A ailable online a www.sciencedi ec .com
Food and Biop oduc s P ocessing
jou nal homepage: www.else ie .com/loca e/ bp
Assessmen o whole egg ac iona ion by
angen ial low il a ion: The p oblem o low-
densi y lipop o ein agg ega es
Gema Pue as, Pa icia Cazón, Manuel Vázquez
⁎
Depa men o Analy ical Chemis y, Food Technology A ea, Facul y o Ve e ina y, Campus Te a, Uni e si y o
San iago de Compos ela, 27002 Lugo, Spain
a icle in o
A icle his o y:
Recei ed 29 Oc obe 2022
Recei ed in e ised o m 4 May 2023
Accep ed 11 May 2023
A ailable online 13 May 2023
Keywo ds:
Fil a ion
Egg p oduc s
Sepa a ion
Choles e ol
Low-densi y lipop o eins
Food ma ix
abs ac
Eggs a e a compelling sou ce o bioac i e compounds. Memb ane sepa a ion is a g een
echnology ha allow componen s o be ob ained while p ese ing hei biological ac-
i i y. The objec i e o his s udy was o assess he applica ion o memb ane sepa a ion
echnology o egg p oduc s. Whole egg plasma, egg yolk plasma and egg whi e we e
angen ial low il a ed e alua ing he e ec o he po e size and memb ane ma e ials.
T ansmission o d y ma e and p o eins om eed o il a e was no iced in all he p o-
cesses. Ne e heless, choles e ol and ca o enoids we e no de ec ed a il a es om
750 kDa o 0.2 µm when mixed es e o polye he sul one we e he memb ane ma e ials.
The in e ac ions be ween egg yolk and egg whi e componen s di icul he sepa a ion.
Choles e ol was pa o low-densi y lipop o eins. These lipop o eins oge he wi h ca -
o enoids we e il a ed wi h modi ied polye he sul one a a minimum po e size o 0.2 µm.
High unce ain y was ob ained due o low-densi y lipop o eins agg ega ion. Th ee kinds
o agg ega es can jus i y hese esul s: low-densi y lipop o eins agg ega ed hemsel es,
low-densi y lipop o eins bound o o omucin and o he smalle egg whi e p o eins
(< 750 kDa), and low-densi y lipop o eins bound only o smalle egg whi e p o eins. The
de ec ed chela e e ec o egg whi e p o eins could be impo an in i o. The e o e, e-
sea ch should be led o assess he in luence o cooking me hod and diges ion o he di -
e en egg componen s on he bioaccessibili y o nu ien s and he gene a ion o bioac i e
pep ides. Mo eo e , he il a ion p ocesses analysed he ein may be use ul echniques o
liquid ood ma ix s udies.
© 2023 The Au ho (s). Published by Else ie L d on behal o Ins i u ion o Chemical
Enginee s. This is an open access a icle unde he CC BY-NC-ND license (h p://c ea i-
ecommons.o g/licenses/by-nc-nd/4.0/).
1. In oduc ion
Hen eggs a e a aluable sou ce o essen ial amino acids and
highly bioa ailable p o eins. I s high con en in hese makes
he egg a benchma k o measu ing he quali y o ood p o-
eins. They also p o ide essen ial a y acids, phospholipids,
choline, sialic acids, mine als, ca o enoids and all i amins
excep i amin C. This composi ion awa d heal h bene icials
o eggs, including an ioxidan p ope ies, an imic obials,
an icance , an ihype ensi e and immunomodula o y ac i -
i ies (Réhaul -Godbe e al., 2019). In addi ion, eggs a e a
mode a e ene gy sou ce wi h a lo o culina y po en ial and
gene ally a ailable a an a o dable p ice (Cza nowska-
Kujawska e al., 2021).
Despi e his g ea nu i ional composi ion, hei high
choles e ol con en o 415 mg / 100 g o edible po ion
(Pue as and Vázquez, 2021a) has ypically domina ed hei
nu i ional e alua ion. Se e al s udies ha e ocused on how
egg in ake modi ied blood choles e ol le els wi h con-
o e sial conclusions. The Eu opean Food Sa e y Au ho i y
h ps://doi.o g/10.1016/j. bp.2023.05.003
0960-3085/© 2023 The Au ho (s). Published by Else ie L d on behal o Ins i u ion o Chemical Enginee s. This is an open access a icle
unde he CC BY-NC-ND license (h p://c ea i ecommons.o g/licenses/by-nc-nd/4.0/).
]]]]
]]]]]]
⁎
Co esponding au ho .
E-mail add ess: [email p o ec ed] (M. Vázquez).
Food and Biop oduc s P ocessing 140 (2023) 99–109
(EFSA) ecognised ha he e is a posi i e dose-dependen
ela ionship be ween in ake o die a y choles e ol wi h blood
low densi y lipop o eins (LDL)-choles e ol concen a ions.
Howe e , such au ho i y decided no o p opose a e e ence
on choles e ol in ake (Pue as and Vázquez, 2019a). Nowa-
days he ecommenda ion o die a y choles e ol consump-
ion is o be as low as possible wi hou comp omising he
nu i ional adequacy o he die (U.S. Depa men o
Ag icul u e and U.S. Depa men o Heal h and Human
Se ices, 2020). Like ha , eggs a e included in a heal hy
die a y pa e n.
Choles e ol in egg is in he yolk exclusi ely. I is 95%
linked o LDL and he emainde is bound o high densi y
lipop o eins (HDL) o lipo i ellin (G i in, 1992). LDL a e
sphe ical nanopa icles wi h a lipid co e o iglyce ides and
choles e ol es e s su ounded by a mono ilm o phospholi-
pids, p o eins and non-es e i ied choles e ol (An on, 2013).
Abou 90% o LDL pa icles had a diame e be ween 17 and
100 nm (Xie e al., 2020) The mono ilm allows he solubili y o
lipids in he aqueous phase. Egg yolk is a na u al oil in wa e
emulsion whe e he con inuous aqueous phase is called
plasma and he dispe sed phase a e insoluble dense s uc-
u es (0.3–2 mm) e e ed o as g anules. Cen i uga ion o
whole egg p oduces he sepa a ion o hese phases. Two egg
p oduc s a e ob ained: he pelle ha encompasses he
g anules and he supe na an ha con ains egg yolk plasma
and albumen (Pue as and Vázquez, 2021a). Egg yolk plasma
con ains LDL and li e ins, while g anules included HDL,
phos i in and LDL g anules (An on, 2013). The liquid na u e
o plasma makes i a good a ge o a memb ane sepa a ion
p ocess.
Mul iple s udies ha e been done o ob ain he bioac i e
compounds o egg yolk and egg whi e independen ly (Li e al.,
2022; Zhang e al., 2021). Howe e , yolk and albumen a e
no mally cooked and consumed oge he ; and whole egg is
he ull sou ce o nu ien s and bioac i e compounds.
Nowadays, he impo an is he whole ood and ood ma ix
s udies a e being explo ed (Mille e al., 2023). As nume ous
nu ien s in he ood ma ix can in e ac in a syne gis ic o
an agonis ic manne , he undamen al nu i ional en i y is
he ood con aining all nu ien s a he han he nu ien in
isola ion (Pé ez-Jiménez e al., 2018). In he p esen , die s
ha ocus on speci ic nu ien s, such as a , o add ess me-
abolic diseases a e deemed subop imal; hus, his o ical
die a y app oaches aimed a isola ed nu ien s ha e become
obsole e (Mille e al., 2023). The e o e, angen ial low il-
a ion (TFF) was applied o whole egg as a ood ma ix case.
