Glycosides Classification

Chemistry of D.purpurea
 Chemistry of D.purpurea
• Steroid cardenolides
– contains 30 glycosides, 6 main ones
– only has 3 aglycones

Aglycones 1y 2y

digitoxigenin A digitoxin

gitoxigenin B gitoxin

gitaloxigenin E gitaloxin

• Purpurea 1y glycosides
– do NOT have acetylated digitoxose third sugar
• but these are found in smaller quantities
– called ABE series
[ii] Digitalose
• found in both species

• only strospeside important as emergency injection for heart attacks

– quickest acting cardiac glycoside
Digitalose

strospeside
Digitalose alpha form
 Assay of Digitalis B.P.
• required to contain not less than 0.3%
total cardenolides calculated as digitoxin
• important N.B for drug dosage
• narrow therapeutic index
• can cause cardiac arrest
• slowly excreted, bound to serum proteins
• long term therapy for patients
• Digitalis B.P. tablets
– crushed dried leaves -> green tablet
– contain 30 glycosides each with different
onset, action and excretion profiles
– in different amounts Factors
• influenced by growing conditions affects on
constituents
• (temp, water, sun, drying process)
– assay for each glycoside as possible – dilute
effects by adding grass

• Two ways:
 [1] Biological assay
• British method - inaccurate but safer
• Tincture of extract of leaves or tablets
• diluted with saline so alcohol <6%v/v
• guinea pigs (6 test, 6 control) x 3 =36
– expensive but can average results
• measure volume injected into vein of leg/foot before heart
stops beating
• monitor heart rate via ECG
– or open chest wall and watch inserted needle with flag on move
– Better to watch ECG – have to differentiate from death from too
large an injected volume
• trained staff required, can calculate potency
• assay acceptable within 80-120% error margin (not that
accurate)
• Disadvantages
– inaccurate, expensive
– injecting material IV (avoiding absorption,
excretion)
– end point is death
– toxicity test not therapeutic assessment

• Advantages
– assessing some biological activity
– safety mechanism
 [2] Chemical assay
• Problem: 30 different glycosides – can measure them
accurately but may not correlate with therapeutic activity
of drug
• Make a tincture (with alcohol)
• decolorize with lead subacetate
• extract glycosides by partition with CHCl3( separatory funnel+ water)
• evaporate to give residue (containing cardiac glycosides)
• hydrolyze with HCl to remove sugars, leaving
aglycone
– residue contains gitoxigenin and digitoxigenin (AB series)
– gitaloxigenin -> gitoxigenin when acid hydrolysed
• Colourimetric assay to separately determine material amounts
(i) total aglycone
• purple color with dinitrobenzoic acid and alkali
(ii) digitoxigenin only
• green color with FeCl3 + acetic acid

• Advantages:
– precise method (standard error 5%)
– unqualified staff, quicker
• Disadvantages:
– doesn’t correlate with biological activity
– only estimating approx 60% therapeutic material

• BOTH methods used in industry