Fatty acids carry out many functions that are necessary for maintaining optimal health, and the fatty acid status of a client provides valuable information that relates to both risk and progression of a range of diseases. Determining fatty acid status assists the practitioner in identifying those individuals who would benefit from omega-3 supplementation. The Igennus Opti-O-3 biomarker test is a safe, cost effective and minimally invasive dried blood spot (DBS) method that can be carried out by the client at home. In addition, the Opti-O-3 offers a personalised approach to condition management by focusing on optimising validated biomarkers of disease risk through identification of the client’s unique fatty acid requirements. In this refresher webinar, Dr Bailey will discuss: - Fatty acid testing: who, why and when? - Why choose the Igennus Opti-O-3: a comparison of laboratories - How to interpret results, including: - New fatty acid reference ranges - Overview of fatty acid functions - Intervention considerations - Personalised intervention

1. An updated practitioner’s guide to the
Opti-O-3 biomarker test
Nina Bailey
BSc (hons) MSc PhD ANutr

2.  What is the Opti-O-3
 The importance of determining baseline omega-3 levels
 Comparison of laboratories
 Reference ranges
 How personalised dosing optimises clinical outcomes
 How to interpret the results: what the biomarkers mean

3. Full fatty acid screen (24 fatty acids):
 Saturated
 Trans fats
 Monounsaturated
 Polyunsaturated
 Omega-6
 Omega-3
Biomarkers:
 Omega-3 index
 Omega-6 to omega-3 ratio
 AA to EPA ratio

4. Primary structural function &
anti-inflammatory
docosanoid production
Resolvins
Protectins
Anti-inflammatory eicosanoid
production
REDUCED INFLAMMATION
Series-3 prostaglandins
Series-3 thromboxanes
Series-5 leukotrienes
Hydroxy fatty acids
Resolvins
DHA
Anti-inflammatory
eicosanoid production
REDUCED INFLAMMATION
Series-1 prostaglandins
Series -1 thromboxanes
DGLA
GLA
LA
EPA
ETA
SDA
ALA
Delta -6 desaturase
Elongase/
desaturaseDelta -5 desaturase
Cyclooxygenase (COX)/lipoxygenase (LOX)
Elongase
Pro-inflammatory eicosanoid
production
INFLAMMATION
Series-2 prostaglandins
Series-2 thromboxanes
Series-4 leukotrienes
Hydroxy fatty acids
AA
COX/LOX
COX
Pro-resolving Lipoxins
Omega-6 to omega-3 ratio

5. Resolvins
Protectins
Omega-3
DHA
ANTI-INFLAMMATORY
Omega-6
DGLA
Omega-3
EPA
PRO-INFLAMMATORY
Omega-6
AA
Lipoxins
Resolvins
Protectins
AA to EPA ratio
Omega-3 index
Using fatty acids as biomarkers of
health and disease
ANTI-INFLAMMATORY

6. Greenland
Inuit
Japanese Western populations Disease biomarker
Optimal Acceptable Suboptimal Poor
0 1.5 3 7 15 20
The AA to EPA ratio as a potential biomarker
of pro-inflammatory to anti-inflammatory balance

7. AA to EPA ratio in health and disease
Fasted blood samples from 1432
[Italian] subjects, who were referred by
their physicians, were analysed to
assess their AA to EPA and total omega-
6 to omega-3 ratios in whole blood and
in RBC membrane phospholipids
Individuals with no diagnosable
conditions had lower AA to EPA ratios
than those with diagnosable health
conditions
Rizzo AM, Montorfano G, Negroni M, Adorni L, Berselli P, Corsetto P, Wahle K, Berra B A rapid method for determining arachidonic:eicosapentaenoic acid ratios in
whole blood lipids: correlation with erythrocyte membrane ratios and validation in a large Italian population of various ages and pathologies. Lipids Health
Dis. 2010 Jan 27;9:7.
AA/EPA and omega-6/omega-3 ratios in whole blood of healthy subjects and
in a group of patients with various pathologies, with and without
consumption of omega-3. Mean ± S.E; Student's t-test: ## p < 0.01 with
omega-3 vs without omega-3; ** p < 0.01 pathological vs healthy

