RESIDUAL SOLVENT METHOD VERIFICATION PROTOCOL OF CHLORPHENAMINE MALEATE Ph.Eur.

RESIDUAL SOLVENT METHOD VERIFICATION PROTOCOL OF CHLORPHENAMINE MALEATE Ph. Eur.

Superseded Protocol No.	Nil
Effective Date

TABLE OF CONTENTS: 

Sr. No.	Subject	Page No.
	Protocol Approval
	Objective
	Scope
	Responsibility
	Product profile
	Methodology
	Verification parameters
	Incident/Deviation
	Summary/Final conclusion/Recommendation
	Abbreviation
	Revision History

1. Protocol Approval :

Prepared By:

Functional Area	Name	Designation	Signature/ Date
Quality Control

Reviewed By:

Functional Area	Name	Designation	Signature/Date
Quality Assurance
Head Quality Control

Approved By:

Functional Area	Name	Designation	Signature/Date
Head QA

Authorized By:

Functional Area	Name	Designation	Signature/Date
Head Quality

• Objective:

The objective of this verification is to provide documentary evidence that analytical methodology used for residual solvents of Chlorphenamine Maleate Ph. Eur. by vendor method is consistent and reliable results within the predetermined acceptance criteria. This verification study is intended to show that the method is suitable for release of Chlorphenamine Maleate for residual solvents of manufacturing batches. The verification shall be done as per the vendor verification.

Analytical method verification will be performed by considering thefollowing parameters:

Parameters	Chlorphenamine Maleate Ph. Eur.
Specificity
Precision
System Precision
Method Precision
Intermediate Precision (Ruggedness)
System Suitability

• Scope :

The scope of this protocol is applicable for the verification of method of residual solvents of Chlorphenamine Maleate Ph. Eur.

• Responsibility of Verification Team:

Departments	Responsibilities
QC	Preparation and Review of Verification protocol.
Perform the verification as per approved protocol and recording of data.
Compilation and checking of data.
Preparation and review of Verification report.
To impart training of protocol to concerned department/persons.
QA	Review and approval of Verification protocol.
Co-ordination with QC to carry out Verification.
Review and approval of Verification report.
Head Quality	Authorization of protocol.

• Product Profile:

Category	Active Product Ingredient
Reason for Verification	First verification
Active Ingredient	Chlorphenamine Maleate Ph. Eur.
Method Reference	API Manufacturer ( In-House)
Specification Limits	Methanol     :  NMT 3000 ppm Isopropanol  :  NMT 5000 ppm O-Xylene       :  NMT 2170 ppm

• Methodology:

Solvents:

Table 1.0: Chemical

Sr. No.	Solvents	Grade
1	Methanol	GC Grade
2	Isopropanol (Isopropyl alcohol)	GC Grade
3	O-Xylene	GC Grade
4	DMSO (Dimethyl sulfoxide)	GC Grade

Residual Solvents (By GC)

      Instrument                   :           Gas Chromatography equipped with FID detector

      Detector                      :           FID

      Column                       :           30 m x 0.32 mm ID x 1.8 µm DB-624, Capillary Column or equivalent

     Column Temperature :           40˚C (Hold 10 minutes) to 250 °C @ 40°C/minutes, hold at 250°C for 5 minutes.

Injector temperature   :           250˚C

Detector temperature             :           260˚C

 Split ratio                     :           10: 1

Carrier gas                  :           Nitrogen @ 35 cm/sec linear velocity

Head space parameters:

Incubation temperature           :           95˚C

Incubation time                       :           15 minutes

Agitation speed                       :           600 RPM

Syringe temperature               :           115°C

Injection volume                      :           1.0 ml

Blank: Dimethyl sulfoxide

Preparation of standard stock solution:

Transfer 1275 µl isopropyl alcohol, 750 µl Methanol and 505 µl O-Xylene into 100.0 ml of volumetric flask containing about 80 ml of Dimethyl sulfoxide (DMSO) and mix. Dilute up to the mark with DMSO and mix.

