REGULATION RTCA 11.03.
39: 06 CENTRAL AMERICAN TECHNICAL REGULATION PHARMACEUTICAL
PRODUCTS. VALIDATION OF ANALYTICAL METHODS FOR THE EVALUATION OF DRUG QUALITY
1. OBJECT This technical regulation aims to establish guidelines for the validation of
physicochemical and microbiological analytical methods used in the quality control of
medicines.
2. SCOPE OF APPLICATION The guidelines of this technical regulation must be applied to all
unofficial and official analytical methods used for the quality control of medicines in order to
comply with current good manufacturing practices and good laboratory practices regulations.
Quality control laboratories using official analytical methods should only verify the linearity and
precision of the system.
3. DEFINITIONS
3.1 Regulatory authority: Entity responsible for the Sanitary Registration and/or Surveillance of each
Member State.
3.2 Specificity; selectivity: The capacity to evaluate, measure, and identify simultaneously or
separately the analytes of interest unequivocally without interferences from impurities,
degradation products, related compounds, excipients, or other foreseeable substances present in
the sample matrix.
3.3 Accuracy; truthfulness: It is the proximity between the test results obtained through that
method and the true value.
3.4 Impurities: Substances foreign to the qualitative-quantitative formula that may originate from
the manufacturing and storage processes, including the degradation of raw materials (active
ingredients and auxiliary pharmaceutical products) and dosage forms. The presence of
contaminants in the finished product is indicative of non-compliance with good manufacturing
practices. These contaminants are considered impurities in some cases, and limits can be
established for them.
3.5 Range: The breadth between the lower and upper concentrations of the analyte (including those
levels), in which the analyte can be determined with an adequate level of precision, accuracy, and
linearity using the method as described.
3.6 Quantification limit: The minimum amount of the analyte in a sample that can be quantitatively
determined with acceptable precision and accuracy. It is a parameter of quantitative analysis for
low levels of compounds in sample matrices and is particularly used for the determination of
impurities and degradation products.
3.7 Detection limit: The minimum amount of analyte in a sample that can be detected by a single
measurement, but not necessarily quantified with an exact value. It is commonly expressed as the
analyte concentration.
3.8 Linearity: The ability to obtain test results that are proportional either directly or through a well-
defined mathematical transformation to the analyte concentration in samples within a given range.
3.9 Analytical method: A specific adaptation of an analytical technique for a selected measurement
purpose, in which the material resources and procedure are identified.
3.10 Viable microorganisms: Microscopic organisms such as bacteria and fungi capable of
reproduction and leading to colony formation.
3.11 Neutralization: Process that blocks the effect of a preservative agent by the addition of a
chemical agent or by dilution.
3.12 Analytical performance parameters; validation test parameters: Validation characteristics that
need to be evaluated and typically correspond to the following list: accuracy, precision, specificity,
detection limit, quantification limit, linearity, and linearity range.
3.13 Precision: Expresses the degree of agreement between a series of individual measurements
obtained from multiple samplings of a single original homogeneous sample or from several samples
obtained by dilution of the sample under established conditions. There are three types of
determination: repeatability, intermediate precision, and reproducibility.
3.14 Analytical procedure: A detailed description of the necessary steps to apply an analytical
method.
3.15 Official analytical procedure: A detailed description of the necessary steps to apply a
standardized and validated analytical method contained in official reference bibliographies, as
harmonized by the Countries of the Central American Region (Resolution 93-2002, COMIECO 24,
September 2002).
3.16 Unofficial analytical procedure: A detailed description of the necessary steps to apply an
analytical method developed by the manufacturer for verifying the quality of their product.
3.17 Pathogenic microorganism detection test: Determines the presence of specific harmful
microorganisms in a sample.
3.18 Antimicrobial effectiveness test: Determines the effectiveness of preservative agents in the
sample.
3.19 Sterility test: Determination of the absence of viable microorganisms in the sample.
3.20 Microbial limit test: It is the count of viable microorganisms present in a non-sterile sample to
determine if it is within established limits.