Two memb ane sepa a ion p ocesses in TFF, also called
c oss- low, we e s udied in ou wo k: ul a il a ion (UF) and
mic o il a ion (MF). Bo h o hem a e p essu e-d i en. A
each p ocess, wo p oduc s a e ob ained om he eed in-
oduced in o he memb ane sys em. Re en a e is he ac-
ion e ained by he memb ane. Fil a e o pe mea e is he
ac ion ha passes h ough he memb ane. I is c oss low
because he eed is pumped pa allelly o he memb ane, and
i is possible o eci cula e he e en a e back o he eed low.
While ‘p essu e-d i en’ means ha he main d i ing o ce o
hese p ocesses is ansmemb ane p essu e (TMP); he
p essu e disc epancy be ween e en a e side and il a e side
(Dhineshkuma and Ramasamy, 2017).
The use o memb ane echnology as a p ocessing and
sepa a ion me hod in ood indus y is gaining wide applica-
ions. I is conside ed a g een echnology. Because mem-
b ane sepa a ion p ocesses a e mo e e icien in economic
and ene ge ic e ms compa ed o high empe a u es ea -
men s o pas eu iza ion and s e iliza ion. In addi ion, hey
could a oid he use o addi i es and chemicals ( o example
o ex end he shel -li e o sol en s o ex ac ion), which is
bo h be e o en i onmen and human heal h.
Fu he mo e, hey p ese e he na u al as e and nu i ional
alue o ood p oduc s in hea -sensi i e componen s
(Dhineshkuma and Ramasamy, 2017), such as eggs. The e-
o e, i has been explo ed as a me hod o ood componen s
ac iona ion. Fo example, egg whi e p o eins ha e been
ob ained using UF, wi h he bene i o e aining hei biolo-
gical ac i i y (Li e al., 2022). O he s udies ha ul a il a ed
eggs we e, o example, o desalina e sal ed duck egg whi es
(Thammasena e al., 2020) o o isola e an ioxidan pep ides
p oduced om egg yolk p o ein enzyma ic hyd olysis (Chay
Pak Ting e al., 2011). In ac , UF is al eady employed in ood
indus y o whole egg and egg whi e concen a ion (Guiga
and Lameloise, 2019). Howe e , o ou knowledge, MF o egg
p oduc s had ne e been s udied un il ou p e ious wo k
(Pue as e al., 2023). A ha publica ion i was demons a ed
ha he egg p oduc s ob ained h ough TFF can be accu a ely
quan i ied by UV–VIS–NIR spec oscopy combined wi h che-
mome ics.
UF e e s o memb ane p ocess wi h po es anging ap-
p oxima ely om 3 o 100 nm. MF conce ns memb anes wi h
po e sizes anging om 0.1 o 10 µm. Bac e ia, yeas s and
milk a globules ha e app op ia e sizes o MF while i us
and soluble p o eins o UF. Colloids and casein micelles a e
sui able o bo h il a ions (Guiga and Lameloise, 2019).
Since UF mainly add esses mac omolecules sepa a ion (such
as p o eins and pep ides) is p e e ably exp essed as mola
mass. The memb anes employed a e classi ied acco ding o
he molecula weigh cu - o (MWCO), in g/mol o Da
(Dal on), ha is de ined as he mola mass o he molecule
ejec ed a 90% (Guiga and Lameloise, 2019). Howe e , he
sepa a ion p inciple is no based on he po e sizes alone, he
cha ge o he molecule/solu es and hei a ini y o he il-
e ing memb ane a e also key aspec s, especially in UF
(Dhineshkuma and Ramasamy, 2017). Hyd ophilic mem-
b anes a e conside ed mo e ad an ageous o e hyd ophobic
o il e an aqueous solu ion o p o eins a neu al pH. Be-
cause nega i ely cha ged p o eins a e mo e epulsi e agains
hyd ophilic memb anes esul ing in less occu ence o
memb ane ouling. This phenomenon indica es he deposi-
ion o solu es/ pa icles on he memb ane su ace and/o
in o he memb ane po es (Dhineshkuma and Ramasamy,
2017). I is desc ibed o be one o he key challenges e-
s ic ing he p ac ical applica ion o UF (Mohammad e al.,
2012). Thus, plasma is a be e a ge han whole egg o egg
yolk. A ouling mi iga ion echnique employs o example in
cell sepa a ion p ocess is al e na ing angen ial low il a-
ion (ATF) (Weinbe ge e al., 2022). In ATF, al e na ing
p essu e and acuum a e applied o he il a e low using a
diaph agm pump. As a esul , he e e se low is used o
educe he ouling o he hollow ibe memb ane
(Ma e, 2022).
The objec i e o his s udy was o assess he applica ion o
TFF o egg p oduc s. Low p essu es, no addi i es, and only
wa e was employed o dilu e whole egg plasma, egg yolk
plasma and egg whi e be o e UF and MF. Choles e ol, d y
ma e , p o ein con en and colou we e analysed. Whole egg
was chosen as he ull sou ce o bioac i e compounds as case
s udy. Yolk and whi e we e employed o unde s and he
mechanisms in ol ed.
100 Food and Biop oduc s P ocessing 140 (2023) 99–109
2. Ma e ials and me hods
2.1. Egg p oduc s p epa a ion
The egg p oduc s employed a e desc ibed a ou p e ious
wo k (Pue as e al., 2023). B ie ly, 11 s a ing egg p oduc s
we e p epa ed om 89 esh eggs. These s a ing p oduc s
we e 7 liquid whole eggs (made wi h 8 o 10 eggs each), 2
liquid egg yolks and 2 liquid egg whi es (made wi h 12 and 13
eggs manually sepa a ed). To simpli y, liquid egg whi e is
called egg whi e hence o h. All samples we e homogenised
using a high-pe o mance homogenize (Ul a Tu ax®, IKA,
S au en, Ge many) a 10,000 pm o 1 min
Nex , liquid egg yolk was dilu ed 1:1 wi h ap wa e be o e
cen i uga ion. Following, liquid whole egg and dilu ed liquid
egg yolk samples we e cen i uged a 12,000 g, 40 min and 8
ºC (So all Con i uge S a os, The mo Fishe Scien i ic,
Wal ham, MA, USA). Condi ions we e se led acco ding o
p e ious s udies (Pue as and Vázquez, 2021a, 2021b) o
maximize g anules sepa a ion and choles e ol in plasma.
These se ings caused some choles e ol p ecipi a ion in-
e i ably due o some LDL p ecipi a ion in g anules. Max-
imum g anules’ yield was p io i ised o minimize memb ane
ouling. A e , plasmas we e sepa a ed om g anules
manually.
2.2. Tangen ial low il a ion
The egg p oduc s il a ed a e iden i ied in Table 1. A o al o
11 uns we e pe o med. Whole egg plasmas we e analysed
om un 1–7, egg yolk plasmas we e employed a uns 8 and
9; while egg whi es we e s udied a uns 10 and 11. Egg
p oduc s we e dilu ed 1:1 (w/w) wi h wa e be o e il a ion.
