Reduced
symptoms
of
inattention
after
dietary
omega-3
fatty
acid
supplementation in boys with and without Attention Deficit/Hyperactivity Disorder
Dienke J. Bos MSc1, Bob Oranje PhD1, E. Sanne Veerhoek MSc1, Rosanne M. Van
Diepen MSc1, Juliette M.H. Weusten MSc1, Hans Demmelmair PhD2, Berthold Koletzko
MD PhD2, Monique G. M. de Sain-van der Velden PhD3, Ans Eilander PhD4, Marco
Hoeksma PhD4, Sarah Durston PhD1
Affiliations
1NICHE
Lab, Department of Psychiatry, Brain Center Rudolf Magnus, University Medical Center
Utrecht, The Netherlands
2Division
of Metabolic and Nutritional Medicine, University of Munich Medical Center, Dr. von
Hauner Children's Hospital, Munich, Germany
3Department
of Medical Genetics, University Medical Center Utrecht, The Netherlands
Wilhelmina Children's Hospital, University Medical Center Utrecht, The Netherlands
4Unilever
Research & Development, Vlaardingen, The Netherlands
Corresponding author
Dienke J. Bos, NICHE Lab, Department of Psychiatry, Brain Center Rudolf Magnus, University
Medical Center Utrecht, HP A.01.126, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands,
T: +31 88 755 9840, E-mail: d.j.bos-2@umcutrecht.nl
Supplemental Material 1: Composition of the intervention product and analysis of
the physiological measures.
Nutrient composition of the intervention product
All children received a daily 10-gram serving of the active- or placebo intervention
product. In the placebo product EPA and DHA were replaced by mono-unsaturated fatty
acids (MUFA); total saturated fatty acid (SAFA) and omega-6 fatty acid content were
matched.
Omega-6 fatty acids are also polyunsaturated fatty acids (PUFA), however these
do not have the currently theorized and investigated properties of omega-3 fatty acids.
Vitamin E content corresponded to the amount of PUFA present in the active or placebo
product (Table S1). The active and placebo product had highly similar sensory
properties, to prevent possible deblinding by the intervention product itself.
Analysis of the Physiological Measures
Lipid extracts were analysed as previously described (Koletzko et al, 1999), at
the Dr. von Haunersches Kinderspital, Division of Metabolic Disorders and Nutrition,
Munich, Germany. Briefly, the extract was dissolved in chloroform-methanol and
phospholipids were isolated by thin layer chromatography. The phospholipid fraction was
transferred into a glass vial for synthesis of fatty acid methyl esters by reaction with 3 M
methanolic hydrochloric acid at 85°C for 45 minutes. The fatty acid methyl esters were
extracted twice into hexane (Merck), the combined extracts were taken to dryness under
a stream of nitrogen, and dissolved in 50 µl hexane containing 2 g/l butylhydroxytoluol as
an antioxidant for storage until gas-chromatography. Gas chromatography was
performed with an Agilent 7890 GC (Agilent, Waldbronn, Germany) equipped with a
split/splitless injector, a BPX-70 column (50m, 0.25 mm inner diameter), and flame
ionization detector. For identification and response quantification of individual fatty acid
methyl esters external calibration was applied using a mixture of fatty acid methyl esters
in hexane (NuCheck Prep., Elysian, USA).
Identification-analyses of organic acids in urine were carried out by gas
chromatography–mass spectrometry (GC–MS) (Agilent), at the University Medical
Center Utrecht. Prior to this GC–MS analysis, the organic acids were extracted with
ethylacetate
and
then
trimethylsilylated
at
60 °C
for
30
min
with
N,N-
bis(trimethylsilyl)trifluoracetamide-/pyridine/trimethylchlorosilane (5:1:0.05 v/v/v). The
gas chromatographic separation was performed on a 25 m X 0.25 mm capillary CP Sil
19CB column (film thickness 0.19 mm) from Varian/Chrompack, Middelburg, The
Netherlands.
References
Koletzko B, Knoppke B, Schenck U von, Demmelmair H, Damli A (1999). Noninvasive
Assessment of Essential Fatty Acid Status in Preterm Infants by Buccal Mucosal
Cell Phospholipid Analysis. J Pediatr Gastroenterol Nutr 29: 467–474.
Table S1: Composition of the intervention product.
Active test product
Placebo product
72
72
FA Content
MUFA (g)
SAFA (g)
PUFA (g)
Trans FA (g)
2.08
2.66
2.66
0.1
4.32
2.59
1.05
0.04
Total n-3 FA (g)
EPA (g)
DHA (g)
1.48
0.65
0.65
0.06
-
0.07
0.06
Total n-6 FA (g)
LA (g)
GLA (g)
DGLA (g)
AA (g)
1.03
1.0
0.03
1.0
1.0
-
Other
Vitamin E (mg)
Protein (g)
6.4
<1
0.52
<1
Energy (kcal)
ALA (g)
Abbreviations: kcal = kilocalory, MUFA = monounsaturated fatty acids, SAFA = saturated
fatty acids, PUFA = polyunsaturated fatty acids, EPA = eicosapentaenoic acid, DHA =
docosahexaenoic aicd, ALA =a-Linolenic acid, LA = Linoleic Acid, GLA = Gammalinolenic Acid, DGLA = Dihomo-gamma-linolenic Acid, AA = Arachidonic Acid, g = gram,
mg = milligram.