TEM-010

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

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

2
Linearity The correlation coefficient (r ) HPLC: equal to or greater than 0.997
for the linear concentration
TOC: equal to or greater than 0.980
range.

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

10.0%.
TOC: The recovery %RSD less than or equal to
15.0%.

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

(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:

result found
% Recovery = × 100%
result expected

(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 Peak Area of Active

(ug/mL)

(50%)

(100%)

(400%)

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

2
Coefficient of Determination (R ) =

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

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

Actual Active Measured responses Average Mean % %

Concentration result recovery RSD

1 2 3 4

(50%)

(100%)

(400%)

1. HPLC Analysis
Compare and summarise results according to acceptance criteria

2. TOC Analysis

Actual Total Measured TOC (ppm) Average Mean % % RSD

Organic TOC (ppm) recovery
Carbon
Concentration
(ppm)

1 2 3 4

(50%)

(100%)

(400%)

Compare and summarise results according to acceptance criteria

Limit of Quantitation

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

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

Linear regression line =

Correlation coefficient (R) =
2
Coefficient of determination (R ) =

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 Acceptable Rinse Volume Final HPLC Limits for

Surface Residue used to rinse Active incorporating
2
Area (cm ) Level of equipment recovery results
Active (mg) Surface

Mixing Vessel eg. 26,664 eg. 11.1325 eg.2500 mL eg. 11.1325mg/2500mL x

2
cm 0.8966 x 1000ug/mg
= 3.993 ug/mL or ppm

Holding Vessel

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 TEM-010
Issue date
Cleaning Validation – Rinsing Test
(Ref.VAL-010)
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 Date Reason for Revision Author Initial for Retrieval

# of Outdated Documents

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