8. AA/EPA ratios in whole blood of patients grouped
according to their specific pathologies. The
horizontal lines indicate the mean value for all the
healthy subjects that do not use omega-3.
Rizzo AM, Montorfano G, Negroni M, Adorni L, Berselli P, Corsetto P, Wahle K, Berra B A rapid method for determining arachidonic:eicosapentaenoic acid ratios in whole blood
lipids: correlation with erythrocyte membrane ratios and validation in a large Italian population of various ages and pathologies. Lipids Health Dis. 2010 Jan 27;9:7.
Individuals with allergic, skin and
neurodegenerative diseases had
higher ratios of AA to EPA compared
to the values for subjects with other
pathologies, possibly due to a higher
turnover of EPA
Subjects who did not take omega-3
supplements and suffered from
allergic, neurodegenerative, skin and
inflammatory diseases had higher
values for AA to EPA ratios than those
with the other diseases (heart,
metabolic, cancer)

9. Silva V, Green P, Singer P: Membrane fatty acid composition of different target populations: importance of baseline on supplementation. Clinical
Nutrition 2014 33:206
The importance of baseline on supplementation efficacy

10. Resolvins
Protectins
Omega-3
DHA
ANTI-INFLAMMATORY
Omega-6
DGLA
Omega-3
EPA
PRO-INFLAMMATORY
Omega-6
AA
Lipoxins
Resolvins
Protectins
AA to EPA ratio
Omega-3 index
Combining biomarkers
ANTI-INFLAMMATORY

11. The Seattle study, the Physicians' Health Study (PHS), the Cardiovascular Health Study (CHS), the Diet and Reinfarction Trial
(DART), the study on the prevention of coronary atherosclerosis by intervention with marine omega-3 fatty acids (SCIMO), and
the GISSI-Prevenzione study. Taken together, these data suggest a desirable target value for the Omega-3 Index of ≥8% and an
undesirable level of ≤4%.
4% 6% 8% 10%
GISSI-P & DART: 9.5%
CHS 8.9%
PHS 3.9%
SCIMO: 3.4%
Seattle: 3.3%
SCIMO: 8.3%
PHS: 7.3%Seattle: 6.5%
LEAST PROTECTION GREATEST PROTECTION
Summary of evidence for the proposed cut points for the Omega-3 Index
Harris WS, Von Schacky C: The Omega-3 Index: a new risk factor for death from coronary heart disease? Prev
Med 2004, 39:212-220.

12. The relative risk for sudden cardiac death (RR SCD) by quartile of the Omega-3 Index compared to other, more traditional
blood-borne risk factors.
Only two risk factors demonstrated statistically significant trends: C-reactive protein and the Omega-3 Index
With the relative risk reduced by approximately 90% in the highest quartiles, the Omega-3 Index is both a strong and
an independent predictor of risk for sudden cardiac death.
CRP = C-reactive protein; Hcy = homocysteine; TC = total cholesterol; LDL = low density lipoprotein cholesterol; HDL = high density lipoprotein
cholesterol; Tg = triglycerides; n-3 Index = Omega-3 Index.
William S Harris, Clemens von Schacky The Omega-3 Index: a new risk factor for death from coronary heart disease? Preventive Medicine,
Volume 39, Issue 1, 2004, 212–220

13. Omega-3 index
 The Omega-3 Index is the result of Dr. William Harris's 30 years of research
in fatty acids and cardiovascular disease
 The Omega-3 Index is a measure of omega-3 fatty acids, EPA+DHA, in red
blood cells, which relates to risk for heart disease
 The omega-3 index is now recognised as a valuable biomarker of risk for
numerous other health conditions and disease
 Baseline levels of omega-3 should be evaluated and considered individually
before generalised supplementation to strengthen the concept of
personalised nutrition

14. Omega-3 index
an early cardiovascular risk indicator
Omega-6 to omega-3 ratio
an established marker of long-term health and chronic illness
AA to EPA ratio
a measure of ’silent’ or chronic inflammation
A personalised plan aims to achieve:
an omega-3 index of more than 8%
an omega-6 to omega-3 ratio of between 3 and 4
an AA to EPA ratio of between 1.5 and 3