Preparation of Standard solution or system suitability solution:

Transfer 5.0 ml standard stock solution into 100.0 ml of volumetric flask and dilute to the mark with DMSO and mix.

Preparation of test solution:

Weight accurately about 500.0 mg Chlorphenamine Maleate sample into 20.0 ml of headspace vial and add 5.0 ml of DMSO and mix.

Procedure:

Take 5.0 ml of standard solution in 6 different 20 ml head space vial. Separately inject equal volumes (1.0 ml) of standard solution (six Injections) and the test solution (duplicate) into the system. Record the chromatograms and measure the peak responses. The relative standard deviation of methanol, isopropyl alcohol and o-xylene peak response from replicate injections is not more than 15 %. Injection sequence should be blank, standard solution, blank and sample solution.

System Suitability:

% RSD    :  Area % RSD for peak due methanol, isopropyl alcohol and o-Xylene is not more than 15%.

Equilibrate the system and column; inject the solution as per following vial order or as per the requirement mentioned. Record the chromatogram and calculate the solvent content.

Table 2.0: Sequence

Sr. No.	Solutions	No of Injection to be injected in Sequence
1	Blank (DMSO)	1
2	Standard solution (Six vials)	6
3	Blank (DMSO) to avoid carry over	1
4	Test Solution	2
5	Standard Solution +Bracketing	1

                            At           Ws             5             5             P

Calculation: =   ———X———–X———-X———-x————-x D X 10⁶

                           As             100               100       Wt         100

Where,

At              :           Average area of the solvent peak in test solution.

As             :           Average area of the solvent peak in standard solution.

Ws                        :           µl of Standard.

Wt             :           Weight taken of test solution (mg).

D               :           Density in g/ml.

P               :           Purity of solvent.

• Verification parameters:

The following parameters to be perform for the verification activity.

• Specificity
  • Precision
  • System Suitability
  • Specificity:

Specificity is the ability to assess unequivocally the Analyte in the presence of components which may be expected to be present.

Specificity of test method should be established by separately injecting blank solution (DMSO), identification solution of Methanol, isopropyl alcohol, o-xylene, test solution and spiked test solution. Identification solution and spiked test solution will be prepared at specification level. Test solution to be prepared as per method of analysis.

Blank: Dimethyl sulfoxide

Preparation of Identification solutions:

Standard stock solution of Methanol :

Transfer 750 µl of Methanol into 100 ml of volumetric flask containing 80 ml of dimethyl sulfoxide (DMSO) and mix. Make up the volume to 100 ml with DMSO and mix.

Identification solution of methanol: Transfer 5.0 ml of methanol standard stock into 100.0 ml volumetric flask and make up the volume with DSMO and mix properly.

Standard stock solution of Isopropyl alcohol:

Transfer 1275 µl of Isopropyl alcohol into 100 ml of volumetric flask containing 80 ml of dimethyl sulfoxide (DMSO) and mix. Make up the volume to 100 ml with DMSO and mix.

Identification solution of Isopropyl alcohol: Transfer 5.0 ml of Isopropyl alcohol standard stock into 100.0 ml volumetric flask and make up the volume with DSMO and mix properly.

Standard stock solution of O-Xylene:

Transfer 505 µl of O-Xylene into 100 ml of volumetric flask containing 80 ml of dimethyl sulfoxide (DMSO) and mix. Make up the volume to 100 ml with DMSO and mix.

Identification solution of O-Xylene: Transfer 5.0 ml of O-Xylene standard stock into 100.0 ml volumetric flask and make up the volume with DSMO and mix properly.

Preparation of standard stock solution:

Transfer 1275 µl isopropyl alcohol, 750 µl Methanol and 505 µl O-Xylene into 100.0 ml of volumetric flask containing about 80 ml of Dimethyl sulfoxide (DMSO) and mix. Dilute up to the mark with DMSO and mix.

