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Omega-3: Ingredients and Stability Monitoring
Dr. Steve Li, MD, M.Sc. (Clin. Pharm)Director of Laboratory Operations
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Outline
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1. Omega-3 and its Product Development
2. Quality Control and Issues with Omega-3 Product Testing
3. Stability Study
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What is omega-3?
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Omega-3 fatty acids
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They are polyunsaturated fatty acids
α-linolenic acid (18:3, n-3; ALA), eicosapentaenoic acid (20:5, n-3; EPA), and docosahexaenoic acid (22:6, n-3; DHA)
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Essential fatty acids
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ω-3 fatty acids:– α-Linolenic acid or ALA (18:3n-3)
• ω-6 fatty acids:– Linoleic acid or LA (18:2n-6)
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Fatty acid synthesis in the human body
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Why do we need it?
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Omega-3 FA and our health
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Omega-3 products
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Common omega-3 dosage forms
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The common active ingredients added in omega-3
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1. Minerals• Calcium• Selenium• Chromiun• Zinc
2. Vitamins• Vitamin D• Vitamin A• Coenzyme Q10
3. Antioxidants• Vitamin E (alpha and
gamma tocopherols)• Vitamin C and
derivatives (Ascobyl palmatate)
4. Others (Antibiotics, Hormones or Biologics)
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Formulated products
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New product development
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• Applications (Enteral, Parenteral and Topical)• Taste and Appearance
• Bioavailability • Multiple health benefits
• Stability
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Omega-3 products
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Meats
Yogurts EggsNutrition Drinks
CheeseMedicinal Formulas
Bread Cosmetics
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Now let’s see if we really do quality control for the products
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3 QbD things useful for omega-3-like products
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1. Can a product be easily tested? a) Is there a reliable method or compendial method
for testing a product?
b) Has a method been verified for a product?
2. Is the formulation compatible?
3. Is a product stable in storage?
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Challenges of quality control methods for testing omega-3 products
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• Oil Based Matrix---- Solubility and Homogeneity
• Continuous Oxidation Process----Stability • Complexity of Formulation----Interfering
Additives or Ingredients for testing
• Less Specific----Less Supportive.
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Test methods for omega-3 fatty acids
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• Fatty Acid Oxidation
• Release Testing of API and Products
• Microbiological Tests
• Heavy Metal Tests
• Pesticides Tests
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Oxidation is the main degradation pathway of omega-3
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Heat & LightOxidation
Degradation
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Omega-3 fatty acid oxidation tests
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• Acid Value: Titration method, colorimetric
• p-Anisidine Value: UV 350 nm• True Anisidine Value: HPLC UV• Peroxide Value: Titration method,
colorimetric
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Acceptable limits for oxidative stability parameters in marine oils
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(Recommended Reporting Threshold) ________________________________________
Acid Value NMT 3 mg KOH/Kg
Peroxide value (PV) NMT 5 mEq/Kg
Anisidine value (AV) NMT 20
TOTOX value NMT 26 (2 x PV + AV )
True Anisidine Value TAVTM70% of Anisidine Remaining______________________________________________________
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Acid Value Test
Titration Process and Color Change
Steve Lianghong Li, NDI ADRL dba Diteba
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Peroxide value test
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Titration Process and Color Change
Titration Process and Color Change
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P-Anisidine value test
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p-Anisidine HPLC-UV method
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Typical Chromatogram of QL(S/N=15, 3.162 µg/mL)
Channel @ W2996 PDA 254.0 nm at 1.2
3.30
5
p-A
nisi
dine
- 4.
855
AU
0.000
0.001
0.002
0.003
0.004
0.005
Minutes
1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00
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Quantitative assay method for omega-3
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• Most common method is GC-FID or GC-MS
• LCMS can be an good alternative.
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Stability Studies
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Unique features of omega-3 containing products and stability testing
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• Most of them are softgel capsules, oil liquid or oil based suspension
• Most common degradation is oxidation and the degradation impurities are mixtures of multiple compounds.
• It is extremely difficult to have a stability indicating method for Omega-3 fatty acids since the degradation products are variable and difficult to be easily identified and quantified using simply analytical methods.
• Most of current methods for testing oxidation products are not specific and directly quantitative for the degradation products.
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Challenges in quality control and stability study of omega-3 products
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• Oil Based Matrix-Homogeneity• Unstable Due to Continuous Oxidation Process • Need More Efficient and Specific QC Analytical
Methods • Formulated Product with Interfering Additives
or Ingredients for the Current Test Methods
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General storage conditions in stability studies
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• Storage ICHWalk-in and Reach-in Chambers
• Storage Conditions25°C/60% RH30°C/65% RH40°C/75% RHPhotostability
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Specifications in stability study
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• Stability studies should include testing of those attributes of the API that are susceptible to change during storage and are likely to influence quality, safety and/or efficacy.
• The testing should cover, as appropriate, the physical, chemical, biological and microbiological attributes. A guide as to the potential attributes to be tested in the stability studies.
• Validated stability-indicating analytical procedures should be applied.
• Whether and to what extent replication should be performed will depend on the results from validation studies
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Stability and testing frequency
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• For APIs with a proposed re-test period or shelf-life of at least 12 months, the frequency of testing at the long-term storage condition should normally be every three months over the first year, every six months over the second year, and annually thereafter throughout the proposed re-test period or shelf-life.
• At the accelerated storage condition , a minimum of three time points, including 0, 3, and 6 month.
• When testing at the intermediate storage condition a minimum of four time points, including the initial and final time points (e.g. 0, 6, 9 and 12 months), from a 12-month study is recommended.
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Typical example of stability program
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ϭϮŵŽŶƚŚƐ
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ϲŵŽŶƚŚƐ
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Typical stability program
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Compatibility studies (3 month)
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Total EPA+DHA CoQ 10p-Anisidine
ValueTAV (%)
Peroxide Value
(mEq/Kg)
Acid Value (mg
KOH/g)0 98.5 102 120 85 6 1 1.003 96.8 96 160 76 10 1.8 1.52
Total EPA+DHA 96Micro Extract Ap-Anisidine
ValueTAV (%)
Peroxide Value
(mEq/Kg)
Acid Value (mg
KOH/g)0 98.5 94 12 85 4 0.5 2.503 96.8 72 160 76 10 1.8 8.00
Time (Month)
Time (Month)
% LC Oxidation
% Total Impurities
Formula 5
% LC Oxidation
% Total Impurities
Formula 1
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Example of stability summary
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% LCMonth Batch 1 Batch 2
0 98.5 98.53 96.8 966 94.7 949 93.3 92.5
12 92.3 8618 92.9 8024 91.6 7636 90.3 75
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Fancy term but common logic
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Quality by Design (QbD)
1. Analytical Methods
2. Compatibility Study (3 Month)
3. Specifications
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QbD principles
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Desired State of Product
ICH Q8-Pharm.Development
ICH Q10 Quality System
ICH Q9-Quality Risk
Management
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Fate of drug product and philosophy
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• Only Very Small % Drug Products Get FDA Approval
Fate of Product
Product Philosophy
Formula
Quality Control
Tox
Application Confirmation
Fate of Product
Product Philosophy
Formula
QbD
Quality Control
Tox
Application Confirmation
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Thank you!
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Dr. Steve Li, MD, M.Sc. (Clin. Pharm)Director of Laboratory Operations
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