U.S. patent application number 13/133929 was filed with the patent office on 2011-12-08 for nutritional compositions with large lipid globule size.
This patent application is currently assigned to N.V. NUTRICIA. Invention is credited to Gunther Boehm, Annemarie Oosting, Gelske Speelmans, Antonie Van Baalen, Eline Marleen Van Der Beek.
Application Number | 20110300204 13/133929 |
Document ID | / |
Family ID | 40756821 |
Filed Date | 2011-12-08 |
United States Patent
Application |
20110300204 |
Kind Code |
A1 |
Van Der Beek; Eline Marleen ;
et al. |
December 8, 2011 |
NUTRITIONAL COMPOSITIONS WITH LARGE LIPID GLOBULE SIZE
Abstract
The present invention relates to a nutritional composition for
infants and/or toddlers comprising a lipid component which has a
large lipid globule size, having a volume weighted mode diameter
above 1.0 .mu.m and/or with a diameter of 2 to 12 .mu.m in a amount
of at least 45 volume % based on total lipid. The composition can
be used to increase bone mineral content and/or bone mass density,
to prevent osteoporosis and/or osteopenia and for the further
prevention of obesity.
Inventors: |
Van Der Beek; Eline Marleen;
(Wageningen, NL) ; Speelmans; Gelske; (Wageningen,
NL) ; Van Baalen; Antonie; (Arnhem, NL) ;
Oosting; Annemarie; (Wageningen, NL) ; Boehm;
Gunther; (Leipzig, DE) |
Assignee: |
N.V. NUTRICIA
|
Family ID: |
40756821 |
Appl. No.: |
13/133929 |
Filed: |
December 11, 2009 |
PCT Filed: |
December 11, 2009 |
PCT NO: |
PCT/NL09/50756 |
371 Date: |
August 25, 2011 |
Current U.S.
Class: |
424/439 ;
424/195.16; 424/195.17; 424/522; 424/523; 424/780; 514/171;
514/560 |
Current CPC
Class: |
A61K 9/1276 20130101;
A23L 33/12 20160801; A23V 2002/00 20130101; A61K 31/688 20130101;
A61P 19/08 20180101; A61K 9/14 20130101; A61K 31/202 20130101; A61K
36/02 20130101; A23L 33/115 20160801; A23L 33/40 20160801; A61P
3/06 20180101; A61K 35/66 20130101; A61K 35/60 20130101; A61P 19/10
20180101; A61K 36/06 20130101; A61K 31/575 20130101; A61K 31/20
20130101; A61K 35/20 20130101; A61P 19/00 20180101; A61K 35/20
20130101; A61K 2300/00 20130101; A61K 35/60 20130101; A23V 2002/00
20130101; A23V 2200/306 20130101; A61K 2300/00 20130101; A23V
2250/18 20130101; A61K 35/66 20130101; A61K 2300/00 20130101; A23V
2002/00 20130101; A23V 2200/306 20130101; A23V 2250/18
20130101 |
Class at
Publication: |
424/439 ;
514/560; 514/171; 424/523; 424/522; 424/195.17; 424/195.16;
424/780 |
International
Class: |
A61K 9/14 20060101
A61K009/14; A61K 31/575 20060101 A61K031/575; A61K 31/20 20060101
A61K031/20; A61P 19/08 20060101 A61P019/08; A61K 36/02 20060101
A61K036/02; A61K 36/06 20060101 A61K036/06; A61K 35/66 20060101
A61K035/66; A61P 19/10 20060101 A61P019/10; A61K 31/202 20060101
A61K031/202; A61K 35/60 20060101 A61K035/60 |
Foreign Application Data
Date |
Code |
Application Number |
Dec 11, 2008 |
NL |
PCT/NL2008/050792 |
Claims
1-14. (canceled)
15. A method of increasing bone mass density and/or increasing bone
mineral content, comprising administering to a human subject in
need thereof a composition comprising 10 to 50 wt. % of vegetable
lipids, based on dry weight of the composition, wherein the
vegetable lipids are comprised in lipid globules having: i) a
volume-weighted mode diameter above 1.0 .mu.m, and/or ii) a
diameter of 2 to 12 .mu.m in an amount of at least 45 volume %
based on total lipid.
16. The method according to claim 15, wherein the lipid globules
comprise: (i) a volume-weighted mode diameter in the range of
1.0-10 .mu.m, and/or (ii) a diameter of 2 to 12 .mu.m in an amount
of at least 55 volume % based on total lipid.
17. The method according to claim 15, wherein the subject is a
human subject between 0 and 36 months of age.
18. The method according to claim 15, wherein the subject is a
human subject 36 months of age or older.
19. The method according to claim 18, wherein the subject is a
human subject above 5 years of age.
20. The method according to claim 19, wherein the composition
further comprises 0.5-10 wt. % phospholipids, based on total
lipid.
21. The method according to claim 20, wherein the phospholipids are
derived from milk lipids.
22. The method according to claim 15, wherein the composition
further comprises glycosphingolipids, cholesterol, or both.
23. The method according to claim 22, wherein the composition
comprises 0.6 to 12 wt. % polar lipids based on total lipid,
wherein the polar lipids are the sum of phospholipids,
glycosphingolipids and cholesterol.
24. The method according to claim 15, wherein the composition has a
fatty acid profile with a linoleic acid to alpha-linolenic acid
weight ratio between 4 and 7.
25. The method according to claim 15, wherein the lipids comprise
at least 16 wt. % palmitic acid based on total fatty acids.
26. The method according to claim 25, wherein over 75 wt. % of the
palmitic acid is in the sn-1 or sn-3 position.
27. The method according to claim 15, wherein the composition
comprises least one lipid selected from the group consisting of
fish oil, marine oil, algal oil, fungal oil and microbial oil.
28. The method according to claim 15, wherein the composition
further comprises non-digestible oligosaccharides.
29. The method according to claim 15, wherein the composition is a
powder suitable for making a liquid composition after
reconstitution with an aqueous solution.
30. The method according to claim 15, wherein the aqueous solution
comprises water.
31. A method of treating osteoporosis and/or osteopenia, comprising
administering to a human subject in need thereof a composition
comprising 10 to 50 wt. % of vegetable lipids, based on dry weight
of the composition, wherein the vegetable lipids are comprised in
lipid globules having: i) a volume-weighted mode diameter above 1.0
.mu.m, and/or ii) a diameter of 2 to 12 .mu.m in an amount of at
least 45 volume % based on total lipid.
32. The method according to claim 31, wherein the lipid globules
comprise: (i) a volume-weighted mode diameter in the range of
1.0-10 .mu.m, and/or (ii) diameter of 2 to 12 .mu.m in an amount of
at least 55 volume % based on total lipid.
33. A method of treating obesity, comprising administering to a
human subject in need thereof a composition comprising 10 to 50 wt.
% of vegetable lipids, based on dry weight of the composition,
wherein the vegetable lipids are comprised in lipid globules
having: i) a volume-weighted mode diameter above 1.0 .mu.m, and/or
ii) a diameter of 2 to 12 .mu.m in an amount of at least 45 volume
% based on total lipid.
Description
FIELD OF THE INVENTION
[0001] The invention relates to the field of infant milk formula
and growing up milks for improving bone health.
BACKGROUND
[0002] Breast-feeding is the preferred method of feeding infants.
However, there are circumstances that make breast-feeding
impossible or less desirable. In those cases infant formulae are a
good alternative. The composition of modern infant formulae is
adapted in such a way that it meets many of the special nutritional
requirements of the fast growing and developing infant.
[0003] However, differences between breast feeding and feeding
infant formulae exist. Breastfeeding in early life is associated
with higher bone mass density and bone mineral content later in
life during childhood and early adolescence compared with those who
were bottle-fed. The implication of this observation is that
osteoporosis prevention programs need to start very early in the
life cycle. Adult degenerative bone disease (osteoporosis), a major
public health problem in the West, has been linked to peak bone
mass attained in young adult life. Following attainment of peak
bone mass, bone mineral content falls and may descend below the
safety level for clinical disease. Most interventions to reduce the
incidence of clinical disease have been in middle life.
[0004] Human milk is the major source of energy for many infants
during the first part of their lives. It has a high content of the
saturated fatty acid palmitic acid (20-25%), which is primarily
located in the sn-2 position of the triacylglycerols (.about.70%).
The n-1, 3 positions of vegetable fats, normally used in infant
formulae, are rich in saturated fatty acids such as palmitate and
stearate and are not appropriate to be used in infant nutrition.
The triglycerides are digested in the infant by lipases which
release the sn-1, 3 fatty acids. When these palmitic- and stearic
acids are released from vegetable triglycerides they tend to create
salts of dietary calcium. Calcium salts of saturated fatty acids
are insoluble and tend to precipitate and to be secreted from the
body. This results in the loss of crucial calcium. Formation of
calcium soaps causes loss in faeces of energy as well as of
calcium, and this loss can be so high that it can influence bone
mineralization, i.e. normal skeletal and bone development of the
infant, as well as other aspects of normal health and development
in infants. Hence, advanced infant formulas include synthetically
structured fats produced to mimic the unique structure and
characteristics of human milk fat. Such structured fats include
Betapole or InFat which provide 22% total palmitic acid of which
43% is at the sn-2 position and 25% palmitic acid, up to 68% of
which are at the sn-2 position, respectively.
[0005] WO 2005/07373 relates to compositions comprising such
synthetically structured triglycerides with high levels of mono- or
polyunsaturated fatty acids at positions sn-1 and sn-3 of the
glycerol backbone, for use in enhancing calcium absorption and in
the prevention and/or treatment of disorders associated with
depletion of bone calcium and bone density, prevention and
treatment of osteoporosis, for the enhancement of bone formation
and bone mass maximization and for the enhancement of bone
formation in infants and young children.
[0006] WO 2007/097523 aims to provide a fat composition as a human
milk substitute comprising a diglyceride in which unsaturated fatty
acids are bonded in the 1,2-positions or 1,3-positions and a
triglyceride containing a large amount of palmitic acid or stearic
acid as a saturated fatty acid in the 2-position of the
triglyceride.
[0007] WO 2005/051092 concerns a lipid preparation comprising a
combination of phosphatidylcholine (PC), phosphatidylethanolamine
(PE), phosphatidylserine (PS) and phosphatidylinositol (PI),
wherein the quantitative ratio between these glycerophospholipids
essentially mimics their corresponding ratio in naturally occurring
human milk fat.
[0008] Other infant formulae reduce the amount of palmitic acid to
levels lower than that observed in human milk. EP 1252824 relates
to a method for increasing the bone mineralization of an infant or
toddler, comprises administering to said human a source of calcium
and a fat blend that is low in palmitic acid.