Preparation of Standard solution or system suitability solution:

Transfer 5.0 ml standard stock solution into 100.0 ml of volumetric flask and dilute to the mark with DMSO and mix.

Preparation of test solution:

Weight accurately about 500.0 mg Chlorphenamine Maleate sample into 20.0 ml of headspace vial and add 5.0 ml of DMSO and mix.

Spiked test solution:

Weight accurately about 500.0 mg Chlorphenamine Maleate sample into 20.0 ml of headspace vial and add 5.0 ml of standard solution and mix.

Tablet 3.0: Sequence

Sr. No	Solution	No of Injection to be injected in Sequence
1	Blank (DMSO)	1
2	Standard Solution (Six vials)	6
3	Identification solution of methanol	1
4	Identification solution of isopropyl alcohol	1
5	Identification solution of O-Xylene	1
6	Test Solution	1
7	Spiked Test Solution	1
8	Standard solution + Bracketing	1

Table 4.0 Specificity data

Sr. No	Sample	RT (min.)	RRT
1	Blank (DMSO)
2	Standard solution	Methanol
Isopropyl Alcohol
O-Xylene
3	Identification solution of Methanol
4	Identification solution of Isopropyl Alcohol
5	Identification solution of O-Xylene
6	Test Solution	Methanol
Isopropyl Alcohol
O-Xylene
Chlorphenamine

Table 5.0 Spiked test solution

Sr. No	Sample	RT (minutes)	RRT
1	Blank
2	Methanol
3	Isopropyl Alcohol
4	O-Xylene
5	Chlorphenamine

Acceptance Criteria:

1. There should be no interference of the blank and solvent content at the same retention time.
2. Solvent peaks should be well resolved from each other and observe the elution of pattern.

• Precision:
  • System Precision:

The system precision is the closeness of agreement between the responses of detector. It is        usually expressed as the standard deviation (SD) or the relative standard deviation (RSD).

Standard solution will be prepared as per method of analysis and injected six replicate injections to be injected in sequence and recorded the area response of main analyte peak. And calculate the % area RSD and % RT RSD of main analyte peak.

Table 6.0 System Precision- Repeatability of Standard Injections

Sr. No.	Methanol	Isopropyl Alcohol	O-Xylene
Peak Area	Retention time (min.)	Peak Area	Retention time (min.)	Peak Area	Retention time (min.)
1
2
3
4
5
6
Mean
SD
% RSD

Acceptance Criteria:

% RSD for peak area and retention time of replicate standard solution injections should be NMT 10.0% and 1.0% respectively.

• Method Precision:

The precision of an analytical method is the degree of agreement among individual test results when the procedure is applied repeatability to multiple samplings of homogenous sample. It is usually expressed as the standard deviation and the relative standard deviation.

Test Procedure:

Prepare the six samples of same batch as per standard analytical procedure and analyzed by spiking the known solvent content of methanol, Isopropyl alcohol and O-xylene at limit level. As such sample will be also analyzed to identify the known solvent content in sample. Obtained known solvent in as such sample will be subtracted in spiked sample to calculate the actually known spiked amount of solvent content.  Record the area on data sheet and calculate the ppm of solvent content. Mean, standard deviation and % relative standard deviation shall be reported.

Table 7.0 Method precision results spiked sample at limit level

Sr. No	Sample wt. (mg)	Methanol	Isopropyl Alcohol	O-Xylene
Peak Area	Results (ppm)	Peak Area	Results (ppm)	Peak Area	Results (ppm)
1
2
3
4
5
6
Mean
SD
%RSD

Acceptance criteria:

Calculate the solvent content in ppm for each analysis and report the mean, SD and % RSD.

% RSD of each of solvent content should be NMT 15.0%.

• Intermediate Precision ( Ruggedness ):

Intermediate precision expresses within laboratory variation with different analysts or equipment or different column/same column on different days using same batch of drug substance as per method of analysis.