K osFlo® Resea ch IIi TFF Sys em (Repligen, Cali o nia, USA)
was used. This sys em employs hollow ibe modules, and i
was se led in a ba ch il a ion con igu a ion. As he aim was
o p ese e nu i ional and unc ional p ope ies o egg p o-
duc s o ganic polyme ic memb anes we e conside ed o e
ino ganic ma e ials. They a e a ailable in a wide ange o
po e sizes, cheape , mo e sensi i e o washing chemicals and
no mally ha e a high packing densi y (Dhineshkuma and
Ramasamy, 2017). The cha ac e is ics o he memb anes
employed a e desc ibed in Table 1. MF was achie ed wi h
h ee po e sizes: 0.1 µm, 0.2 µm and 0.65 µm. While UF was
pe o med wi h a MWCO o 750 kDa. Assuming he simples
shape o a sphe e, his MWCO is equi alen o a p o ein
diame e o abou 12 nm minimum (E ickson, 2009). Th ee
kinds o hyd ophilic o ganic memb anes we e s udied:
Polye he sul one (PES), modi ied polye he sul one (mPES)
and mixed es e (ME, consis ing o cellulose ace a e and
cellulose ni a e). Acco ding o he hollow ibe inne lumen
and hei e ec i e leng h, i e su ace a eas we e analysed
om 500 cm
2
o 2600 cm
2
.
Th ee pa ame e s we e calcula ed o quan i y he mem-
b ane sepa a ion: ansmission a e (T ), concen a ion ac o
(CF) and weigh educ ion a io (WRR). T ansmission a e was
exp essed as a pe cen age acco ding o he ollowing
equa ion:
= ×T Concen a ion
Concen a ion
(%) 100
i
Fil a e
Re en a e
(1)
Whe e i was he composi ion pa ame e s udied because
ansmission a es we e calcula ed o each composi ion
pa ame e s udied (see below). The concen a ions o he
composi ion pa ame e s s udied we e measu ed a he end
o he TFF in he il a e and he e en a e (Dhineshkuma
and Ramasamy, 2017).
CF is he a io be ween he concen a ion in he e en a e
di ided by ini ial concen a ion (Guiga and Lameloise, 2019).
They we e also calcula ed o each composi ion pa ame e :
=CF Concen a ion
Concen a ion
i
Re en a e
Ini ial
(2)
Whe eas WRR is he a io o he ini ial weigh di ided by inal
weigh . This e m is no mally calcula ed as olume educ ion
a io (VRR) (Guiga and Lameloise, 2019). In his wo k, ini ial
and e en a e samples we e weigh ed:
=WRR Weigh
Weigh
Ini ial
Re en a e
(3)
Ini ial concen a ion o weigh conside ed we e p e ious
dilu ion.
2.3. Composi ion pa ame e s
The composi ion pa ame e s s udied on he ini ial eed
(plasmas and egg whi e), e en a e and il a e we e d y
ma e (DM), p o eins and choles e ol. DM con en was de-
e mined by g a ime ic me hod. An aliquo o 2–5 g o
sample was weigh ed, d ied o 24 h in an o en a 105 ºC and
Table 1 – Iden i ica ion and cha ac e iza ion o he il a ion sys ems s udied.
Run Sys em Cha ac e is ics
No. Sample Po e size / MWCO Memb ane ma e ial E ec i e Fibe Leng h (cm) Su ace a ea (cm
2
) Fibe inne lumen (mm)
1 WEP 750 kDa mPES 41.5 1600 0.5
2 WEP 750 kDa mPES 41.5 1600 0.5
3 WEP 0.1 µm ME 20 720 0.63
4 WEP 0.2 µm PES 20.8 500 1
5 WEP 0.2 µm PES 20.8 500 1
6 WEP 0.2 µm mPES 65 2600 0.5
7 WEP 0.65 µm mPES 41.5 1075 0.75
8 EYP 750 kDa mPES 41.5 1600 0.5
9 EYP 0.65 µm mPES 41.5 1075 0.75
10 EW 750 kDa mPES 41.5 1600 0.5
11 EW 0.65 µm mPES 41.5 1075 0.75
WEP: whole egg plasma. EYP: egg yolk plasma. EW: egg whi e. MWCO: molecula weigh cu -o . mPES: modi ied polye he sul one. ME: mixed
es e . PES: polye he sul one.
101 Food and Biop oduc s P ocessing 140 (2023) 99–109
hen d ied sample was weigh ed. DM con en was exp essed
as g d y ma e pe 100 g esh sample.
P o ein con en was measu ed wi h he mic o olume
spec opho ome e Nanod op 2000 (The mo Scien i ic™,
Wal ham, MA, USA). The me hod ollowed is desc ibed
elsewhe e based on he Wa bu g and Ch is ian me hod
(Wa bu g and Ch is ian, 1942). B ie ly, i was an es ima ion o
p o ein based on he maximum abso p ion a 280 nm ha
mos p o eins exhibi due p ima ily o he p esence o y -
osine and yp ophan. As nucleic acids also abso b a ha
wa eleng h, a p opo ion is emo ed by calcula ion. Thei
con ibu ion is elimina ed by sub ac ing a p opo ion o he
abso bance a 260 nm, whe e nucleo ides abso b much mo e
s ongly (Layne, 1957). P o ein con en was exp essed as mg
pe ml o sample. An enzyma ic ki me hod (Enzy ec™, R-
Biopha m AG, Da ms ad , Ge many) was employed o cho-
les e ol quan i ica ion based on p e ious s udies (Pue as
and Vázquez, 2019b). The sizes o he samples o analysis
we e 0.5 g o egg yolk, 1 g o liquid whole egg, 1.5–2 g o
plasmas, 1–2.5 g o e en a es and 5 g o il a es. These
quan i ies we e decided based on p elimina y s udies o
wo k wi hin he de ec ion limi s s ablished by manu ac u e .
Choles e ol con en was exp essed as mg pe 1 g o egg p o-
duc . All he composi ion de e mina ions we e done a leas
in duplica e, excep o p o ein con en ha measu emen s
we e in iplica e.
Ini ial pH o plasmas and egg whi es we e measu ed wi h
C ison pH 25 + pH-me e (C ison Ins umen s S. A.,
Ba celona, Spain).
2.4. Colou pa ame e s
Spec opho ome e Jasco V670 (Jasco Inc., Hachioji, Tokyo,
Japan) was employed o acqui e he VIS spec a om 380 o
780 nm o each egg p oduc . The cu e e employed was made
o Qua z Sup asil ® 300 wi h a 1 mm ligh pa h (Hellma
GmbH & Co. KG, Mulheim, Ge many). Samples we e mea-
su ed a 22–24 °C. The spec al da a we e collec ed wi h
Spec a Manage ™ II so wa e (Jasco Inc., Hachioji, Tokyo,
Japan). This so wa e calcula ed he CIE pa ame e s L* ,
a* and b* es ablished on he 2 Deg ee s anda d obse e wi h
ligh sou ce D65. Duplica e alues we e achie ed o each
sample. The pa ame e L* exp esses he deg ee o ligh ness
(0 = black, 100 = whi e). The coo dina e a* measu es edness
(−100 = g een, 100 = ed) and b* e lec s yellowness (−100 =
blue, 100 = yellow) (Pue as e al., 2023).
2.5. S a is ical analysis
Signi ican di e ences be ween he il a ion esul s we e
de e mined by one-way analyses o a iance (ANOVA) and
pos -hoc Tukey’s Hones ly Signi ican Di e ence (HSD) es .
Signi ican di e ence a p < 0.05 was de ined.