SUMMARY OF THE INVENTION
[0009] The inventors surprisingly found that the lipid globule size
in infant formulae affects the body composition later in life.
Specific selection of the lipid globule size in infant formulae
results in an increased bone mineral content (BMC) and increased
bone mass density (BMD) later in life. Specific selection of the
lipid globule size in infant formulae also results in an increased
bone mass density and/or increased bone mineral content later in
life. When early in life an infant formula of the present invention
that comprised lipids with a larger lipid globule size than present
in conventional infant formulae, was administered, it was observed
that later in life the body composition was changed, resulting in
increased bone mineral content and increased bone mass density.
[0010] The important difference between the two formulae was the
size of the lipid globules, whereas the fatty acid profile was
similar in both formulae and the amount of palmitic and stearic
acid present at sn1 and sn3 positions in the fat was also similar.
Both formulae further enabled a similar good growth and development
early in life. Surprisingly the increase in BMD and/or BMC remained
later in life when both groups received the same diet for a long
period, indicating that early nutrition has an effect on BMD and/or
BMC extending beyond the period in which it is actually
administered. Early diet of the present invention has a programming
effect on BMD and/or BMC.
[0011] Standard infant milk formulae have vegetable fat as lipid
component. The lipid is homogenized in order to create a stable
emulsion and the lipid globules are small, with a volume-weighted
mode diameter of about 0.3-0.6 .mu.m. Less than 55 volume %,
typically less than 35 volume %, based on total lipid has a size
between 2 and 12 .mu.m. The lipid globules are for a large part
covered with milk proteins, in particular with casein.
[0012] The present invention relates to infant formulae or growing
up milks for toddlers comprising vegetable fats with a lipid
globule size larger than that of standard infant formulae. The
present composition comprises lipid globules with a lipid
volume-weighted mode diameter of above 1.0 .mu.m, preferably
between 1.0 and 10 .mu.m, and/or with at least 45, more preferably
at least 55 volume % with a diameter of 2 to 12 .mu.m based on
total lipid. This can be achieved upon homogenizing of the lipid
component comprising vegetable fat at lower pressures, preferably
in the presence of polar lipids, in order to coat the enlarged
lipid globules and make them more stable. It was found that the
thus obtained oil in water emulsion is stable for at least 48 h.
Especially when the formulae is dried to a powder and subsequently
reconstituted with water to a ready to drink formula shortly before
use, no disadvantageous effects regarding stability are
observed.
[0013] It has now surprisingly been found that the size of the
lipid globule administered early in life is one of the
determinative factors which affect body composition, in particular
bone mineral content and bone mass density and/or lean body mass,
later in life. The present invention therefore can be used for food
compositions intended for infants and/or toddlers in order to
increase bone mineral content and/or increase bone mass density.
The present invention therefore can be used for food compositions
intended for infants and/or toddlers in order to prevent or reduce
the risk for osteoporosis later in life, for the enhancement of
bone formation and bone mass maximization and for the enhancement
of bone formation in infants and young children.
[0014] The present invention also allows to formulate infant milk
formulae with high levels of palmitic and stearic acid, as observed
in human milk and with the use of natural lipids, i.e. without the
use of synthetically made triglycerides, which are more expensive
and subject to strict food legislations. The use of the
synthetically made lipids with palmitic acid in the sn-2 position
has the additional disadvantage of having direct diet effects
regarding body weight, lean body mass and increasing fat mass.
Surprisingly, it was found that while increasing BMD and BMC,
advantageously fat mass and relative fat mass was decreased later
in life using the lipid globules of the present invention.
DETAILED DESCRIPTION
[0015] The present invention thus concerns a method [0016] for
increasing bone mass density and/or increasing bone mineral content
and or [0017] preventing osteoporosis and/or osteopenia
[0018] said method comprising administering to a human subject a
nutritional composition comprising [0019] a) 10 to 50 wt. %
vegetable lipids based on dry weight of the composition, and [0020]
b) lipid globules [0021] i) with a volume-weighted mode diameter
above 1.0 .mu.m, preferably between 1.0 and 10 .mu.m, and/or [0022]
ii) with a diameter of 2 to 12 .mu.m in an amount of at least 45,
more preferably at least 55 volume % based on total lipid.
[0023] For sake of clarity it is noted that the lipid globules as
defined under b) comprise vegetable lipids as defined under a) or
in other words that a) and b) overlap.
[0024] The present invention can also be worded as the use of lipid
for the manufacture of a nutritional composition for increasing
bone mass density and/or increasing bone mineral content, said
nutritional composition comprising [0025] a) 10 to 50 wt. %
vegetable lipids based on dry weight of the composition, and [0026]
b) lipid globules [0027] i) with a volume-weighted mode diameter
above 1.0 .mu.m, preferably between 1.0 and 10 .mu.m, and/or [0028]
ii) with a diameter of 2 to 12 .mu.m in an amount of at least 45,
more preferably at least 55 volume % based on total lipid.
[0029] The present invention can also be worded as the use of lipid
for the manufacture of a nutritional composition for preventing
osteoporosis and/or osteopenia, said nutritional composition
comprising [0030] a) 10 to 50 wt. % vegetable lipids based on dry
weight of the composition, and [0031] b) lipid globules [0032] i)
with a volume-weighted mode diameter above 1.0 .mu.m, preferably
between 1.0 and 10 .mu.m, and/or [0033] ii) with a diameter of 2 to
12 .mu.m in an amount of at least 45, more preferably at least 55
volume % based on total lipid.
[0034] The present invention can also be worded as a nutritional
composition comprising [0035] a) 10 to 50 wt. % vegetable lipids
based on dry weight of the composition, and [0036] b) lipid
globules [0037] i) with a volume-weighted mode diameter above 1.0
.mu.m, preferably between 1.0 and 10 .mu.m, and/or [0038] ii) with
a diameter of 2 to 12 .mu.m in an amount of at least 45, more
preferably at least 55 volume % based on total lipid
[0039] for use in increasing bone mass density and/or increasing
bone mineral content.
[0040] The present invention can also be worded as a nutritional
composition comprising [0041] a) 10 to 50 wt. % vegetable lipids
based on dry weight of the composition, and [0042] b) lipid
globules [0043] i) with a volume-weighted mode diameter above 1.0
.mu.m, preferably between 1.0 and 10 .mu.m, and/or [0044] ii) with
a diameter of 2 to 12 .mu.m in an amount of at least 45, more
preferably at least 55 volume % based on total lipid
[0045] for use in preventing osteoporosis and/or osteopenia.
[0046] In one embodiment, the present method further is for
preventing obesity, or in other words, the nutritional composition
is further for prevention of obesity or the nutritional composition
is for further use in prevention of obesity.
[0047] Bone Mass Density, Bone Mineral Content, Osteoporosis
[0048] The present composition is preferably administered to a
human subject with an age below 36 months, preferably below 18
months, more preferably below 12 months, even more preferably below
6 months.
[0049] Bone mass density (BMD) refers to the amount of matter per
cubic centimeter of bones. Herein the term "bone mass" refers to
the mass of bone mineral. In adults a low BMD is a strong predictor
for osteoporosis and/or osteopenia. In infants a higher BMD is
related to increased length, and lower risk fracture. Early optimal
growth is predicting increased length in adulthood. Bone mineral
content (BMC) refers to bone mineral content as a measure of bone
strength. During growth BMC is a more relevant parameter than BMD,
because it factors out most of the component of bone accumulation
that is associated with change in bone size. So, in infancy, when
assessing bone parameters, BMC is the more relevant parameter.
[0050] The terms "bone mineralization" and "bone mass accretion"
are being used interchangeably within this application. Thus within
the present specification and claims, they should be considered as
synonyms. "Bone mineralization" should also be considered
synonymous with increasing, enhancing or improving "bone strength",
"bone mineral density", "bone mineral content", "bone mass", "bone
accretion", etc.
[0051] BMD and BMC are typically determined by ultrasound,
radiographic absorptiometry, single energy X-ray absorptiometry
(SXA), peripheral dual energy X-ray absorptiometry (PDXA, dual
energy X-ray absorptiometry (DEXA), single photon absorptiometry
(SPA), dual energy radioactive photon absorptiometry (DPA) and
quantitative computerized tomography (QCT). Preferably BMD and/or
BMC is measured by DEXA.
[0052] In the context of this invention, increase in BMD is defined
as an increase of at least 2%, preferably at least 4%, when
compared to a control not receiving the nutrition of the present
invention. For example as determined in a comparative study in an
animal model as described in example 2.
[0053] In the context of this invention, increase in BMC is defined
as an increase of at least 5%, preferably at least 7%, when
compared to a control not receiving the nutrition of the present
invention. For example as determined in a comparative study in an
animal model as described in example 2.
[0054] The term "osteopenia," as used herein, refers to decreased
bone mass below a threshold that compromises the structural
integrity of the skeletal bone. An `osteopenic` condition is a
condition in which the bone mass density is decreased compared to a
young normal control value. "Young normal" known as the "T-score"
compares BMD to optimal or peak density of a 30-year old healthy
adult and determines the fracture risk, which increases as BMD
falls below young normal levels. The World Health Organization
(WHO) has set the values for interpreting T-scores and defined
osteoporosis and osteopenia based on these values: Osteopenia, on
the other hand, is defined as a T-score between -1 and -2.5.
[0055] Osteoporosis is a disease of bone that leads to an increased
risk of fracture. In osteoporosis the bone mass density (BMD) is
reduced, bone microarchitecture is disrupted, and the amount and
variety of non-collageneous proteins in bone is altered.
Osteoporosis is defined by the World Health Organization (WHO) as a
bone mass density with a T-score below -2.5.
[0056] Obesity
[0057] Obesity in the present invention relates to an excess of
body fat mass. Fat mass is also known as adipose tissue or fat
tissue. An adult human person suffers from obesity if over 25 wt. %
(for a man) or over 30 wt. % (for a woman) of body weight is fat
mass. Obesity is sometimes referred to as adiposity.
[0058] Suitable ways to determine % body fat mass are underwater
weighing, skin fold measurement, bioelectrical impedance analysis,
computed tomography (CT/CAT scan), magnetic resonance imaging
(MRI/NMR), ultrasonography and dual energy X-ray absorptiometry
(DEXA). A preferred method is DEXA measurement. In the context of
this invention body fat mass is determined by DEXA.
