ARTEMETHER SOP


ARTEMETHER SOP

1.0  OBJECTIVE:
To lay down a procedure of analytical report for the active raw material of Artemether from the Pharmacopoeial specifications.
2.0  SCOPE:
This SOP shall be applicable in Q.C laboratory.
3.0  RESPONSIBILITY:
3.1  Q.C Analyst.
4.0  ACCOUNTABILITY:
4.1  Q.C Manager.
5.0  PROCEDURE:
5.1  Characters:
5.1.1        Appearance:
5.1.1.1  Take about 1.0g of the test sample on a clean and dry watch glass, spread and observe visually.
5.1.1.2  The sample should be white crystalline powder, odorless and slightly bitter in taste.
5.1.2        Solubility:
5.1.2.1  Material and equipment:
5.1.2.1.1        Glassware (5 test tubes, spatula).
5.1.2.1.2        Chloroform.
5.1.2.1.3        Ethyl acetate.
5.1.2.1.4        Ethanol.
5.1.2.1.5        Acetone.
5.1.2.1.6        Purified water.
5.1.2.2  Sample:
5.1.2.2.1        Small quantity.
5.1.2.3  Method:
5.1.2.3.1        Take 5 test tubes and add small quantity of sample for testing solubility according to B.P specifications.
5.1.2.3.2        Add chloroform in test tube 1, ethyl acetate in test tube 2, ethanol in test tube 3, acetone in test tube 4 and purified water in test tube 5 in a small volume and observe the solubility of the sample.
5.1.2.4  Observations:
5.1.2.4.1        The sample in test tube 1 containing with chloroform is clear.
5.1.2.4.2        The sample in test tube 2 containing with ethyl acetate is clear.
5.1.2.4.3        The sample in test tube 3 containing with ethanol is clear.
5.1.2.4.4        The sample in test tube 4 containing with acetone is clear.
5.1.2.4.5        The sample in test tube 5 containing with purified water is not clear.
5.2  Identification tests:
5.2.1        Specific optical rotation:
5.2.1.1  Material and equipment:
5.2.1.1.1        Glassware (according to the requirement).
5.2.1.1.2        Polarimeter.
5.2.1.2  Sample:
5.2.1.2.1        10.0mg of sample.
5.2.1.3  Method:
5.2.1.3.1        Prepare test solution 10.0mg/ml in ethanol.
5.2.1.3.2        Operate the Polarimeter according to SOP No. BM/QCEO/SOP030-00.
5.2.1.3.3        Determine optical rotation on the test solution.
5.2.1.3.4        Fill the Polarimeter tube with blank solution and determine the observed optical rotation.
5.2.1.3.5        Similarly, fill the Polarimeter tube with sample solution and determine the observed optical rotation.
5.2.1.3.6        Note down the values in annexure-1.
5.2.1.3.7        Calculate the specific optical rotation with reference to anhydrous substance by using formula:
[α]λ T = α/lc
5.2.1.4  Observation:
5.2.1.4.1        Between +168o to +173o.
5.3  Assay:
5.3.1        Apparatus:
5.3.1.1  HPLC apparatus.
5.3.1.2  Analytical weighing balance.
5.3.1.3  Glassware (according to the requirement).
5.3.1.4  Magnetic stirrer.
5.3.2        Material and reagents:
5.3.2.1  Methanol.
5.3.2.2  Purified water.
5.3.2.3  50.0mg of artemether RS.
5.3.2.4  50.0mg of artemether.
5.3.3        Requirements:
5.3.3.1  Mobile phase:
5.3.3.1.1        Mix volume of purified water and methanol in a ratio (12:88). Filter through 0.45μm nylon membrane filter and degas.
5.3.3.2  Standard solution:
5.3.3.2.1        Take a 25.0ml volumetric flask and transfer a suitable quantity of 50.0mg of artemether RS to it.
5.3.3.2.2        Dissolve it in sufficient quantity of mobile phase by using magnetic stirrer operate according to SOP No. BM/QCEO/SOP007-00.
5.3.3.2.3        Dilute it up to the mark with mobile phase.
5.3.3.3  Sample solution:
5.3.3.3.1        Take a 25.0ml volumetric flask and transfer a suitable quantity of 50.0mg of artemether sample to it.
5.3.3.3.2        Dissolve it in sufficient quantity of mobile phase by using magnetic stirrer operate according to SOP No. BM/QCEO/SOP007-00.
5.3.3.3.3        Dilute it up to the mark with mobile phase.
5.3.3.4  Chromatographic system:
5.3.3.4.1        Mode: Liquid chromatography.
5.3.3.4.2        Detector: UV 210nm.
5.3.3.4.3        Column: C18, (4.6mm x 25mm).
5.3.3.4.4        Flow rate: 1.0ml/min.
5.3.3.4.5        Attenuation: 7 or as required.
5.3.3.4.6        Injection size: 20.0μL.
5.3.3.5  Analysis:
5.3.3.5.1        Samples:
5.3.3.5.1.1  Standard solution and sample solution.
5.3.3.5.2        Calculate the percentage of Artemether (C16H26O5) in the portion of the Artemether taken:
Result= (rU/rS) x (CS/CU) x 100.
rU= peak response from the sample solution.
rS= peak response from the standard solution.
CS= concentration of the standard solution (mg/mL).
CU= concentration of the sample solution (mg/mL).
5.3.3.5.3        Acceptance criteria:
5.3.3.5.3.1  98.0%-102.0% (on dried basis).
5.3.4        Procedure:
5.3.4.1  Equilibrate the column and detector with mobile phase at specified flow rate until a constant signal is received.
5.3.4.2  Inject a sample and standard solution of 20μl through the injector, or use an auto-sampler.
5.3.4.3  Begin the gradient program.
5.3.4.4  Record the spectrum.
5.3.4.5  Analyze as directed in the monograph.