15. Combining the AA to EPA ratio with the omega-3 index gives
us a comprehensive overview of health status

16. Plasma vs red blood cell membranes (RBC)

17. Fatty acids can be measured in various blood fractions and tissues including
plasma, erythrocytes and adipose tissue
PUFAs and trans fatty acids in adipose tissue correlated with intake measured by
FFQs
Availability of adipose tissue limits its use and blood is therefore more widely
used!
Saturated fatty acids and MUFAs in plasma and erythrocytes do not directly reflect
intake, because these 2 classes of fatty acids can be made by endogenous
synthesis from carbohydrates
In contrast, PUFAs and trans fatty acids in plasma and erythrocytes are a direct
reflection of dietary intake – FOCUS on PUFA
 Plasma fatty acids = recent intake
 Erythrocytes =long-term intake

18. • Fatty acid levels from red blood cells (RBCs), plasma and plasma phospholipids (PL)
obtained from 20 healthy volunteers tested weekly over 6 weeks
• The average intake of oily fish (tuna or other non-fried fish) was 3 ± 2.9 times per
month, and 5 subjects reported taking stable doses of fish oil supplements (providing
between 300 and 1800 mg of EPA + DHA per day)
• All subjects reported stable dietary and supplementation habits throughout the study
Harris WS, Thomas RM. Biological variability of blood omega-3 biomarkers. Clin Biochem.2010 Feb;43(3):338-40.
Variability in plasma and RBC omega-3 index isolated whole blood

19. Considerable fluctuations are
observed in plasma vs RBC
Within-subject coefficient of variation
(CV) for the omega-3 index in RBCs vs
plasma was 4.1%±1.9% and
15.9%±6.4%, respectively
Thus, the RBC test had the lowest
biological variability, which means
that a single measurement (not
several) is all that is needed to
determine the true omega-3 index
Harris WS, Thomas RM. Biological variability of blood omega-3
biomarkers. Clin Biochem. 2010 Feb;43(3):338-40.

20. Fasting vs Fed
Plasma fatty acids are prone to considerable fluctuations
Consuming a meal with fat/omega-3 will not affect the omega-3 index
within RBC
Consuming a meal with fat/omega-3 will affect the omega-3 index within
plasma
Plasma omega-3 index rises when omega-3s are consumed prior to
testing
A non-omega-3 fat-containing meal will dilute plasma omega-3 with non-
omega-3 fatty acids, thereby lowering the omega-3 index
Harris WS, Thomas RM. Biological variability of blood omega-3
biomarkers. Clin Biochem. 2010 Feb;43(3):338-40.

21. Effects of a single dose of EPA + DHA (3.4 g) taken
with breakfast on the percentage of change from
baseline (hour 0) in EPA + DHA levels in plasma
(expressed as a percentage of total fatty acids, top;
and expressed as concentrations, middle) and in RBCs
(expressed as a percentage of total fatty acids;
bottom) over 24 hours (n = 20)
Mean estimate and 95% Dunnett-adjusted confidence
intervals are shown. *Difference from baseline, P <
.05. DHA, docosahexaenoic acid; EPA,
eicosapentaenoic acid; RBC, red blood cell.
Harris WS, Varvel SA, Pottala JV, Warnick GR, McConnell JP. Comparative effects of an acute dose of fish oil on omega-3 fatty acid levels in red blood cells
versus plasma: implications for clinical utility. J Clin Lipidol. 2013 Sep-Oct;7(5):433-40

22. Whole blood vs blood spot

23. Whole blood Blood spot
Requires phebotomist (client inconvenience) Client can take test (no phebotomist required)
Invasive Minimally invasive
More analytical steps Less analytical steps
Higher costs Lower costs
Inconvenience to client Convenient for practitioner and client
Can measure plasma or RBC Measures plasma and RBC (fasting required)
Whole blood vs blood spot

24. Dried blood spot (DBS)
Minimally invasive
Convenient (can be performed by
the client)
Data derived from RBC & plasma
Whole blood
Invasive (requires a phlebotomist)
Higher cost than DBS
Data derived from RBC only
High correlation between RBC omega-3 and DBS omega-3 (R=0.96, p<0.0001)