[0059] Lipid Component
[0060] The present composition comprises lipid. The lipid provides
preferably 30 to 60% of the total calories of the composition. More
preferably the present composition comprises lipid providing 35 to
55% of the total calories, even more preferably the present
composition comprises lipid providing 40 to 50% of the total
calories. When in liquid form, e.g. as a ready-to-feed liquid, the
composition preferably comprises 2.1 to 6.5 g lipid per 100 ml,
more preferably 3.0 to 4.0 g per 100 ml. Based on dry weight the
present composition preferably comprises 10 to 50 wt. %, more
preferably 12.5 to 40 wt. % lipid, even more preferably 19 to 30
wt. % lipid.
[0061] Lipids include polar lipids (such as phospholipids,
glycolipids, sphingomyelin, and cholesterol), monoglycerides,
diglycerides, triglycerides and free fatty acids. Preferably the
composition comprises at least 85 wt. % triglycerides based on
total lipids.
[0062] The lipid of the present invention comprises vegetable
lipids. The presence of vegetable lipids advantageously enables an
optimal fatty acid profile, high in (poly)unsaturated fatty acids
and/or more reminiscent to human milk fat. Using lipids from cow's
milk alone, or other domestic mammals, does not provide an optimal
fatty acid profile. Preferably the present composition comprises at
least one, preferably at least two lipid sources selected from the
group consisting of linseed oil (flaxseed oil), rape seed oil (such
as colza oil, low erucic acid rape seed oil and canola oil), salvia
oil, perilla oil, purslane oil, lingonberry oil, sea buckthorn oil,
hemp oil, sunflower oil, high oleic sunflower oil, safflower oil,
high oleic safflower oil, olive oil, black currant seed oil, echium
oil, coconut oil, palm oil and palm kernel oil. Preferably the
present composition comprises at least one, preferably at least two
lipid sources selected from the group consisting of linseed oil,
canola oil, coconut oil, sunflower oil and high oleic sunflower
oil. Commercially available vegetable lipids are typically offered
in the form a continuous oil phase. When in liquid form, e.g. as a
ready-to-feed liquid, the composition preferably comprises 2.1 to
6.5 g vegetable lipid per 100 ml, more preferably 3.0 to 4.0 g per
100 ml. Based on dry weight the present composition preferably
comprises 10 to 50 wt. %, more preferably 12.5 to 40 wt. %
vegetable lipid, even more preferably 19 to 30 wt. %. Preferably
the composition comprises 50 to 100 wt. % vegetable lipids based on
total lipids, more preferably 70 to 100 wt. %, even more preferably
75 to 97 wt. %. It is noted therefore that the present composition
also may comprise non-vegetable lipids. Suitable and preferred
non-vegetable lipids are further specified below.
[0063] Lipid Globule Size
[0064] According to the present invention, lipid is present in the
composition in the form of lipid globules, emulsified in the
aqueous phase. The lipid globules comprise a core and a coating.
The core comprises vegetable fat and preferably comprises at least
90 wt. % triglycerides and more preferably essentially consists of
triglycerides. The coating comprises phospholipids and/or polar
lipids. Not all phospholipids and/or polar lipids that are present
in the composition need necessarily be comprised in the coating,
but preferably a major part is. Preferably more than 50 wt. %, more
preferably more than 70 wt. %, even more preferably more than 85
wt. %, most preferably more than 95 wt. % of the phospholipids
and/or polar lipids that are present in the composition are
comprised in the coating of lipid globules. Not all vegetable
lipids that are present in the composition need necessarily be
comprised in the core of lipid globules, but preferably a major
part is, preferably more than 50% wt. %, more preferably more than
70 wt. %, even more preferably more than 85 wt. %, even more
preferably more than 95 wt. %, most preferably more than 98 wt. %
of the vegetable lipids that are present in the composition are
comprised in the core of lipid globules.
[0065] The lipid in the present composition according to the
invention is present in the form of lipid globules, emulsified in
the aqueous phase. The lipid globules of the present invention have
[0066] 1. a volume-weighted mode diameter above 1.0 .mu.m, more
preferably above 3.0 .mu.m, more preferably 4.0 .mu.m or above,
preferably between 1.0 and 10 .mu.m, more preferably between 2.0
and 8.0 .mu.m, even more preferably between 3.0 and 8.0 .mu.m, most
preferably between 4.0 and 8.0 .mu.m and/or [0067] 2. a size
distribution in such a way that at least 45 volume %, preferably at
least 55 volume %, even more preferably at least 65 volume %, even
more preferably at least 75 volume % has a diameter between 2 and
12 .mu.m. More preferably at least 45 volume %, preferably at least
55 volume %, even more preferably at least 65 volume %, even more
preferably at least 75 volume % has a diameter between 2 and 10
.mu.m. Even more preferably at least 45 volume %, preferably at
least 55 volume %, even more preferably at least 65 volume %, even
more preferably at least 75 volume % has a diameter between 4 and
10 .mu.m
[0068] The percentage of lipid globules is based on volume of total
lipid. The mode diameter relates to the diameter which is the most
present based on volume of total lipid, or the peak value in a
graphic representation, having on the X-as the diameter and on the
Y-as the volume (%). The volume of the lipid globule and its size
distribution can suitably be determined using a particle size
analyzer such as a Mastersizer (Malvern Instruments, Malvern, UK),
for example by the method described in Michalski et al, 2001, Lait
81: 787-796.
[0069] Polar Lipids
[0070] The present invention preferably comprises polar lipids.
Polar lipids are amphipathic of nature and include
glycerophospholipids, glycosphingolipids, sphingomyelin and/or
cholesterol. More preferably the composition comprises
phospholipids (the sum of glycerophospholipids and sphingomyelin).
Polar lipids in the present invention relate to the sum of
glycerophospholipids, glycospingolipids, sphingomyelin and
cholesterol. The presence of polar lipids helps to maintain the
lipid globules emulsified in the aqueous composition. This is
especially important when the lipid globule size is large.
[0071] The present composition preferably comprises
glycerophospholipids. Glycerophospholipids are a class of lipids
formed from fatty acids esterified at the hydroxyl groups on
carbon-1 and carbon-2 of the backbone glycerol moiety and a
negatively-charged phosphate group attached to carbon-3 of the
glycerol via an ester bond, and optionally a choline group (in case
of phosphatidylcholine, PC), a serine group (in case of
phosphatidylserine, PS), an ethanolamine group (in case of
phosphatidylethanolamine, PE), an inositol group (in case of
phosphatidylinositol, PI) or a glycerol group (in case of
phosphatidylglycerol, PG) attached to the phosphate group.
Lysophospholipids are a class of phospholipids with one fatty acyl
chain. Preferably the present composition contains PC, PS, PI
and/or PE, more preferably at least PC. Preferably the
glycerophospholipids comprise negatively charged phospholipids in
particular PS and/or PI. Negatively charged glycerophospholipids
advantageously improve the stability of the oil in water
emulsion.
[0072] The present composition preferably comprises
glycosphingolipids. The term glycosphingolipids as in the present
invention particularly refers to glycolipids with an amino alcohol
sphingosine. The sphingosine backbone is O-linked to a charged
headgroup such as ethanolamine, serine or choline backbone. The
backbone is also amide linked to a fatty acyl group.
Glycosphingolipids are ceramides with one or more sugar residues
joined in a .beta.-glycosidic linkage at the 1-hydroxyl position.
Preferably the present composition contains gangliosides, more
preferably at least one ganglioside selected from the group
consisting of GM3 and GD3.
[0073] The present composition preferably comprises sphingomyelin.
Sphingomyelins have a phosphorylcholine or phosphorylethanolamine
molecule esterified to the 1-hydroxy group of a ceramide. They are
classified as phospholipid as well as sphingolipid, but are not
classified as a glycerophospholipid nor as a glycosphingolipid.
[0074] Sphingolipids are in the present invention defined as the
sum of sphingomyelin and glycosphingolipids. Phospholipids are in
the present invention defined as the sum of sphingomyelin and
glycerophospholipids. Preferably the phospholipids are derived from
milk lipids. Preferably the weight ratio of
phospholipids:glycosphingolipids is from 2:1 to 10:1, more
preferably 2:1 to 5:1.
[0075] Preferably the present composition comprises phospholipids.
Preferably the present composition comprises 0.2 to 20 wt. %
phospholipids based on total lipid, more preferably 0.5 to 20 wt. %
phospholipids based on total lipid, more preferably 0.5 to 10 wt.
%, more preferably 1 to 10 wt. %, even more preferably 3 to 8 wt.
%. Preferably the present composition comprises 0.1 to 10 wt. %
glycosphingolipids based on total lipid, more preferably 0.5 to 5
wt. %, even more preferably 2 to 4 wt %. Preferably the present
composition comprises 0.3 to 20 wt. % (glycosphingolipids plus
phospholipids) based on total lipid, more preferably 0.5 to 20 wt.
% (glycosphingolipids plus phospholipids) based on total lipid,
more preferably 1 to 10 wt. %.
[0076] The present composition preferably comprises cholesterol.
The present composition preferably comprises at least 0.005 wt. %
cholesterol based on total lipid, more preferably at least 0.02 wt.
%, more preferably at least 0.05 wt. %., even more preferably at
least 0.1 wt. %. Preferably the amount of cholesterol does not
exceed 10 wt. % based on total lipid, more preferably does not
exceed 5 wt. %, even more preferably does not exceed 1 wt. % of
total lipid.
[0077] Preferably the present composition comprises 0.3 to 25 wt. %
polar lipids based on total lipid, wherein the polar lipids are the
sum of phospholipids, glycosphingolipids, and cholesterol, more
preferably 0.6 to 25 wt. % polar lipids based on total lipid, more
preferably 0.6 to 12 wt. %, more preferably 1 to 10 wt. %, even
more preferably 3 to 10 wt. % polar lipids based on total lipid,
wherein the polar lipids are the sum of phospholipids,
glycosphingolipids, and cholesterol.
[0078] Preferred sources for providing the phospholipids,
glycosphingolipids and/or cholesterol are egg lipids, milk fat,
buttermilk fat and butter serum fat (such as beta serum fat). A
preferred source for phospholipids, particularly PC, is soy
lecithin and/or sunflower lecithin. The present composition
preferably comprises phospholipids derived from milk. Preferably
the present composition comprises phospholipids and
glycosphingolipids derived from milk. Preferably also cholesterol
is obtained from milk. Polar lipids derived from milk include the
polar lipids isolated from milk lipid, cream lipid, butter serum
lipid (beta serum lipid), whey lipid, cheese lipid and/or
buttermilk lipid. The buttermilk lipid is typically obtained during
the manufacture of buttermilk. The butter serum lipid or beta serum
lipid is typically obtained during the manufacture of anhydrous
milk fat from butter. Preferably the phospholipids,
glycosphingolipids and/or cholesterol are obtained from milk cream.
