Quantitative Evaluation of β-Carotene and Xanthophyll in Some Medicinal Plants from Kumaon Himalayas
Antioxidant phytochemicals such as β-carotene and xanthophyll contents were estimated in three different medicinal plants, widely distributed in Kumaon Himalayas using a reverse-phase HPLC system. Maximum β-carotene content (481.57 mg/100 g) was found in the leaves of Asplenium dalhousiae and maximum xanthophyll content (678.61 mg/100 g) was found in the leaves of Osmanthus fragrans. The aim of this study was to characterize the antioxidant value of the medicinal plants with particular attention to carotenoids and xanthophyll.
to cite this article:
Kundan Prasad, S. Gupta and G. Bisht, 2014. Quantitative Evaluation of β-Carotene and Xanthophyll in Some Medicinal Plants from Kumaon Himalayas. Research Journal of Phytochemistry, 8: 205-209.
Received: July 20, 2014;
Accepted: August 13, 2014;
Published: November 13, 2014
Melia azedarach (Meliaceae) is a wild plant native to India, southern
states of America, Africa, South Europe and warmer parts of the globe. It has
been widely used for its analgesic, emetic, antiseptic and anthilmintic properties.
The plant resembles neem in having the medicinal properties (Rastogi
and Mehrotra, 1995; Chopra et al., 1956).
Asplenium dalhousiae (Aspleniaceae) is a wild plant native to India,
Mule, Hauchuca and Baboquivari mountains of southern Arizona. It has been widely
used in traditional medicines for spleen ailments jaundice and diuretic. Rhizomes
of the plant are used in abscesses (Dhar et al.,
1974). Osmanthus fragrans (Oleaceae) is a wild plant native to India,
E. Asia, China, Japan and Himalayas. O. fragrans, has been used in the
treatment of dysmenorrhoea, rheumatism and bruises. Flowers of plant are used
by the Chinese to impart a pleasant flavour to tea, wine and sweet dishes such
as lotus seed soap, pastries and steamed pears (Srivastava
and Kapoor, 1985).
Carotenoids are important nutritionally as antioxidants in the prevention of
antherosclerosis and in the prevention of age related macular degeneration (AMD)
(Palozza and Krinsky, 1992; Dwyer
et al., 2001; Moeller et al., 2000).
These natural pigments are more acceptable to consumers as they have always
been present in natural foods and are readily metabolized. The metabolites are
good for human health. The hydrocarbon carotenoids have provitamin A. Activity
and the oxygenated carotenoids or xanthophyll are possibly linked to a lower
risk of cancer (Beecher and Khachik, 1984). Some workers
have reported the possibility that certain carotenes or their isomers have anticancer
potential (Beems, 1987; Hennekens,
1986). β-Carotene has been proved to prevent peroxidation caused by
singlet oxygen and also by scavenging free radicals (Di
Mascio et al., 1991; Krinsky, 1989). Carotenoids
are mainly responsible for the prevention of the deleterious effect of singlet
oxygen (Cadenas, 1989).
MATERIALS AND METHODS
Chemicals: Standard of xanthophyll and β-carotene were procured from
Sigma Chemical Co. St Louis, USA. Individual standard was accurately weighed,
developed and diluted with HPLC grade ethanol. Petroleum ether, methanol, ethyl
acetate and anhydrous sodium sulphate and other chemicals and reagents used
in this study were purchased form Merck Chemical Co. Mumbai, India.
Plant material: Plant material (sample) was collected from Nainital
District, Uttaranchal and authentic identification was done in Botany Department,
Kumaun University, Nainital. The plant meterial was dried in shade after collection.
The dried plant material was powdered separately in a electrical mill to 60
mesh size. The fine plant material powder so obtained was used for further vitamins
Extraction and analysis of β-carotene and xanthophyll: The procedure
described by Kurilich et al. (1999) was followed
for the analysis of β-carotene as well as xanthophyll. The 300-500 mg of
sample was taken in a test tube and added to it 10 mL of ethanol containing
0.1 g of BHT. The test tube along with the sample was placed in a water bath
at 700C for 15 min. After removing the tubes from the water bath, 180 μL
of 80% KOH was added to each tube. The sample was vortexed and then saponified
at 700C for 30 min. Saponification was essential for maximum extraction of carotene
and their esters. The samples were placed directly on ice bath and 2.5 mL of
de-ionized water and 2.5 mL hexane/toluene mixture (10:8). Then, the tubes were
vortexed and then centrifuged at 2100 rpm for 5 min. The upper layer hexane/toluene
fraction was then transferred to a separate test tube. The hexane/toluene extraction
was repeated for two more times. The combined hexane/toluene fractions were
dried using a Speed-vac concentrator. The residue was reconstituted in 200-400
μL THF. The solution was filtered on a 0.2 μ nylon filter and 20 μL
of the filtered solution was injected in the Shimadzu high performance liquid
chromatograph. The mobile phase consisted of acetonitrile:methanol:THF (52:40:8)
(v/v/v) at a flow rate of 2.0 mL min-1. The absorbance was recorded
at 450 nm for β-carotene and lutein. The retention time for the standard
β-carotene was recorded as 6.192 min for xanthophyll at 2.350 min and for
vitamin A at 4.400 min.
RESULTS AND DISCUSSION
In this study, we have observed that amongest three medicinal plants the range
of β-carotene in leaves varied from 110.68-481.57 mg/100 g (Table
1) and xanthophyll in leaves varied from 124.58-678.61 mg/100 g (Table
2) at dry weight basis.
||Standard peaks of β-carotene and xanthophyll
In the leaves of M. azedarach, both β-carotene and xanthophyll
content was much higher than the roots of the same plant. The maximum β-carotene
was observed in A. dalhausiae and minimum in M. azedarach leaves.
The maximum xanthophyll content was observed in O. fragrans and minimum
in M. azedarach leaves.
Khachik and Beecher (1987) have reported carotene
137.60-179.82 mg/100 g in carrots, 47.38-53.33 mg/100 g in sweet potatoes and
208.00-232.01 mg/100 g in pumpkin at dry weight basis. Plants of A. dalhausiae
have more carotene level than carrot, sweet potatoes and pumpkin.
The chromatographic separation of β-carotene and xanthophyll standard
is presented in Fig. 1. Typical chromatograms of β-carotene
and xanthophyll extracted from medicinal plants are shown in Fig.
2. Standards of β-carotene and xanthophyll were purchased from sigma
and each individual standard was accurately weighed, dissolved and diluted with
ethanol to obtain concentration about 100 μg mL-1.
Identification of the β-carotene and xanthophyll was performed with reference
to standard. The quantification of these carotenoids in samples were compared
with known quantity of standard peak area.
Thus, the study concludes that M. azedarach, A. dalhausiae and
O. fragrans are excellent sources of antioxidants especially β-carotene
and xanthophyll. The distribution of carotenoids and their related geometrical
isomers in common medicinal plants has important application for the health
of Asian people in addition to the basic needs of developing countries.
The significance of xanthophyll and E/Z isomers is becoming increasingly evident
in eye health and more specifically in relation to the prevention of AMD and
other blinding disorders.
||Standard peaks of β-Carotene and xanthophyll extracted
from (a) O. fragrans, (b) A. dalhousiae, (c) M. azedarach
(leaves) and (d) M. azedarach (roots)
Authors are thankful to Dr. J. Singh, Department of Biochemistry, I.I.V.R.
(B.H.U.), Varanasi for Providing HPLC facilities for this study.
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