EXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE

1. EXTRACTION, ISOLATION &
ESTIMATION OF EPHEDRINE
gerardiana &
– BIOLOGICAL SOURCE:
It consist of the dried young stems of Ephedra
E.Nebrodensis E.sinica E.equisetina family
Ephedraceae.
n-methyl ephedrine,
– CHEMICAL CONSTITUENT:
It contains ephedrine, nor-ephedrine,
pseudo-ephedrine, etc

2. EPHEDRINE
CHARACTERISTICS OF EPHEDRINE
Chemical Formula:
C10 H15 NO
Molecular weight:
165.23 grams/mole
Melting point: 37-39 0C
Boiling point:253-255 0C

3. EXTRACTION
•Take weighed quantity of dried young stems of ephedra and powder to
obtain coarsesizedparticles.
•Extract with petroleum ether.
•Filter and discard ether extract.
•Dry the marc and moisten with dilute ammonia.
•Extract this marc with aqueousalcohol.
• Filter and evaporate filtrate to obtained syrupy mass.
•Extract this masswith 3 portions of aqueous alcohol.
•Add sufficient quantity of sodium carbonate solution to make the extract
alkaline.
•Filter and evaporate filtrate to obtained dry residue of ephedrine.

4. – ESTIMATION:
– TLC
Testsolution:
•Extract about 5g of freshly powered drug with 50ml methanol
by shakingfor 30min at 500 in aconical flask.
•Filter and concentrate the filtrate under vacuum to about 5ml.
– Strandard solution.
• Dissolve 5mg ephedrine in 10mlmethanol

5. – Solvent system:
Chloroform: Toluene : Ethanol[3:6:2]
– Procedure:
Apply separately 10µl eachof test and reference solution on apercoated
silica gel 60 F254TLCplate of uniform thickness of0.2mm
Develop the plate in the solvent systemto adistance of 8cm and dry in a
current of hotair.

6. – Visualization
spray the air dried plate with ninhydrin reagent and heat at 1100 for
5min note the Rfvalue.
Rfvalue : 0.15 color ofband reddish purple (ephedrine)
– HPTLC:
solvent system:
Ethylacetate: Ethanol : Diethyamine
7 : 3 : 0.2

7. – TESTSOLUTION:
accurately weighed about 5g of powered drug & extract in a Soxhlet
apparatus 50ml for 4hr. Filter and adjust the volume to 50ml in a
volumetric flask. Pipette out 1ml of this solution and dilute to 10ml.
– STANDARD SOLUTON
prepare 1mg/ml stock solution of ephedrine in methanol pipette out
aliquots of 0.5 to 3ml in increments of 0.5ml into 10ml volumetric flask
and makeup the volume eachflask withmethanol.

8.  CALIBRATION CURVE
Apply 10µl of each concentration of standard solution in triplicate
on TLC plate. Develop the plate in the solvent system to a
distance of 8cm, dry in a current of hot air and scan plate in the
TLC scanner at 210 nm. Record the peak area for each
concentration of ephedrine and construct thecalibration curve.

9. Rf
Wavelength: 210nm
HPTLC DENSITOMETER
A:Ephedrine standard B:Test solution of
ephedra gerardiana stem.
500
450
400
350
300
250
200
150
100
50
0
0.0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
B
A
AU

10. ESTIMATION OFEPHEDRINE INTHEDRUGS:
– Apply 10µl of the test solution in triplicate on aTLCplate. Develop the
plate in the solvent system to a distance of 8cm and record the
chromatogram as described under the calibration curve. Determine the
amount of ephedrine in the sample from the calibration curve of
ephedrine.
-maximum content of ephedrine wasfound tobe 2.60%.

11. – USES:
It is usedasbronchodilator in asthma ephedrine used in to correct the
10w blood pressurecondition.
– DOSE:
ephedrine HCLor Sulphate 25-50mg, 6-8 times aday orally and parentally
I.V0.1ML%solution , 2-3times aday.