The composition preferably comprises phospholipids,
glycosphingolipids and/or cholesterol from milk of cows, mares,
sheep, goats, buffalos, horses and camels. It is most preferred to
use a lipid extract isolated from cow's milk. The size of the lipid
globules of the present invention are more comparable to that of
human milk which are coated with a milk fat globule membrane
comprising polar lipids and membrane proteins. The use of polar
lipids from milk fat advantageously comprises the polar lipids from
milk fat globule membranes, which are more reminiscent to the
situation in human milk. The concomitant use of polar lipids
derived from domestic animals milk and trigycerides derived from
vegetable lipids therefore enables to manufacture lipid globules
with an architecture more similar to human milk, while at the same
time providing an optimal fatty acid profile. Suitable commercially
available sources for milk polar lipids are BAEF, SM2, SM3 and SM4
powder of Corman, Salibra of Glanbia, and LacProdan MFGM-10 or PL20
from Arla. Preferably the source of milk polar lipids comprises at
least 4 wt. % phospholipids based on total lipid, more preferably 7
to 75 wt. %, most preferably 20 to 70 wt. % phospholipids based on
total lipid. Preferably the weight ratio phospholipids to protein
is above 0.10, more preferably above 0.20, even more preferably
above 0.3. Preferably at least 25 wt. %, more preferably at least
40 wt. %, most preferably at least 75 wt. % of the polar lipids is
derived from milk.
[0079] Fatty Acid Composition
[0080] Herein LA refers to linoleic acid and/or acyl chain (18:2
n6); ALA refers to .alpha.-linolenic acid and/or acyl chain (18:3
n3); LC-PUFA refers to long chain polyunsaturated fatty acids
and/or acyl chains comprising at least 20 carbon atoms in the fatty
acyl chain and with 2 or more unsaturated bonds; DHA refers to
docosahexaenoic acid and/or acyl chain (22:6, n3); EPA refers to
eicosapentaenoic acid and/or acyl chain (20:5 n3); ARA refers to
arachidonic acid and/or acyl chain (20:4 n6); DPA refers to
docosapentaenoic acid and/or acyl chain (22:5 n3); PA refers to
palmitic acid and/or acyl chain (16:0); SA refers to stearic acid
and/or acyl chain (18:0).
[0081] Preferably the composition comprises PA and/or SA. PA is a
major component of human milk lipids. Preferably the composition
comprises at least 16 wt. %, more preferably at least 19 wt. %
based on total fatty acids, even more preferably at least 20 wt. %.
Preferably the composition comprises less than 35 wt. % based on
FA, more preferably less than 30 wt. %. A too high content of PA
results in excessive calcium soap formation and has a negative
effect on BMD and/or BMC. Preferably the palmitic acid in the
lipids is for over 75 wt. %, more preferably 90 wt. % in the sn-1
or sn-3 position. The present invention also allows to formulate
infant milk formulae with high levels of palmitic and stearic acid,
as observed in human milk and with the use of natural lipids, i.e.
without the use of synthetically made triglycerides with PA or SA
on the sn-2 position, which are more expensive and subject to
strict food legislations. The use of the synthetically made lipids
with palmitic acid in the sn-2 position has the additional
disadvantage of having direct diet effects by increasing body
weight, lean body mass and fat mass early in life.
[0082] A high weight ratio of dietary LA to ALA is associated with
a lower bone mass density. Therefore, LA preferably is present in a
sufficient amount in order to promote a healthy growth and
development, yet in an amount as low as possible to prevent a
decrease in BMD. The composition therefore preferably comprises
less than 15 wt. % LA based on total fatty acids, preferably
between 5 and 14.5 wt. %, more preferably between 6 and 10 wt. %.
Preferably the composition comprises over 5 wt. % LA based on fatty
acids. Preferably ALA is present in a sufficient amount to promote
a healthy growth and development of the infant. The present
composition therefore preferably comprises at least 1.0 wt. % ALA
based on total fatty acids. Preferably the composition comprises at
least 1.5 wt. % ALA based on total fatty acids, more preferably at
least 2.0 wt. %. Preferably the composition comprises less than 10
wt. % ALA, more preferably less than 5.0 wt. % based on total fatty
acids. The weight ratio LA/ALA preferably is well balanced in order
to improve BMD, while at the same time ensuring a normal growth and
development. Therefore, the present composition preferably
comprises a weight ratio of LA/ALA between 2 and 15, more
preferably between 2 and 7, more preferably between 4 and 7, more
preferably between 3 and 6, even more preferably between 4 and 5.5,
even more preferably between 4 and 5.
[0083] Preferably the present composition comprises n-3 LC-PUFA,
since n-3 LC-PUFA improve peak bone mass density. More preferably,
the present composition comprises EPA, DPA and/or DHA, even more
preferably DHA. Since a low concentration of DHA, DPA and/or EPA is
already effective and normal growth and development are important,
the content of n-3 LC-PUFA in the present composition, preferably
does not exceed 15 wt. % of the total fatty acid content,
preferably does not exceed 10 wt. %, even more preferably does not
exceed 5 wt. %. Preferably the present composition comprises at
least 0.2 wt. %, preferably at least 0.5 wt. %, more preferably at
least 0.75 wt. %, n-3 LC-PUFA of the total fatty acid content.
[0084] As the group of n-6 fatty acids, especially arachidonic acid
(AA) and LA as its precursor, counteracts the group of n-3 fatty
acids, especially DHA and EPA and ALA as their precursor, the
present composition comprises relatively low amounts of AA. The n-6
LC-PUFA content preferably does not exceed 5 wt. %, more preferably
does not exceed 2.0 wt. %, more preferably does not exceed 0.75 wt.
%, even more preferably does not exceed 0.5 wt. %, based on total
fatty acids. Since AA is important in infants for optimal
functional membranes, especially membranes of neurological tissues,
the amount of n-6 LC-PUFA is preferably at least 0.02 wt. % more
preferably at least 0.05 wt. %, more preferably at least 0.1 wt. %
based on total fatty acids, more preferably at least 0.2 wt. %. The
presence of AA is advantageous in a composition low in LA since it
remedies LA deficiency. The presence of, preferably low amounts, of
AA is beneficial in nutrition to be administered to infants below
the age of 6 months, since for these infants the infant formulae is
generally the only source of nutrition.
[0085] Preferably, in addition to the vegetable lipid, a lipid
selected from fish oil (preferably tuna fish oil) and single cell
oil (such as algal, microbial oil and fungal oil) is present. These
sources of oil are suitable as LC-PUFA sources. Preferably as a
source of n-3 LC-PUFA single cell oil, including algal oil and
microbial oil, is used.
[0086] Process for Obtaining Lipid Globules
[0087] The present composition comprises lipid globules. The lipid
globule size can be manipulated by adjusting process steps by which
the present composition is manufactured. A suitable and preferred
way to obtain larger lipid globule sizes is to adapt the process of
homogenization. In standard infant milk formula the lipid fraction
(usually comprising vegetable fat, a small amount of polar lipids
and fat soluble vitamins) is mixed into the aqueous fraction
(usually comprising water, skimmed milk, whey, digestible
carbohydrates such as lactose, water soluble vitamins and minerals
and optionally non-digestible carbohydrates) by homogenization. If
no homogenization was to take place, the lipid part would cream
very quickly, i.e. separate from the aqueous part and collect at
the top. Homogenization is the process of breaking up the fat phase
into smaller sizes so that it no longer quickly separates from the
aqueous phase but is maintained in a stable emulsion. This is
accomplished by forcing the milk at high pressure through small
orifices. However, the present inventors found that homogenization
at a lower pressure than usually applied in the preparation of
infant formula resulted in the larger lipid globules of the present
invention, while maintaining a sufficiently stable emulsion.
[0088] The process comprises the following steps:
[0089] 1 Mixing Ingredients
[0090] The ingredients of the composition are mixed, e.g.
preferably blended. Basically a lipid phase, comprising the
vegetable lipids, and an aqueous phase are added together. The
ingredients of the aqueous phase may comprise water, skimmed milk
(powder), whey (powder), low fat milk, lactose, water soluble
vitamins and minerals. Preferably the aqueous phase comprises
protein, digestible carbohydrates, and polar lipids, more
preferably phospholipids. Preferably the aqueous phase comprises
non-digestible oligosaccharides. Preferably the aqueous phase is
set at a pH between 6.0 and 8.0, more preferably pH 6.5 to 7.5.
Preferably the polar lipids, in particular the phospholipids are
derived from milk. The presence of polar lipids advantageously
results in more stable lipid globules. This is especially important
in case of larger lipid globules.
[0091] Preferably the lipid phase comprises 50 to 100 wt. %
vegetable lipids based on total weight of the lipid phase. Instead
of in the aqueous phase, the polar lipids, more preferably the
phospholipids, may also be present in the lipid phase or in both
phases. Alternatively the polar lipids may be added separately to
an aqueous and lipid phase. Preferably, the weight ratio of
phospholipid to total lipid is from 0.2 to 20 wt. %, more
preferably from 0.5 to 10 wt. %, even more preferably 3 to 8 wt. %
0.2.
[0092] The aqueous and lipid phase are preferably heated before
adding together, preferably at a temperature of 40.degree. C. to
80.degree. C., more preferably 55.degree. C. to 70.degree. C., even
more preferably 55.degree. C. to 60.degree. C. The mixture is also
kept at this temperature and blended. A suitable way for blending
is using an Ultra-Turrax T50 for about 30-60 s at 5000-10000 rpm.
Subsequently demi-water may be added to this blend, to obtain the
desired dry matter %. A desired dry matter % is for example 15%.
Alternatively, the lipid phase is injected to the aqueous phase
immediately prior to homogenization.
[0093] Minerals, vitamins, and stabilizing gums may be added at
various points in the process depending on their sensitivity to
heat. Mixing can for instance be performed with a high shear
agitator. In the process of the present invention, skimmed milk
(casein) is preferably not present in this step and added to the
composition after homogenization of the fat fraction into the
aqueous fraction (comprising compounds such as whey, whey protein,
lactose).
[0094] 2 Pasteurization
[0095] Preferably the mixture is then pasteurized. Pasteurization
involves a quick heating step under controlled conditions which
microorganisms cannot survive. A temperature of 60 to 80.degree.
C., more preferably 65 to 75.degree. C., held for at least 15 s,
usually adequately reduces vegetative cells of microorganisms.
Several pasteurization methods are known and commercially feasible.
Alternatively this step can also be performed before mixing as in
step 1 and/or be replaced by the heating step to 60.degree. C. in
step 1.