3. Resul s and discussion
3.1. Fil a ion pa ame e s and composi ion esul s
In his wo k, we in es iga ed TFF as a me hod o egg com-
pounds sepa a ion in he whole egg plasma ma ix. Egg yolk
plasma and egg whi e we e also analysed o de ec di e -
ences be ween ood ma ices and o comp ehend he me-
chanisms in ol ed. The sepa a ion p ocess was analysed
h ough changes in DM, p o eins, choles e ol and colou
pa ame e s. Ou app oach aimed o p ese e nu i ional and
unc ional p ope ies o egg p oduc s. The e o e, o ganic
hyd ophilic memb anes we e analysed and no addi i es
we e employed, only wa e was added as diluen . Mean TMP
and il a e luxes a e desc ibed in Table 2. To a oid i e e -
sible memb ane ouling, il a ion was s opped when p es-
su e a he eed ose while il a e weigh did no . Excep o
un 9 whe e his memb ane esis ance was no de ec ed, and
sepa a ion was s opped when e en a e weigh was
minimum. As a esul , mean TMP achie ed in his wo k
(maximum almos 45 kPa in MF) we e close o o he s udies
whe e a minimum o 50 kPa was employed in MF
(Weinbe ge and Kulozik, 2021a). All TFF de eloped wi h si-
mila luxes (p > 0.05); excep o un 9 ha achie ed he
highes lux. Egg yolk plasma MF ( un 9) was accomplished
quickly wi h low esis ance.
The ac iona ion e ec o TFF, can be analysed by
ansmission a es. As his pa ame e conside s concen a-
ions a il a es (Eq. (1)). Mean ansmission a es a e shown
in Fig. 1. In con as , o isualise he concen a ion e ec o
TFF on he egg p oduc s s udied, CF and WRR we e included
in Fig. 2. A black dash-do ed line ma ks a limi a 1. Ac-
co ding o Eq. (2), CF was abo e ha black line when he
concen a ion a he e en a e was o e he ini ial con-
cen a ion, hen TFF main accomplishmen was concen a-
ion. Whe eas, when he concen a ion a he e en a e was
lowe han ini ial concen a ion, CF was below he black line;
and did no achie e o concen a e a he e en a e. The g ey
dash-do ed line ma ks a CF alue o 0.5. CF a ound ha line
supposed ha inal concen a ion a e en a e was close o
ha o he dilu ed sample in oduced in he sys em; because
hey we e dilu ed 1:1 be o e TFF.
MF a un 3 (0.1 µm - ME) and un 4 (0.2 µm – PES) achie ed
he highes a e age TMP (Table 2). Howe e , hese TMP we e
no linked o he highes ansmission a es (Fig. 1). On he
con a y, om un 5, i was de ec ed ha p o ein and DM
ansmission a es inc eased (p < 0.05) when a lowe TMP
was applied (Table 2). Same TFF sys em was employed a un
4 and 5 (Table 1). This e ec has been desc ibed be o e
(Weinbe ge and Kulozik, 2021b). P o ein ac iona ion was
Table 2 – Resul s om ope a ional a iables.
Run key da a
No MWCO / Po e size &
ma e ial
TMP (kPa) Fil a e lux
(Lh
−1
m
−2
)
1 750 kDa mPES 35.51 ± 17.24
b
2.46 ± 1.36
b
2 750 kDa mPES 35.4 ± 9.03
b
2.95 ± 1.33
b
3 0.1 µm ME 44.61 ± 15.17
a
4.07 ± 1.75
b
4 0.2 µm PES 42.26 ± 13.93
a
2.62 ± 1.16
b
5 0.2 µm PES 33.16 ± 11.24
b
3.34 ± 1.01
b
6 0.2 µm mPES 26.27 ± 13.72
bc
7.99 ± 7.07
b
7 0.65 µm mPES 29.64 ± 13.44
b
3.30 ± 2.00
b
8 750 kDa mPES 32.68 ± 8.41
b
2.40 ± 2.38
b
9 0.65 µm mPES 1.24 ± 1.86
e
24.81 ± 37.42
a
10 750 kDa mPES 18.34 ± 7.45
cd
6.72 ± 1.04
b
11 0.65 µm mPES 10.76 ± 4.41
de
4.56 ± 0.86
b
MWCO: molecula weigh cu -o . mPES: modi ied
Polye he sul one. ME: Mixed Es e . PES: Polye he sul one. TMP:
mean ansmemb ane p essu e. Means wi h di e en le e s
wi hin he column di e ed signi ican ly om each o he ac-
co ding o Tukey’s es (p < 0.05). Runs 1–7 employed whole egg
plasma; uns 8 and 9 employed egg yolk plasma; uns 10 and 11
employed egg whi e.
102 Food and Biop oduc s P ocessing 140 (2023) 99–109
nega i ely a ec ed by high TMP due o deposi laye com-
pac ion and low p o ein ansmission. In his wo k, he lowe
TMP desc ibed a un 5, le c ossed mo e p o eins (and DM)
and his educed CF o i s e en a e, being all a ound 1 (Fig. 2).
Di e ences be ween CF om un 4 and 5 we e s a is ically
signi ican (p < 0.05). As choles e ol did no pass PES mem-
b ane, bo h CF we e o e one.
On he o he side, mPES memb anes allowed he il a ion
o choles e ol, and mo e p o eins and DM han PES; in
ag eemen wi h he highe ansmission a es (p < 0.05)
achie ed a uns 1, 2 and 6 despi e po e sizes we e below
(750 kDa) o equal (0.2 µm) o uns 3–5. Choles e ol did no
c oss h ough UF memb anes (750 kDa). I appea ed a 0.2 µm
il a es. To unde s and he a ia ions be ween whole egg
plasma UF ( un 1 and 2) in ansmission a es (Fig. 1), al hough
no di e ences a ope a ional pa ame e s we e de eloped
(Table 2), an analysis o uns 8 and 10 ga e he answe . A un
8 plasma om egg yolk was ul a il a ed wi h he same
memb ane o 750 kDa made o mPES. Re en a e om un 8
had almos ini ial plasma composi ion: all CF we e sligh ly
abo e 1 (Fig. 2) and ansmission a es we e close o ze o
(Fig. 1). Mainly wa e om dilu ion c ossed h ough he
memb ane; a concen a ion p ocess was de eloped. A un 10,
egg whi e was il e ed h ough he same 750 kDa sys em. In
his case, wa e , p o eins and DM om egg whi e c ossed
h ough he memb ane as shown WRR o 3.26 (Fig. 2) and
ansmission a es a ound 27% (Fig. 1). Hence DM and p o eins
de ec ed a il a es om uns 1 and 2, we e mainly om egg
whi e. A un 1, mo e egg whi e p o eins il a ed, p oducing
highe ansmission a es (Fig. 1), e en like un 10 (p o ein
ansmission a e: p = 0.42), and lowe CF (Fig. 2). In con as , i
seems ha whole egg plasma employed a un 2 had mo e egg
yolk compa ed o ha o un 1. Consequen ly, a e UF, e-
en a e was mo e concen a ed, p oducing highe CF (Fig. 2);
e en p o ein CF was equal o un 8 (p = 0.63). Bu no highe
ansmission a es we e de ec ed (Fig. 1). Di e ences be ween
bo h uns we e mainly due o he biological in insic a ia-
bili y be ween ini ial eggs, ood om animal o igin.
DM and p o eins ansmission a es o un 6 (0.2 µm) we e
highe (p < 0.05) han un 7 (0.65 µm); in con as , choles e ol
ansmission a es, mean TMP and mean il a e luxes we e
no signi ican ly di e en (p > 0.05) al hough un 6 had
nea ly 2.5 imes mo e su ace a ea (Table 1). These uns can
be compa ed o egg yolk plasma ( un 9) and egg whi e ( un
11) MF. Fil a e and e en a e om egg yolk plasma we e
close o each an equilib ium o concen a ions (Table 3),
p oducing ansmission a es close o 100% (Fig. 1) and CF
a ound 0.5 (Fig. 2). Final concen a ion a e en a e was nea
o ha o he plasma dilu ed in oduced in he hollow ibe s.
