CLINICAL AND PRE-CLINICAL ANALYSIS
MOUNTAIN INSTRUMENTAL LABORATORIES
FT. COLLINS, CO
RML has been performing state of the art analysis of pharmaceutical compounds
(API), formulated materials, and biosamples since 1980. We have multiple
LCMS and GCMS systems and, more importantly, the
experience to know how to use them to best advantage. We specialize in the
development of analyses for difficult compounds whose stability may be
problematic. We also perform VOC (volatiles) analysis. Our
particular specialty is in the bioanalysis, in biofluids and solid tissues, of
Taxol, rapamycin, and other
FDA and ICH Guidelines (1, 2, 3) require
that pharmaceutical agents be tested in vivo as well as in vitro.
RML provides both cGLP and non-GLP analyses for both research/development work
and for clinical trials assays. We offer both a "Mini-validation"
and a full, cGLP validation for new and mature drugs. This allows the
costs for high quality testing to be appropriate for the intended use.
The complete cGLP method validation is
in complete accord with FDA and ICH Guidances. In particular, we have
followed for many years the quality requirements of both entities. We
include the processing of incurred samples (4) in the validation, and have done so
since well before they were required. It just seemed to be logical to do
so as a way to check our quality internally. Now that such testing is
mandatory, it is not a change for us.
In general, FDA Guidances require that
the test be appropriate to the intended use. If the method is to be used
for regulatory submissions, this validation must be performed. This includes,
per FDA Guidance for Industry on "Bioanalytical Method Validation" (May 2001),
Matrix-based standard curves with a minimum of six non-zero concentrations
covering the expected range of the assay for the analyte(s) in question.
2. Adequate accuracy
Adequate record of controls, system suitability results, specificity,
sensitivity, linearity, and robustness.
4. Proof of stability
of the method and samples at ambient temperature, both processed and in the
autosampler, and following multiple freeze-thaw cycles.
5. Proof of the
independence of the method on matrix effects. This includes both matrix
effects on the LCMS process itself and on the extraction. It also will be
important to validate the method as the tissue or species is changed.
The complete validation report
includes the above, plus statements of how and why the final method exists, any
CAPA reports, and plots of system suitability and control results for the
multiple assays performed during the validation process (5).
If, however, the method is to provide
analyses for concept evaluation, formulation development, or other pre-clinical
efforts, the not all of the above may be needed. Then, a
"Mini-validation" may be more appropriate. The cost is significantly less
and, for research purposes, can be just as useful.
In this stage, we develop the general
method and determine the linearity, sensitivity, appropriate internal standard,
extraction, and overall sample handling. The work is performed with the
intent of making the actual validation go more smoothly. It is also at
this stage in the development process that metabolism, analyte stability, and
formulation development takes place.
Information derived form the stress
testing work is used here to assist in avoiding problems in the development of
the fully validated method.
At RML we can establish the
stress testing protocol for a particular drug, drug product, or device.
Using HPLC/MS/MS and other technologies,
we can the identify the degradants and metabolites that are formed. Note that this is not
always routine or simple.
Of particular interest is the use of our quadrupole -
linear ion trap system (API-4000Q) which allows us to search for the various
metabolites and degradants that have a major fragment in common. This
greatly improves our ability to detect low levels of such related degradants,
contaminants, and metabolites.
If additional information
on our services is needed, please contact either Dr. Patricia Sulik or
Dr. Robert Lantz.
for Industry (ICH). Many documents are available from FDA and ICH.
Of particular interest are Q1A (R2) and Q3A on Stability Testing and Impurities.
2. Guidance for Industry "Bioanalytical
Method Validation" May 2001.
This is the most important document for bioanalytical method validation.
The AAPS has a number of other articles that supplement this.
3. Bansal, S. and DeStefano, A. "Key
Elements of Bioanalytical Method Validation for Small Molecules," AAPS Journal,
9 (1) E109-E114 (2007).
4. Rocci, M. et al "Confirmatory Reanalysis
of Incurred Bioanalytical Samples," AAPS Journal 9 (3) E336-E343 (2007).
Gonzalez, G. and Herrador, M.A. "A Practical Guide to Analytical Method
Validation, Including Measurement Uncertainty and Accuracy Profiles," Trends in
Analytical Chemistry 26 (3) 227-238 (2007).
Rocky Mountain Instrumental
108 Coronado Ct.
Ft. Collins, CO 80525
17 APRIL 2012