Novel Approaches to Omega-3 Stability Testing Performing the p-Anisidine Value (TAV) Test by a New HPLC-UV Method Dr. Steve Li, MD, M.Sc. (Clin.Pharm) Director of Laboratory Operations Diteba, a Nutrasource Diagnostics Inc. company [email protected]www.nutrasource.ca
In the last decade, nutritional oil products such as omega-3s have grown to a multi-billion dollar industry globally. As a result, new analytical testing challenges have arisen from the current trend toward producing more attractive and more appetizing products created for a wider consumer base.
In formulating these "new and improved," better looking and better tasting products, different color and flavor additives are used, which can interfere with the most popular analytical procedure for determining the secondary oxidation of nutritional oil products, the p-anisidine value test.
In order to overcome these analytical challenges, a new alternative method for testing these additive-laden products has been established and is ready for use in this fast growing marketing segment.
Dr. Steven Li provides details about the latest innovation in omega-3 testing and its application in the omega-3 industry, at GOED Exchange 2014.
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Transcript
Novel Approaches toOmega-3 Stability Testing
Performing the p-Anisidine Value (TAV) Test by a New HPLC-UV Method
Dr. Steve Li, MD, M.Sc. (Clin.Pharm)Director of Laboratory Operations
• Free radical formation starts the chain reaction of degradation to unsaturated fatty acid.
• The major initial oxidation products, ROOH hydroperoxide, form compounds responsible for off-flavors and odors.
Such secondary products include aldehyde and ketones, such as hexanal, pentanal, and malonaldehyde.
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Oxidation Pathways of Oil Autooxidation
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Traditional p-Anisidine Value Test
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1. The anisidine value is defined as 100 times the optical density measured in a 1 cm cell of a solution containing 1 g of the substance to be examined in 100 ml of a mixture of solvents and reagents.
2. Test solution (a). Dissolve 0.500 g of the substance to be examined in iso-octane and dilute to 25.0 ml with the same solvent.
3. Test solution (b). To 5.0 ml of test solution (a) add 1.0 ml of a 2.5 g/l solution of p-anisidine R in glacial acetic acid R, shake and store protected from light.
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Traditional P-Anisidine Value Test
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4. Reference solution. To 5.0 ml of isooctane add 1.0 ml of a 2.5 g/l solution of p-anisidine in glacial acetic acid, shake and store protected from light.
5. Measure the absorbance of test solution (a) at the maximum at 350 nm using isooctane as the compensation liquid. Measure the absorbance of test solution (b) at 350 nm exactly 10 min after its preparation using the reference solution as the compensation liquid.
The p-anisidine value (p-A.V.) is given by the formula:
25 (1.2 As – Ab)pAV = -------------------------
m
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Why Do We Have Problems with the p-Anisidine Value Test?
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The Test Has No Problem to Evaluate and Compare the Oxidation Level of Simple
Marine Oil.
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Marine Oil Products Changed When they Became Important Natural Health Products
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The Challenge Comes From the Advanced Formulated Products Containing Marine Oil
p-Anisidine HPLC-UV MethodRecommended Test Criteria
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Flavoured Marine oil TAV p-anisidine
1. Marine oil containing Vitamin A yes no
2. Marine oil containing Vitamin A + D yes no
3. Marine oil containing orange Flavour yes no
4. Marine oil containing low level of orange flavour yes no
5. Marine oil containing Tangerine lime (concentration unknown) yes no
Biomedical Research Findings Support Its Usage with Important Indications
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1. Cardiovascular diseases (CVDs) are the number one cause of death globally
2. The most common disorders: hypertension (arteries), heart attacks (coronary arteries) and strokes (brain arteries). The changes are all related to blood vessels, a build-up of fatty deposits on the inner walls of the blood vessels, blood clots and bleeding
3. Importantly, these conditions are reversible and preventable4. Omega-3s EPA and DHA can help by lowering blood
triglycerides , preventing fatty deposits and reducing blood pressure
p-Anisidine Value test is to determine the secondary oxidation compounds in the oil products. This spectrophotometric test is