International Journal of Pure and Applied Mathematics

Volume 119 No. 18 2018, 2309-2317

ISSN: 1314-3395 (on-line version)
url: http://www.acadpubl.eu/hub/
Special Issue
http://www.acadpubl.eu/hub/

PREDICTION OF SUITABLE LEAFY VEGETABLES AND MEDICINAL PLANTS

FOR REDUCING FLUORIDE TOXICITY
1
Dr. G. Viswanathan, Dr. V. Dhanapal, 3 Dr. M. P. Pachamuthu, 4 Dr. V. Vijayakumar, 5 Dr. P.
2

Subhapriya,6 Dr. M. Malathi magalingam

1
Assistant Professor, Department of Chemistry, Bannari Amman Institute of Technology,
Sathyamangalam, Tamil Nadu, India
2
Assistant Professor (II) & Head, Department of Chemistry, Bannari Amman Institute of
Technology, Sathyamangalam, Tamil Nadu, India
3
Assistant Professor, Department of Chemistry, Bannari Amman Institute of Technology,
Sathyamangalam, Tamil Nadu, India
4
Assistant Professor, Department of Chemistry, Bannari Amman Institute of Technology,
Sathyamangalam, Tamil Nadu, India
5
Assistant Professor, Department of Chemistry, Bannari Amman Institute of Technology,
Sathyamangalam, Tamil Nadu, India
6
Assistant Professor, Department of Chemistry, Bannari Amman Institute of Technology,
Sathyamangalam, Tamil Nadu, India
Corresponding author 1 viswakind@gmail.com,
2
vdhanachemist@gmail.com,3 pachamuthu@bitsathy.ac.in4 vijayakumarv@bitsathy.ac.in,5 subhap
riyap@bitsathy.ac.in,6 malathimahalingamm@bitsathy.ac.in

ABSTRACT
Intake of drinking water and diet with more calcium with rich in bioavailability can
reduce the absorption of fluoride from gut. Commonly available leafy vegetables and some
medicinal plants are the good source of dietary calcium. Hence the present study is proposed, to
account the total and bio-available calcium level in various leafy vegetables and five Cassia
species. Total and bio-available calcium levels are determined through wet digestion and in-vitro
digestion method, respectively. Among the selected leafy vegetables; Dwarf-copper, Block-
nightshade, Curry leaves, Cabbage, Drumstick leaves and Mint leaves contain high total calcium
with rich bioavailability. Among the five Cassia species; Cassia tora, Cassia alata and Cassia

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angustifolia shows high total and bio-available calcium level. Hence, the frequent usage of these
selected plant materials through daily diet can reduce the risk of fluoride toxicity in endemic
regions.
Key words: fluorosis; bioavailability; calcium; leafy vegetables; cassia species.
1. Introduction
Prolong intake of fluoride more than the recommended level through water and food may
cause dental and skeletal fluorosis [1]. Dental fluorosis is characterized by lusterless, opaque
white patches in the enamel, which may become stained yellow to dark brown, and in severe
forms cause marked pitting and brittleness of teeth. Skeletal fluorosis is associated with limited
movement of the joints, skeletal deformities, and intense calcification of ligaments, muscle wasting
and neurological deficits [2].
Bioavailability is generally defined as “the fraction of administered dose of drug that
reaches the systemic circulation”. Fluoride is almost completely absorbed from drinking water
through gastro-intestinal tract in the absence of Ca, Mg and Al ions [3]. But, the bioavailability
of fluoride exposed through various food items varies between 2% to 79%. Fluoride
bioavailability was governed by various factors such as pH and mineral content of the food.
Calcium intake reduces the absorption of fluoride from gut, by forming calcium-fluoride
insoluble complex [4]. Some of the earlier studies reported that, the serum fluoride level is
greatly decreased when sodium fluoride is administered along with high-calcium-diet [5, 6]. The
risk of fluorosis is lower in children intake adequate level of calcium. Therefore, the intake of
food items with high bioavailable calcium helps to reduce the absorption of fluoride and
minimize the fluorosis risk [6]. Moreover, calcium is essential for normal growth and
development of the skeleton as well as for teeth, nerve, and muscle and enzyme functions.
Apart from drinking water, milk, dairy products, some of the commonly available leafy
vegetables and few medicinal plants are good and cheapest sources of dietary calcium. Also the
intake of leafy vegetables and medicinal plants through diet can impart various health benefits.
Calcium presents in plant materials primarily complexes with some anti-nutrients such as
oxalate, phytate, fiber and other phenolic compounds. This reduces the solubility of calcium in
the gastrointestinal tract and turn it indigestible [7]. Acidic medium in stomach can increase the
solubility of calcium complexes and enhances the calcium absorption [8].