[0096] 3 Homogenization
[0097] Subsequently the optionally pasteurized mixture is
homogenized. Homogenization is a process which increases emulsion
uniformity and stability by reducing the size of the lipid globules
in the formula. This process step can be performed with a variety
of mixing equipment, which applies high shear to the product. This
type of mixing breaks the lipid globules into smaller globules. The
mixture obtained is preferably homogenized in two steps at high
temperature and pressure, for example 60.degree. C. at 30 to 100
and 5 to 50 bar respectively, with a total pressure of 35 to 150
bar. Alternatively, the mixture obtained is preferably homogenized
in two steps at a lower temperature, between 15 and 40.degree. C.,
preferably about 20.degree. C. at 5 to 50 and 5 to 50 bar
respectively, with a total pressure of 5 to 100 bar. This is
remarkably lower than standard pressures, which typically are 250
to 50 bar, respectively, so a total pressure of 300 bar.
[0098] It will be dependent on the specific homogenizer used, which
pressure to apply. A suitable way is to use a pressure of 100 bar
in the first step and 50 bar in the second step in a Niro Suavi NS
2006 H Homogenizer at a temperature of 60.degree. C. A suitable way
is to use a pressure of 5 bar in the first step and 20 bar in the
second step in a Niro Suavi NS 2006 H Homogenizer at a temperature
of 20.degree. C. Subsequently optionally other ingredients, not
being lipid, may be added.
[0099] 4 Sterilization
[0100] Subsequently, the emulsion obtained in step 3 is preferably
sterilized. Preferably sterilization takes place in-line at ultra
high temperature (UHT) and/or in appropriate containers to obtain a
formula in the form of a sterile liquid. A suitable way for UHT
treatment is a treatment at 120-130.degree. C. for at least 20 s.
Alternatively, the emulsion obtained in step 3 is concentrated by
evaporation, subsequently sterilized at ultra high temperature and
subsequently spray dried to give a spray dried powder which is
filled into appropriate containers.
[0101] Alternatively this sterilization step is performed before
the homogenization step. Preferably the composition obtained by the
above process is spray dried afterwards.
[0102] The difference on coating of the lipid globules can further
be derived from the zeta potential. Zeta potential (.zeta.
potential) measures the difference in milliVolts (mV) in
electrokinetic potential between the tightly bound layer around the
surface and the distant zone of electroneutrality and is a measure
of the magnitude of the repulsion or attraction between particles
in a dispersion. Its value is also related to the stability of
colloidal dispersions. A high absolute zeta potential will confer
stability, i.e. the solution or dispersion will resist
aggregation.
[0103] Digestible Carbohydrate Component
[0104] The composition preferably comprises digestible
carbohydrate. The digestible carbohydrate preferably provides 30 to
80% of the total calories of the composition. Preferably the
digestible carbohydrate provides 40 to 60% of the total calories.
When in liquid form, e.g. as a ready-to-feed liquid, the
composition preferably comprises 3.0 to 30 g digestible
carbohydrate per 100 ml, more preferably 6.0 to 20, even more
preferably 7.0 to 10.0 g per 100 ml. Based on dry weight the
present composition preferably comprises 20 to 80 wt. %, more
preferably 40 to 65 wt. % digestible carbohydrates.
[0105] Preferred digestible carbohydrate sources are lactose,
glucose, sucrose, fructose, galactose, maltose, starch and
maltodextrin. Lactose is the main digestible carbohydrate present
in human milk. The present composition preferably comprises
lactose. The present composition preferably comprises digestible
carbohydrate, wherein at least 35 wt. %, more preferably at least
50 wt. %, more preferably at least 75 wt. %, even more preferably
at least 90 wt. %, most preferably at least 95 wt. % of the
digestible carbohydrate is lactose. Based on dry weight the present
composition preferably comprises at least 25 wt. % lactose,
preferably at least 40 wt. %.
[0106] Non-Digestible Oligosaccharides
[0107] Preferably the present composition comprises non-digestible
oligosaccharides with a degree of polymerization (DP) between 2 and
250, more preferably 3 and 60. The non-digestible oligosaccharides
advantageously improve mineral absorption, bone composition and
architecture. The underlying mechanisms are via an increased
solubility of minerals is presumed to be via an increased bacterial
production of short-chain fatty acids in the intestine, and/or an
enlargement of the intestinal absorption surface by promoting
proliferation of enterocytes mediated by these short chain fatty
acids. Therefore the non-digestible oligosaccharides are presumed
to enhance the BMD and/or BMC increasing effects of the larger
lipid globules of the composition according to the present
invention.
[0108] The non-digestible oligosaccharide is preferably selected
from the group consisting of fructo-oligosaccharides (such as
inulin), galacto-oligosaccharides (such as
transgalacto-oligosaccharides or beta-galacto-oligisaccharides),
gluco-oligosaccharides (such as gentio-, nigero- and
cyclodextrin-oligosaccharides), arabino-oligosaccharides,
mannan-oligosaccharides, xylo-oligosaccharides,
fuco-oligosaccharides, arabinogalacto-oligosaccharides,
glucomanno-oligosaccharides, galactomanno-oligosaccharides, sialic
acid comprising oligosaccharides and uronic acid oligosaccharides.
Preferably the composition comprises gum acacia on combination with
a non-digestible oligosaccharide.
[0109] Preferably the present composition comprises
fructo-oligosaccharides, galacto-oligosaccharides and/or
galacturonic acid oligosaccharides, more preferably
galacto-oligosaccharides, most preferably
transgalacto-oligosaccharides. In a preferred embodiment the
composition comprises a mixture of transgalacto-oligosaccharides
and fructo-oligosaccharides. Preferably the present composition
comprises galacto-oligosaccharides with a DP of 2-10 and/or
fructo-oligosaccharides with a DP of 2-60. The
galacto-oligosaccharide is preferably selected from the group
consisting of transgalacto-oligosaccharides, lacto-N-tetraose
(LNT), lacto-N-neotetraose (neo-LNT), fucosyl-lactose, fucosylated
LNT and fucosylated neo-LNT. In a particularly preferred embodiment
the present method comprises the administration of
transgalacto-oligosaccharides ([galactose].sub.n-glucose; wherein n
is an integer between 1 and 60, i.e. 2, 3, 4, 5, 6, . . . , 59, 60;
preferably n is selected from 2, 3, 4, 5, 6, 7, 8, 9, or 10).
Transgalacto-oligosaccharides (TOS) are for example sold under the
trademark Vivinal.TM. (Borculo Domo Ingredients, Netherlands).
Preferably the saccharides of the transgalacto-oligosaccharides are
.beta.-linked.
[0110] Fructo-oligosaccharide is a non-digestible oligosaccharide
comprising a chain of .beta. linked fructose units with a DP or
average DP of 2 to 250, more preferably 10 to 100.
Fructo-oligosaccharide includes inulin, levan and/or a mixed type
of polyfructan. An especially preferred fructo-oligosaccharide is
inulin. Fructo-oligosaccharide suitable for use in the compositions
is also already commercially available, e.g. Raftiline.RTM.HP
(Orafti).
[0111] Uronic acid oligosaccharides are preferably obtained from
pectin degradation. Uronic acid oligosaccharides are preferably
galacturonic acid oligosaccharides. Hence the present composition
preferably comprises a pectin degradation product with a DP between
2 and 100. Preferably the pectin degradation product is prepared
from apple pectin, beet pectin and/or citrus pectin. Preferably the
composition comprises transgalacto-oligosaccharide,
fructo-oligosaccharide and a pectin degradation product. The weight
ratio transgalacto-oligosaccharide:fructo-oligosaccharide:pectin
degradation product is preferably (20 to 2):1:(1 to 3), more
preferably (12 to 7):1:(1 to 2).
[0112] Preferably, the composition comprises of 80 mg to 2 g
non-digestible oligosaccharides per 100 ml, more preferably 150 mg
to 1.50 g, even more preferably 300 mg to 1 g per 100 ml. Based on
dry weight, the composition preferably comprises 0.25 wt. % to 20
wt. %, more preferably 0.5 wt. % to 10 wt. %, even more preferably
1.5 wt. % to 7.5 wt. %. A lower amount of non-digestible
oligosaccharides will be less effective in preventing BMC and/or
BMD, whereas a too high amount will result in side-effects of
bloating and abdominal discomfort.
[0113] Protein Component
[0114] The present composition preferably comprises proteins. The
protein component preferably provides 5 to 15% of the total
calories. Preferably the present composition comprises a protein
component that provides 6 to 12% of the total calories. More
preferably protein is present in the composition below 9% based on
calories, more preferably the composition comprises between 7.2 and
8.0% protein based on total calories, even more preferably between
7.3 and 7.7% based on total calories. Human milk comprises a lower
amount of protein based on total calories than cow's milk. The
protein concentration in a nutritional composition is determined by
the sum of protein, peptides and free amino acids. Based on dry
weight the composition preferably comprises less than 12 wt. %
protein, more preferably between 9.6 to 12 wt. %, even more
preferably 10 to 11 wt. %. Based on a ready-to-drink liquid product
the composition preferably comprises less than 1.5 g protein per
100 ml, more preferably between 1.2 and 1.5 g, even more preferably
between 1.25 and 1.35 g.
[0115] The source of the protein should be selected in such a way
that the minimum requirements for essential amino acid content are
met and satisfactory growth is ensured. Hence protein sources based
on cows' milk proteins such as whey, casein and mixtures thereof
and proteins based on soy, potato or pea are preferred. In case
whey proteins are used, the protein source is preferably based on
acid whey or sweet whey, whey protein isolate or mixtures thereof
and may include .alpha.-lactalbumin and .beta.-lactoglobulin. More
preferably, the protein source is based on acid whey or sweet whey
from which caseino-glyco-macropeptide (CGMP) has been removed.
Removal of CGMP from sweet whey protein advantageously reduces the
threonine content of the sweet whey protein. A reduced threonine
content is also advantageously achieved by using acid whey.
Optionally the protein source may be supplemented with free amino
acids, such as methionine, histidine, tyrosine, arginine and/or
tryptophan in order to improve the amino acid profile. Preferably
.alpha.-lactalbumin enriched whey protein is used in order to
optimize the amino acid profile. Using protein sources with an
optimized amino acid profile closer to that of human breast milk
enables all essential amino acids to be provided at reduced protein
concentration, below 9% based on calories, preferably between 7.2
and 8.0% based on calories and still ensure a satisfactory growth.
If sweet whey from which CGMP has been removed is used as the
protein source, it is preferably supplemented by free arginine in
an amount of from 0.1 to 3 wt. % and/or free histidine in an amount
of from 0.1 to 1.5 wt. % based on total protein.
[0116] Casein is advantageously present. During digestion of casein
casein phosphopeptode (CPP) is released which improves BMD and/or
BMC. CPP improves calcium absorption in the small intestine.
Preferably the composition comprises at least 3 wt. % casein based
on dry weight. Preferably the casein is intact and/or
non-hydrolyzed.
