A DIETARY FACTOR ESSENTIAL FOR GUINEA PIGS*
VIII. THE ISOLATION OF THE ANTISTIFFNESS FACTOR
FROM CANE JUICE
BY WILLEM J. VAN WAGTENDONK AND ROSALIND
WULZEN
(From the Department of Chemistry and the Department of Zoology,
Oregon State College, Corvallis)
(Received for publication, April 23, 1946)
The existence of a fat-soluble dietary factor essential for guinea pig
nutrition was well established through investigations by
Wulzen and
Bahrs (l-3) and has been confirmed by Anderson and Caldwell.’ The
isolation of a highly active fraction from raw cream, capable of alleviating
an induced stiffness in guinea pigs, was described by van Wagtendonk and
Wulzen (4). It seems apparent from physiological studies that the factor
has a regulatory effect on the phosphorus metabolism. One of the most
prominent changes found was a sharp decrease in the easily hydrolyzable
phosphorus fraction in the liver and kidneys during the deficiency. This
fraction responded immediately to the administration of the antistiffness
factor to deficient animals in that the values returned to normal after a
short time of treatment (5). Similar changes were observed in the concentration
of the acid-soluble phosphorus in the muscle. The concentrations
of creatine phosphate and adenosine tri- and diphosphate are
lower in the deficient animal (6). As a result of the deranged phosphorus
metabolism other, probably secondary, changes are (1) an increase in the
concentration of inorganic phosphorus and calcium in the blood, (2) an
increase in calcium in the body tissues (7), and (3) an abnormal distribution
of the protein nitrogen in the blood (
.
Only 3 mg. of an oil, curative in a 0.1 y dosage, were obtained from
55 gallons of raw cream (4). Since much larger amounts of raw cream
would have been needed for a successful isolation of the antistiffness factor,
it was decided to test other possible sources of raw material for the presence
of this factor. It was found that crude cane molasses and crude unheated
cane juice were good sources of the factor, the latter being around 100
times as active. Due to the unavailability of crude cane juice, cane molasses
was first used for the extraction of the factor. Later cane juice
became available and was used exclusively.
We were able to extract and purify a crystalline compound which in a
daily dose of 0.002 y would relieve the stiffness induced by the skim milk
diet in 5 days. The low level of the easily hydrolyzable phosphorus
returned to normal values in the same time.
... (details in pdf at above link)
DISCUSSION
From this and other previously reported investigations (4-7) it becomes
apparent that the compound isolated from the cane molasses and the cane
juice is able to prevent and to cure the symptoms characteristic for the
deficiency. In view of the extremely small dosages required we feel justified
in classifying the compound as an essential metabolite. Work towards the
elucidation of its structure is in progress. It has not yet been possible to
establish whether this compound is also present in the previously isolated
fraction from raw cream.
SUMMARY
A procedure for the isolation of a crystalline factor present in molasses
and raw cane juice, which cures an induced stiffness in guinea pigs, has been
described. The abnormal low level of the easily hydrolyzable P in the liver
of deficient animals can be returned to a normal value by the administration
of minute dosages of the antistiffness factor. The smallest curative dosage
is 0.002 y.