![– 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.](https://image.slidesharecdn.com/extractionisolationestimationofephedrine-1-181128145304/85/Extraction-isolation-estimation-of-ephedrine-5-320.jpg)
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.V0.1ML%solution , 2-3times aday.