25. Why choose Igennus?

26. Why choose Igennus?
How it the omega-3 index determined?
William Harris who validated the Omega-3 index, is the founder of OmegaQuant Analytics
The number and type of fatty acids will affect the omega-3 index
Laboratory Genova Genova Genova Biolab Igennus OmegaMetrix OmegaQuant
UK or USA based UK UK UK UK UK USA USA
Fatty acid analysis Plasma RBC Blood spot RBC Blood spot RBC Blood spot
Phlebotomist required Yes Yes No Yes No Yes No
Total number of fats analysed 33 23 7 32 24 23 24
Expressed as mol/L % total fat % area mol/L % total % total % total

27. Laboratory Genova Genova Genova Biolab Igennus OmegaMetrix OmegaQuant
UK or USA based UK UK UK UK UK USA USA
Fatty acid analysis Plasma RBC Blood spot RBC Blood spot RBC Blood spot
Phlebotomist required Yes Yes No Yes No Yes No
Total number of fats analysed 33 23 7 32 24 23 24
Expressed as mol/L % total fat % area mol/L % total % total % total
Saturated fat 14 8 0 31 6 4 6
Capric 10:0
Lauric acid 12:0
Myristic acid 14:0
Pentadecanoic acid 15:0
Palmitic acid 16:0
Margaric acid 17:0
Stearic acid 18:0
Nonadecanoic 19:0
Arachidic acid 20:0
Heneicosanoic acid 21:0
Behenic acid 22:0
Tricosanoic acid 23:0
Lignoceric acid 24:0
Hexacosanoic 26:0
Monounsaturated fat 6 4 0 6 4 3 4
Myristoleic acid n-5 14:1
Palmitoleic acid n-7 16:1
Vaccenic acid n-7 18:1
Oleic acid n-9 18:1
Eicosenoic acid n-9 20:1
Euric cid n-6 22:1
Nervonic acid n-9 24:1
Polyunsaturated fat n-9 1 0 0 1 0 0 0
Mead acid n-9 20:3

28. Laboratory Genova Genova Genova Biolab Igennus OmegaMetrix OmegaQuant
UK or USA based UK UK UK UK UK USA USA
Fatty acid analysis Plasma RBC Blood spot RBC Blood spot RBC Blood spot
Phlebotomist required Yes Yes No Yes No Yes No
Total number of fats analysed 33 23 7 32 24 23 24
Expressed as mol/L % total fat % area mol/L % total % total % total
Polyunsaturated fat n-6 7 6 4 7 7 7 7
Linoleic acid (LA) 18:2
Gamma-linolenic acid (GLA) 18:3
Eicosadienoic acid (EDA) 20:2
Dihomo-gamma linolenic acid (DGLA) 20:3
Arachidonic acid (AA) 20:4
Docosadienoic acid 22:2
Docosatetraenoic acid n-6 22:4
Docosapentaenoic acid n-6 22:5
Polyunsaturated fat n-3 4 4 3 4 4 4 4
Alpha-linolenic acid (ALA) 18:3
Stearidonic acid (SDA) 18:4
Eicosatetraenoic acid (ETA) 20:4
Eicosapentaenoic acid (EPA) 20:5
Docosapentaenoic (DPA) 22:5
Docosahexaenoic acid (DHA) 22:6
Trans fat 1 1 ?? 3 3 5 3
Trans palmitoleic acid n-7 16:1
Trans vaccenic acid n-7 18:1
Trans oleic acid n-9 18:1
Trans linoleic acid n-6 18:2
Cis/trans linoleic acid 18:2
Trans/cis linoleic acid n-6 18:2
Total trans

29. Laboratory Igennus OmegaQuant
UK or USA based UK USA
Fatty acid analysis Blood spot Blood spot
Phlebotomist required No No
Total number of fats analysed 24 24
Expressed as % total % total
Saturated fat 6 6
Myristic acid 14:0
Palmitic acid 16:0
Stearic acid 18:0
Arachidic acid 20:0
Behenic acid 22:0
Lignoceric acid 24:0
Monounsaturated fat 4 4
Palmitoleic acid n-7 16:1
Oleic acid n-9 18:1
Eicosenoic acid n-9 20:1
Nervonic acid n-9 24:1
Polyunsaturated fat n-6 7 7
Linoleic acid (LA) 18:2
Gamma-linolenic acid (GLA) 18:3
Eicosadienoic acid (EDA) 20:2
Dihomo-gamma linolenic acid (DGLA) 20:3
Arachidonic acid (AA) 20:4
Docosatetraenoic acid n-6 22:4
Docosapentaenoic acid n-6 22:5
Polyunsaturated fat n-3 4 4
Alpha-linolenic acid (ALA) 18:3
Eicosapentaenoic acid (EPA) 20:5
Docosapentaenoic (DPA) 22:5
Docosahexaenoic acid (DHA) 22:6
Trans fat 3 3
Trans palmitoleic acid n-7 16:1
Trans oleic acid n-9 18:1
Trans linoleic acid n-6 18:2

