GABAPENTIN SOP

GABAPENTIN SOP

1.0  OBJECTIVE:

To lay down a procedure for the active raw material of the Gabapentin 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  White or almost white.

5.1.1.2  Crystalline powder.

5.1.2        Solubility:

5.1.2.1  Material and equipment:

5.1.2.1.1        Glassware (5 test tubes, 1 spatula).

5.1.2.1.2        Ethanol (96%).

5.1.2.1.3        Methylene chloride.

5.1.2.1.4        Dilute acids (e.g. dil. HCl, dil. H₂SO₄).

5.1.2.1.5        Dilute alkali hydroxide (e.g. dil. NaOH).

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 purified water in test tube 1 and observe.

5.1.2.3.3        Add Ethanol (96%) in test tube 2 and observe.

5.1.2.3.4        Add methylene chloride in test tube 3 and observe.

5.1.2.3.5        Add dil. HCl in test tube 4 and observe.

5.1.2.3.6        Add dil. NaOH in test tube 5 and observe.

5.1.2.4  Observations:

5.1.2.4.1        The sample in test tube 1 containing with purified water is sparingly soluble.

5.1.2.4.2        The sample in test tube 2 containing with Ethanol (96%) is slightly soluble.

5.1.2.4.3        The sample in test tube 3 containing with methylene chloride is practically insoluble.

5.1.2.4.4        The sample in test tube 4 and 5 containing with dil. HCl & dil. NaOH is dissolved. ^(7.1)

5.2  Assay:

5.2.1        Apparatus:

5.2.1.1  HPLC apparatus.

5.2.1.2  Glassware (according to the requirement).

5.2.2        Material and reagents:

5.2.2.1  Monobasic ammonium phosphate.

5.2.2.2  Phosphoric acid.

5.2.2.3  Sodium perchlorate.

5.2.2.4  Perchloric acid.

5.2.2.5  Acetonitrile.

5.2.2.6  Purified water.

5.2.3        Sample:

5.2.3.1  14.0mg.

5.2.4        Diluent:

5.2.4.1  2.32g/L of monobasic ammonium phosphate in purified water. Adjust with phosphoric acid to a pH of 2.0.

5.2.5        Buffer:

5.2.5.1  0.58g/L of monobasic ammonium phosphate and 1.83g/L of sodium perchlorate in purified water. Adjust with Perchloric acid to a pH of 1.8.

5.2.6        Mobile phase:

5.2.6.1  Acetonitrile and buffer (24:76).

5.2.7        Standard solution:

5.2.7.1  14.0mg/ml of USP Gabapentin RS in diluent.

5.2.8        System suitability solution:

5.2.8.1  2.3mg/ml of USP gabapentin RS from the standard solution in diluent.

5.2.9        Sample solutions:

5.2.9.1  14.0mg/ml of Gabapentin in diluent.

5.2.10    Chromatographic system:

5.2.10.1          Mode: Liquid chromatography.

5.2.10.2          Detector: UV 215nm.

5.2.10.3          Column: 4.6mm x 25cm; packing L1.

5.2.10.4          Temperature: 40^o.

5.2.10.5          Flow rate: 1.0ml/min.

5.2.10.6          Injection volume: 20μL.

5.2.11    System suitability:

5.2.11.1          Samples: Standard solution and system suitability solution.

5.2.11.2          Suitability requirements:

5.2.11.2.1    Column efficiency: NLT 1900 theoretical plates for the gabapentin peak, system suitability solution.

5.2.11.2.2    Relative standard deviation: NMT 2.0% for the gabapentin peak, standard solution.

5.2.12    Analysis:

5.2.12.1          Samples: Standard solution and sample solution.

5.2.13    Procedure:

5.2.13.1          Equilibrate the column and detector with mobile phase at specified flow rate until a constant signal is received.

5.2.13.2             Separately inject equal volumes (about 20μl) of the standard solution and the sample solution into the chromatogram.

5.2.13.3             Record the spectrum.

5.2.13.4             And measure the responses for the analyte peaks.

5.2.13.5             Calculate the percentage of gabapentin (C₁₉H₁₇NO₂) in the portion of gabapentin taken:

Result = (r_U/r_S) x (C_S/C_U) x 100.

r_U = peak area from the sample solution.

r_S = peak area from the standard solution.

C_S = concentration of USP gabapentin RS in the standard solution (mg/ml).

C_U = concentration of gabapentin in the sample solution (mg/ml).

5.2.14    Acceptance criteria:

5.2.14.1    98.0% to 102.0% on anhydrous basis. ^(7.2)

6.0  REVISION LOG:

Revision No.	Effective Date	Reason
00		New SOP

7.0  REFERENCES:

7.1  The British Pharmacopoeia. Vol I., Official Monograph / Gabapentin: 2015, pp. 1053-1054.

7.2  USP38NF33 Volume-4 Official Monograph/Gabapentin: 2015, pp.: 3630-3631.

7.3  USP38NF33 Volume-1 Official Monograph/ Chromatography: 2015, pp.: 424-434.

8.0  ANNEXURES:

Annexure 1: Observations and calculations of HPLC method.

Annexure: 1

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 chromatogram. 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
Vol	Volume
g	Grams
ml	Milliliter
oC	Degree centigrade
mg	Milligram
M	Molar
%	Percentage
R	Reagent
μm	Micron/ micrometer
g/L	Gram per liter
CRS	Chemical reference solution
m	Meter
θ	Theta
mm	Millimeter
ml/min	Milliliter per minute
nm	Nanometer
μL	Microliter
Min	Minute
λ	Lamda