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ANALYTICAL METHOD VALIDATION
PROTOCOL NO.: JLPL/QC/AMV/INJ/312 EFFECTIVE DATE: 05.01.2019
REVISION NO.: R0 NEXT REVIEW DATE : 05.01.2021
PRODUCT NAME GRACIDE INJECTION 1gm
LABEL CLAIM Each ml Contains :
Ceftriaxone Sodium USP
Eq to Ceftriaxone Anhydrous 1gm
APPROVAL
DESIGNATION PERSON NAME SIGNATURE / DATE
EXECUTIVE QA RAMINDER KAUR
MANAGER QC RANDEEP KAUR
MANAGER QA NAVDEEP SINGH
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ANALYTICAL METHOD VALIDATION
1. OBJECTIVE
1.1. Validation is assurance of reproducibility of results with documented evidence under given set of conditions for
particular method. Analytical method validation is carried out to ensure that there is no interference of excipients
in the analytical method, it should be linear for minimum to maximum assay, it is compatible from person to
person, The standard deviation is within the limit i.e. it should be precise, can sense minor change in assay value
i.e. accurate, sensitive and it can work in smallest concentration.
2. SCOPE:
2.1. To validate method which are used in testing of GRACIDE INJECTION 1GM
3. RESPONSIBILITY:
3.1. Validation team consisting of QA, QC, & Production
4. COPY DISTRIBUTED TO:
4.1. Quality Assurance Department
4.2. Quality Control Department.
5. REVALIDATION:
5.1. Analytical Method validation is subjected to revalidation due to following changes.
5.2. Change in formula.
5.3. Change of supplier of active materials or synthesis of drug substance.
5.4. Changes in analytical procedure.
5.5. Change of major part of Instruments.
6. ADVANTAGE OF VALIDATION
6.1. It gives documented evidence about the quality.
6.2. Variation is minimized and time consumption and retesting is Controlled.
6.3. Re processing is minimized.
6.4. It develops confidence in mind of chemist.
6.5. It is mandatory for international certification
6.6. It is a very good tool for training new entrants.
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ANALYTICAL METHOD VALIDATION
6.7. It is also tool for legal protection matters especially Retrospective Validation.
7. ASSAY: (ADD METHOD OF ANALYSIS OR STP HERE )
8. VALIDATION PARAMETERS
8.1. The method for the quantitative determination of GRACIDE INJECTION has been validated. In this method
validation study the following validation parameters were evaluated.
8.1.1. Specificity
8.1.2. Linearity and Range
8.1.3. Ruggedness
8.1.4. Accuracy (By recovery)
8.1.5. Precision
8.2. SPECIFICITY
8.2.1. Prepare a placebo sample of (A) without adding Active ingredient
8.2.2. Analyze the Placebo sample (A) & the other sample where active ingredients (WS) are added (B) by the
method of analysis mentioned above.
8.2.3. The method is said to be specific if excipients are not interfering in the analytical Method for the determination
of % of active ingredients in the Product.
8.3. OBSERVATIONS FOR SPECIFICITY FOR CEFTRIAXONE SODIUM :
S. No. SAMPLE NAME AREA IN HPLC
SAMPLE 1
1 4596412
SAMPLE 2
2 4595987
8.3.1. Acceptance criteria – Placebo interference is not more than 1.0 %
8.3.2. Conclusion: From the above results and the supporting data it is concluded that the method is specific, since
there is no interference of the excipients used in the product for the determination of active ingredient.
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ANALYTICAL METHOD VALIDATION
ANALYST QC (NAME ) MANAGER QC (NAME )
9. LINEARITY AND RANGE:
9.1. The Linearity of an analytical method is its ability to elicit test result that are directly, or by a well defined
mathematical transformation proportional to the concentration of Analyte in samples within a given range.
9.2. Methodology:
9.3. Experimental
9.4. Preparation of sample of CEFTRIAXONE SODIUM FOR INJECTION
9.4.1. As per method of analysis sample taken for analysis is CEFTRIAXONE SODIUM FOR INJECTION
9.4.2. Take the Powder of sample to get 80%, 90%, 100%, 110% and 120% of stated amount of Ceftriaxone sodium
For injection respectively in method of analysis in linearity test.
9.4.3. Analyze these samples by the above-mentioned analytical method.
9.4.4. Plot a graph of concentration against Area.
9.4.5. If the graph obtained for each active is a straight line and Correlation coefficient is not less than 0.999 then the
method is said to be Linear.