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Bio-available calcium level varying greatly among every plant materials may be due
to the presence of various levels of anti-nutrients. Hence, it is necessary to estimate the total as
well as bio-available calcium levels in commonly available leafy vegetables and medicinal
plants. In-vitro digestion experiment is conducted to estimate the bioavailability of calcium by
simulating the human digestive system via a two-step digestion that includes a gastric digestion
and intestinal digestion.

2. Materials and methods

2.1 Collection of leafy vegetables and five Cassia species
About 200 g of fresh and raw leafy-vegetables were procured from markets in three
different areas of Erode District and about 200 g of fresh and raw leaves of five Cassia species,
Cassia tora, Cassia alata, Cassia angustifolia, Cassia auriculata, Cassia fistula were collected
in three different areas of Erode District using clean polyethylene bags. All the collected samples
were cleaned with double-distilled water and stored in refrigerator at 10°C.
2.2 Estimation of total calcium level in plant materials
About 4 g to 5 g of accurately weighed edible portion of homogeneous plant sample was
taken into a 250 ml long-necked conical flask. About 5 ml of concentrated sulphuric acid was
added to the sample and boiled. After the digestion, the content was mixed with 5 ml of
concentrated nitric acid and boiled. Then the content was treated with 5 ml of 60% perchloric
acid and heated up to the brown fumes are completely expelled out. At the end of the digestion
process the content was cooled and mixed with 20 ml of double-distilled water. Then the content
was filtered and made up to 100 ml. Calcium level in the sample solution was measured through
Atomic Absorption Spectrophotometer.
The optimum conditions for the estimation of calcium in Atomic Absorption
Spectrophotometer were, the wave length set at 422.7 nm, slit width 0.7 nm, relative noise 1.0
and oxidizing lean, blue flame obtained from air-acetylene mixture was used for the aspiration of
samples. In order to reduce the ionization interferences and chemical interferences 0.1% of KCl
and 0.1% of LaCl3 were used respectively. Then the sample solution was introduced into Perkin-
Elmer Flame Atomic Absorption Spectrophotometer (Perkin Elmer AA – 100, USA), after the
calibration of the instrument using appropriate calcium standards and control. Calcium level of
the unknown sample was directly read out from the digital display of the meter. The level of

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calcium in sample was calculated from the reading shown in digital display of the meter by
multiplying with appropriate dilution factor [9].
2.3 Estimation of bio-available calcium level in five Cassia species
Estimation of bio-available calcium levels was carryout through in-vitro analysis
described by Miller et al, which was well correlate with human in-vivo studies. This method has
the advantage that includes enzymatic digestion of plant materials under simulated

Vernacular Common name Botanical name Total Bioavailable

name calcium calcium
(mg/100 g) (mg/100g)
Usei-thagarai Coffee Cassia Cassia tora 3200 ± 205 192 ± 12
Seemai-agathi Ringworm senna Cassia alata 267 ± 20 24 ± 2
Tirunelveli-senna Senna Cassia angustifolia 187 ± 14 41 ± 3
Avarai Avaram senna Cassia auriculata 133 ± 12 9±1
Sarakkondrai Indian laburnum Cassia fistula 211 ± 17 8±1
gastrointestinal condition, followed by measuring soluble calcium which diffused from the
sample [10, 11].

3. Results
3.1 Total and bio-available calcium levels in five Cassia species and leafy vegetables
The levels of total and bio-available calcium in commonly available five Cassia species
are shown in Table 1.
Table 1. Total and bioavailable calcium levels in the five selected Cassia species
Values represent mean level with standard deviation
It is worth mentioning here, that among the five Cassia species, Cassia tora shows the
highest total calcium level (3412 mg/100g). Even though Coffee Cassia (Cassia tora - Usei-
thagarai) contains a large amount of total calcium, it has very low bioavailability (≈ 23%).
However, in general, this small fraction of bio-available calcium in Cassia tora is more than that
of the calcium intake through a cup of milk. Ringworm-senna (Cassia alata – Seemai-agathi)
and Avaram-senna (Cassia auriculata - Avarai) are showing considerable level of total and bio-
available calcium.