Standard solution will be prepared as per method of analysis and injected six replicate injections to be injected in sequence and recorded the area response of main analyte peak. And calculate the % area RSD and % RT RSD of main analyte peak.

Table 8.0 System Precision- Repeatability of Standard Injections

Sr. No.	Methanol	Isopropyl Alcohol	O-Xylene
Peak Area	Retention time (min.)	Peak Area	Retention time (min.)	Peak Area	Retention time (min.)
1
2
3
4
5
6
Mean
SD
% RSD

Acceptance Criteria:

% RSD for peak area and retention time of replicate standard solution injections should be NMT 10.0% and 1.0% respectively.

Test Procedure:

Prepare the six samples of same batch as per standard analytical procedure and analyzed by spiking the known solvent content of methanol, Isopropyl alcohol and O-xylene at limit level. As such sample will be also analyzed to identify the known solvent content in sample. Obtained known solvent in as such sample will be subtracted in spiked sample to calculate the actually known spiked amount of solvent content.  Record the area on data sheet and calculate the ppm of solvent content. Mean, standard deviation and % relative standard deviation shall be reported.

Table 9.0 Intermediate precision results spiked sample at limit level

Sr. No	Sample wt. (mg)	Methanol	Isopropyl Alcohol	O-Xylene
Peak Area	Results (ppm)	Peak Area	Results (ppm)	Peak Area	Results (ppm)
1
2
3
4
5
6
Mean
SD
%RSD

Acceptance criteria:

Calculate the solvent content in ppm for each analysis and report the mean, SD and % RSD. % RSD of each of solvent content should be NMT 15.0%.

Table 10.0 Pooled results of analyst – I & analyst – II

Sr. No	Sample wt. (mg)	Methanol	Isopropyl Alcohol	O-Xylene
Peak Area	Results (ppm)	Peak Area	Results (ppm)	Peak Area	Results (ppm)
Analyst I
1
2
3
4
5
6
Analyst II
1
2
3
4
5
6
Mean
SD
%RSD

Acceptance criteria:

The pooled % RSD of each of solvent content of analyst I & analyst II should be NMT 15.0%.

• System Suitability:

System suitability tests are based on concept that the equipment, electronics, analytical operations and sample to be analyzed, system suitability test provide the added assurance that on specific occasion the method is given accurate and precise results.

The system suitability should be as per below mention criteria in Table 11.0.

Table 11.0

Parameters	System Suitability Criteria	Limit
Area % RSD	The area %RSD of six replicate injection of each of solvent content in standard solution	NMT – 15.0%

• Incident/Deviation:

Any Incident or Deviation observed during Analytical Method verification should be recoded and reported in verification Report.

• Summary/Conclusion/recommendation:

Final Conclusion should be drawn from analytical method verification for its use to analyze the residual solvent of Chlorphenamine Maleate by in-house method and if any differences shall be observed between methodology and experiments than accordingly will be revised and update our methodology.

1. Abbreviations

RES                :           Residual solvent

            VER                :           Verification

            P                      :           Protocol

            RSD                :           Related standard deviation

            HPLC              :           High performance liquid chromatography

            DAD                :           Diode array detector

            mL                   :           Milliliter

            mg                   :           Milligram

            PPM                :           Parts per million

            min.                 :           Minutes

            QA                   :           Quality Assurance

            QC                  :           Quality Control

            %                     :           Percentage

            ºC                    :           Degree centigrade

            hrs                   :           Hours

            µm                   :           Micrometer

            µl                     :           Microlitre

            BP                   :           British Pharmacopoeia

            RSD                :           Relative standard deviation

            RT                   :           Retention time

            NLT                 :           Not less than

NMT                :           Not more than

Ws                   :           Working standard

Vol                   :           Volume

DMSO             :           Dimethyl sulfoxide

API                  :           Active product ingredient

Revision History :

Revision No.	Details of changes	Reason for change
00	Nil	New Document