This mean ha almos e e y compound a plasma egg yolk
was able o c oss h ough he memb ane o 0.65 µm - mPES.
The e o e, his MF o plasma egg yolk was poo ly selec i e.
In con as , some egg whi e componen s we e e ained a
0.65 µm. A un 11, ini ial weigh was educed mo e han 4
imes wi h a TMP o 10.76 kPa only, bu ansmission a es
we e no o e 50% (Fig. 1); hen i did no achie e an equili-
b ium concen a ion wi h il a e.
Jux aposed o egg whi e ( un 11) o egg yolk plasma MF
( un 9), when whole egg plasma was mic o il a ed wi h
mPES and a minimum po e size o 0.2 µm ( un 6 and 7), CF o
DM and choles e ol we e o e 1.2 (Fig. 2). A concen a ion
p ocess was de eloped. Specially egg yolk lipids, because CF
o p o eins we e abou 1 and 1.1 o un 6 and 7, espec i ely.
In Table 3, plasma om whole egg had simila DM o egg yolk
plasma (p = 1), abou 21%; and double DM compa ed o egg
whi e. Unlike DM, whole egg plasma s a ed wi h simila
p o eins (p = 0.21) o egg whi e (a ound 86 mg/ml) and nea ly
wice p o eins compa ed o egg yolk plasma (abou 47 mg/
ml). Fil a ion o plasma was hinde ed by he p esence o
p o eins om egg whi es. Random agg ega ion o egg whi e’s
p o eins was isualized, caused by only 0.5% o added egg
yolk (Yao e al., 2014). In ou wo k, hese agg ega es de-
c eased ansmission a es compa ed o egg yolk plasma
(Fig. 1) and p o oked di e ences in mean TMP (Table 2).
3.2. Colou esul s
L*, a* and b* colou pa ame e s we e de e mined in egg
whi es, plasmas, wa e diluen , e en a es and il a es.
Values om e en a es we e compa ed o hose o ini ial
samples. While il a es we e compa ed o wa e . Resul s a e
shown in Table 4. Means ha ing same le e wi hin he
column did no di e signi ican ly om each o he acco ding
Fig. 1 – T ansmission a es o d y ma e (DM) in ed,
p o eins in blue and choles e ol in g een. S anda d
de ia ions a e gi en as e o ba s (some a e oo small o be
obse ed). Fo each pa ame e (N ≥ 2), means wi h di e en
le e s di e ed signi ican ly om each o he acco ding o
Tukey’s es (p < 0.05). Runs 1–7 employed whole egg
plasma; uns 8 and 9 employed egg yolk plasma; uns 10
and 11 employed egg whi e. Run 1, 2, 8 and 10 employed
memb anes o 750 kDa made o mPES (modi ied
polye he sul one). Run 3: 0.1 µm o ME (mixed es e ). Run 4
and 5: 0.2 µm o PES (polye he sul one). Run 6: 0.2 µm o
mPES. Run 7, 9 and 11: 0.65 µm o mPES.
103 Food and Biop oduc s P ocessing 140 (2023) 99–109
o Tukey’s es , hen p was o e 0.05. Ini ial egg whi e had
some compounds ha ga e a so yellow appea ance wi h a
b* alue a ound 3. Ins ead, egg yolk plasma had ca o enoids
ha dec eased ligh ness (L*) and inc eased edness (a*) and
yellowness (b*). When hese compounds we e dilu ed wi h
egg whi e, like i was in whole egg, a* and b* alues de-
c eased, and ligh ness inc eased as seen in Table 4.
Egg yolk and whi e e en a es we e da ke han ini ial
samples (p < 0.05). Whe eas whole egg e en a es had simila
b igh ness o ini ial whole egg plasma (p > 0.05). Only e-
en a es wi h WRR o e 3 ( un 6 and 7) we e signi ica i ely
da ke han ini ial plasma (p < 0.01). Egg whi e e en a es
we e no signi ican ly di e en in edness and yellowness
(p > 0.05) due o he g ea di e ences wi h he o he s.
Howe e , when hey we e es ed sepa a ely h ough Tukey
HSD, signi ica i e di e ences we e de ec ed (p < 0.01). Egg
whi e e en a es we e mo e han h ee old yellowish (b*
a ound 12) and wice edde (a* o e 0) han ini ial albumen
despi e he al eady men ioned il a ion o egg whi e com-
ponen s (Fig. 1). The e we e egg whi e colou ed componen s
ha we e concen a ed by il a ion. Concen a ion p ocess
should ha e been highe a 750 kDa (UF) han a 0.65 µm (MF).
Because some o hose componen s il a ed a highe po e
size, al hough some we e s ill e ained a 0.65 µm. Di e -
ences in il a es we e no de ec ed due o wa e dilu ion. All
il a es om egg whi e had a* nega i e alues and b* a ound
1. They we e signi ican ly di e en o wa e (p < 0.05).
Fil a e om egg yolk plasma UF ( un 8) was no sig-
ni ican ly di e en o wa e (p > 0.05). Then ca o enoids we e
concen a ed a e en a e. This concen a ion esul ed in a
e en a e as yellow as ini ial yolk (simila b*) (p > 0.05), bu
edde (highe a*) (p < 0.01). On he o he side, a 0.65 µm
Fig. 2 – Concen a ion ac o s (CF) o d y ma e (DM) in ed, p o eins in blue and choles e ol in g een. S anda d de ia ions
a e gi en as e o ba s (some a e oo small o be obse ed). Fo each composi ion pa ame e (N ≥ 2), means wi h di e en
le e s di e ed signi ican ly om each o he acco ding o Tukey’s es (p < 0.05). ME: Mixed Es e . PES: Polye he sul one.
mPES: modi ied Polye he sul one. WRR: weigh educ ion a io.
Table 3 – Composi ion esul s o e en a es, il a es and ini ial samples.
Run key da a D y ma e (%) P o eins (mg/ml) Choles e ol (mg/g)
No Po e size/ MWCO &
ma e ial
Re en a e Fil a e Re en a e Fil a e Re en a e Fil a e
1 750 kDa mPES 20.24 ± 0.02
d
2.90 ± 0.01
g
70.72 ± 0.30
g
19.26 ± 0.36
e
4.55 ± 0.17
0
d
2 750 kDa mPES 28.43 ± 0.01
b
3.00 ± 0.0003
97.27 ± 0.60
b
16.69 ± 0.19
6.12 ± 0.12
cd
0
d
3 0.1 µm ME 28.97 ± 0.01
ab
1.58 ± 0.01
j
106.99 ± 0.92
a
8.50 ± 0.14
h
5.51 ± 0.08
e
0
d
4 0.2 µm PES 30.15 ± 0.01
a
1.66 ± 0.01
i
109.41 ± 0.98
a
9.20 ± 0.07
h
5.88 ± 0.06
de
0
d
5 0.2 µm PES 20.68 ± 0.00
d
1.86 ± 0.001
h
78.81 ± 0.61
e
10.48 ± 0.50
g
4.14 ± 0.09
g
0
d
6 0.2 µm mPES 27.24 ± 0.03
b
8.28 ± 0.01
b
89.42 ± 0.36
c
35.94 ± 0.16
a
6.42 ± 0.12
bc
1.32 ± 0.03
b
7 0.65 µm mPES 28.64 ± 0.01
ab
6.62 ± 0.03
c
100.15 ± 0.73
b
32.93 ± 0.72
b
6.69 ± 0.03
ab
0.88 ± 0.02
c
8 750 kDa mPES 23.53 ± 0.01
c
0.46 ± 0.003
k
49.88 ± 0.22
h
2.16 ± 0.01
i
7.14 ± 0.13
a
0
d
9 0.65 µm mPES 10.69 ± 0.00
g
10.09 ± 0.002
a
27.42 ± 0.34
i
25.20 ± 0.39
d
3.07 ± 0.11
h
2.89 ± 0.04
a
10 750 kDa mPES 14.01 ± 0.01
e
4.08 ± 0.01
e
105.81 ± 0.28
a
27.89 ± 0.79
c
na na
11 0.65 µm mPES 9.80 ± 0.05
g
4.61 ± 0.01
d
74.52 ± 0.98
31.97 ± 1.24
b
na na
Whole egg plasma 21.14 ± 0.77
d
86.58 ± 2.95
d
3.77 ± 0.21
g
Egg yolk plasma 21.26 ± 0.02
d
46.82 ± 0.89
h
6.16 ± 0.16
cd
Egg whi e 11.82 ± 0.56
85.10 ± 2.36
d
na
MWCO: molecula weigh cu -o . mPES: modi ied Polye he sul one. ME: Mixed Es e . PES: Polye he sul one. na: no applicable. Means wi h
di e en le e s wi hin he column di e ed signi ican ly om each o he acco ding o Tukey’s es (p < 0.05). Runs 1–7 employed whole egg
plasma; uns 8 and 9 employed egg yolk plasma; uns 10 and 11 employed egg whi e.
