Scientific Summary
Sn-2 palmitate Oil benefits
for Infant Health
Contents
Executive Summary
01
Introduction
04
INFAT® Improves Infant’s Comfort
09
Summary
10
References
11
01
Executive Summary
H
uman breast milk provides the optimal
nutrition for infants. Designed to provide
perfectly balanced nutrition, human breast
milk naturally meets the needs of growing infants in
the first months of life. In human milk, and in most
infant formulas, about 50% of energy is supplied
to newborns as fat. More than 98% of this fat is in
the form of triglycerides, which contain saturated
and unsaturated fatty acids esterified to a glycerol
backbone glycerol back bone in certain location
called sn-1, sn-2 and sn-3. The special triglycerides
structure in human milk, with its specific fatty acids positioning, is one of the keys to its perfection.
Palmitic acid (C16:0) is the predominant saturated
fatty acid, comprising 17-25% of the fatty acids
in human milk. This fatty acid has a preferential
positioning in human milk for the sn-2 position
(about 70-75%). This is highly conserved across
populations, regardless of the mother’s nutritional
diet and is called sn-2 palmitate.
As part of the efforts of developing infant formulas
that are close to human milk, sn-2 palmitate from
vegetable sources was developed.
INFAT® is Advanced Lipids’ sn-2 palmitate fat
ingredient, a clinically proven component used
in infant formula that mimics the composition,
structure and benefits of human milk fat.
P
O
O
P
P
O
Vegetable oil
Sn-2 Palmitate
P
Palmitic Acid
O
Oleic Acid
Patented Enzymatic Process
02
Newly published clinical and pre-clinical studies of
INFAT® point to several sn-2 palmitate benefits on
infant health and well-being:
life. This study strengthen earlier findings on the
effect of SN-2 palmitate on bone development
published by Kennedy et al 19993.
Fat absorption: A clinical study on healthy
term newborns demonstrated that INFAT®
affects the fatty acids absorption even on top of
prebiotics. INFAT® was shown to enhance fatty
acids absorption compared to a standard formula1
and was closer to breastfeeding.
Controlled Intestinal Health: A preclinical
study revealed the potential protective role of INFAT® in a spontaneous colitis mice model, showing
that a low sn-2 palmitate diet increased intestinal
damage while a high sn-2 palmitate diet limited the
damage, though both diets contained the same
palmitic acid content4. This data suggests a crucial
role for INFAT® in limiting intestinal inflammation.
Increased Bone Strength: A clinical study
on term newborns demonstrated that the bone
strength (expressed as speed of sound) was
significantly higher in newborns fed for 12 weeks
with infant formula containing INFAT®, compared
to control formula. The bone strength of newborns
fed with INFAT® formula was similar to that of
breastfed newborns2. Thus, INFAT® may have a
beneficial role in bone development during early
Beneficial Gut Flora: A pilot clinical study on
healthy term newborns demonstrated that INFAT®
may affect the intestinal microbiota composition
during the first weeks of life by increasing the
Lactobacillus and Bifidobacteria abundance5.
These findings suggest that the lipid structure may
play a role in the gut microbiota colonization.
03
IMPROVED
COM
FO
RT
Less
Crying
Fat & Calcium
Absorption
Softer
Stools
TH
HEAL
RO
W
TH
ENH
Softer Stools: A clinical study demonstrated
that the percentage of infants with hard stools
were 23.8% in the control group (standard
vegetable oil) while only 14.3% in the INFAT®
and 0% in the breastfed groups. Statistical
significance was shown only between the control
and the breastfed groups6. The data suggests
INFAT® may contribute to the comfort of formulafed infants.
MU
Intestinal
Health
IM
YG
Bone
Strength
N IT
Y
Healthy Gut
Bacteria
A
E
NC
D
Reduced Infant Crying: Two clinical studies
demonstrated that INFAT® affects the infant crying
pattern during the first weeks of life even on
top of prebiotics. Comparable to breastfeeding,
INFAT® was shown to reduce crying duration and
frequency, mainly during afternoon and evening
hours6. Therefore, INFAT® has a beneficial effect
on the well-being of formula-fed infants as well
as of their parents.
Introduction
H
uman milk provides the optimal nutrition
for infants. Designed to provide balanced
nutrition, human milk naturally meets the
needs of growing infants in the first months after
birth. In human milk, and in most infant formulas,
approximately 50% of the energy is supplied to
newborns as fat, of which more than 98% is in the
form of triglycerides7; three fat components called
fatty acids, attached together. Triglyceride synthesis
occurs in the mammary gland, where the fatty acids
are specifically positioned to sn1, sn2 or sn3 positions
on the glycerol backbone to yield the structurespecific triglycerides that are found in human milk.
Palmitic acid (C16:0) is the predominant saturated
fatty acid, comprising 17-25% of the fatty acids in
mature human milk, with approximately 70-75% of
it esterified to the sn2 position of the triglyceride (sn2
palmitate)8,9, pointing at the preferential positioning
of palmitic acid (16:0) at the sn-2 position. This
position differs from other human tissue and plasma
lipids, and vegetable oils common in human diets
and in the fat blends used in the manufacture of
infant formula10. The significance of this fatty acid
and its position is evident from the fact that this
positioning is conserved in all women, regardless
of their ethnic origin or nutrition, unlike other fatty
acids in human milk.
