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Digoxin Assay and Biological Testing Methods

The document outlines the assay method for digoxin, detailing the preparation of solutions and the measurement of absorbance to determine its content. It also discusses biological testing methods using animals, particularly cats, to evaluate cardiac activity and the statistical comparison of sample potency. The testing involves administering measured quantities of digoxin to assess its effects until a defined endpoint is reached.

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6 views6 pages

Digoxin Assay and Biological Testing Methods

The document outlines the assay method for digoxin, detailing the preparation of solutions and the measurement of absorbance to determine its content. It also discusses biological testing methods using animals, particularly cats, to evaluate cardiac activity and the statistical comparison of sample potency. The testing involves administering measured quantities of digoxin to assess its effects until a defined endpoint is reached.

Uploaded by

vkdine61
Copyright
© All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd

DIGOXIN

Assay of digoxin
 Weigh accurately about 40mg,dissolve in sufficient
ethonal(95%)to produce 50ml and dilute 5ml of this
solution to 100ml with same solvent.

 To 5ml of the resulting solution add 3ml of “alkaline picric


acid solution” allow to stand in subdued light for 30min
and measure the absorbance of the solution at the
maximum at about 495 nm,using the blank as a mixture of
5ml of ethanol(95%) and 3ml of alkaline picric acid solution.

 Calculate the content of C41H64O14 from the absorbance


obtained by repeating the operation using digoxinRS in
place of the substance under examination.
BIOLOGICAL CONSIDARATION
 In order to complete the analytical picture of
digoxin it is necessary to mention briefly some of
the typical processes used in biological testing.

 Three species of animals have been used to


evaluate cardiac activity, these are the frog,cat
and pigeon.

 For many years the testing of cardiac activity in


cats was an official procedure.
BIOLOGICAL TEST IN RATS
 In this, four cats were used as controls and were
given measured quantities of reference standard
substance, while an additional four animals were
simultaneously given measured quantities of
sample preparation being evaluated.
 According to the body weight of the cats
suitable solvent, (usually aqueous alcohol) was
used to dissolve the sample and standard.
 After that, administration of sample and
standard solution is done by injecting in to the
femoral vein at the rate of 1ml per 5min.
 This titration continued until the animal
died.

 By a statistical comparison of the


quantity of the sample necessary to
produce death with the quantity of
standard necessary to reach the same
“end point” one could compute the
potency of the sample material in terms
of the standard.

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