CIPROFLOXACIN HYDROCHLORIDE



CIPROFLOXACIN HYDROCHLORIDE


1.0  OBJECTIVE:
To lay down a procedure of analytical report for the active raw material of the Ciprofloxacin HCl from the Pharmacopoeial specifications.
2.0  SCOPE:
This SOP shall be applicable in Q.C laboratory.
3.0  RESPONSIBILITY:
3.1  Q.C Analysts.
4.0  ACCOUNTABILITY:
4.1  Q.C Manager.
5.0  PROCEDURE:
5.1  Characters:
5.1.1        Appearance:
5.1.1.1  Pale yellow.
5.1.1.2  Crystalline.
5.1.1.3  Slightly hygroscopic powder.
5.1.2        Solubility:
5.1.2.1  Material and equipment:
5.1.2.1.1        Glassware (test tubes, spatula).
5.1.2.1.2        Purified water.
5.1.2.1.3        Methanol.
5.1.2.1.4        Anhydrous ethanol.
5.1.2.1.5        Acetone.
5.1.2.1.6        Ethyl acetate.
5.1.2.1.7        Methylene chloride.
5.1.2.2  Sample:
5.1.2.2.1        Small quantity.
5.1.2.3  Method:
5.1.2.3.1        Take 6 test tubes and add small quantity of sample for testing solubility according to B.P specifications.
5.1.2.3.2        Add purified water, methanol, anhydrous ethanol, acetone, ethyl acetate and methylene chloride in each test tube separately 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 water is soluble.
5.1.2.4.2        The sample in test tube 2 containing with methanol is slightly soluble.
5.1.2.4.3        The sample in test tube 3 containing with anhydrous ethanol is very slightly soluble.
5.1.2.4.4        The sample in test tube 4 containing with acetone is practically insoluble.
5.1.2.4.5        The sample in test tube 5 containing with ethyl acetate is practically insoluble.
5.1.2.4.6        The sample in test tube 6 containing with methylene chloride is practically insoluble.
5.2  Identification tests:
5.2.1        Chlorides test:
5.2.1.1  Material and equipment:
5.2.1.1.1        Glassware (test tube, stirrer, spatula).
5.2.1.1.2        Filter paper strip.
5.2.1.1.3        Diphenylcarbazide solution.
5.2.1.1.4        Potassium dichromate.
5.2.1.1.5        Sulphuric acid.
5.2.1.1.6        Purified water.
5.2.1.2  Sample:
5.2.1.2.1        0.1g.
5.2.1.3  Method:
5.2.1.3.1        Introduce into the test tube 0.1g of sample.
5.2.1.3.2        Add 0.2g of potassium dichromate and 1ml of sulfuric acid.
5.2.1.3.3        Place a filter paper strip impregnated with 0.1ml of diphenylcarbazide solution over the opening of the test tube.
5.2.1.3.4        The impregnated paper must not come into contact with the potassium dichromate.
5.2.1.4  Observations:
5.2.1.4.1        The paper will turn violet-red.
5.3  Other tests:
5.3.1        Solution S:
5.3.1.1  Material and equipment:
5.3.1.1.1        Glassware (1 beaker, 1 stirrer, 1 spatula).
5.3.1.1.2        Carbon dioxide-free water.
5.3.1.2  Sample:
5.3.1.2.1        0.5g.
5.3.1.3  Preparation of Solution S:
5.3.1.3.1        Dissolve 0.50g in carbon dioxide-free water and dilute to 20ml with the same solvent.
5.3.2        pH:
5.3.2.1  Material and equipment:
5.3.2.1.1        Glassware (according to the requirement).
5.3.2.1.2        pH meter.
5.3.2.2  Sample:
5.3.2.2.1        Solution S.
5.3.2.3  Method:
5.3.2.3.1        Firstly clean the pH meter with clean dry cloth
5.3.2.3.2        Operate the pH meter according to SOP for operation of pH meter i.e.
5.3.2.3.3        Rejuvenate the electrode before use according to SOP instructions, if there is any need of.
5.3.2.3.4        Calibrate the electrode of the pH meter
5.3.2.3.5        Perform the test on solution S.
5.3.2.3.6        Take a beaker of 100.0ml and add solution S in it. Such that it immersed electrodes in it completely.
5.3.2.3.7        Maintain the temperature of sample at 25oC±2oC.
5.3.2.3.8        Dip the electrode along with temperature sensor into the sample.
5.3.2.3.9        When dipping electrode into sample, it must be completely immersed in it.
5.3.2.3.10    Stir the probe gently in the sample to create a homogeneous sample.
5.3.2.3.11    Allow the reading to stabilize for a time.
