This document discusses the extraction, isolation, and estimation of ephedrine from Ephedra plant species. It describes how to extract ephedrine from dried Ephedra stems using solvent extraction. The extracted ephedrine can then be isolated and purified using column chromatography. Estimation of the ephedrine content is done using thin layer chromatography and high performance thin layer chromatography to develop calibration curves. These methods allow for quantitative analysis of ephedrine in samples.

1. EXTRACTION, ISOLATION
& ESTIMATION OF
EPHEDRINE
Presented by : Heera Karemore
Assistant Professor

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

3. 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

4. EXTRACTION
•T
akeweighed 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 makethe extract
alkaline.
•Filter andevaporatefiltrate to obtained dry residueof ephedrine.

5. – ESTIMATION:
– TLC
Testsolution:
•Extract about 5g of freshly powered drug with 50ml
by shakingfor 30min at 500in aconical flask.
methanol
•Filter and concentratethe filtrate undervacuumto about 5ml.
– Strandardsolution.
• Dissolve 5mgephedrine in 10mlmethanol

6. – Solvent system:
Chloroform: T
oluene : Ethanol[3:6:2]
– Procedure:
Apply separately 10µl eachof test andreferencesolution on apercoated
silicagel60F254TLCplate of uniform thicknessof0.2mm
Developthe plate in the solvent systemto adistanceof 8cmanddry in a
current of hotair.

7. – 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

8. – 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 anddilute to 10ml.
– ST
ANDARD 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
andmakeup the volumeeachflaskwith methanol.

9.  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 andconstruct thecalibration curve.

10. 0.0 0.1
Wavelength: 210nm
0.2 0.3 0.4 0.5 0.6
HPTLC DENSITOMETER
A:Ephedrine standard B:Test solution of
ephedra gerardiana stem.
0.7 Rf
500
450
400
350
300
250
200
150
100
50
0
B
A
AU

11. ESTIMATION OFEPHEDRINEINTHEDRUGS:
– 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%.

12. – USES:
It isusedasbronchodilator in asthmaephedrineusedin to correct the
10w blood pressurecondition.
– DOSE:
ephedrine HCLor Sulphate25-50mg,6-8times adayorally andparentally
I.V 0.1ML%solution , 2-3times aday
.