[0117] Preferably the composition comprises calcium. Calcium is the
major cation of bone mineral. Preferably the composition comprises
at least 200 mg calcium based on 100 g dry weight, more preferably
at least 300 mg, even more preferably at least 350 mg/100 g dry
weight. Preferably the composition comprises less than 1500 mg
calcium per 100 g dry weight, more preferably less than 1000 mg
even more preferably less than 800 mg/100 g dry weight.
[0118] Preferably the composition comprises phosphate. Phosphate is
the major anion of bone mineral. Preferably the composition
comprises at least 100 mg phosphate based on 100 g dry weight, more
preferably at least 150 mg, even more preferably at least 200
mg/100 g dry weight. Preferably the composition comprises less than
1000 mg phosphate per 100 g dry weight, more preferably less than
500 mg even more preferably less than 350 mg/100 g dry weight.
[0119] Preferably the weight ratio calcium to phosphate is between
2.5 and 1.0, more preferably between 2.0 and 1.5. A balanced
calcium phosphate ratio beneficially effects BMD and/or BMC in
infants.
[0120] Preferably the composition comprises vitamin D. Vitamin D
regulates the calcium and phosphorus levels in the blood by
promoting their absorption from food in the intestines, and by
promoting re-absorption of calcium in the kidneys, which enables
normal mineralization of bones. It is also needed for bone growth
and bone remodeling by osteoblasts and osteoclasts. Preferably the
composition comprises at least 3 .mu.g vitamin D based on 100 g dry
weight, more preferably at least 5 .mu.g, even more preferably at
least 8 .mu.g/100 g dry weight. Preferably the composition
comprises less than 100 .mu.g vitamin D per 100 g dry weight, more
preferably less than 50 .mu.g, even more preferably less than 20
.mu.g/100 g dry weight.
[0121] Nutritional Composition
[0122] The present composition is particularly suitable for
providing the daily nutritional requirements to a human with an age
below 36 months, particularly an infant with the age below 24
months, even more preferably an infant with the age below 18
months, most preferably below 12 months of age. The present
composition comprises a lipid, a protein and a digestible
carbohydrate component wherein the lipid component preferably
provides 30 to 60% of total calories, the protein component
preferably provides 5 to 20% of the total calories and the
digestible carbohydrate component preferably provides 25 to 75% of
the total calories. Preferably the present composition comprises a
lipid component providing 35 to 50% of the total calories, a
protein component provides 6 to 12% of the total calories and a
digestible carbohydrate component provides 40 to 60% of the total
calories. The amount of total calories is determined by the sum of
calories derived from protein, lipids and digestible
carbohydrates.
[0123] The present composition is not human breast milk. The
present composition comprises vegetable lipids. The compositions of
the invention preferably comprise other fractions, such as
vitamins, minerals according to international directives for infant
formulae.
[0124] Preferably the composition is a powder to be reconstituted
with water. It was surprisingly found that the size and the coating
with polar lipids of the lipid globules remained the same after the
drying step and subsequent reconstitution. The presence of larger
lipid globules may have a slightly negative effect on the long term
stability of the liquid composition. However, separation of the
lipid and aqueous layers was not observed within 48 h, which is
much longer than the time between reconstituting the powder to a
ready to drink liquid and the consumption of it, which will be less
than 24 h and typically within 1 h. The composition being in a
powder form has therefore an additional advantage in the present
invention with large lipid globules.
[0125] In order to meet the caloric requirements of the infant, the
composition preferably comprises 50 to 200 kcal/100 ml liquid, more
preferably 60 to 90 kcal/100 ml liquid, even more preferably 60 to
75 kcal/100 ml liquid. This caloric density ensures an optimal
ratio between water and calorie consumption. The osmolarity of the
present composition is preferably between 150 and 420 mOsmol/l,
more preferably 260 to 320 mOsmol/l.
[0126] Preferably the composition is in a liquid form, with a
viscosity below 35 mPa.s, more preferably below 6 mPa.s as measured
in a Brookfield viscometer at 20.degree. C. at a shear rate of 100
s.sup.-1. Suitably, the composition is in a powdered from, which
can be reconstituted with water to form a liquid, or in a liquid
concentrate form, which should be diluted with water. When the
composition is in a liquid form, the preferred volume administered
on a daily basis is in the range of about 80 to 2500 ml, more
preferably about 450 to 1000 ml per day.
[0127] Infant
[0128] Bone growth is very fast during infancy. Hence, the present
composition is therefore advantageously administered to a human of
0-36 months, more preferably to a human of 0-18 months, more
preferably to a human of 0-12 months, even more preferably to a
human of 0-6 months.
[0129] Preferably the composition is to be used in infants which
are prematurely born or which are small for gestational age. These
infants experience after birth a catch up growth, which is an extra
risk for developing a too low BMD and/or BMC later in life.
[0130] Application
[0131] The present composition is preferably administered orally to
the infant. According to the present invention the BMD and/or BMC
increase, particularly at the age above 5 years, particularly above
13 years, more particularly above 18 years.
[0132] The inventors surprisingly found that when mice were fed,
during infancy and childhood, a food composition comprising
enlarged lipid globules, a different and significant effect on body
composition later in life was observed compared to mice which
during infancy and childhood had been fed a food composition having
a similar fatty acid composition, but a smaller lipid globule size.
At day 42, a day corresponding to childhood in a human setting, no
difference was observed in growth (weight) between the two groups,
but from day 42 both groups were fed a Western style diet which was
high in fat and high in palmitic acid. Surprisingly at day 70 and
126, which are a time points corresponding to early adulthood and
adulthood respectively in humans, the mice, which had previously
consumed the food composition of the present invention before
turning to the Western style diet, had a significantly increased
bone mineral content and increased bone mass density than mice
which had received a control composition during infancy. This
indicates that early nutrition has an effect on BMD and/or BMC
extending beyond the period in which it is actually administered.
In one embodiment the effect on BMD and/or BMC occurs later in
life. With later in life is meant an age exceeding the age at which
the diet is taken, preferably with at least one year.
[0133] The important difference between the two formulae was the
size of the lipid globules. The fatty acid profile was similar in
both formulae and the amount of palmitic and stearic acid present
at sn1 and sn3 positions in the fat was also similar. Both formulae
further enabled a similar good growth and development early in
life. Despite the improved fat absorption, an increased fat mass
was not observed, even an increase in lean body mass was observed.
The present inventors believe that the difference in lipid globule
architecture, in particularly the size, between the composition of
the present invention and conventional infant formulae on bone
health cannot be explained by an effect on improved calcium
absorption via a decrease of palmitic and/or stearic acid calcium
soap formation as known from the prior art with structured lipids.
Furthermore, the use of such lipids exerted a different effect on
body composition, such as body weight, lean body mass and fat mass,
as shown in example 4.
[0134] The present invention therefore can be used for food
compositions intended for infants and/or toddlers in order to
increase bone mineral content and/or increase bone mass density.
The present invention therefore can be used for food compositions
intended for infants and/or toddlers in order to prevent or reduce
the risk for osteoporosis later in life, for the enhancement of
bone formation and bone mass maximization and for the enhancement
of bone formation in infants and young children. Also
qualifications like `enhances bone strength` or `for stronger
bones` and the like are encompassed by the use or method according
to the present invention.
[0135] The present invention also allows to formulate infant milk
formulae with high levels of palmitic and stearic acid, as observed
in human milk and with the use of natural lipids, i.e. without the
use of synthetically made triglycerides, which are more expensive
and subject to strict food legislations
[0136] In this document and in its claims, the verb "to comprise"
and its conjugations is used in its non-limiting sense to mean that
items following the word are included, but items not specifically
mentioned are not excluded. In addition, reference to an element by
the indefinite article "a" or "an" does not exclude the possibility
that more than one of the element is present, unless the context
clearly requires that there be one and only one of the elements.
The indefinite article "a" or "an" thus usually means "at least
one".
EXAMPLE 1
Process for Preparing an IMF with Larger Lipid Globule Size
Example 1A
[0137] Infant formulae were prepared by dissolving using
demineralised whey powder, lactose, whey protein concentrate, skim
milk powder, galacto-oligosaccharides, minerals and vitamin pre-mix
in demineralised water to a dry weight content of 22.5 g/100 g and
heating the water phase at 65.degree. C.
[0138] The oil blend was prepared using over 98 wt. % vegetable
oils, an oil comprising LC-PUFA, oil soluble vitamins and
antioxidants. Both the water phase and the oil blend were heated to
65.degree. C. prior to mixing. The oil blend was added to the water
phase and blended with an Ultra-Turrax T50 for about 30-60 s at
5000-1000 rpm. The dry weight of this mixture was about 26%. The
product was UHT treated for 30 s at 125.degree. C. and subsequently
cooled to 20.degree. C.
[0139] For infant formula 1 the homogenization pressure was 200 and
50 bar, respectively in a Niro Suavi NS 2006 H homogenizer. For
infant formula 2 this mixture was homogenized in two steps at a
pressure of 5 and 20 bar respectively in a Niro Suavi NS 2006 H
homogenizer. The products were dried to a powder by spray drying.
Long chain inulin was blended dry into the powder. The amount of
vegetable glycerophospholipids was 0.2 wt. % based on total fat for
diet 1 and 2.
[0140] The size of the lipid globules was measured with a
Mastersizer 20000 (Malvern Instruments, Malvern UK) and shown in
Table 1. It was checked with confocal laser scanning microscopy
that the lipid globules were not coated with phospholipids, before
spray drying. As fluorescent probes Annexin V Alexa Fluor 488 (In
Vitrogen molecular probes) for labeling phospholipids, and Nile Red
(Sigma-Aldrich) for labeling triglycerides, were used. After
labeling the milk samples Vectrahield mounting medium (Vector
laboratories inc., Burliname USA) for reducing particle movement
and photo-bleaching was added. Observations were made using a Zeiss
Laser Scanning Microscope with excitation wavelengths of
488/543/633 nm and emission filters set at band pass 505-530, and
band pass 560-615.
TABLE-US-00001 TABLE 1 Lipid globule characteristics of different
milks Volume Mode Volume % with a diameter IMF diameter .mu.m
between 2 and 12 .mu.m 1, Standard IMF 0.5 5.1 2, Experimental IMF
4.0 72.2 (large lipid globules)
[0141] After 5 months storage at room temperature the size of the
lipid globules in diet 1 had not changed, with a volume mode
diameter of 0.5. Also the volume mode diameter of diet 2 was rather
stable being 4.8 .mu.m.