104 Food and Biop oduc s P ocessing 140 (2023) 99–109
(MF), ca o enoids passed h ough he memb ane. Re en a e
was as ed as ini ial plasma (p > 0.05) and il a e ob ained
a* like e en a e (a ound 9). The desc ibed equilib ium con-
cen a ion was also measu able a colou pa ame e s. Fil a e
was sligh ly yellowe han e en a e, bu bo h we e nea ly
hal yellowish o ini ial plasma. Be ween e en a e and il-
a e, ca o enoids we e sepa a ed a he e enly.
This colou sepa a ion was nei he de ec ed a whole egg
plasma il a ion. Despi e ca o enoids also il a ed a 0.2 and
0.65 µm wi h mPES memb anes ( un 6 and 7), e en a es we e
edde (p < 0.01) and e en yellowe (only un 6) han ini ial
plasma. As a o emen ioned, he in e ac ions be ween whole
egg plasma componen s no desc ibed o egg whi e o egg yolk
plasma may be he eason. Fu he mo e, di e ences be ween
il a ion p oduc s om uns 6 o 7 could indica e ha ca -
o enoids and o he colou ed compounds om egg whi e may
sepa a e di e en a 0.2 µm o 0.65 µm wi h mPES memb anes.
The low ansmission a es p e iously desc ibed o PES
and ME memb anes ( un 3–5) explained he small di e ences
de ec ed a colou Tukey HSD es . Due o concen a ion,
e en a es we e edde han ini ial plasma (a* p < 0.01). A
ligh -yellow colo a ion (b* a ound 1) was de ec ed a il a es.
This colo a ion was al eady desc ibed a egg whi e il a es
and no de ec ed a egg yolk plasma UF ( un 8). I seems ha
some egg whi e-colou ed compounds il a ed a all he
memb anes s udied.
Colou di e ences be ween e en a es om un 1 and 2
suppo ed he hypo hesis o an ini ial whole egg plasma wi h
mo e egg yolk a eed 2. Because e en a e 2 was edde and
yellowe han ini ial plasma (p < 0.01). While he colou o e-
en a e 1 was no signi ican ly di e en o plasma (p > 0.05).
The lack o in e na ional s anda ds o egg colou mea-
su emen s (Milo ano ic e al., 2021) was also e iden in his
wo k. Howe e , he objec i e me hod employed o colou
assessmen was p ecise and allowed samples compa a ions.
Al hough i canno be used o measu e ini ial whole egg and
egg yolk due o hei low ansmi ance, i was sui able o
plasmas and albumen. The alues om egg whi e di e s
om li e a u e mainly in L* and b* (Milo ano ic e al., 2021).
Plasma om egg yolk o whole egg di e s wi h li e a u e no
only because o disc epancies in s anda ds measu emen s,
bu also because cen i uga ion condi ions change colou
pa ame e s as ca o enoids sepa a e di e en ly (Pue as and
Vázquez, 2021b). In ou wo k only ca o enoids ha emained
in plasma we e s udied.
3.3. In e ac ion o whole egg compounds
The componen s o egg yolk plasma we e mainly isola ed
be ween 750 kDa and 650 nm. While egg whi e p o eins we e
ob ained along he di e en po e sizes. Small po e size
memb anes made o mPES we e use ul o concen a ed
choles e ol wi h ca o enoids and o he p o eins. Then, e-
sul s con i med ha cen i uga ion and il a ion condi ions
did no dis up LDL because ee choles e ol was no a
plasmas nei he a il a ion p oduc s. Choles e ol was pa
o LDL pa icles. LDL and ca o enoids we e indi ec ly app e-
cia ed. LDL h ough choles e ol quan i ica ion and ca -
o enoids om colou pa ame e s. Ca o enoids a e molecules
ha could ha e c ossed memb anes o 750 kDa. Thei main
absence a hose il a es oge he wi h hei p esence a
hose in which LDL was de ec ed (b* alues o e 20 a uns 6,
7 and 9) sugges hey we e bound. P o eins o LDL p esen a
la ge p opo ion o hyd ophobic amino acids and, conse-
quen ly, ha e a high abili y o in e ac ing by his way
(Spe oni e al., 2005). O he s udies ha emo ed choles e ol
om egg yolk wi h chela es o abso ben s also de ec ed
ca o enoids ex ac ion (Pue as and Vázquez, 2019a).
LDL in egg yolk plasma c ossed memb anes o 650 nm cu -
o almos eely. I nea ly achie ed an equilib ium con-
cen a ion a bo h sides o he memb ane. The e o e, LDL did
no sepa a e selec i ely. These esul s showed ha LDL ag-
g ega es had a molecula size mainly below 0.65 µm, as de-
sc ibed in li e a u e (Xie e al., 2020).
Howe e , choles e ol concen a ions in e en a es om
un 7 and 9 (Table 3) we e s a is ically di e en (p < 0.01).
This di e ence highligh ed a possible LDL agg ega ion a
whole egg plasma ha a oided LDL il a ion eely. Fi s ly, i
was desc ibed he o ma ion o la ge agg ega es o LDL (o e
1000 nm) when pH was 8 (Spe oni e al., 2005). The a e age
pH measu ed a ini ial eed we e: 8.38 ± 0.12 o whole egg
plasma, 6.20 ± 0.29 o egg yolk plasma and 9.29 ± 0.06 o
Table 4 – Colou esul s o he e en a es, il a es and ini ial samples.
