Add the following:

Fish Oil Containing Omega-3 Acids Delayed-Release Capsules

DEFINITION

Fish Oil Containing Omega-3 Acids Delayed-Release Capsules are enteric-coated Capsules that contain NLT 95.0% and NMT 105.0% of the labeled amount of Fish Oil Containing Omega-3 Acids where Fish Oil Containing Omega-3 Acids is the purified, winterized, and deodorized fatty oil obtained from fish of the families Engraulidae, Carangidae, Clupeidae, Osmeridae, Scombroidae, and Ammodytidae. The omega-3 acids are defined as the following: alpha-linolenic acid (C18:3 n

3), moroctic acid (C18:4 n

3), eicosatetraenoic acid (C20:4 n

3), eicosapentaenoic acid (EPA) (C20:5 n

3), heneicosapentaenoic acid (C21:5 n

3), docosapentaenoic acid (C22:5 n

3), and docosahexaenoic acid (DHA) (C22:6 n

3). It contains NLT 28.0% (w/w) of total omega-3 acids, expressed as free acids, consisting of NLT 13.0% of EPA and NLT 9.0% of DHA. Suitable antioxidants in appropriate concentrations may be added.

IDENTIFICATION

• A. The oil contained in the Capsules meets the requirements for the following test: The retention times of the peaks for eicosapentaenoic acid methyl ester and docosahexaenoic acid methyl ester obtained in the chromatogram of Test Solution 2 in the test for Content of EPA and DHA correspond to those for the respective compounds in the chromatogram of Standard Solution 1. The sum of the area for EPA and DHA methyl esters is NLT 22% of the total detected area for the methyl esters, and no other peak has an area higher than 20% of the total detected area for the methyl esters. The chromatogram of Test Solution 2 exhibits at least 15 additional peaks at the retention times of the methyl esters of unsaturated fatty acids exhibited in Standard Solution 2.

STRENGTH

• Content of Fish Oil: Weigh NLT 10 Capsules in a tared weighing bottle, carefully open the Capsules, without loss of shell material, and transfer the combined Capsule contents to a 100-mL beaker. Remove any adhering substance from the emptied Capsules by washing with several small portions of 2,2,4-trimethylpentane. Discard the washings, and allow the empty Capsules to dry in a current of dry air until the 2,2,4-trimethylpentane is completely evaporated. Weigh the empty Capsules in the original tared weighing bottle, and calculate the average net weight per Capsule.

Acceptance criteria: 95.0%–105.0% of the labeled amount

• Content of EPA and DHA

(See Fats and Fixed Oils

401

, Omega-3 Fatty Acids Determination and Profile.)

Analysis

Samples: Standard Solution 1, Standard Solution 2, Test Solution 1, and Test Solution 2

Identify the retention times of the relevant fatty acid methyl esters peaks by comparing the chromatogram of Standard Solution 2 with the reference chromatogram supplied with the USP Fish Oil RS. Identify the retention time for the internal standard peak from Test Solution 2 by comparing with that of Test Solution 1.

Calculate the percentage of EPA or DHA in the portion of fish oil containing omega-3 acids taken from the Capsules:

Result = (RU/RS) × (WS/WU) × F × 100

RS	=	= ratio of peak responses of either EPA or DHA relative to the internal standard from Standard Solution 2
WS	=	= weight of either USP Docosahexaenoic Acid Ethyl Ester RS or USP Eicosapentaenoic Acid Ethyl Ester RS used to prepare Standard Solution 1 (mg)
WU	=	= weight of the fish oil containing omega-3 acids taken to prepare Test Solution 2 (mg)
F	=	= factor to express the content of DHA (0.921) and EPA (0.915) as free fatty acids
RU	=	= ratio of the peak response of either EPA or DHA to the corrected peak response of the internal standard from Test Solution 2, calculated as follows:

RU = 1/[(rU2/rT2)

(rU1/rT1)]

rU2	=	= response of the peak at the locus of the internal standard from Test Solution 2
rT2	=	= peak response of EPA or DHA from Test Solution 2
rU1	=	= response of any peak at the locus of the internal standard from Test Solution 1
rT1	=	= peak response of EPA or DHA from Test Solution 1. [Note—If no peak is found at the locus of the internal standard, RU = rT2/rU2. ]

Acceptance criteria: NLT 13.0% (w/w) of EPA and NLT 9.0% (w/w) of DHA

• Content of Total Omega-3 Acids

(See Fats and Fixed Oils

401

, Omega-3 Fatty Acids Determination and Profile.)