9.5. OBSERVATIONS: FOR Ceftriaxone sodium
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ANALYTICAL METHOD VALIDATION
CEFTRIAXONE SODIUM INJECION
4600000
4599000
4598000
4597000
4596000
4595000
4594000
SAMPLE 1 SAMPLE 2 SAMPLE 3 SAMPLE 4
Series1
9.6. Conclusion - From the above results, supporting data and the graph it is concluded that the method is Linear.
ANALYST QC (NAME ) MANAGER QC (NAME )
10. RUGGEDNESS:
10.1. Take samples of CEFTRIAXONE SODIUM INJECTION containing 100% concentration of active ingredient.
10.2. Get the samples analyzed from two different validated analysts in duplicate.
10.3. Compare the results obtained from the two different analysts.
10.4. If the variance is not more than 0.5% then the method is said to be RUGGED.
S. NO. SAMPLE ASSAY RESULTS BY TWO OF ANALYST
VARIANCE
ANALYST -1 ANALYST-2
1 A SAGAR ARORA PAWANDEEP 0.92%
2 B SAGAR ARORA PAWANDEEP 0.94%
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ANALYTICAL METHOD VALIDATION
11. Observation: The variance is NMT 1.0 %.
11.1. Note: Variance can be calculated by excel sheet.
11.2. Conclusion: From the above results and the supporting data it is concluded that the method is rugged.
ANALYST QC (NAME ) MANAGER QC (NAME )
12. ACCURACY (BY RECOVERY):
12.1. Accuracy of an analytical method is the closeness of test results obtained by that method to the true value.
12.2. Accuracy may often be expressed as percent recovery, by the assay, of known added amount of analyte.
12.3. The accuracy of the method shall be determined by adding known amount of Ceftriaxone sodium for Injection
in the sample.
12.4. Prepare three samples in triplicate containing known amount of ceftriaxone sodium for Injection . add extra
(Overage) 2%, 3% and 10% of respectively. Label them as A, B and C.
12.5. If the results obtained for A, B and C are sufficiently different and sensitive to the addition of 2%, 3% and 10 %
of active ingredient and percentage recovery is between 98% to 101% then the method is said to be accurate.
12.6. OBSERVATION TABLE:-FOR API (Ceftriaxone Sodium)
Concentration level Mass Observed Results Assay initial % % Recovery
+2 % of API 1020mg 101.65% 99.58% 2.07%
+3 % of API 1030mg 103.17% 100.14% 3.03%
+10 % of API 1100mg 109.91% 99.71% 10.20%
12.7. Limit for % recovery – 98.0 % to 101.0 %
12.8. Conclusion: From the above data difference in assay of sample A, sample B and sample C are within the
acceptable limit and sufficiently different and sensitive to the addition of extra active ingredient and hence, it is
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ANALYTICAL METHOD VALIDATION
concluded that the method is accurate
13.
ANALYST QC (NAME ) MANAGER QC (NAME )
14. PRECISION:
14.1. Precision of an analytical method is the degree of agreement among individual test results when method is
applied repeatedly to multiple sampling of a homogenous sample.
14.2. The precision of analytical method is usually expressed as standard deviation or relative standard deviation of a
series of measurement.
14.3. ICH documents recommended that repeatability should be assessed using a minimum of nine determinations
covering the specified range for the procedure i.e. three concentration and three replicate of each concentration
or using a minimum of 6 determination at 100% of the test concentration.
14.4. Take sample of Ceftriaxone Sodium For Injection Analyze these samples six times by the above mentioned
analytical method. Find out the standard deviation and relative standard deviation from the mean value. If the
relative standard deviation is within limit then the method is said to be precise.
14.5. The relative standard deviation from the mean value. If the relative standard deviation is NMT 2.0% then the
method is said to be precise.
15. Observations: - For CEFTRIAXONE SOIUM FOR INJECTION 1GM
15.1. RELATIVE STANDARD DEVIATION: NMT 2.0% Supporting data is attached herewith.
15.2. CONCLUSION: From the above results and supporting data it is concluded that the method is said to be
precise.
ANALYST QC (NAME ) MANAGER QC (NAME )
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ANALYTICAL METHOD VALIDATION
16. FINAL CONCLUSION:
16.1. From all the above data and graph it is concluded that the method of CEFTRIAXONE SODIUM FOR
INJECTION is independent of the Excipients and is linear, rigid, precise and accurate under normal working
conditions and hence the method is analytically validated.
17. ABBREVIATIONS:
17.1. QC: Quality Control
17.2. QA: Quality Assurance
17.3. RSD: Relative Standard Deviation
17.4. AMV: Analytical Method Validation
17.5. NMT: Not More Than.
(Sign / Date) (Sign / Date)
(Sign / Date)
Executive QC Manager QC Manager QA
Prepared By Checked By Approved By
(RAMINDER KAUR ) (RANDEEP KAUR) (NAVDEEP SINGH )
