a SciTechnol journal Research Article
Madziva et al., J Food Nutr Disor 2014, 3:1
http://dx.doi.org/10.4172/2324-9323.1000130
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Journal of Food &
Nutritional Disorders
International Publisher of Science,
Technology and Medicine
Encapsulated Folic Acid
in Cheddar Cheese Reduce
Methionine -Induced Hyper-
Homocysteinemia in Male
BALB/C Mice
Honest Madziva
1
, Michael Phillips
2
and Kasipathy
Kailasapathy
3
*
Abstract
Methionine is the precursor of homocysteine. Elevated plasma
homocysteine (hyperhomocysteinemia) is associated with
occlusive vascular disease. Nutritional interventions are thought to
be an effective and safe means of reducing the associated risk of
disease through dietary intake of folates. We compared the impact
of free and encapsulated (alginate-pectin) folic acid on methionine-
induced hyperhomocysteinemia in male Balb/c mice. Cheddar
cheese was used as the food carrier (folic acid incorporated into
Cheddar cheese either in free form or as encapsulated capsules)
and the evaluation was conducted over a 12-wk period. Mice were
fed six experimental diets:
(i) Met 10 [L-methionine (10 g/Kg) only intake],
(ii) Met 20F [L-methionine ( 20 g/kg) with free folic acid (2 mg/kg)],
(iii) Met 20E [L-methionine (20 g/kg) with encapsulated folic acid
(2 mg/kg)],
(iv) Met 10F [L-methionine (10 g/kg) with free folic acid (2 mg/kg)],
(v) Met 10E [L-methionine (10 g/kg) with encapsulated folic acid
(2mg/kg)],
(vi) Control (Cheddar cheese without added folic acid).
Supplementation of the diet with Met 10 and Met 20F increased
plasma homocysteine to 1.8 and 6.1 times the control, respectively.
The hyperhomocysteinemia caused by the dietary addition of Met
10 was counteracted by the encapsulated folic acid (Met 10E)
while free folic acid (Met 10F) showed a higher result. Similarly,
encapsulated folic acid caused substantial reduction in plasma
homocysteine in mice fed with Met 20E compared to Met 20F. The
results show that Cheddar cheese incorporated with encapsulated
folates can be used as a functional food vehicle to reduce
hyperhomocysteinemia.
Keywords
Folic acid; Alginate-pectin microcapsules; Homocysteine; Cheddar
cheese; Microencapsulation
*Corresponding author: Dr. Kasipathy Kailasapathy, School of Biosciences,
Taylor’s University, Lakeside Campus, No. 1 Jalan Taylors, 47500 Subang
Jaya, Malaysia, Tel: +61 2 45 701 653; Fax: +61 2 45 701 954; E-mail:
Kailasapathy.Kasipathy@taylors.edu.my
Received: July 01, 2013 Accepted: December 31, 2013 Published: January
06, 2013
genetic defects and or an inadequate status of folate and its cofactors.
Less than optimal folate nutrition has been implicated as a risk factor
in a number of negative health outcomes, including congenital neural
tube defects (NTD) [1,2], anaemia during pregnancy [3], low infant
birth weight [4], colorectal cancer and cardiovascular disease [5].
In addition, a growing body of sound epidemiological evidence has
shown a strong association of elevated homocysteine with incidence
of vascular disease [6-8]. he association remains strong even ater
adjustment for major determinants of homocysteine, such as age and
renal function, folate, vitamin B12, and vitamin B6 status, suggest
that homocysteine is an independent risk factor for occlusive vascular
disease [9,10]. he data provide the basis for a hypothesis that
elevated blood homocysteine is a cause of vascular disease and has
generated great interest because of the possibility that lowering blood
homocysteine through nutritional interventions might prove to be a
safe and efective means of reducing associated risk of disease. Plasma
homocysteine is inversely correlated with blood folate levels and
taking folic acid either as a supplement or in fortiied food has been
shown to lower total plasma homocysteine (tHcy) concentration [11].
Most naturally occurring folate derivatives in foods are highly
sensitive to temperature, oxygen, light and their stability is afected
by food processing conditions. Some foods have been either enriched
or fortiied with folic acid, for example, since 1995 in Australia and in
1996 in New Zealand, certain foods have been fortiied with folic acid.
However, during the enrichment or fortiication food processing
operations, the susceptibility of folic acid to cleavage under acidic
conditions, exposure to light and high temperature has been long been
established. he degree and rate of destruction is largely inluenced by
the pH of the medium, reducing agents in the bufer, folate derivatives,
types of bufer, and the food matrix components [12]. 5-Methyl-
tetrahydrofolate is the predominant food folate [12] and is readily
oxidised to 5-methyl-5, 6-dihydrofolate [13]. In this oxidised form it
may represent a substantial amount of the food folate. 5-methyl-5, 6
dihydrofolate is rapidly degraded under the acidic conditions in the
gastric environment. hus folic acid needs protection against not only
during food processing operations, but also during gastrointestinal
transit, especially through the acidic stomach.
In earlier work, we reported a microencapsulation method [14]
for folic acid using food grade polymers, alginate and pectin, to
increase its stability since like all folates, it is in danger of oxidative
degradation, which is enhanced by oxygen, light, heat and a shit
in pH either way from neutral. he protective efect of the micro
capsules and their applicability was evaluated in Cheddar cheese as a
food matrix for the delivery of folic acid [15]. In the present study, the
efects of folic acid (free and encapsulated) incorporated into Cheddar
cheese on plasma homocysteine levels and the vascular consequences
of hyperhomocysteinemia in mice were evaluated.
Materials and Methods
All the chemicals and reagents were purchased from Sigma,
Castle Hill, Australia, unless otherwise speciied.
Cheddar cheese production
Cheddar cheeses were made as reported by Madziva et al.
Introduction
Elevations of plasma total homocysteine (tHcy) may occur due to