Example 1 B
[0142] An infant formula was prepared comprising per kg powder 4800
kcal, 248 g lipid, 540 g digestible carbohydrates, 55 g
non-digestible oligosaccharides and 103 g protein. The composition
was prepared using BAEF powder (Corman, Goe, Belgium), a vegetable
oil blend, demineralised whey powder, lactose, non-digestible
oligosaccharides (galacto-oligosaccharides and long chain
fructo-oligosaccharides in a weight ratio of 9/1). Also vitamins,
minerals, trace elements as known in the art were used.
[0143] The amount of BAEF was such that 7.24 wt. % phospholipids
(from BAEF) based on total lipids were present in the composition.
Based on a small amounts of phospholipids in the oil blend, the
total amount of phospholipids was 7.39 wt. % based on total lipid.
BAEF also supplied a small amount of cholesterol (about 0.08 wt. %
based on total lipid of the infant formula) and glycosphingolipids
(about 1.65% glycosphingolipids based on total lipid of the infant
formula). The BAEF powder was mixed with galacto-oligosaccharides,
lactose, vitamin pre-mixtures and mineral premixes in water, at
room temperature, by stirring. Potassium hydroxide was used to set
the pH at 6.8-7.0. The dry weight matter of the mixture was about
27%. The mixture was heated to 60.degree. C. The vegetable oil
blend was also heated to 60.degree. C. and added to the water phase
and blended with an Ultra-Turrax T50 for about 30-60 s at
5000-10000 rpm. Subsequently demi-water was added to achieve a dry
matter content of about 15%.
[0144] Subsequently the oil-water mixture was homogenised at a
pressure of 100 bar in a first step and 50 bar in a second step in
a Niro Suavi NS 2006 H Homogenizer. The temperature was 60.degree.
C. Subsequently demineralized whey powder was added to arrive at a
final dry matter content of 18%. The product was UHT treated at
125.degree. C. for 30 s. The product was dried to a powder by spray
drying. Maltodextrin together with long chain inulin was blended
dry into the powder.
[0145] The size of the lipid globules was measured with a
Mastersizer 20000 (Malvern Instruments, Malvern UK). The volumetric
mode diameter was 7.3 .mu.m. A second, much smaller peak was
present at 0.52 .mu.m. The volume % of lipid globules with a size
between 2 and 12 m was 71% based on total lipid volume. It was
checked with confocal laser scanning microscopy that the larger
lipid globules of the present invention were coated with
phospholipids, before spray drying and after reconstitution of the
spray dried powder with water. In both cases the lipid globules
were covered with a layer of phospholipids. As fluorescent probes
Annexin V Alexa Fluor 488 (In Vitrogen molecular probes) for
labeling the phospholipids, and Nile Red (Sigma-Aldrich) for
labeling triglycerides, were used. After labeling the milk samples
Vectrahield mounting medium (Vector laboratories inc., Burliname
USA) for reducing particle movement and photo-bleaching was added.
Observations were made using a Zeiss Laser Scanning Microscope with
excitation wavelengths of 488/543/633 nm and emission filters set
at band pass 505-530, and band pass 560-615. Also the size of the
lipid globules was the same before drying and after reconstitution
of the spray dried powder with water.
[0146] As a control the lipid globules of a standard infant formula
(Nutrilon 1) did not show labeling with phospholipids as observed
with the confocal laser scanning microscopy. Instead the globules
were covered with protein, as determined with the fluorescent
protein stain Fast Green FCF. The volumetric modal diameter of the
lipid globules in this standard infant milk formula was measured to
be 0.5 .mu.m. A second much smaller peak was present at 8.1 .mu.m.
The volume % of lipid globules with a size between 2 and 12 m was
34% based on total lipid volume.
TABLE-US-00002 TABLE 2 Lipid globule characteristics of different
milks Volume % with a Volume Mode diameter between .zeta. potential
diameter .mu.m 2 and 12 .mu.m (mV) Standard infant 0.5 34 -22.4
milk formula (Nutrilon 1) Infant milk 7.3 71 -16 formula of the
invention Human milk 5.3 98 -13.8
[0147] Also human milk was analyzed and showed a volumetric modal
diameter of the lipid globules of 5.3 .mu.m. The volume % of lipid
globules with a size between 2 and 12 m was 98% based on total
lipid volume. The lipid globules were covered with a layer of
phospholipids.
[0148] The zeta potentials and volume weighted mean diameters were
also measured. The results are shown in table 2.
EXAMPLE 2
Programming Effect of Lipid Globule Size on Adult Body
Composition
[0149] Offspring of C57/BL6 dams were weaned from day 15 on. The
experimental weaning diets were continued until day 42. From day 42
to day 126 all pups were fed the same diet based on AIN-93G diet
with an adjusted lipid fraction (containing 10 wt. % lipid of which
50 wt. % lard and 1% cholesterol, based on total lipid), which is
representative for a Western style diet.
[0150] The experimental diets that were used for weaning were:
[0151] 1) an infant milk formula (IMF) based control diet. This
diet comprised 282 g standard IMF per kg, IMF 1 of example 1A, i.e.
small lipid globules. The rest of the diet was AIN-93G protein,
carbohydrates and fibre. All lipid present in the diet was derived
from the IMF. [0152] 2) an IMF based diet of the present invention.
This diet differed from diet 1 in that it comprised 282 g IMF 2 of
example 1A, i.e. comprised lipid globules larger than the control.
All lipid present in the diet was derived from the IMF.
[0153] At day 42, all mice switched to a "Western style diet"
comprising 10 wt. % lipid until day 98. The fatty acid composition
of the two experimental was the same with calculated linoleic acid
(LA) of 14% based on total fatty acids, with alpha linolenic acid
(ALA) of 2.6 in based on total fatty acids and with LA/ALA of 5.4.
The amount of DHA was 0.2 wt. % and the amount of ARA was 0.35 wt.
%. The fatty acid composition of the Western style diet shown in
table 5.
[0154] The mice were weighed twice a week. The food intake was
determined once a week during the entire experiment. To determine
body composition (i.e., BMC, BMD, fat mass (FM) and fat-free mass
(FFM)) DEXA scans (Dual Energy X-ray Absorbiometry) were performed
under general anesthesia at 6, 10 and 14 weeks of age, 42, 70, and
98 days after birth respectively, by densitometry using a PIXImus
imager (GE Lunar, Madison, Wis., USA). At the age of 98 days the
male mice were sacrificed.
[0155] Results:
[0156] No effect on growth (body weight) and food intake was
observed during the experimental period between the groups.
Moreover, the development of body weight and fat mass (determined
with DEXA) was not significantly different at day 42 (end of the
diet intervention period). There was a direct diet effect on BMC.
Mice receiving diet 2 with large lipid globules showed a higher
BMC.
[0157] A subsequent treatment with a Western style diet between day
42 and day 98 of all groups resulted in clear differences in body
composition at the end of the experiment (day 98), see Table 3.
There was no difference in effects on BMD at day 98. However, on
day 98 the BMC was higher in mice receiving the lipid with large
lipid globules (diet 2 versus diet 1). This is indicative that the
effects on BMC are maintained later in life when receiving a diet
with large lipid globules early in life and that this increased
bone mineral content is accompanied by at least the same quality of
bone as expressed as BMD. Interestingly, the fat mass and relative
fat mass developed later in life was reduced in the mice which had
received the diet with the larger lipid globules during their
infancy and childhood, compared to mice which had received the
control diet.
TABLE-US-00003 TABLE 3 Bone Mineral Content, Bone Mass Density, Fat
Mass and relative Fat Mass. Parameter Day Diet 1 Diet 2 Body weight
g 42 25.84 (046) 26.10 (0.30) (s.e.) 70 30.93 (0.69) 30.19 (0.55)
98 33.98 (0.99) 33.60 (0.68) Lean Body mass 42 20.83 (0.30) 20.52
(0.21) g (s.e.) 70 20.34 (0.37) 22.19 (0.69) 98 22.89 (0.43) 22.73
(0.42) Bone mineral 42 0.461 (0.009) 0.475 (0.011)* content g 70
0.475 (0.007) 0.537 (0.018)* Mean (s.e.) 98 0.538 (0.009) 0.551
(0.012)* Bone Mass 42 0.049 (0.001) 0.050 (0.001) Density
g/cm.sup.2 70 0.054 (0.001) 0.055 (0.001) Mean (s.e.) 98 0.057
(0.001) 0.058 (0.001) Fat mass g 42 4.42 (0.19) 4.19 (0.13) Mean
(s.e.) 70 5.94 (0.39) 5.58 (0.27) 98 8.35 (0.67) 7.29 (0.37) Fat %
of body 42 17.43 (0.49 16.98 0.48 weight 70 22.40 (0.99 20.07 0.80
Mean (s.e.) 98 26.31 (1.33) 24.18 (0.83) *P < 0.05
[0158] These results demonstrate that the BMC and/or BMD in later
life clearly is increased by an early in life diet with increased
lipid globule size alone. It is concluded that food comprising
lipid globules with an altered lipid architecture program and/or
imprint the body early in life in such a way that later at life a
healthier body composition has developed, with increased BMD and/or
BMC, which prevents and/or reduces the risk for osteoporosis.
EXAMPLE 3
Programming Effect of Lipid Globule Size on Adult Body
Composition
[0159] Offspring of C57/BL6 dams were weaned from day 15 on. The
experimental weaning diets were continued until day 42. From day 42
to day 126 all pups were fed the same diet based on AIN-93G diet
with an adjusted lipid fraction (containing 10 wt. % lipid of which
50 wt. % lard and 1% cholesterol, based on total lipid), which is
representative for a Western style diet.
[0160] The experimental diets that were used for weaning were:
[0161] 1) an infant milk formula (IMF) based control diet. This
diet comprised 282 g standard IMF (Nutrilon 1) per kg, with the
lipid globule size as mentioned in example 1A. The rest of the diet
was AIN-93G protein, carbohydrates and fibre. All lipid present in
the diet was derived from the IMF. [0162] 2) an IMF based diet of
the present invention. This diet differed from diet 1 in that it
comprised 282 g IMF of example 1A, i.e. comprised lipid globules
larger than the control. All lipid present in the diet was derived
from the IMF.
[0163] At day 42, all mice switched to a "Western style diet"
comprising 10 wt. % lipid until day 126. The composition of the
diets is given in table 4. The fatty acid composition of the two
experimental and cafeteria diet is shown in table 5. The fatty acid
profile of the two experimental diets was very similar.
