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 anti-proliferative drugs.

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), the following:

1.    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 and precision.

3.    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.


1.    Guidances 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).

5.    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 Laboratories
108 Coronado Ct.
Ft. Collins, CO 80525

970-266-8108  VOICE

303-530-1169  FAX