5.3.2.3.12    Record the observed values of pH & temperature in the respective Annexure-1.
5.3.2.3.13    Wash the electrodes again after use and store the electrode in storage solution as recommended in SOP of cleaning i.e.. Calibrate the instrument before use
5.3.2.4  Observation:
5.3.2.4.1        3.5 to 4.5 for solution S.
5.4  Assay:
5.4.1        Apparatus:
5.4.1.1  HPLC apparatus.
5.4.1.2  Glassware (according to the requirement).
5.4.1.3  Spectrophotometer (detector).
5.4.2        Material and reagents:
5.4.2.1  Ciprofloxacin hydrochloride sample.
5.4.2.2  Base-deactivated octadecylsilyl silica gel.
5.4.2.3  Acetonitrile.
5.4.2.4  2.45g/L solution of phosphoric acid.
5.4.2.5  Triethylamine.
5.4.3        Requirements:
5.4.3.1  Sample:
5.4.3.1.1        25.0mg (sample to be examined).
5.4.3.1.2        25.0mg (Ciprofloxacin hydrochloride CRS) for reference solution.
5.4.3.2  Test solution:
5.4.3.2.1        Take 100ml beaker and dissolve 25.0mg of the substance to be examined in the mobile phase and dilute to 50ml with the mobile phase.
5.4.3.3  Reference solutions:
5.4.3.3.1        Reference solution (a):
5.4.3.3.1.1  Take 100ml beaker and dissolve 25.0mg of the ciprofloxacin hydrochloride CRS in the mobile phase and dilute to 50.0ml with the mobile phase.
5.4.3.4  Column:
5.4.3.4.1        Size:
5.4.3.4.1.1  Length=0.25m,
5.4.3.4.1.2  θ=4.6mm.
5.4.3.4.2        Stationary phase:
5.4.3.4.2.1  Base-deactivated octadecylsilyl silica gel for chromatography (5μm).
5.4.3.4.3        Temperature:
5.4.3.4.3.1  40oC.
5.4.3.5  Mobile phase:
5.4.3.5.1        Mix 13 volumes of acetonitrile and 87 volume of a 2.45g/L solution of phosphoric acid previously adjusted to pH 3.0 with triethylamine.
5.4.3.6  Flow rate:
5.4.3.6.1        1.5ml/min.
5.4.3.7  Detection:
5.4.3.7.1        Spectrophotometer at 278nm.
5.4.3.8  Injection:
5.4.3.8.1        10μL of the test solution and reference solution (a).
5.4.3.9  Run time:
5.4.3.9.1        2.3 times the retention time of ciprofloxacin.
5.4.4        Method of analysis:
5.4.4.1  Firstly prepare the test solution, reference solution and mobile phase according to the requirements.
5.4.4.2  The solutions must be free from solid particles.
5.4.4.3  Prepare the apparatus.
5.4.4.4  The mobile phase solvent mixtures must be deaerated prior to use either by boiling or by applying a partial vacuum to the solvent reservoir.
5.4.4.5  Equilibrate the column with the prescribed mobile phase, flow rate and at temperature specified until a suitable baseline is achieved.
5.4.4.6  Test solution of the mixture to be separated is now introduced into the mobile phase with the help of an injector just before entering the separating column.
5.4.4.7  As the eluate leaves the column it enters a detector, where it is continuously monitored at the specified λ.
5.4.4.8  The electrical signal obtained from detector is amplified and routes to recorder which record the developed chromatogram.
5.4.4.9  Calculate the percentage content of Ciprofloxacin HCl (C17H19ClFN3O3).
5.4.5        Limit:
5.4.5.1  98% to 102% (anhydrous substance).


6.0  REVISION LOG:
Revision No.
Effective Date
Reason
00

New SOP

7.0  REFERENCES:
7.1  The British Pharmacopoeia. Vol I., Official Monograph /Ciprofloxacin hydrochloride: 2015, pp. 563-565.
8.0  ANNEXURES:
Annexure 1: pH measurement.
Annexure 2: Observations and calculations of HPLC method.

Annexure: 1
pH measurement
Sr.#
Temperature
pH

















Result: _________________________________________________________________
















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
HCl
Hydrochloric acid
Q.C
Quality control
B.P
British pharmacopoeia
g
Grams
ml
Milliliter
oC
Degree Celsius
mm
Millimeter
g/L
Gram per liter
mg
Milligram
m
Meter
μm
Micron/ micrometer
θ
Theta
λ
Lamda
ml/min
Milliliter per minute
nm
Nanometer
μL
Microliter
%
Percentage
Vol
Volume
CRS
Chemical reference substance
QCA
Quality control active ingredient
F
Format



No comments:

Post a Comment