1.What is freez thaw study and its importance 2.Which guidelines explains freez thaw study.
In the Bioequivalence studies (in-vivo) on human volunteers, the Bioanalytical method (usually LC-MS/MS methods) used to quantify the drug (analyte) in human matrix (plasma, serum, blood etc.) should be validated according ICH -M10 guidelines and also as per regulatory requirements of that specific country. As a part of Bioanalytical method validation to assess the impact of repeatedly removing biological samples (in the matrix of plasma, serum, blood etc.) from frozen storage (deep freezers at -20 or -80 deg. C), the stability of the analyte should be assessed after multiple cycles of freezing and thawing. Low and high stability QC standards should be thawed and analysed according to the same procedures as the study samples (in the matrix of plasma, serum, blood etc). Stability QCs should be kept frozen for at least 12 hours between the thawing cycles. Stability Quality Control samples (similar to internal standards with known concentration of analyte) for freeze-thaw stability should be assessed using freshly prepared Calibration standards and QCs or QCs for which stability has been proven. The number of freeze-thaw cycles validated should equal or exceed that of the freeze-thaw cycles undergone by the study samples, but a minimum of three cycles should be conducted.
Individual Regulatory authorities have adopted the ICH M10 guideline such as USFDA, EMA, MHRA & other countries such as Brazil (ANVISA) etc.
Reference: BIOANALYTICAL METHOD VALIDATION -ICH (M10), Feb 2019, Draft version
The guideline does not describe the method of Freeze-Thaw stability study . At what temp. for how much period , the sample should be stored ? How much time to allow for thaw and when to test ? Please give details if you have .
The number of freeze-thaw cycles validated should be equal or exceed that of the freeze-thaw cycles undergone by the study samples, but a minimum of three cycles should be conducted.
The experimental conditions during freez-thaw cycles validation should be the same as those used in regular analysis of plasma / biological samples, such as temperature, time to allow for thawing plasma samples and time required for its analysis. Generally the temperature of freezing should be the same as that of all study samples, Calibration curve (CC) and QC samples being used to stored during routine analysis of clinical (Bioequivalece) study samples. This is based on stability of drug (analyte) in plasma samples (either -20 or -80 deg C).
Therefore, all the conditions used during subject samples analysis (human volunteers plasma samples) must be the same as those used in Bioanalytical method validation.
Thanks @sunilrbudhkar for your prompt reply . .
Is it for plasma products only or can we apply this stability study for any product ?
This is applicable to biological samples of Clinical studies, mainly Bioavailability, Bioequivalence studies (also called pharmacokinetic Pk studies).These are Plasma, Serum, Whole blood samples which are frozen at low temperatures at -20 or -80 deg C due to stability characteristics of the drugs / analytes in such biological matrix. Therefore, these samples are required to be thawed at room temperature before taken for analysis.