Omega-3: Ingredients and Stability Monitoring

Dr. Steve Li, MD, M.Sc. (Clin. Pharm)Director of Laboratory Operations

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

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)

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)

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)

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

Omega-3 products

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Meats

Yogurts EggsNutrition Drinks

CheeseMedicinal Formulas

Bread Cosmetics

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?

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.

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

Oxidation is the main degradation pathway of omega-3

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Heat & LightOxidation

Degradation

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

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______________________________________________________

Acid Value Test

Titration Process and Color Change

Steve Lianghong Li, NDI ADRL dba Diteba

Peroxide value test

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Titration Process and Color Change

Titration Process and Color Change

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

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.

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.

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

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

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

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.

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

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

Fancy term but common logic

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Quality by Design (QbD)

1. Analytical Methods

2. Compatibility Study (3 Month)

3. Specifications

QbD principles

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Desired State of Product

ICH Q8-Pharm.Development

ICH Q10 Quality System

ICH Q9-Quality Risk

Management

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

Thank you!

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Dr. Steve Li, MD, M.Sc. (Clin. Pharm)Director of Laboratory Operations

stevel@diteba.com

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