Professional Documents
Culture Documents
Techniques and Method in in Production
Techniques and Method in in Production
Semester V
Amity Institute of Biotechnology
Amity University Rajasthan
Jaipur Submitted by:
Mechanism of action: -
The artemisinin radical binds to membrane proteins of parasite and
alkylation reactions eventually cause destruction of the parasite.
Tread Names: -
• ARTECOPE
• Larither
• Larimef
• Larinate
• RAPITHER - AB INJECTION (IM Only)
Material and method
• Extraction technique.
• Purification technique.
• Analysis technique.
Extraction technique: -
Objective : - To Extract plant leaves content from A.Annua.L
plant leaves.: -
• Take 25g of grinded leaves for the sample.
Put them in thimble.
• Add 200 ml of the hexane in the distilling pot.
• Set the temperature of water bath to 70-80’C
• Continue the process for 4 hrs.
• Make the total volume of the
extracted solution 250ml.
• Distillation of the extracted solution in
rotary evaporator . at 750C and 50 rpm.
Result:-
Crude (Extracted material) weight was 420 mg.
Purification Technique: -
Objective: -Separation of Reaction Mass of method for the preparation
of artemether from artemesinin.
Materials:- Column, Silica gel, TLC, Chromatogram Immersion Devise,
TLC plate heater.
Method: -
• Initial set up of column.
• Made slurry with 60 g of silica and N hexane. Packed the column with the slurry.
• Added reaction mass 3g , silica and N hexane mixture as slurry on the surface.
• Purification of component.
• Charged the column with the ethyl acetate and N hexane in the ratio of (2:98).
• Collected the fraction, Change the conical flask for each 100-150ml collection.
• TLC analysis of the fraction of reaction mass.
RESULT:-
Alpha Artemether :- 1.78g
Beta Artemether: - .65g
furnished pure alpha and beta Artemether 2.43 g .
Analysis Technique: -
Objective: - Sulphated ash analysis of Artemisinin.
Material:-
Conc. Sulphuric acid, Muffle furnace.
Method: -
• Weight of blank crucible was X1=120g.
• Conditioning.
• Burn the sample by heating the crucible along with addition of conc. Sulphuric acid.
• put the crucible in the muffle furnace for 3 hr.
Calculation:-
Weight of left content:- X2-X1= 120.012g-120g=.012g=12mg
Weight of left content 12mg
Sulphated ash % =
………………………. = …….. × 100 = 1.2%
Weight of sample used 1000mg
Result:-
The sulphated ash content of 1g artemisinin was 1.2%.
Objective : - To find out Bulk density and Tap density.
Material:-
Tapping instrument, 25ml measuring cylinder,10-15g of artemisinin.
Method: -
Sample taken for Bulk and Tap density was 5mg.
Calculation and Result: -
Grams of sample taken 5
Bulk density=…………………………………= ………..= .47g/ml
Volume occupied by sample 10.5
Grams of sample taken 5
Tap density= ………………………………… = …… =.754 g/ml
Volume occupied by sample after tapping 8.7
Sample preparation:-
Take 3.2 g of artemisinine and dissolve it in 100% methanol.
Method: -
• Select mercury lamp.
• Select the wavelength 599 by pressing key “waveln” and key enter
press.
• For “SORT” feed the concentration by pressing “conc” added the
value in which the sample solution prepared 3.20%. Press the “enter”
key.
• After setting this, put the sample firstly “blank” (Methanol) in cell
quartz cell & then the pressing key “Auto zero”.
• After this put the sample in quartz cell. Press “Enter”.
Standard preparation:-
Take 10mg artemisinin and dissolve in the 10 ml hexane.
Method: -
• Software linomat 5 setup.
• Linomat auto sampler set up.
• HPTLC plates development.
• Scanning of TLC plate.
Standard graph Sample graph
The analysis and calculation of samplegraph in win cats give the artemisinin
content was .8mg/ml.
• Object:- Impurity analysis of Sulphamethoxy pyrazine by using Gas
chromatography.
Material.
Diluents:-
DMSO:Water(90:10)
Standard stock solution:-
200mg benzene add in 100ml flask containing 50ml diluents. Conc. 2000ppm.
Preparation of standard.
1.0ml standard stock solution. Add in 100ml diluents. Conc. 0.2ppm.
Transfer 1.0ml of standard solution in dry head space vial and seal it.
Test solution.
100mg of sample add in 1ml diluents.
Method:-
• procedure for operation Totalchrom navigator.
• Operation of Instrument Option of Totalchrom navigator.
• Operation of Sequence set up.
• GC Instrument Setup.
• operating of Head space sampler-Turbomatrix -40 .
• Click the RUN option of GC total chrome Navigator
Result: -
Calculation And Result:-
AT WS 1 1 1
ppm of benzene =…….....×……….×………×…….×….......×106
AS 100 100 100 WT
WS=200mg, WT=100mg, AT=1995.33, AS=2003.25
1995.33 200 1 1 1
ppm of benzene =…….....×……….×………×…….×….......×106
2003.25 100 100 100 100