smrama
☆

India,
2022-05-31 13:41
(29 d 03:27 ago)

Posting: # 23032
Views: 426

## Bioanalytical method validation [Bioanalytics]

Hi all,
This forum has been a good guidance to me, to most of my queries and has been helpful to decide the uncertain situations.

But there is situation which currently I needed some guidance with and below are the details,

In long-term stability experiment, only one out of six LQC was within 85-115%
      % Bias QC1   +16.99 QC2   +19.44 QC3   -15.59 QC4   -27.04 QC5    +1.21 QC6   +16.90 % CV   19.13 % Bias  1.99

The %CV at LQC was not within 15%. Still, the results were accepted since the % bias was within 15%.

The analytical investigator said that the results can be accepted as per in-house sop, and the “Bioanalytical Method Validation Guidance for Industry” of USFDA and “Guideline on bioanalytical method validation” of EMA if the accuracy (% nominal) at each level be ± 15%.

Please clarify if this approach is acceptable.

Edit: Tabulators changed to spaces and BBcoded; see also this post #6[Helmut]
Ohlbe
★★★

France,
2022-05-31 19:30
(28 d 21:39 ago)

@ smrama
Posting: # 23034
Views: 357

## Bioanalytical method validation

Dear smrama,

» In long-term stability experiment, only one out of six LQC was within 85-115%
»       % Bias» QC1   +16.99» QC2   +19.44» QC3   -15.59» QC4   -27.04» QC5    +1.21» QC6   +16.90» % CV   19.13» % Bias  1.99
»
» The %CV at LQC was not within 15%. Still, the results were accepted since the % bias was within 15%.

Mmmm, not an easy question. That's not a common situation and I would not refer blindly to the acceptance criteria in the SOP.

Some questions:
- what were the results at the HQC level: did they pass or fail ? Did you get a high variability also for the HQC ?
- how was the CV at the LQC level during the Q&A batches: borderline, or passing easily ?
- were the 6 values obtained from 6 separate tubes, or were they pipetted from the same tube ? What was the volume of plasma in each tube and the capacity of the tube ?

If the 6 LQC were pipetted from a single tube: one hypothesis is that the sample was insufficiently mixed after thawing and before pipetting. This can happen if the tube was overfilled (not enough air above the sample for the vortex mixer to efficiently mix it). It has been described as a reason for ISR failure.

Overall: there is something wrong here and I would repeat the test. No regulatory authority would blame you for repeating the analysis in this particular situation: the test passes according to your SOP, which is very different from repeating a failed test until it passes. Make sure you document your decision and why you took it.

Regards
Ohlbe
qualityassurance
☆

Jordan,
2022-06-01 11:47
(28 d 05:21 ago)

@ smrama
Posting: # 23035
Views: 334

## Bioanalytical method validation

» The analytical investigator said that the results can be accepted as per in-house sop, and the “Bioanalytical Method Validation Guidance for Industry” of USFDA and “Guideline on bioanalytical method validation” of EMA if the accuracy (% nominal) at each level be ± 15%.

IMHO (definitely others may have different opinion than me) first of all your in house SOP should be updated to include acceptance criteria for each run with respect to calibration curve and quality control samples during validation. So for QC following criteria should be applied for each and every run during validation. "The accuracy values of the QC samples should be within ±15% of the nominal values. At least 67% of the QC samples and at least 50% at each concentration level should comply with this criterion."

Scientifically experiment should not be accepted as QCs are deviating from their nominal value which clearly indicate the stability issues or might be preparation error. As per data it seems mostly it is preparation error since the deviation of stability QCs are not in trend.

Regards,
Ohlbe
★★★

France,
2022-06-01 13:14
(28 d 03:54 ago)

@ qualityassurance
Posting: # 23036
Views: 321

## Bioanalytical method validation

Dear Qualityassurance,

» IMHO (definitely others may have different opinion than me) first of all your in house SOP should be updated to include acceptance criteria for each run with respect to calibration curve and quality control samples during validation.

We should differentiate QCs for acceptance of runs and stability samples. Even in an analytical run for the determination of stability you should include quality control samples (either fresh or within their previously established storage period), used for the acceptance of the run following the usual criteria (67% of the samples within 15% of the nominal concentration). If the run is valid: then you can look at the results of the stability samples.

If you don't have QC samples in your stability runs: if your stability samples fail, you won't know if this is because you have a stability issue or because the run failed.

It is standard practice to have acceptance criteria for the accuracy of the stability samples (as this is what the guidelines define), but I can't remember seeing acceptance criteria for their precision. But I may not have paid a particular attention and have missed it.

Regards
Ohlbe
smrama
☆

India,
2022-06-02 12:50
(27 d 04:18 ago)

@ Ohlbe
Posting: # 23039
Views: 293

## Bioanalytical method validation

Thank you
smrama
☆

India,
2022-06-02 12:51
(27 d 04:18 ago)

@ qualityassurance
Posting: # 23040
Views: 289

Thank you