TEM-010 Issue date

Cleaning Validation – Rinsing Test (Ref.VAL-010)

File Location: Date Printed: Page 1 of 9

Project number: Protocol Number: Product/Active: Process Line:

RINSING RECOVERY STUDIES

1. Test Description

This test is to be conducted to document the validation of Total Organic Carbon (TOC) analysis method for use in measuring samples for cleaning validation. A parallel analysis of rinse samples will be carried out and compared using TOC and High Performance Liquid Chromatography (HPLC) analysis. Standard solutions will be applied to stainless steel plates, dried and the residue removed by rinsing using an appropriate solvent (usually water). Assessment of Linearity, Accuracy (recovery), LOQ, LOD and precision of the rinsing method will be determined.

2. Test Objective

1. Determine the linearity and precision of a series of standards rinsed from a stainless

steel plate or container and measured by TOC and HPLC over a known concentration range.

2. Determine the accuracy (recovery) of a series of standards rinsed from a stainless steel plate or container and measured by TOC and HPLC over a known concentration range.

3. Determine the Limit of quantitation and limit of detection of a series of standards measured by TOC and HPLC over a known concentration range.

4. Determine correlation between HPLC and TOC analysis.

5. Determine the final HPLC and TOC rinsing limits based on recovery studies.

3. Acceptance Criteria

Test Objective Measured Response Acceptance Criteria

Linearity The correlation coefficient (r2)

for the linear concentration range.

HPLC: equal to or greater than 0.997

TOC: equal to or greater than 0.980

Precision % RSD HPLC: The recovery %RSD less than or equal to 10.0%.

TOC: The recovery %RSD less than or equal to 15.0%.

TEM-010 Issue date

Cleaning Validation – Rinsing Test (Ref.VAL-010)

File Location: Date Printed: Page 3 of 9

(ii) Carefully pour the required volume of rinse solvent into the container making certain that all the surface area is contacted by the solvent (the rinse solvent should be at the same temperature as used in the manufacturing environment). The volume of rinse solution used in the recovery experiments should be proportional to the rinse volume used in the manufacturing area. Analyze the resultant rinsate samples by HPLC and TOC. Residue recovery will be calculated as follows:

% Recovery = result found

result expected× 100%

(iii) Tabulate results as detailed in results section and plot linear regression and document results of linear regression analysis for both HPLC and TOC analysis.

5. Results

Linearity - Rinsing Active Residue

1. HPLC Analysis

Active Concentration (ug/mL)

Peak Area of Active

(50%)

(100%)

(400%)

Plot Concentration v’s Peak Area (calculate linear regression)

Coefficient of Determination (R2) =

TEM-010 Issue date

Cleaning Validation – Rinsing Test (Ref.VAL-010)

File Location: Date Printed: Page 5 of 9

Accuracy - Rinsing active residue

For each measured response over the concentration range examined in the Linearity test calculate % recovery and % RSD for each of the triplicate samples. where % recovery = result found x 100 result expected

1. HPLC Analysis

Compare and summarise results according to acceptance criteria

2. TOC Analysis

Actual Total Organic Carbon Concentration (ppm)

Measured TOC (ppm)

Average TOC (ppm)

Mean % recovery

% RSD

1 2 3 4

(50%)

(100%)

(400%)

Compare and summarise results according to acceptance criteria

Limit of Quantitation

Actual Active Concentration

Measured responses

Average result

Mean % recovery

% RSD

1 2 3 4

(50%)

(100%)

(400%)

TEM-010 Issue date

Cleaning Validation – Rinsing Test (Ref.VAL-010)

File Location: Date Printed: Page 7 of 9

Plot Average Recovery (Active) ppm HPLC V’s Average recovery (Total Product Carbon) ppm TOC

Linear regression line =

Correlation coefficient (R) =

Coefficient of determination (R2) =

Determining Final HPLC and TOC Rinsing Limits Based on Recovery Studies HPLC Rinsing Limits Determine the lowest and most conservative active rinse recovery value obtained during the method validation study for HPLC analysis. Lowest HPLC rinse recovery value = ____________

The final limits for the active residue for rinse solutions are:

Equipment Equipment Surface Area (cm

2)

Acceptable Residue Level of Active (mg)

Rinse Volume used to rinse equipment Surface

Final HPLC Limits for Active incorporating recovery results

Mixing Vessel eg. 26,664 cm

2

eg. 11.1325 eg.2500 mL eg. 11.1325mg/2500mL x 0.8966 x 1000ug/mg

= 3.993 ug/mL or ppm

Holding Vessel

TEM-010 Issue date

Cleaning Validation – Rinsing Test (Ref.VAL-010)

File Location: Date Printed: Page 9 of 9

6. Comments

7. Conclusion

8. Attachments

Validation Discrepancy Forms - nil

Completed Operational Qualification Test protocol:

Position Signature Date

Documented by:

Checked by:

Approved by:

DOCUMENT REVISION HISTORY

Revision #

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