Analysis: Proceed as directed in the test for Content of EPA and DHA.

Calculate the percentage of the total omega-3 acids in the portion of fish oil containing omega-3 acids taken from the Capsules:

Result = EPA + DHA + [An

3(EPA + DHA)]/(AEPA + ADHA)

EPA	=	= content of EPA from the test for Content of EPA and DHA [% (w/w)]
DHA	=	= content of DHA from the test for Content of EPA and DHA [% (w/w)]
An3	=	= sum of the areas of the peaks corresponding to C18:3 n3, C18:4 n3, C20:4 n3, C21:5 n3, and C22:5 n3 methyl esters from Test Solution 2
AEPA	=	= area of the peak corresponding to the EPA methyl ester from Test Solution 2
ADHA	=	= area of the peak corresponding to the DHA methyl ester from Test Solution 2

Acceptance criteria: NLT 28.0% (w/w) of total omega-3 acids, expressed as free acids

PERFORMANCE TESTS

• Weight Variation

2091

: Meet the requirements

• Disintegration and Dissolution

2040

: Meet the requirements for Disintegration, Delayed-Release (Enteric-Coated) Soft Shell Capsules

CONTAMINANTS

• Fats and Fixed Oils, Trace Metals

401

: NMT 0.1 ppm each of Pb, Cd, As, and Hg

• Limit of Dioxins, Furans, and Polychlorinated Biphenyls

Analysis: Determine the content of polychlorinated dibenzo-para-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) by method No. 1613 revision B of the Environmental Protection Agency. Determine the content of polychlorinated biphenyls (PCBs) by method No. 1668 revision A of the Environmental Protection Agency.

Acceptance criteria: The sum of PCDDs and PCDFs is NMT 2.0 pg/g of WHO toxic equivalents. The sum of PCDDs, PCDFs, and dioxin-like PCBs (polychlorinated biphenyls, non-ortho IUPAC congeners PCB-77, PCB-81, PCB-126, and PCB-169, and mono-ortho IUPAC congeners PCB-105, PCB-114, PCB-118, PCB-123, PCB-156, PCB-157, PCB-167, and PCB-189) is NMT 10.0 pg/g of WHO toxic equivalents.

SPECIFIC TESTS

• Fats and Fixed Oils, Acid Value

401

: NMT 3

• Fats and Fixed Oils, Anisidine Value

401

: NMT 20.0

• Fats and Fixed Oils, Peroxide Value

401

: NMT 5.0

• Fats and Fixed Oils, Total Oxidation Value (TOTOX)

401

: NMT 26, calculated:

Result = (2 × PV) + AV

PV	=	= peroxide value
AV	=	= anisidine value

• Fats and Fixed Oils, Unsaponifiable Matter

401

: NMT 1.5%

• Stearin: 10 mL remains clear after cooling at 0

for 3 h.

• Absorbance

Sample solution: 0.24 mg/mL in isooctane

Acceptance criteria: NMT 0.70, determined at 233 nm

ADDITIONAL REQUIREMENTS

• Packaging and Storage: Preserve in tight containers, and store at room temperature, protected from light.

• Labeling: The label states the amount of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) in mg/Capsule.

• USP Reference Standards

USP Docosahexaenoic Acid Ethyl Ester RS

all cis-4,7,10,13,16,19-Docosahexaenoic ethyl ester.
C24H36O2 356.55

USP Eicosapentaenoic Acid Ethyl Ester RS

all cis-5,8,11,14,17-Eicosapentaenoic ethyl ester.
C22H34O2 330.51

USP Fish Oil RS

USP Methyl Tricosanoate RS

USP35

Auxiliary Information— Please check for your question in the FAQs before contacting USP.

Topic/Question	Contact	Expert Committee
Monograph	Natalia Davydova Scientific Liaison 1-301-816-8328	(DS2010) Monographs - Dietary Supplements
Reference Standards	RS Technical Services 1-301-816-8129 rstech@usp.org

USP35–NF30 Page 1297

Pharmacopeial Forum: Volume No. 36(6) Page 1594