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The levels of total and bio-available calcium in leafy vegetables commonly used in diet
and chosen for the present study are shown in Table 2. Bakphul, curry leaves, cauliflower leaves,
pnarnava leaves and dwarf-copper show the total calcium level more than 500 mg/100g. Even

Common name Botanical name Total Ca Bio-available Ca

(mg/100 g) (mg/100 g)
Curry leaves Murraya koenigii 824 ± 42 272 ± 14
Cauliflower leaves Brassica Oleracea 615 ± 32 86 ± 4
Tropical-amaranth Amaranthus polygonoides 248 ± 23 52 ± 5
Bakphul leaves Sesbania grandiflora 1152 ± 53 115 ± 5
Coriander leaves Coriandrum sativum 158 ± 21 33 ± 4
Black night-shade Solanum nigrum 417 ± 30 179 ± 13
Mint leaves Mentha spicata 204 ± 16 55 ± 4
Dwarf copper Alternanthera sessilis 517 ± 31 212 ± 13
Spinach Spinacia oleracea 131 ± 11 10 ± 1
Tamarind Tamarindus indica 101 ± 12 13 ± 2
Punarnava Boerhavia diffusa 637 ± 36 172 ± 10
Red spinach Amaranthus gangeticus 366 ± 24 7±1
Spiny pigweed Amaranthus viridis 327 ± 19 7±1
Betel leaves Piper betle 221 ± 21 84 ± 8
Common-purslane Portulaca oleracea 112 ± 12 13 ± 1
Drumstick leaves Moringa Olifera 443 ± 30 128 ± 9
Susni sag Marsilea minuta 367 ± 27 103 ± 8
Cabbage Brassica oleracea 42 ± 4 13 ± 1
Values represent mean level with standard deviation
Table 2. Total and bio-available calcium levels in leafy vegetables
though, tropical-amaranth, coriander, spinach, red-spinach, spiny-pigweed and common-
purslane show considerable level of total calcium with low bioavailability. Among the selected
leafy vegetables, dwarf-copper (Ponnanakanni-keerai) and block-nightshade (Manathakkali-
keerai) contain more than 40% of bio-available calcium. Also, curry, cabbage, drumstick leaves
and mint leaves contain high total and rich bio-available calcium level.

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4. Discussion
Commonly, plants are the good source of calcium and vitamins. Consumption of plant
sources with high calcium and rich bioavailability is the suitable way to minimize the risk of
fluorosis. Frequent intake of dietary calcium through leafy vegetables and medicinal plants helps
to get required amount of calcium for reducing the risk of fluorosis and for maintaining good
health. Calcium is absorbed from the small intestine by both active and passive mechanisms.
Active transfer mechanism of calcium absorption is predominant when the calcium intake level
is low or moderate. Active transfer needs the action of the active-form of vitamin D [12].
Normally, calcium is soluble from calcium complexes during digestion and is released probably
in ionized form for absorption. If calcium intake level increase then the active transfer
mechanism becomes saturated and an increased proportion of calcium is absorbed via passive
diffusion [13].
Many of the leafy vegetables and cassia species show high level of total calcium, but
their bio-available calcium level varies from total calcium levels. Calcium in plants may
primarily complexes with oxalate, phytate, fiber, lactate, fatty acids, protein and other
compounds. Oxalate is present in large amount in most of the commonly consumed plant
materials [14, 15]. Calcium oxalate complexes are unavailable for the absorption from the gut
[16]. Soluble oxalates in medicinal plants and other foods can also reduce the absorption of
minerals. The insoluble form of calcium (calcium-oxalate) may not be absorbed through any
absorption process.
It has been suggested that the intake of dietary calcium through leafy vegetables like
Black- night-shade, Dwarf-copper, Curry, Cabbage, Drumstick leaves and Mint leaves can also
impart some other important beneficial effects to human health. Also these common leafy
vegetables are frequently used as diet components. Intake of Cassia species like Cassia tora
containing total calcium with rich bioavailability through diet can deliver more calcium.
Moreover, it has been suggested that, the intake of dietary calcium from plant materials consists
of high bio-available calcium with low anti-nutrient level can reduce the risk of kidney stone
formation [15].
5. Conclusion
Among the five cassia species Cassia tora, Cassia alata and Cassia angustifolia show
considerable levels of total and bio-available calcium. Also, the intake of leafy vegetables like

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black night-shade, dwarf-copper, curry, cabbage, drumstick and mint leaves contain high total
and bio-available calcium through diet can deliver enough calcium to minimize the absorption of
fluoride.
Hence, it is recommended to the people residing in fluoride endemic areas to consume
these selected plants along with their diet. To keep away from some health defects, plants with
high calcium level with low bio-availability should be avoided due to the presences of
considerable level of anti-nutrients such as oxalate, phytate and tannin.
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