The triglycerides with the structure OPO (18:1n-9—
16:0—18:1n-9) are the major triglyceride species
in human milk and represent an estimated ~12%
of total triglyceride species8. Palmitic acid (16:0) is
the major saturated fatty acid mainly esterified at
sn-2 position (middle) and Oleic acid (18:1n-9) is
the major unsaturated fatty acid mostly esterified
at the triglyceride sn-1,3 positions (outer).
The development of structured triglycerides
enables mimicking both the composition as well as
the structure of human milk fat in infant formulas.
Structured TG are achieved through an enzymatic
process. The resulting product contains 17-25%
palmitic acid with above 40% of it located at the
sn-2 position.
Clinical and pre-clinical studies have provided
increasing evidence that this specific positioning
of palmitic acid on the triglycerides in human milk
has a significant holistic effect on optimal infant
development and well-being that is related to the
increased absorption of both palmitic acid and
calcium, softer stools, increased bone strength,
increased beneficial gut flora, controlled intestinal
health, and reduced infant crying. All of these
contribute to the benefits of infant wellbeing.
05
INFAT® Enhances Nutrient
Absorption
Infants require a bio-available source of energy
and nutrients to meet the requirements of their
rapid growth, development and expanding
skeletal mass. Human milk provides the well
balanced nutrition satisfying the infants need. For
formula-fed infants, the availability of nutrients
and energy depends on the composition of the
formula. The nutritional environment of infants
involves a high fat diet with frequent feedings;
therefore, efficient fat absorption is required. In
breastfed infants, this is achieved using complex
fat globule lipids and TG structures that enable
the efficient absorption without requiring a high
metabolic effort from the infant.
and the fatty acids and calcium absorption.
Figure 1 shows the correlation between the level
of palmitic acid in the milk or formula triglyceride
sn-2 position and infant fatty acid and calcium
absorption calculated as a modified Cohen’s
effect size (f2)20 using data from published studies.
C16 absorption
Linear (C16 absorption)
Calcium absorption
Linear (Calcium absorption)
Triglyceride digestion by endogenous lipases leads to
hydrolysis of fatty acids from the triacylglyceride sn1,3 linkages, releasing two unesterified fatty acids
and one sn-2 monoglyceride from each triglyceride12.
The absorption efficiency is influenced by the type
of fatty acid and its position on the triglyceride
molecule. Palmitic acid is absorbed efficiently from
human milk as sn2 monoacylglycerol13, and it is
conserved through digestion and absorption14.
Clinical studies in term3, 15-17 and preterm
infants18, 19 have demonstrated the correlation
between the level of palmitic in the sn2 position
Palmitic acid at sn-2 position
of total palmitic acid (%)
Figure 1: Modified Cohen’s effect size of the effect
of palmitic acid position in formula triglycerides on the
absorption of palmitic acid and calcium in term and preterm
infants based on available publications.
06
INFAT® was recently shown to enhance fat absorption and reduce calcium soaps formation in
infant formulas containing prebiotics (GOS)1. In
a multicenter clinical study on Chinese term infants, consumption of infant formula with INFAT®
resulted in better nutrient absorption, specifically
fat absorption. Comparable with breastfeeding,
the formula with INFAT® reduced fat excretion,
primarily in form of saponified fat or insoluble calcium soaps (Figure 2)1.
Soaped palmitic acid excreted (gr)
The results show that progressively increasing ratio
of palmitic acid at the sn-2 position of the formula
triglyceride leads to a dose-dependent increase in
palmitic acid and calcium absorption (r=0.95 and
r=0.78 for palmitic acid and calcium, respectively).
The reduction in fecal calcium and saponified
palmitic acid (calcium soaps) is accompanied by
a decrease in the incidence of hard stools3, 17, 21, 22.
Control
INFAT®
BF
Figure 2: Stool fatty acids in 24 hr stool collection at 6
weeks postnatal. The significance was calculated for the two
groups by the Mann Whitney test. Different letters indicate
statistical significance (p<0.05) between groups.
07
INFAT® Supports
Healthy
Bone
Development
INFAT® Promotes
Development
of Beneficial
Intestinal Flora
The absorption of nutritional factors, such as
minerals, fats, carbohydrates, and proteins, is
significantly important for normal infant growth
and development and may contribute to early
bone mineral accretion23. Kennedy et al., showed
in a randomized, controlled, double-blind study
higher body bone mass in infants after 12 weeks
of feeding with sn2 palmitate formula3 by using
dual-energy X-ray absorptiometry (DEXA) to
assess bone mineralization. Litmanovitz et al.,
showed in a randomized, controlled, doubleblind study, that bone speed of sound (SOS) of
healthy term newborns fed for the first 12 weeks
of life with sn2 palmitate infant formula was
significantly higher than that of newborns fed
with regular formula and comparable to that of
breastfed newborns2 (figure 3).