3.21 Growth promotion test: Determines the ability of the culture medium to allow the growth of
specific viable microorganisms.
3.22 Microbiological recovery test: Determines the neutralization caused by preservative agents or
other agents that cause inhibition of the growth of viable microorganisms.
3.23 Validation: Establishment of documentary evidence that an analytical procedure will lead with
a high degree of certainty to obtaining precise and accurate results within the previously established
specifications and quality attributes.
3.24 Validation of an analytical procedure: Procedure to establish documentary evidence that
scientifically demonstrates that an analytical method has the performance characteristics that are
suitable to meet the requirements of the intended analytical applications. It involves demonstrating
the determination of sources of variability and systematic and random error of a procedure, not
only within calibration but in the analysis of real samples.
[Link] PROCEDURES SUBJECT TO VALIDATION The following chemical, physical, and
microbiological analytical procedures must be validated:
4.1 Category I: Quantitative tests of the content of the active ingredient(s), constitute chemical or
microbiological procedures that measure the analyte(s) present in a specific sample.
4.2. Category II: Tests for determining the content of impurities or limit values for impurity control.
They can be quantitative tests or a qualitative test to determine if the impurity is present in the
sample above or below a specified limit value. Either of the two aims to reflect the true amount of
impurities in the sample. The validation parameters required for a quantitative test are different
from those of a qualitative compliance test.
4.3 Category III: Physical-chemical performance tests. These are testing procedures that measure
characteristics specific to the performance of the drug, such as the dissolution test. The validation
characteristics are different from those of other tests, although they may overlap.
4.4 Category IV: Identification tests. Tests conducted to ensure the identity of an analyte in a sample.
This is typically done by comparing a property of the sample against that of a reference standard,
such as spectra, chromatographic behavior, chemical reactivity, and microcrystalline tests. 4.5
Microbiological tests: Tests conducted to ensure the microbial quality of the drug. The merit
parameters for each test category are detailed in Tables 1 and 2.
Table 1. Performance parameters of physico-chemical analytical procedures and microbiological
potency.
Impurity
Impurity Physical-
Active Content
Category Parameter Content Chemical Identification
Ingredient(s) Qualitative-
Quantitative Performance
Limit
Performance
I Yes * * * *
Parameter
II Accuracy Yes Yes * Yes Yes
II Precision Yes Yes Yes Yes Yes
II Specificity Yes Yes Yes Yes Yes
II Detection Limit No No Yes Yes No
Quantification
II No Yes No Yes No
Limit
II Linearity Yes No No Yes No
II Range Yes Yes Yes No No
*Requirement may vary depending on the nature of the assay.
Table 2. Performance parameters of microbiological procedures
Microbial limit and detection of Antimicrobial
Test type Sterility
pathogenic microorganisms effectiveness
Effectiveness of the growth
YES YES YES
medium (Growth promotion
Effectiveness of the preservative
YES YES YES
agent neutralization method
5. DOCUMENTATION TO BE SUBMITTED FOR THE REVIEW OF VALIDATED ANALYTICAL METHODS
For the review by the Regulatory Authority of validated methods, the validation study report
must contain the following documentation:
a. Detailed description of the analytical procedure.
b. Description of the performance parameters evaluated according to tables 1 and 2.
c. Evaluation and statistical calculations for the verification of the performance parameters. d.
Summary of the instrumental results obtained (areas or printed absorptions).
e. Summary of the validation results must be in Spanish or properly translated. f. Conclusions
must be delivered in Spanish or properly translated.
6. REJECTION OF THE SUBMITTED DOCUMENTATION The documentation will be rejected if
technical or scientific inconsistencies are detected, inadequate statistical support for
conclusions, or failures in the experimental design or conduct of the validation. Once the
Regulatory Authority communicates the rejection of validation documentation, the interested
party has the time established by the legislation of each Central American country to address
the non-conformity.