Hi everybody,

Suppose I am performing Bioanalytical Method Validation (by LC-MS) as per USFDA guidelines (May 2001), and wish to first prepare a standard curve with 6 non-zero concentrations.

1. What would be the minimum number of injections of EACH standard concentration?

2. Are the results of single injection of each standard concentration acceptable as per USFDA guidelines?

3. Is unweighted regression analysis accepted as per USFDA guidelines?

4. Can we include the zero sample (matrix sample processed with the internal standard) in the statistical analysis?

5. If weighted regression is compulsary, what are the weighting factors to be tried in order to justify the statistical goodness of fit?

Thanks in advance.