30. What’s in the kit?
1 x test information sheet
1 x Spot Saver card
2 x finger-prick lancets
1 x resealable plastic bag with moisture-absorbent gel sachet
1 x sterile alcohol cleansing pad
1 x return envelope
1 x personal information form

31. Taking the test

32. Taking the test

33. Taking the test

35. Taking the test

36. Fatty acid stability in transit
 Pre-treating blood slot cards with Butylated hydroxytoluene (BHT) prevents
PUFA degradation for up to 8 weeks
 BHT-mediated protection of PUFA is most likely due to free radical scavenging
by BHT
 The phenol group in BHT is thought to donate a proton to free radicals, thus
neutralising the free radicals and preventing them from accepting hydrogen
protons from the methylene groups in PUFA and thereby preventing
degradation
 Additionally, storing dried blood spots in sealable containers/bags further
prevents PUFA loss
Metherel AH, Hogg RC, Buzikievich LM, Stark KD. Butylated hydroxytoluene can protect polyunsaturated fatty acids in dried blood spots from degradation for up
to 8 weeks at room temperature. Lipids Health Dis. 2013 Feb 20;12:22.

37. Weight % of fatty acids in dried, untreated (BHT) blood spots over 28 days
Metherel AH, Hogg RC, Buzikievich LM, Stark KD. Butylated hydroxytoluene can protect polyunsaturated fatty acids in dried blood spots from degradation for up
to 8 weeks at room temperature. Lipids Health Dis. 2013 Feb 20;12:22.

38. Weight % of fatty acids in dried blood spots treated with BHT (5 mg/ml) over 28 days
Metherel AH, Hogg RC, Buzikievich LM, Stark KD. Butylated hydroxytoluene can protect polyunsaturated fatty acids in dried blood spots from degradation for up
to 8 weeks at room temperature. Lipids Health Dis. 2013 Feb 20;12:22.

39. The nutraceutical approach to managing inflammation: shift the balance!
AA, EPA and DHA contents of cell membranes can be altered through consumption of
omega-3 (marine products/marine oils)
 Improves cardiovascular health
 Improves cognitive function
 Improves immune function
 Improves inflammatory resolution
 Reduces disease risk
How much omega-3 do we need?
Knowledge of a person’s baseline omega-3 allows us to calculate a mg/kg dose of
EPA/DHA to raise their omega-3 to a predetermined level
Flock MR, Skulas-Ray AC, Harris WS, Etherton TD, Fleming JA, Kris-Etherton PM Determinants of erythrocyte omega-3 fatty acid content in response to
fish oil supplementation: a dose-response randomized controlled trial. J Am Heart Assoc. 2013 Nov 19;2(6):e000513. doi: 10.1161/JAHA.113.000513.

40. • It is suggested that whilst dietary intervention with fish oil results in
the incorporation of EPA and DHA into cell membranes, the omega-3
index must reach the level suggested to be optimal (≥8% in the case
of cardiovascular patients) to obtain clinical efficacy
• From a therapeutic stance, dietary intervention for clinical outcomes
must therefore focus on ensuring that an ideal omega-3 index is
achieved
Kagan I, Cohen J, Stein M, Bendavid I, Pinsker D, Silva V, Theilla M, Anbar R, Lev S, Grinev M, Singer P:
Preemptive enteral nutrition enriched with eicosapentaenoic acid, gamma-linolenic acid and
antioxidants in severe multiple trauma: a prospective, randomized, double-blind study. Intensive care
medicine 2015.

