arsenic, phlorhizin or ether, etc., glycosuria sets in and when glucose escapes unburnt from the system, the retardation of the oxidation of the fats and proteins due to the oxidation of glucose cannot take place and thus we get acidosis in all these cases. I n the same paper, it was shown that small quantities of alcohol might be useful in decreasing acidosis in diabetes, because alcohol being a substance which is readily oxidised can retard the oxidation of fats and thus may lead to the more complete but slow oxidation of fats. Xylose, gluconic acid, propionic acid, citric acid, glutaminic acid, glutaric acid, etc., have been found to decrease acidosis and formation of acetone bodies in diabetes. We are of opinion that the action of these substances is similar to that of alcohol as they are all reducing agents and are likely to reduce the velocity of the oxidation of fats and hence are likely to act as anti-ketogenic substances hy checking the formation of acetone bodies. “Chemie der Zelle und Gewebe”, 13, 119 (1926).
1680
C.
C. PALIT AND
Irj.
R. DHAR
We are trying t o obtain quantitative and comparative results on the velocity of oxidation of fats, proteins, and carbohydrates by air and thus establish definitely whether fats or carbohydrates are oxidised more readily in the system. Summary and Conclusions Experimental results show that nitrogenous substances like urea, hippuric acid, and glycine can be oxidised by air at the ordinary temperature in presence of reducing agents. In presence of alkali, hippuric acid and glycine can be oxidised even in the absence of a catalyst and the greater the amount of alkali, the greater is the amount of oxidation. Experimental results have been obtained showing that in presence of (2) caustic aoda or sodium bicarbonate, potassium stearate and potassium oleate have been oxidised. The amount of oxidation is increased in presence of different metallic hydroxides. ( 3 ) From a mixtureof carbohydrates, and potassium stearate or potassium oleate, both the carbohydrates and fats are oxidised by passing air a t the ordinary temperature and in presence of caustic alkali or sodium bicarbonate, the amount of oxidation is increased and the greater the amount of alkali, the greater is the amount of oxidation. In presence of ferrous hydroxide or cerous hydroxide, the amount of oxidation of fats and carbohydrates is increased from a mixture. (4) The experimental results conclusively prove that the amount of oxidation of carbohydrates is greatly decreased by the presence of fats. Also the oxidation of fats is reduced by the presence of carbohydrates. Moreover, in presence of carbohydrates or fats the oxidation of urea, hippuric acid and glycine is appreciably decreased. In presence of urea, the oxidation of fat or carbohydrate is also retarded. ( 5 ) These experimental results have thrown considerable light on the metabolism in the animal body in normal health and in diabetes. (6) We are of opinion that alkaline treatment should prove efficacious in gout, diabetes, beri-beri, rickets and other metabolism diseases, because in presence of even sodium bicarbonate, the amount of oxidation of fats, carbohydrates, and nitrogenous substances is greatly increased. (I)
Chemistry Department
University of Allahabah, Allahdad, India. June. 1987.