6.0  REVISION LOG:
Revision No.
Effective Date
Reason
00

New SOP

7.0  REFERENCES:
7.1  Manufacturer specifications.
8.0  ANNEXURES:
Annexure 1: Specific optical rotation observations and calculations.
Annexure 2: Observations and calculations of HPLC method.

Annexure: 1
Specific optical rotation observations and calculations
Specific optical rotation
Instrument: ___________________                                              Date: _______________
Model: _______________________        Length of Polarimeter tube: ________________
Sample: ________________________________g.
Solvent: ________________________________ml.
Concentration of sample solution: ____________g/ml.
Blank solution:
Sr.#
Blank solution
Temperature
Optical rotation
(α)












                                                                                                 Average: _______________
Optical rotation of blank solution: _______________
Sample solution:
Sr.#
Sample solution
Temperature
Optical rotation
(α)












                                                                                                 Average: _______________
Optical rotation of sample solution: ______________
Optical rotation of substance = Blank solution - Sample solution.



Specific optical rotation of sample solution by using formula:
[α]λ T = α/lc






                                                                      Result: ________________
Remarks: ___________________________________________________________



Annexure: 2
Observations and calculations of HPLC method
Analysis on HPLC
Instrument: ___________________                                           Date: _________________
Model: ___________________
Column size:
Length=
θ=
Stationary phase:

Temperature:

Mobile phase:

Flow rate:

Injection size:

Detector:

Wavelength:
λ=

Sample solution: _______________________
Reference standard solution: ______________
Impurities: ____________________________
(calculate each component calculation separately)
OBSERVATIONS:
Attach spectrum.






CALCULATIONS:
1.      Retention time:                                                                                n= no. of peak
Retention time of unretained peak (tm)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Height of peak of interest
(h)n
Width of peak of interest
(w)n
Area of peak of interest
A=1/2(h x w)




















2.      Retention volume:
Flow rate= _______________ml/min.
No. of peaks
Retention time of peak of interest
(tr)n
Retention volume = retention time x flow rate












3.      Retention factor:
Retention time of unretained peak (tm)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Retention factor of a component
k= (tr-tm)/tm














4.      Separation factor (α):
No. of peaks
Retention factor of a component
(kn)
Relative retention of two adjacent peaks
α = k2/k1












5.      Resolution:
Retention time of unretained peak (tm)= _____________
No. of peaks
Retention time of peak of interest
(tr)n
Width of peak of interest
(w)n
Resolution
Rs = 2 (tr2-tr1)
        (w1-w2)
















6.      Efficiency:
No. of peaks or components
Retention time of peak of interest
(tr)n
Width of peak of interest
(w)n
Efficiency
(No. of theoretical plates)
N= 16 (tr/w)2



















7.      Height equivalent to a theoretical plate (HETP):
Length of column = ________________________
No. of peaks or components
No. of theoretical plates
(N)
Height equivalent to a theoretical plate HETP = L/N












8.      Symmetry factor (tailing factor):
No. of peaks or components
Distance from the peak max. to leading edge of the peak
(f)
Width w
Symmetry factor
At 5%
At 10%
As = w5%
       2f
As = w10%
       2f
























9.      Response factor & Relative response factor:
Conc. (mg/ml)= ___________________
No. of peak
Peak area
Response factor = (peak area/conc.)
Relative response factor = (response factor of impurity/response factor of API)


















10.  Relative standard deviation (%RSD):
Use formula of relative standard deviation where it is required i.e.,


11.  Percentage of content:
Percentage content = (rU/rS) x (CS/CU) x 100.



rU= peak response of substance from the sample solution.
rS= peak response of substance from the standard solution.
CS= concentration of substance in the standard solution (mg/mL).
CU= concentration of substance in the sample solution (mg/mL).

RESULTS:
________________________________________________________________________________________________________________________________________________


9.0  ABBREVIATIONS:
Abbreviation
Expanded Form
SOP
Standard operating procedure
&
And
No.
Number  
Ltd.
Limited
QCA
Quality control active ingredient
F
Format
Q.C
Quality control
ml
Milliliter
L
Length
g/L
Gram per liter
mg/ml
Milligram per milliliter
μg/ml
Microgram per milliliter
UV/VIS
Ultra violet/ Visible
μg
Microgram
mg
Milligram
RS
Reference standard
UV
Ultra violet
USP
United states pharmacopoeia
nm
Nanometer
mm
Millimeter
cm
Centimeter
μm
Micron
oC
Degree centigrade
ml/min.
Milliliter per minute
μL
Microliter
NMT
Not more than
%
Percentage
NF
National formulary
o
Degree (angle)
l
Length
c
Concentration (g/ml)
g/ml
Gram per milliliter
α
Alpha
λ
Lambda


No comments:

Post a Comment