3. FOOD ChEMISTRy & bIOTEChNOLOGy 3.1. Lectures

Chem. Listy, 102, s265–s1311 (2008) Food Chemistry & Biotechnology
3.2. Posters
P01 DETERMINATION OF β-CAROTENE IN THE
GELATIN CAPSuLE
ZUZAnA KOLAJOVá a , BAYAnMUnKH
ALTAnGEREL a , DAnIELA KRAMářOVá a , OTAKAR
ROP a and IGnáC HOZA a
a Department of Food Engineering Faculty of Engineering,
Faculty of Technology, TBU in Zlin, Czech Republic,
bayanmunkh_mn@yahoo.com
Introduction
β-carotene is the most well known of the carotenoids,
a phytonutrients family that represents one most of the widespread
groups of naturally occurring pigments. Steenbock
suggested that the plant pigment carotene was responsible for
the vitamin A activity and that carotene could not be vitamin
A, it may be converted metobolitically to the actual vitamin in
the 1919 1 . β-carotene consists of a chain of 40 carbon atoms,
with conjugated double bonds and a ring structure at each
end of the chain. Depending on the positions of the molecular
groups attached to the carbon chain, naturally occurring beta
carotene may be: cis-trans and all trans isomers. The most
common carotenoid in all green plants is β-carotene. Animal
and our body cannot synthesis β-carotene and they depend on
feed for their supply. There are many studies showing β-carotene
can show role of antioxidant potency. Also several studies
have been focused on the content of β-carotene in food
products, food supplements and biological fluids. 2–4
One of the vitamin supplement package products is
gelatin capsule. Gelatin capsule has a very rapid dissolution
property, usually 90 seconds in water at 37 °C. So, this protein
is a very useful raw material for emulsifier in food, pharmaceutical
and others products. Previous studies show that
gelatin is used as a carrier, coating or separating agent for
other substances. Supplemental intake of β-carotene probably
should not exceed 3–15 mg per day. β-carotene supplements
are found on the Generally Recognized as Safe (GRAS) list
issued by the Food and Drug Administration.
The purpose of this study was to find new method for
determination of β-carotene and isolation process, of course.
Experimental
High-performance liquid chromatography is generally
accepted as the modern method of choice to separate, identify,
and quantify caroteniods. Specially, it combined with
ECD, UV-VIS and fluorescence detector are the common
method for determination and identification of β-carotene in
vegetables and other samples.
C h e m i c a l s a n d M a t e r i a l s
β-carotene was obtained from personal prepared tablet
materials (at TBU in Zlin). Methanol (purity: 99.8 %), acetonitrile
(purity: 99.9 %) were obtained from Merck KGa
(Darmstadt, Germany). Phosphoric acid (purity: 85 %) was
s575
obtained from Chemapol (Prague, Czech Republic). All solvents
were of HPLC grade. Deionized water, purified by Aqua
osmotic system (Aquaosmotic, Tisnov, Czech Republic) was
used for solve sample preparation stages. Hexan (purity:
99.9 %) was obtained from Penta (Prague, Czech Republic),
was used in extraction of sample. Ethanol was obtained from
Chemapol (Prague, Czech Republic), used to solve sample
after evaporation.
E q u i p m e n t a n d C h r o m a t o g r a p h i c
C o n d i t i o n s
HPLC separation was performed using HPLC with ECD
detection (Coulochem III, ESA). Potentials of the cells for
detector were 500 and 600 mV.
β-carotene was separated on the Supelcosil LC18 DB
(250 × 4.6 mm,5 μm) with a mobile phase of methanol/acetonitrile/phosphoric
acid (70 : 29.5 : 0.5, v/v/v) at a flow rate of
1.1 ml min –1 . The separation was carried out ambient temperature.
In this experiment we used isocratic elution.
S a m p l e P r e p a r a t i o n
Filling from gelatin capsule was dissolved into 10 ml
of deionized water and after that extracted using 10 ml of
n-Hexan solution and extraction process was repeat six times.
After extraction, sample solution was evaporated to dryness
in rotary evaporator at 35–40 °C. After evaporation, the residue
was dissolved in 10 ml of ethanol and was filtered trough
0.45 μm filter (nylon) before injected into column. Sample
(20 μl) was injected into a column that has been equilibrated
with solvent mixture of mobile phase.
Results
The total carotenoid content of the tested gelatin capsule
of vitamin supplement (β-carotene, extract from blueberries)
was 6 mg (Table I).
Table I
β-carotene content in the gelatin capsules
name Weight β-Carotene Extraction of
[g] [mg] blueberries [mg]
0.3156 6 150
Fig. 1. Calibration curve of β-Carotene