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© 2013 AOAC INTERNATIONAL
This appendix contains three complementary documents for the
validation of dietary supplements and botanical methods:
Part I: AOAC Guidelines for Single-Laboratory Validation of
Chemical Methods for Dietary Supplements and Botanicals
Part II: AOAC Guidelines for Validation of Botanical
Identification Methods
Part III: Probability of Identification: A Statistical Model for the
Validation of Qualitative Botanical Identification Methods
PART I
AOAC Guidelines for Single-Laboratory Validation
of Chemical Methods for Dietary Supplements
and Botanicals
Contents
1 Introduction
1.1 Definitions
1.1.1 Validation
1.1.2 Method of Analysis
1.1.3 Performance Characteristics of a Method of Analysis
2 Single-Laboratory Validation Work
2.1 Preparation of the Laboratory Sample
2.2 Identification
2.3 Method of Analysis or Protocol
2.3.1 Optimization
2.3.2 Reference Standard
2.3.3 Ruggedness Trial
2.3.4 Specific Variables
a. Analyte Addition
b. Reextraction of the Extracted Residue
c. Comparison with Different Solvents
d. Comparison with Results from a Different Procedure
e. System Suitability Checks
3 Performance Characteristics
3.1 Applicability (Scope)
3.2 Selectivity
3.3 Calibration
3.3.1 External Standard Method
3.3.2 Internal Standard Method
3.3.3 Standard Addition Method
3.4 Reliability Characteristics
3.4.1 Accuracy
3.4.2 Repeatability Precision (s
r
, RSD
r
)
3.4.3 Measurement Uncertainty
3.4.4 Reproducibility Precision (s
R
, RSD
R
)
3.4.5 Intermediate Precision
3.4.6 Limit of Determination
3.4.7 Reporting Low-Level Values
3.4.8 Dichotomous Reporting
3.5 Controls
3.5.1 Control Charts
3.5.2 Injection Controls
3.5.3 Duplicate Controls
3.6 Confirmation of Analyte
3.7 Stability of the Analyte
4 Report (as applicable)
4.1 Title
4.2 Applicability (Scope)
4.3 Principle
4.4 Reagents
4.5 Apparatus
4.6 Calibration
4.7 Procedure
4.8 Calculations
4.9 Controls
4.10 Results of Validation
4.10.1 Identification Data
4.10.2 Performance Data
Appendix K: Guidelines for Dietary Supplements and
Botanicals
Under a 5-year contract (2003–2008) with the National Institutes
of Health-Office of Dietary Supplements, through the U.S. Food
and Drug Administration, AOAC undertook an effort to validate
methods for dietary supplement ingredients of interest. As part of
the initiative, AOAC adapted and revised the traditional
Official
Methods
SM
process to include single-laboratory validation (SLV).
Methods were first validated within a single laboratory to test
their suitability and ruggedness without the complications of a
multilaboratory collaborative study. SLVs proved to be an excellent
debugging tool for complex methods; problems found within one
laboratory could be dealt with so that a stronger method went on to
the collaborative study. The SLV process, thus, became a step in
preparation for the collaborative study.
The SLV guidelines were approved by the AOAC Official
Methods Board and Board of Directors in December 2002.