IN assay by HPLC method validation,recovery(accuracy) parameter required or not?
Yes it is required!!! Please check ICH Q2R1.
Yes it is required!!! Please check ICH Q2R1. I want to give one input here, if you are dealing with family validation (Method validation for 2 or three strengths) and if there is no major difference in fromulation except some variation in excepients quantities then you can perform full validation parameter for one strength and for the other strength you can go with limited parameters like Specificity and precision repeatability, but a comprehensive report of full validation should be attached with the report of other strengths as a ready reference.
Dear Srinivas,
Assay
Drug Substance
Several methods of determining accuracy are available:
a) application of an analytical procedure to an analyte of known purity (e.g. reference material);
b) comparison of the results of the proposed analytical procedure with those of a second well-characterized procedure, the accuracy of which is stated and/or defined.
c) accuracy may be inferred once precision, linearity and specificity have been established.
Drug Product
Several methods for determining accuracy are available:
a) application of the analytical procedure to synthetic mixtures of the drug product components to which known quantities of the drug substance to be analysed have been added.
b) in cases where it is impossible to obtain samples of all drug product components , it may be acceptable either to add known quantities of the analyte to the drug product or to compare the results obtained from a second, well characterized procedure, the accuracy of which is stated and/or defined (independent procedure,accuracy may be inferred once precision, linearity and specificity have been established.(Ref ICH Q2R1)
Dear sir,
Actually we are validating the api producut.please avise me how i can perform the recovery in assay validation with any example.
Rgds
Vasu
[quote=srinivas2202]Dear sir,
Actually we are validating the api producut.please avise me how i can perform the recovery in assay validation with any example.
Rgds
Vasu[/quote]
Dear Srivas,
Analytical methods for API should be validated unless the method employed is included in the relevant pharmacopoeia or other recognised standard reference. Methods should be validated to include consideration of characteristics included within the any recognized guidelines on validation of analytical methods. The degree of analytical validation performed should reflect the purpose of the analysis and the stage of the API production process.
Thanks
Actually we are performing Accuracy like this for API Assay validation
From linearity exercise you can calculate C i.e y-intercept
Y=mX+C
where m= slope
C- Y intercept
Y-Response
X-Concentration
Therefore
X=(Y/m)-C
You know X (theoritical ) value from the theoritical dilution or from the weight taken and you can calculate the X (Experimental) value from the above formula from the graph.
Accuracy = X(Experimental)/X(theoritical) X 100
As an additional acceptance criteria You can add Residual sum squares also .
ThanQ sir.
actually i asked not about calculation part.which compound is spiking with standard solution when standard and sample as same concentration.(i mean impurity or sample)
Rgds
vasu
HI,
Yes its really need to the validation its very important to the recovery process the accuracy level is increased that should be the increase the recovering of the addictors level.
Thank you…
if known imp limit is NMT0.1% and total rs is nmt 0.5% and sample conc.is 1mg/ml.then how i will prepare 80%,100%,120% recovery solution by spiking the imp.is there any criteria for that.
It is the riboflavin test requered to demonstrate recovery of the sprayballs??
there is a regulation to explain how to do that?:
suppose i am doing validation for api, no impurity is using in the analysis. Then how we perform recovery
[quote=prabhakar]Actually we are performing Accuracy like this for API Assay validation
From linearity exercise you can calculate C i.e y-intercept
Y=mX+C
where m= slope
C- Y intercept
Y-Response
X-Concentration
Therefore
X=(Y/m)-C
You know X (theoritical ) value from the theoritical dilution or from the weight taken and you can calculate the X (Experimental) value from the above formula from the graph.
Accuracy = X(Experimental)/X(theoritical) X 100
As an additional acceptance criteria You can add Residual sum squares also .[/quote]
When you do this,
- would you do linearity and accuracy test on the same run, can you do it on separate runs as well?
- if the same run, would you use accuracy samples in generating linearity plot?
- if the linearity pass (eg. r>0.999), is there any possibility accuracy can fail (provided, samples and testing have no errors)?
thanks,
ranjith
call me on 9676749937.
Bujji Reddy Kanchi.
1.why to to seperately. both linearity and accuracy in single sample set.
consider linearity solutions are standards for accuracy samples for the recovery calculation.
2.No need to establish linearity(correlation coefficient) for accuracy study.
3.Once linearity passess. accuracy will definitely pass.
Its better not to conduct for Validation for Related substances test. If no impurities is available.
Forced degradation study is enough for that method.
Always spike the impurity stock solutions rather than impurity standard solutions.
Usually There is no possibility of having standard and Test sample having same concentrations.
Good Question
For eg:
Test preparaton(1mg/ml)= 100mg /100 ml (which gives 1mg/ml concentration)
Impurity Standard(0.001mg/ml) = 1 mg /100 ml (this is stock solution)
Dilute 10 ml of stock to 100ml with diluent.
Preparation of 80% Recovery solution =
weigh 100 mg of test sample into a 100mL volumetric flask. add 8 ml of stock solution then make up to 100ml with diluent.
Preparation of 100% Recovery solution =
weigh 100 mg of test sample into a 100mL volumetric flask. add 10 ml of stock solution then make up to 100ml with diluent
Preparation of 150% Recovery solution =
weigh 100 mg of test sample into a 100mL volumetric flask. add 15 ml of stock solution then make up to 100ml with diluent.
Acceptance Criteria :85% to 115% to the theoretical amount added
[quote=bujjikanchi]Good Question
For eg:
Test preparaton(1mg/ml)= 100mg /100 ml (which gives 1mg/ml concentration)
Impurity Standard(0.001mg/ml) = 1 mg /100 ml (this is stock solution)
Dilute 10 ml of stock to 100ml with diluent.
Preparation of 80% Recovery solution =
weigh 100 mg of test sample into a 100mL volumetric flask. add 8 ml of stock solution then make up to 100ml with diluent.
Preparation of 100% Recovery solution =
weigh 100 mg of test sample into a 100mL volumetric flask. add 10 ml of stock solution then make up to 100ml with diluent
Preparation of 150% Recovery solution =
weigh 100 mg of test sample into a 100mL volumetric flask. add 15 ml of stock solution then make up to 100ml with diluent.
Acceptance Criteria :85% to 115% to the theoretical amount added[/quote]
What will be the validation range? 80% to 150% of standard concentration level? Can you report if you found impurity at a level outside this? How about using lower level at LOQ rather than 80%. This can extend the validation range.