The human gut is the natural habitat for a large and
dynamic microorganisms’ community24. The colonization begins immediately after birth and is influenced by the mode of delivery, the infant’s diet,
hygiene levels and medications25. The intestinal microflora is an essential “organ” that is involved in
numerous important functions for the human host,
including protection against pathogens and provision of enhanced metabolic capabilities.
Bone speed of sound (m/sec)
Lactobacilli counts (CFU/gr x1E+09)
INFAT® was shown to increase numbers of Lactobacilli and Bifidobacteria in the stool of healthy term
infants fed for 6 weeks postnatal compared to a
control infant formula5 (Figure 4). The infants in the
INFAT® group exhibited also a notable increase in the
counts of beneficial bacteria at week six from birth
compared to start of feeding.
Control INFAT®
BF
Figure 3: Ultrasound Speed of Sound (SOS) of the tibia of
term infants 3 months of age after 12 weeks of feeding.
Study groups:
Control
INFAT®
BF
Baseline
6 weeks
Figure 4: The effect of the type of feeding on the lactobacillus counts at baseline and 6 postnatal weeks.
08
INFAT® Controls
Intestinal
Inflammation Damage
An animal knock out model for intestinal inflammation (Muc2 deficient mice), which is a well-described model of enterocolitis, was used to address
the possible role of INFAT® on intestinal inflammation. Muc2 deficient mice (Muc2-/-) lack mucin2,
which is a major component of the mucus layer
that provides a barrier for the intestinal epithelial
cells from the intraluminal contents26, and forms a
physical barrier protecting the underlying epithelium against luminal substances and microbes27-29.
The deficiency of mucins in the Muc2-/- mice affects the protective capacities of the mucus layer30, and as a consequence, bacteria are in direct
contact with the intestinal epithelial cells31. This
leads to the development of spontaneous colitis
in Muc2-/- mice32-34. In this study mice fed diet with
INFAT® demonstrated a lower extent of intestinal
inflammatory damage compared with mice fed
the control diet4 (Figure 5).
Figure 5: Morphology of the distal colon of Muc2-/- mice fed AIN93G, control diet (low sn2 palmitate),
or sn2 palmitate (INFAT). Distal colonic sections of mice fed with the different diets were stained with
hematoxylin and eosin. Shown are representative sections for each diet group.
INFAT® improves infant’s comfort
One of the major concerns of parents is their baby’s
comfort and wellbeing. Various parameters may
indicate comfort to the newborn infants. Those
parameters include stool characteristics, such as
consistency, frequency and volume, as well as
crying duration and frequency during the day.
The reduction in calcium and fatty acid absorption
is accompanied by increased calcium soaps and,
consequently, hard stools21. Infants receiving the
stools compared to the control regular vegetable
oil formula group3, 6, 35.
A double blind clinical study revealed that feeding
term infants an infant formula with INFAT® for 12
weeks from birth reduced the number of infants with
hard stools (Figure 6), the number of crying infants,
and their crying duration during the day and night,
and especially in the afternoon and evening hours
compared to a control formula (Figure 7). Lately, this
was shown also on top of prebiotics (data not shown).
Infants with hard stools (%)
Total Daily Crying Duration (min)
high sn2 formula had softer stools and fewer hard
09
Control INFAT®
BF
Figure 6: The percent of infants with hard stools at age
of 12 weeks. Hard stools were evaluated by using parents
questionnaires based on 3 consecutive days.
Control INFAT®
BF
Figure 7: Total daily crying duration at 12 weeks postnatal.
Crying was evaluated by calculating the mean crying
duration per day based on parents’ reports of crying periods
of more than 5 minutes.
Summary
Human breast milk is the optimal nutrition for
infants, by providing a perfectly balanced nutrition
meeting the naturally needs of growing infants
in the first months of life. In human milk, and
in most infant formulas, about 50% of energy
is supplied to newborns as fat. More than 98%
of this fat is in the form of triglycerides, which
contain saturated and unsaturated fatty acids
esterified to a glycerol backbone. The special
triglycerides structure in human milk, with its
specific fatty acids positioning, is one of the keys
to its perfection.
Palmitic acid (C16:0) is the predominant saturated
fatty acid, comprising 17-25% of the fatty acids in
human milk. Surprisingly, 70-75% of the palmitic
fatty acids are esterified to the sn-2 position of
the triglyceride (sn-2 palmitate). This preferential
positioning is highly conserved across populations,
regardless of the mother’s nutritional diet.
INFAT® is Advanced Lipids’ sn-2 palmitate fat
ingredient, a clinically proven component used
in infant formula that mimics the composition,
structure and benefits of human milk fat.
Studies over the last two to three decades have
provided increasing evidence that this specific
positioning of palmitic acid on triglycerides in
human milk has a significant holistic effect on
optimal infant development and well-being. It was
shown to contribute to the increased absorption
of both palmitic acid and calcium, softer stools,
increased bone strength, increased beneficial gut
flora, controlled intestinal health, and reduced
infant crying.
11
References
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ennedy, K., et al., Double-blind, randomized trial of
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12
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