Run key da a Re en a e Run Fil a e
No Po e size/ MWCO &
ma e ial
L* a* b* No L* a* b*
1 750 kDa mPES 68.81 ± 0.32
cd
3.64 ± 0.06
e g
58.97 ± 0.13
bc
1 96.40 ± 0.02
bc
0.05 ± 0.00
b
1.15 ± 0.01
de
2 750 kDa mPES 62.58 ± 0.23
d
5.32 ± 0.05
ce
69.56 ± 0.13
a
2 96.22 ± 0.11
bc
0.02 ± 0.01
b
1.05 ± 0.00
de
3 0.1 µm ME 63.37 ± 0.02
d
5.98 ± 0.02
cde
58.68 ± 0.04
bc
3 95.54 ± 0.08
de
0.04 ± 0.01
b
1.24 ± 0.01
de
4 0.2 µm PES 60.52 ± 0.25
de
5.88 ± 0.00
cde
60.47 ± 0.20
ac
4 95.90 ± 0.06
cd
0.00
b
1.46 ± 0.01
d
5 0.2 µm PES 59.13 ± 0.25
de
5.46 ± 0.01
cde
57.10 ± 0.13
bc
5 96.41 ± 0.01
bc
0.00
b
0.86 ± 0.01
e
6 0.2 µm mPES 51.92 ± 0.25
e
7.04 ± 0.02
bc
63.42 ± 0.22
ab
6 87.14 ± 0.03
-1.87 ± 0.01
e
29.67 ± 0.02
b
7 0.65 µm mPES 39.81 ± 0.08
7.70 ± 0.04
bc
51.18 ± 0.04
c
7 84.40 ± 0.06
g
-0.37 ± 0.01
d
22.23 ± 0.01
c
8 750 kDa mPES 39.91 ± 0.95
13.25 ± 0.03
a
57.75 ± 0.95
bc
8 96.74 ± 0.13
ab
0.00
b
0.19 ± 0.01
9 0.65 µm mPES 24.49 ± 0.02
g
9.44 ± 0.48
b
30.46 ± 0.49
d
9 40.48 ± 0.44
h
9.10 ± 0.18
a
39.36 ± 0.57
a
10 750 kDa mPES 74.43 ± 0.06
bc
1.28 ± 0.03
gh
15.70 ± 0.06
e
10 95.32 ± 0.25
e
-0.18 ± 0.01
c
1.07 ± 0.01
de
11 0.65 µm mPES 79.86 ± 0.19
b
0.67 ± 0.01
h
11.65 ± 0.01
e
11 95.44 ± 0.08
de
-0.25 ± 0.02
cd
1.35 ± 0.01
de
Whole egg plasma 66.43 ± 3.41
d
3.36 ± 0.71
53.12 ± 4.16
c
Wa e 96.89 ± 0.03
a
-0.02 ± 0.01
b
0.13 ± 0.03
Egg yolk plasma 61.88 ± 5.64
d
7.58 ± 1.55
bd
63.54 ± 1.34
ab
Egg whi e 93.91 ± 0.46
a
-0.31 ± 0.06
h
3.79 ± 0.25
MWCO: molecula weigh cu -o . mPES: modi ied Polye he sul one. ME: Mixed Es e . PES: Polye he sul one. Means wi h di e en le e s
wi hin he column di e ed signi ican ly om each o he acco ding o Tukey’s es (p < 0.05). Runs 1–7 employed whole egg plasma; uns 8 and
9 employed egg yolk plasma; uns 10 and 11 employed egg whi e.
105 Food and Biop oduc s P ocessing 140 (2023) 99–109
egg whi e. Subsequen ly, a highe amoun o hese la ge
agg ega es in un 7 could explain di e ences; besides minu e
amoun s o hem in un 9 could jus i y ansmission a es
below 100% a un 9. These la ge agg ega es could ha e
o med a gel/cake laye on op o he memb ane by adso p-
ion and subsequen comp ession by smalle pa icles. This
cake laye led o memb ane blockage (Nikolay e al., 2020).
Secondly, he p esence o egg whi e p o eins seems o be
essen ial o explain di e ences be ween un 7 and 9. The
o ma ion o a complex be ween a yolk componen wi h
o omucin was desc ibed as he cause o he dec ease o he
albumen oaming abili y when e en small quan i ies o egg
yolk we e p esen ed (Lomakina and Míko á, 2006). And as
a o emen ioned, o he au ho s epo ed hese agg ega es.
O omucin is he bigges egg whi e p o ein. I is a sul a ed
glycop o ein ha consis s o a ca bohyd a e poo subuni (β-
o omucin) and a ca bohyd a e- ich subuni (ß-o omucin).
These subuni s ha e molecula weigh s below 750 kDa
(Omana e al., 2010). Howe e , o omucin size showed la ge
a ia ions acco ding o di e en condi ions. Fo example,
s o age ime has demons a ed o a y o omucin size be-
ween 1 and 1000 nm (Shan e al., 2020). While he e ec o
di e en pH and ionic s eng hs de ec ed a ia ions a ound
3–1124 nm (Sun e al., 2018). When Sun e al. (2018) analysed
o omucin hyd odynamic size a high sal concen a ion
(150 mM NaCl), i was below 25 nm. Because he sal s de-
c eased he epulsi e o ces and he e o e educed o omucin
pa icle size. Whe eas when condi ions we e like hose em-
ployed in ou wo k (neu al pH and low ionic s eng h) mo e
han 70% o o omucin was o e 0.65 µm. A his pH, o o-
mucin had ne nega i e cha ge om aspa ic and glu amic
acids in p o ein pa , and sialic acid and sul a e in he glycan
pa . These nega i e cha ges cause epulsion ha could
s e ch o omucin chains. In ou s udy, sal s we e no added,
so o omucin a e age size may ha e been o e 0.65 µm.
Subsequen ly, much o he o omucin did no il a e a none
o he il a ions s udied. Then, i was p obably ha his
glycop o ein o med agg ega es as well wi h LDL. These ag-
g ega es would explain he lowe ansmission a es de-
sc ibed o whole egg plasma ( uns 6–7) compa ed o egg yolk
plasma ( un 9) (Fig. 1, p < 0.05).
Mo eo e , o omucin had shown hypocholes e olemic ac ion
in i o (Nagaoka e al., 2002). I was de ec ed ha his egg whi e
p o ein inhibi ed choles e ol up ake which could signi ican ly
educe se um choles e ol in a s. As pa o he mechanism o
choles e ol lowe ing induced by o omucin, i was desc ibed he
dec ease o solubili y o choles e ol micelles ha may inhibi
choles e ol abso p ion h ough he di ec in e ac ion be ween
choles e ol mixed micelles and o omucin in jejunal epi helium
(jejunal e ec ) (Tu e al., 2020). This associa ion also co obo a e
he agg ega ion de ec ed a he memb ane sepa a ion p ocesses.
This e ec can explain o he s udies we e die a y choles e ol
con ained in whole egg was no well abso bed and did no
acu ely a ec plasma o al choles e ol concen a ion. Howe e ,
same esea ch de ec ed ha iglyce ides in plasma ose a e
whole egg consump ion (Kim and Campbell, 2018). And igly-
ce ides a e pa o LDL pa icles (An on, 2013). The e o e, u he
esea ch is needed o unde s and he a e o LDL along he di-
ges i e ac .
On he o he side, i canno be disca ded he in e ac ion o
LDL wi h o he egg whi e p o eins o ming p o ein ag-
g ega es. Egg whi e p o eins consis o i e majo p o eins,
o albumin (54%), o o ans e in (12%), o omucoid (11%), ly-
sozyme (3.5%), o omucin (3.5%) and se e al mino p o eins
(A amescu e al., 2008). O albumin in i s na u al s a e
usually exis s in hyd ophilic o m and has poo binding
abili y wi h hyd ophobic bioac i e componen s. Bu unde
app op ia e condi ions, i could un old easily, exposing i s
in e nal hyd ophobic g oups (Liu e al., 2022). These could
bind wi h LDL pa icles inc easing hei size and p e en ing
hem o c oss he memb ane o p oducing a pa ial o com-
ple e po e blocking.