41. Distribution of the percentage of red blood cell (RBC) EPA+DHA values (omega‐3 index) in the study
population at baseline. Lines at 8% and 4% indicate proposed low‐ and high‐risk horizons respectively, and the
dotted line at 4.3% is the population average
7%
2%
4.3%
Flock MR, Skulas-Ray AC, Harris WS, Etherton TD, Fleming JA, Kris-Etherton PM Determinants of erythrocyte omega-3 fatty acid content in response to
fish oil supplementation: a dose-response randomized controlled trial. J Am Heart Assoc. 2013 Nov 19;2(6):e000513. doi: 10.1161/JAHA.113.000513.

42. Omega-3 dosing – ‘one size fits all?’
 40 individuals with a baseline omega-3 index <5% (black
bar) and post treatment (white bar) after a 6-week
intervention with omega-3 EPA & DHA (0·5 g/d)
• The mean omega-3 index increased from 4·37% to
6·80% and inter-individual variability in response was
high (varied by a factor of up to 13 inter-individually)
(Kohler et al. 2010)
Köhler A, Bittner D, Löw A, von Schacky C. Effects of a convenience drink fortified with n-3 fatty acids on the n-3 index. Br J Nutr. 2010 Sep;104(5):729-36.

43. The Opti-O-3 results and recommendations

44. Interventions use a ‘one size fits all’ approach irrespective of personal
requirements
Factors that influence the omega-3 index
 age
 physical activity
 gender
 body weight
We are metabolically unique and using biomarkers to identify personal
requirements for omega-3 can optimise clinical outcomes
omega-3 baseline levels and body weight will determine the dose needed to
optimise the omega-3 index

45. Example 1

48. Example 2

51. Example 3

54. Opti-O-3 biomarker summary Average Low High
Omega-3 index 5.47 1.21 17.91
Omega-6 to omega-3 ratio 5.26 0.96 14.70
AA to EPA ratio 9.27 0.42 54.43
In 1992, the BNF Task Force on Unsaturated Fatty Acids suggested a desirable population intake for EPA and
DHA of 0.5% of energy, which equates to about 8g/week (1.14g/day) for women and 10g/week (1.42g/day) for
men, equivalent to 2-3 medium servings of oil-rich fish per week
•Current recommendations are set at 450mg EPA and DHA daily (2 portions fish weekly of which one should be
oily)
•Mean consumption of oily fish (all age groups) was below the recommended one portion (140g) per week
(rolling programme for 2008 and 2009 to 2011 and 2012)
•Mean consumption in adults aged 19 to 64 years was 54g per week (52g for men and 54g for women) and for
adults aged 65 years and over mean consumption was 90g per week (103g for men and 81g for women)
https://www.gov.uk/government/statistics/national-diet-and-nutrition-survey-results-from-years-1-to-4-combined-of-the-rolling-programme-for-2008-and-2009-to-2011-and-2012
British Nutrition Foundation. Unsaturated fatty acids nutritional and physiological significance: the report of the British Nutrition Foundation's task force. New York: Chapman & Hall, 1992.

55. Using reference ranges
 Laboratories provide reference ranges using in-house
data
 In-house data is skewed because it fails to provide
information gathered from ‘normal’ population
 There are no validated reference ranges for individual
fatty acids
 However...........

