Introduction
a. Any Method is Good as Long as it is Validated
i. Validate yourself
ii. Compendium of Methods (i.e. Association of Official
Analytical Chemists or AOAC)
a. Buy the whole compendium (expensive)
b. Go to their website to buy method (cheaper)
b. Proximate Analysis
Analytical Method Validation
(Overview Only)
Analytical Method Validation
Documented Proof that the Analytical
Method is Suitable Under Actual
Conditions of Use
3 Types of Analytical Methods
Based on its Use
1. Quantitative Method
*You need to Quantify a Specific Substance
2. Qualitative Method
*You need to Identify a Specific Substance
3. Limit Test
*You need to Determine if a Specific
Substance is Below a Specific Limit
How to Quantitative Method
Prove that the method can Tablet Tablet Table
with 450 with 500 with 550
reliably (i.e. accurate, mg Vit C mg Vit C mg Vit C
precise) quantify expected
analyte concentrations in Accuracy
your sample
Precision
i.e. 500 mg Vit C tablet is
actually 450 to 550 mg Disclaimer
The exact protocol
will depend on the regulatory body
within a specific industry
(i.e AOAC, USP)
How do you Validate the Accuracy?
1. Spiking
2. Running your method in parallel with a
validated/official method
i. when you don’t know the conc of analyte in your sample
ii. when the official method is not perfect
How do you Validate for Precision?
1. Repeatability
*do is again and again
2. Intermediate Precision
*do it again but change a parameter (i.e. different
analyst, equipment, days)
3. Reproducibility
*do it in another laboratory
Qualitative Method
Qualitative Test for
Dextrose How to Validate?
Identify all substances that
are there
a. Check if no false
positive
b. Check if no false
negative
Limit Test
a. Check for False Positive
b. Check for False Negative
c. Check for Limit of Detection (LOD)*
How to Check for Limit of Detection (LOD)?
a. Check the Lowest b. Check the Lowest
Concentration where there is Concentration where the
Discernible Color Change (i.e. RSD is acceptable (i.e. 33%)
colorimetric test)
c. Check the Lowest Conc where
the Signal to Noise Ratio is
acceptable (i.e. 3:1)
https://www.ema.europa.eu/en/documents/scientific-guideline/ich-q-2-
r1-validation-analytical-procedures-text-methodology-step-5_en.pdf
https://manufacturingchemist.com/news/article_page/Adequately_
establishing_contamination_risk_in_drug_products/143752
What is Signal to Noise?
https://www.frague.at/your-signal-noise-
https://www.sielc.com/compound-
ratio/
caffeine.html
Things to Qualify Before Doing Method Validation
Qualification Steps
1. Instrument Qualification
This is Important
2. Reagent Qualification When You Do
Method Transfer
3. Analyst Qualification
Equipments should be qualified
based on its intended use?
Example
a. Check for Accuracy
b. Check for Linearity
c. Check for Precision
Example of Equipment
Qualification
a. Volumetric Glassware (accurate/precise)
a. Volumetric Flask
b. Volumetric Pipette
c. Analytical Balance
b. UV-VIS (linear/precise)
Materials Should be Qualified
Based on its Use
“Use
American Chemical Society (ACS)
Reagent Grade”
Amazon Price: 365 US Dollars
HCl Reagent Specifications
(Sigma Aldrich)
Analyst Should be Qualified Based
on his intended use
a. Does he know how to use glassware?
b. Does he know how to use the
balance? Quantitative Transfer?
c. Does he know how to use instrument?
Quality Control
Part of
Verifying the WHY ANALYTICAL METHOD registration
suitability of VALIDATION application
analytical
method
For submission to Compendia
If you don’t want to validate
“Use Compedial Method”
e.g. AOAC, USP, BP
Find the Analytical Protocol Suitable for Determining
the Analyte in a Particular Matrix
USP Official Methods
Official Methods of Analysis
How do we make sure we are
doing the compedial method right?
a. Qualify Instrument (USP, AOAC manual)
b. Qualify Materials (USP, AOAC manual)
c. Qualify Analyst (wala)
Tip: Use Controls when you are doing
analysis
Method Official
Do you need to do method
validation?
“Yes, you need to do at least a partial
validation (Method Verification)”
As per ISO if conc is +/- 20% expected value
If not official do If official do
validation verification
Accuracy at Accuracy at
400 mg Vit C
500 mg Vit C
500 mg Vit C
600 mg Vit C
In doing method validation/verification, who
decides on the specs ( i.e. specifications fo
accuracy and precision)
It actually depends on the risks involve, so
you decide
1. What will happen if 5% of your product is off specs?
2. Does the regulatory body have certain specifications?
References
1. Pure Appl. Chem., Vol. 74, No. 5, pp.
835–855, 2002
2. http://www.aoac.org/imis15_prod/AOAC_
Docs/LPTP/alacc_guide_2008.pdf
3. ISO 17025
Proximate Analysis
Proximate Analysis in Food
(Approximate)
Estimation of the Main Parameters Measured in
Components in Food Using Proximate Analysis
Reasonable Rapid and
Acceptable Measurement. 1. Moisture
Without the Use of
Sophisticated Equipment 2. Fat
3. Protein
4. Carbohydrate
5. Loss on Drying
6. Residue on Ignition
https://www.slideserve.com/dalit/introdu
ction-to-food-analysis