TABLE-US-00004 TABLE 4 composition of experimental diets per kg
Diet 1, Diet 2, IMF of Western Control IMF the invention style diet
Kcal 3922 3922 4016 Lipid (g) 70 70 100 Phospholipids (g) 0.12 5.16
n.d. Cholesterol (g) 0.00 0.06 1 Digestible 644 644 600
Carbohydrates (g) Lactose (g) 145.9 145.9 0 Sucrose, glucose (g) 85
85 150 Maltodextrin (g) 360 360 450 Fiber (g) 47.5 47.5 47.5
Protein 179 179 179 n.d. = not determined
[0164] The mice were weighed twice a week. The food intake was
determined once a week during the entire experiment. To determine
body composition (i.e., fat mass (FM) and fat-free mass (FFM)) DEXA
scans (Dual Energy X-ray Absorbiometry) were performed under
general anesthesia at 6, 10 and 14 weeks of age, 42, 70, 98 and 126
days after birth respectively, by densitometry using a PIXImus
imager (GE Lunar, Madison, Wis., USA). At the age of 126 days the
male mice were sacrificed and organs were dissected and weighed
(i.e. fat tissues, liver, Muscle tibialis). Blood was analyzed for
leptin, resistin, and (fasting) insulin.
TABLE-US-00005 TABLE 5 Fatty acid composition of the experimental
diets Diet 1, Diet 2, IMF of Western Control IMF the invention
style diet C12:0 9.4 8.7 5.3 C14:0 4.4 5.3 2.7 C16:0 18.7 21.3 23.1
C18:0 3.5 5.2 9.0 C18:1 n-9 39.9 37.7 40.5 C18:2 n-6 (LA) 15.7 12.6
11.9 C18:3 n-3 (ALA) 2.4 2.1 1.3 Others 6.0 7.1 6.7 n-6 16.0 12.9
11.9 n-3 2.4 2.1 1.3 n-6/n-3 6.58 6.12 9.1 LA/ALA 6.46 6.00 9.15
SFA 39.3 44.4 41.9 MFA 42.1 39.8 42.3 PUFA 18.3 14.9 13.2
[0165] Results:
[0166] No significant difference regarding growth (body weight) and
food intake was observed during the experimental period between the
two groups.
[0167] A subsequent exposure to a Western style diet between day 42
and day 126 of all groups resulted in clear differences in body
composition at the end of the experiment (day 126), see Table 6.
Both the BMC and BMD developed later in life were increased in the
mice which had received the diet with the larger lipid globules
during their infancy and childhood, compared to mice which had
received the control diet. The overall body weight was comparable
between the two groups. The experimental group had an increased
lean body mass.
TABLE-US-00006 TABLE 6 Body weight, lean body mass, bone mineral
content and bone mass density in time in mice receiving control or
experimental diet during infancy. Diet 1, Diet 2, IMF of Parameter
Day Control IMF the invention Bodyweight g 42 23.50 (0.45) 24.24
(0.51) Mean (s.e.) 70 29.88 (0.46) 30.16 (0.77) 98 33.32 (0.57)
33.69 (0.95) 126 34.47 (0.80) 34.15 (1.16) Lean body mass g 42
18.96 (0.34) 19.96 (0.40)* Mean (s.e.) 70 21.31 (0.42) 22.32 (0.48)
98 22.22 (0.49) 23.91 (0.45)* 126 23.30 (0.43) 24.19 (0.53)* Bone
Mineral Content g 42 0.364 (0.005) 0.383 (0.009) Mean (s.e.) 70
0.436 (0.007) 0.474 (0.013)* 98 0.468 (0.011) 0.501 (0.013)* 126
0.482 (0.000) 0.523 (0.012)* Bone Mass Density g/cm.sup.2 42 0.046
(0.000) 0.048 (0.001)* Mean (s.e.) 70 0.051 (0.001) 0.053 (0.001)
98 0.052 (0.001) 0.055 (0.001) 126 0.052 (0.001)) 0.055 (0.001) Fat
mass g Mean 42 3.78 (0.13) 3.77 (0.21) (s.e.) 70 7.84 (0.35) 7.13
(0.65) 98 10.68 (0.53) 9.19 (0.79)* 126 10.48 (0.67) 9.11 (0.90)*
Fat % of body weight 42 16.59 (0.45) 15.83 (0.68) Mean (s.e.) 70
26.89 (1.07) 23.81 (1.61) 98 32.38 (1.42) 27.25 (1.67)* 126 30.78
(1.42) 26.67 (1.77)* *P < 0.05 compared to control group
[0168] The amount of palmitic acid is 18.7% in control and 21.3% in
the experimental group, so it is even higher in the experimental
group. About 25% of the fat in the experimental diet is derived
from cow's milk fat, having about 37.8% of the palmitic acid in the
sn-2 position thus having about 72.2% palmitic acid in the sn-1 and
sn-3 position. The vegetable fat has about 7.5% of its palmitic
acid residues in the sn-2 position and thus 92.5% in the sn-1 and
sn-3 position. So, in the control diet 18.7*0.925=17.3% of the
palmitic acid residues are present on the sn-1 and sn-3 positions,
based on total fatty acids and in the experimental group
(0.25*0.772*21.3)+(0.75*0.925*21.3)=18.1% of the palmitic acid is
present in the sn-1 and sn-3 positions based on total fatty acids.
Since palmitic acid residues at the sn-1 and sn-3 position are
associated with decreased calcium absorption, decreased fat
absorption and decreased BMD and/or BMC, it is very surprising that
an increased BMD and/or BMC was observed when the experimental diet
was taken during early growth.
[0169] These results demonstrate that the BMC and/or BMD in later
life clearly is increased by an early in life diet with increased
lipid globule size. It is concluded that food comprising lipid
globules with an altered lipid architecture program and/or imprint
the body early in life in such a way that later at life a healthier
body composition has developed, with increased BMD and/or BMC,
which prevents and/or reduces the risk for osteoporosis. When
comparing the outcome of experiment 2 and 3, the effects at day 98
are relatively higher in experiment 3, indicative for a further
improved effect when the lipid globules are coated with polar
lipids, more preferably milk derived polar lipids.
[0170] Interestingly, at the same time the development of fat mass
(being not significantly different at day 42, the end of the diet
intervention period) the fat mass and relative fat mass developed
later in life was reduced in the pups which had received the diet
with the larger lipid globules during their infancy and childhood,
compared to pups which had received the control diet.
EXAMPLE 4
[0171] In parallel an experiment was performed wherein the effects
of an IMF with standard vegetable lipid was compared with IMF
wherein the lipid component comprises structured triglycerides with
an increased amount of palmitic acid in the sn-2 position. From
literature it is known that upon using such lipids, less free
palmitic acid is formed, resulting in less formation of insoluble
calcium palmitate, thereby increasing the bioavailability of
calcium and palmitic acid. The experimental set up was similar as
in example 3. The tested diets were based on AIN93-G comprising the
same carbohydrate and protein component. The diet comprised 7%
lipid, wherein diet 1 comprised the palm oil, coconut oil, rapeseed
oil, sunflower oil, and high oleic acid sunflower oil. In diet 2
part about 70 wt % of the fat was Betapol 45 (Lipid Nutrition, The
Netherlands) a lipid in which about 45% of the palmitic acid is
esterified in the sn-2 position of the triglyceride instead of the
7.5% typical for vegetable fats. The fatty acid composition of the
diets is very similar, see Table 7.
TABLE-US-00007 TABLE 7 Fatty acid composition of the diets Diet 1,
Diet 2, Control IMF betapol C12:0 11.5 11.5 C14:0 4.6 4.3 C16:0
17.1 17.1* C18:0 3.0 2.8 C18:1 n-9 36.0 38.7 C18:2 n-6 (LA) 14.0
14.0 C18:3 n-3 (ALA) 2.6 2.6 Others 11.2 9.3 *Increased fraction
position at the sn-2 position of the lipid.
[0172] Results are shown in table 8. The diet comprising more
palmitic acid on the sn-2 position increased body weight, lean body
mass and fat mass on day 42. These effects were maintained later in
life. An increase later in life on bone mineral content and bone
mineral density was also observed. This differs from the effect of
the large lipid globules of example 2 and 3, where a concomitant
decrease in fat mass and no effect on overall body weight was
observed later in life and wherein the direct diet effects (i.e.
effects on day 42) on body weight, lean body mass and fat mass were
much less.
TABLE-US-00008 TABLE 8 Bone Mineral Content, Bone Mass density, Fat
mass and relative fat mass. Day Diet 1 Diet 2 Body weight g 42 23.8
(0.55) 24.7 (0.63) Mean (s.e.) 70 26.8 (0.72) 30.3 (0.86)* 98 28.1
90.67) 31.8 (1.49)* Lean body mass g 42 18.0 (0.81) 20.7 (0.41)
Mean (s.e) 70 20.0 (0.33) 22.7 (0.54)* 98 21.3 90.48) 22.9 (0.77)*
Bone mineral content g 42 0.410 (0.013) 0.411 (0.010) Mean (s.e.)
70 0.504 (0.008) 0.535 (0.014) 98 0.557 (0.016) 0.583 (0.18) Bone
mineral density g/cm2 42 0.045 (0.001) 0.046 (0.001) mean (s.e.) 70
0.053 (0.001) 0.054 (0.001) 98 0.053 (0.001) 0.055 (0.001) Fat mass
g 42 3.7 (0.22) 4.3 (0.14) Mean (s.e.) 70 5.1 (0.22) 5.9 (0.44) 98
5.1 (0.23) 5.4 (0.62) Fat % of body weight 42 17.0 (0.39) 17.6
(0.57) Mean (s.e.) 70 20.3 (0.57) 20.4 (0.91) 98 19.2 (0.78) 18.8
(1.33) *p < 0.05
EXAMPLE 5
Infant Nutrition with Larger Lipid Globules Size
[0173] An infant formula comprising per kg powder 4810 kcal, 255 g
lipid, 533 g digestible carbohydrates, 58 g non-digestible
oligosaccharides (galacto-oligosaccharides and long chain
fructo-oligosaccharides in a weight ratio of 9/1), 96 g protein,
and vitamins, minerals, trace elements as known in the art.
[0174] The lipid composition is such that 0.57 wt. % of the lipid
is composed of phospholipids. The composition comprises about 0.17
wt. % glycosphingolipids based on total lipid. The composition
comprises about 0.006 wt. % cholesterol based on total lipids. As a
source of phospholipids, glycosphingolipids and cholesterol SM-2
powder (Corman, Goe, Belgium) is used. About 97-98% of the lipid is
vegetable lipid, the rest being milk fat, fish oil and microbial
oil. The amount of LC-PUFA is about 0.64 wt. % based on total fatty
acids. The LA/ALA ratio is 5.2.
[0175] Homogenization was performed similar as in example 1. The
volumetric mode diameter was above 1 .mu.m. The volume % of lipid
globules with a size between 2 and 12 m was above 45% based on
total lipid volume.
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