56. Genova blood spot

57. Genova blood spot
0.41 - 4.06?

58. Q1 Q2 Q3 Q4 Q5
Low Average High
Saturated fat
Myristic acid 14:0 0.17 0.47 0.62 0.85 2.01
Palmitic acid 16:0 15.5 21.55 23.01 24.38 29.26
Stearic acid 18:0 1.45 13.17 14.48 15.62 23.07
Arachidic acid 20:0 0.05 0.14 0.16 0.18 0.75
Behenic acid 22:0 0.15 0.36 0.43 0.51 1.13
Lignoceric acid 24:0 0.19 0.51 0.64 0.76 2.36
Monounsaturated fat
Palmitoleic acid n-7 16:1 0.1 0.69 0.92 1.28 3.51
Oleic acid n-9 18:1 12.38 17.96 21.03 32.97 32.97
Eicosenoic acid n-9 20:1 0.08 0.18 0.22 0.26 0.85
Nervonic acid n-9 24:1 0.07 0.4 0.51 0.65 1.69
Polyunsaturated fat n-6
Linoleic acid (LA) 18:2 11.08 16.83 18.56 21.15 28.74
Gamma-linolenic acid (GLA) 18:3 0.02 0.13 0.18 0.27 0.97
Eicosadienoic acid (EDA) 20:2 0.10 0.16 0.19 0.22 0.98
Dihomo-gamma linolenic acid (DGLA) 20:3 0.39 0.99 1.23 1.48 2.47
Arachidonic acid (AA) 20:4 2.5 8.56 10.05 11.38 16.51
Docosatetraenoic acid n-6 22:4 0.12 0.64 0.85 1.13 2.58
Docosapentaenoic acid n-6 22:5 0.03 0.14 0.17 0.23 1.53
Polyunsaturated fat n-3
Alpha-linolenic acid (ALA) 18:3 0.16 0.34 0.41 0.51 1.4
Eicosapentaenoic acid (EPA) 20:5 0.2 0.86 1.4 2.44 10.68
Docosapentaenoic (DPA) 22:5 0.41 0.89 1.11 1.45 3.97
Docosahexaenoic acid (DHA) 22:6 0.96 2.49 3.35 4.37 8.89
Trans fat
Trans palmitoleic acid n-7 16:1 0.11 0.19 0.23 0.28 0.84
Trans oleic acid n-9 18:1 0.01 0.09 0.12 0.18 0.54
Trans linoleic acid n-6 18:2 0.07 0.16 0.19 0.23 1.7

60. Q1 Q2 Q3 Q4 Q5
RESULT Low Average High
Saturated fat
Myristic acid 14:0 0.53
Palmitic acid 16:0 24.2
Stearic acid 18:0 17.81
Arachidic acid 20:0 0.14
Behenic acid 22:0 0.43
Lignoceric acid 24:0 0.57
Monounsaturated fat
Palmitoleic acid n-7 16:1 0.88
Oleic acid n-9 18:1 17.26
Eicosenoic acid n-9 20:1 0.20
Nervonic acid n-9 24:1 0.43
Polyunsaturated fat n-6
Linoleic acid (LA) 18:2 20.37
Gamma-linolenic acid (GLA) 18:3 0.23
Eicosadienoic acid (EDA) 20:2 0.16
Dihomo-gamma linolenic acid (DGLA) 20:3 1.37
Arachidonic acid (AA) 20:4 10.05
Docosatetraenoic acid n-6 22:4 0.76
Docosapentaenoic acid n-6 22:5 0.12
Polyunsaturated fat n-3
Alpha-linolenic acid (ALA) 18:3 0.27
Eicosapentaenoic acid (EPA) 20:5 0.79
Docosapentaenoic (DPA) 22:5 0.74
Docosahexaenoic acid (DHA) 22:6 2.31
Trans fat
Trans palmitoleic acid n-7 16:1 0.14
Trans oleic acid n-9 18:1 0.06
Trans linoleic acid n-6 18:2 0.17
Base line fatty acids

61. Q1 Q2 Q3 Q4 Q5
RESULT Low Average High
Saturated fat
Myristic acid 14:0 0.33
Palmitic acid 16:0 22.32
Stearic acid 18:0 14.62
Arachidic acid 20:0 0.15
Behenic acid 22:0 0.34
Lignoceric acid 24:0 0.57
Monounsaturated fat
Palmitoleic acid n-7 16:1 0.69
Oleic acid n-9 18:1 20.43
Eicosenoic acid n-9 20:1 0.29
Nervonic acid n-9 24:1 0.51
Polyunsaturated fat n-6
Linoleic acid (LA) 18:2 19.04
Gamma-linolenic acid (GLA) 18:3 0.10
Eicosadienoic acid (EDA) 20:2 0.22
Dihomo-gamma linolenic acid (DGLA) 20:3 1.07
Arachidonic acid (AA) 20:4 9.16
Docosatetraenoic acid n-6 22:4 0.38
Docosapentaenoic acid n-6 22:5 0.10
Polyunsaturated fat n-3
Alpha-linolenic acid (ALA) 18:3 0.37
Eicosapentaenoic acid (EPA) 20:5 3.58
Docosapentaenoic (DPA) 22:5 1.82
Docosahexaenoic acid (DHA) 22:6 3.58
Trans fat
Trans palmitoleic acid n-7 16:1 0.13
Trans oleic acid n-9 18:1 0.06
Trans linoleic acid n-6 18:2 0.15
6-month fatty acids