Al hough o omucin accoun s o a ound 3.5% o egg whi e
p o eins and he o he majo p o eins a e below 750 kDa
(A amescu e al., 2008), p o ein ansmission a es a uns
10 and 11 we e only 26% and 43%, espec i ely. I is desc ibed
ha o omucin is usually p esen in egg whi e as a complex
wi h lysozyme and o he egg whi e p o eins (Omana e al.,
2010). Then, o omucin in e ac ed wi h o he egg whi e p o-
eins, a oiding hei il a ion. I hese complexes we e si-
mila o bigge han memb ane po e size, hey could ha e
p oduced a pa ial o comple ed po e blocking and gel/cake
laye o ma ion. Bo h mechanisms could p oduce memb ane
ouling and s op il a ion (Nikolay e al., 2020).
Fig. 3 summa izes LDL agg ega es ha could be ound a
whole egg plasma: pa icles o LDL agg ega ed hemsel es,
LDL bound o o omucin and o he smalle egg whi e p o eins
(< 750 kDa), and LDL bound only o smalle egg whi e p o-
eins. LDL was also bound o ca o enoids p esen ed in
plasmas. All hese in e ac ions we e plausible due o he
la ge p opo ion o hyd ophobic amino acids abo e-
men ioned p esen a LDL apop o eins. Hyd ophobic o ces
we e p edominan o e elec os a ic in e ac ions because a
pH s udied all main p o eins excep lysozyme we e nega-
i ely cha ged (A amescu e al., 2008; Chalamaiah e al.,
2017; Na idghasemizad e al., 2015). Then, elec os a ic e-
pulsion be ween main p o eins and memb anes was
achie ed as desi ed. Because i a oided p o ein adso p ion o
memb ane and educed il e cake o ma ion (Nikolay e al.,
2020). Mo eo e , i has been sugges ed ha hyd ophobic in-
e ac ions a e he d i ing o ce o p o ein binding. Because
a p o ein−p o ein in e aces nonpola esidues a e ypically
concen a ed a he cen e and cha ged esidues a e localized
a he im (Zhou and Pang, 2018).
Fig. 3 – Types o LDL agg ega es a whole egg plasma: pa icles o LDL agg ega ed hemsel es, LDL bound o o omucin and
o he smalle egg whi e p o eins (< 750 kDa), and LDL bound only o smalle egg whi e p o eins. Ca o enoids we e mainly
bound o LDL.
106 Food and Biop oduc s P ocessing 140 (2023) 99–109
The andom o ma ion o hese LDL agg ega es oge he
wi h di e ences in su ace a eas jus i ied he dispa i ies
desc ibed a uns 6 and 7. The e o e, egg componen s il a-
ion canno be p edic ed wi h he condi ions employed.
Addi ion o sal s o pH modi ica ions could dec ease ag-
g ega ion. Consequen ly, u he s udies could be led chan-
ging condi ions. Bu p o ein ac i i y should be moni o ed. In
addi ion, hese agg ega es explained he low ansmission
a es de ec ed a 0.1 µm ME and 0.2 µm PES memb anes ( uns
3–5). Those memb anes mus lack in po es wi h a size en-
ough o LDL and LDL agg ega es il a ion. I has been shown
ha cu -o desc ibed by manu ac u e can di e con-
side ably om e ec i e cu -o and po e size dis ibu ion
(Nikolay e al., 2020).
The possible chela e e ec o egg whi e p o eins should be
deeply s udied. This could be s udied in i o using dynamic
gas ic models ha ha e al eady been used in ood and
pha maceu ical esea ch (Réhaul -Godbe e al., 2019).
These models could assess he in luence o cooking me hod
and diges ion on he bioaccessibili y o nu ien s and he
gene a ion o bioac i e pep ides. Bu di e ences be ween
whole egg, egg yolk and egg whi e in ake mus be conside ed.
Hollow- ibe memb anes wi h small po e sizes (5–10 kDa cu -
o ) a e al eady employed o calcula e he bioaccessibili y o
nu ien s in di e en in i o diges ion app oaches. Dialysis
h ough hese a i icial memb anes is used o simula e ab-
so p i e in es inal compa men s (jejunum and ileum)
(Ma ze, 2017). Howe e , i is known ha ood ma ix plays a
key ole in nu ien bioaccessibili y. The e m ‘ ood ma ix’
ema ks ha nu ien s a e con ained in a medium whe e
hey may in e ac wi h o he componen s and s uc u es also
p esen . The e o e, he physical s a e o he ma ix is es-
sen ial in he elease, mass ans e , accessibili y and bio-
chemical s abili y o many ood componen s (Oli ei a e al.,
2018). Fo example, Nimala a ne e al. (2015) demons a ed
ha bioaccessibili y o ca o enoids (lu ein and zeaxan hin)
was lowe om sc ambled egg yolks compa ed o boiled egg
yolks. Chemical and s uc u al changes o p o eins and li-
pop o eins in egg yolk du ing di e en cooking condi ions
we e unde lined as a p obable eason. Conside ing ou e-
sul s, a en ion can be d awn o possible di e ences be ween
LDL agg ega es. Ne e heless, as yolks we e sepa a ed be o e
s i ing, he e ec o egg whi e p o eins was no s udied.
This wo k has demons a ed physical di e ences be-
ween whole egg plasma choles e ol and egg yolk plasma
choles e ol in hei ma ices. Consequen ly, on he one hand,
i e lec ed he impo ance o ood ma ix wi h egg choles-
e ol as an example. On he o he hand, UF and MF may be
use ul echniques o unde s and o he liquid ood ma ices.
4. Conclusion
Egg p oduc s we e s udied h ough di e en memb ane se-
pa a ion p ocesses. The in e ac ions be ween egg yolk and
egg whi e componen s made sepa a ion di icul . Choles e ol
was pa o LDL pa icles. These pa icles oge he wi h ca -
o enoids we e only il a ed wi h mPES memb anes and a
minimum po e size o 0.2 µm. High unce ain y was ob ained
due o LDL agg ega ion. Th ee ypes o agg ega es can jus i y
he esul s: pa icles o LDL agg ega ed hemsel es, LDL
bound o o omucin and o he smalle egg whi e p o eins
(< 750 kDa), and LDL bound only o smalle egg whi e p o-
eins. As pH adjus men s o sal addi ion could educe ag-
g ega ion, addi ional esea ch could be pe o med o
implemen TFF o he sepa a ion o whole egg componen s.
Fu he s udies a e needed o unde s and egg whi e p o eins
in e ac ions wi h LDL and hei agg ega es composi ion. UF
and MF can be use ul echniques o unde s and liquid ood
ma ices.
Funding
This wo k was suppo ed by he Spanish Na ional Plan o
Scien i ic and Technical Resea ch and Inno a ion. A
Uni e si y P o esso Educa ion g an (FPU 16/05128) by he
Spanish Minis y o Educa ion, Cul u e and Spo o au ho
Gema Pue as is g a e ully acknowledged.
CRediT au ho ship con ibu ion s a emen
Gema Pue as: In es iga ion, Me hodology, W i ing – o iginal
d a , Visualiza ion. Pa icia Cazón: Valida ion, W i ing – e-
iew & edi ing. Manuel Vázquez: Concep ualiza ion, Fo mal
analysis, W i ing – e iew & edi ing, Supe ision.
Decla a ion o Compe ing In e es
The au ho s decla e ha hey ha e no known compe ing i-
nancial in e es s o pe sonal ela ionships ha could ha e
appea ed o in luence he wo k epo ed in his pape .
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