62. RESTORE with pure EPA
MAINTAIN with EPA, DHA
and GLA
Minimum 3-6 months
Therapeutic role of Pharmepa®
RESTORE & MAINTAIN™
 AA to EPA ratio
 Inflammatory regulation
 Symptoms of inflammatory illness
 Optimum brain, cell, heart, immune
and CNS function
 Optimum wellbeing
 Omega-3 index
 Omega-6 to 3 ratio
 Long-term general and cellular
health
 Heart, brain and eye health
 Prevent chronic illness and protect
against inflammatory disease

63. 1 RESTORE 2 RESTORE 3 RESTORE 4 RESTORE 1 MAINTAIN 2 MAINTAIN 3 MAINTAIN 4 MAINTAIN
EPA (mg) 500 1000 1500 2000 250 500 750 1000
DHA (mg) 0 0 0 0 83 167 250 333
GLA (mg) 0 0 0 0 20 40 60 80
Vitamin E (mg) 5 10 15 20 3 6 9 12
Vitamin D (g) 0 0 0 0 10 20 30 40
EPA/DHA ratio - - - 3 3 3 3
Total Omega-3 (mg) 500 1000 1500 2000 333 666 1000 1333
RESTORE & MAINTAIN mixed doses
0
500
1000
1500
2000
2500
1 RESTORE 2 RESTORE 3 RESTORE 4 RESTORE 1 MAINTAIN 2 MAINTAIN 3 MAINTAIN 4 MAINTAIN
GLA (mg) DHA (mg) EPA (mg)
Dose(mg)

64. 0
500
1000
1500
2000
2500
3000
GLA (mg) DHA (mg) EPA (mg)
1 RESTORE
1 MAINTAIN
1 RESTORE
4 MAINTAIN
2 RESTORE
1 MAINTAIN
2 RESTORE
2 MAINTAIN
2 RESTORE
3 MAINTAIN
1 RESTORE
3 MAINTAIN
1 RESTORE
2 MAINTAIN
2 RESTORE
4 MAINTAIN
1 RESTORE 1 RESTORE 1 RESTORE 1 RESTORE 2 RESTORE 2 RESTORE 2 RESTORE 2 RESTORE
1 MAINTAIN 2 MAINTAIN 3 MAINTAIN 4 MAINTAIN 1 MAINTAIN 2 MAINTAIN 3 MAINTAIN 4 MAINTAIN
EPA (mg) 750 1000 1250 1500 1250 1500 1750 2000
DHA (mg) 83 167 250 333 83 167 250 333
GLA (mg) 20 40 60 80 20 40 60 80
Vitamin E (mg) 8 11 14 17 13 16 19 22
Vitamin D (mg) 10 20 30 40 10 20 30 40
EPA/DHA ratio 9 6 5 5 15 9 7 6
Total Omega-3 (mg) 833 1167 1500 1833 1333 1667 2000 2333
Dose(mg)
RESTORE & MAINTAIN mixed doses

65. Benefits of the Opti-O-3
 Knowledge of baseline levels will guide the practitioner recommendations—
unsurprisingly, low baseline values may require a larger dose than a high baseline
value!
 The omega-3 index and AA to EPA ratio are invaluable for assessing both baseline
risk and the change in risk (as function of intake – retesting is advisable ≥6 months)
 Dose response studies show us that high doses of omega-3 are required to reduce
the AA to EPA ratio and achieve an omega-3 index ≥8%, especially where the
baseline levels are suboptimal ≤4%
Use in conjunction with Pharmepa RESTORE & MAINTAIN
 Pre-loading with pure EPA for 6 months (RESTORE) is advised to reduce the AA to EPA
ratio, before the introduction of EPA/DHA with GLA (MAINTAIN) for long-term
support of the omega-3 index

66. Education Technical
Sophie Tully
Nutrition Education Manager
sophiet@igennus.com
Dr Nina Bailey
Head of Nutrition
ninab@igennus.com