Nutritional Profile of Chekurmanis (Sauropus androgynus), A Less Explored Green Leafy Vegetable

Abstract

Chekurmanis (Sauropus androgynus) belonging to Euphorbiaceae, is a perennial shrub, growing wildly in Southeast Asia. The leaves of chekurmanis are highly nutritious, being a very rich source of micronutrients and protein. In this study, chekurmanis leaves were evaluated for protein and micronutrient content at different stages of maturity. Maturity of the leaves did not alter the rich content of protein which remained around 22.0 g/100 g. Total dietary fibre content of the leaves ranged between 34 and 36% at different stages of maturity. Fully matured leaves contained a significantly higher amount of calcium, as compared to the tender leaves. The iron content of these leaves ranged from 3.89 to 4.50 mg /100 g, while the zinc content of the same was between 1.26 and 1.48 mg/100 g. The bioaccessibility of iron and zinc was significantly higher in the tender leaves. The tender and partially mature leaves of chekurmanis contained 74 and 69 mg/100 g of niacin, respectively. The β-carotene content of the leaves ranged from 7400 to 9250 μg/100 g, while that of vitamin E ranged between 17.6 to 15.6 mg/100 g. Chekurmanis leaves contain an alkaloid, the content of which was significantly higher in mature leaves (1740 mg/ 100 g) as compared to tender leaves (1439 mg/100 g). The alkaloid content of chekurmanis leaves was significantly reduced upon pressure cooking. This is the first report on the nutrient content of chekurmanis leaves at different stages of maturity, as also on the bioaccessibility of minerals.

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Nutritional Profile of Chekurmanis (Sauropus androgynus),
A Less Explored Green Leafy Vegetable
Kalpana Platel and Krishnapura Srinivasan
(Department of Biochemistry, CSIR – Central Food Technological Research Institute,
Mysore – 570 020, India)
e-mail: ksri.cftri@gmail.com
(Received 27th March, 2017)
Abstract
Chekurmanis (Sauropus androgynus) belonging to Euphorbiaceae, is a perennial
shrub, growing wildly in Southeast Asia. The leaves of chekurmanis are highly
nutritious, being a very rich source of micronutrients and protein. In this study,
chekurmanis leaves were evaluated for protein and micronutrient content at different
stages of maturity. Maturity of the leaves did not alter the rich content of protein
which remained around 22.0 g/100 g. Total dietary fibre content of the leaves ranged
between 34 and 36% at different stages of maturity. Fully matured leaves contained a
significantly higher amount of calcium, as compared to the tender leaves. The iron
content of these leaves ranged from 3.89 to 4.50 mg /100 g, while the zinc content of
the same was between 1.26 and 1.48 mg/100 g. The bioaccessibility of iron and zinc
was significantly higher in the tender leaves. The tender and partially mature leaves
of chekurmanis contained 74 and 69 mg/100 g of niacin, respectively. The
β
-carotene content of the leaves ranged from 7400 to 9250 µg/100 g, while that of
vitamin E ranged between 17.6 to 15.6 mg/100 g. Chekurmanis leaves contain an
alkaloid, the content of which was significantly higher in mature leaves (1740 mg/
100 g) as compared to tender leaves (1439 mg/100 g). The alkaloid content of
chekurmanis leaves was significantly reduced upon pressure cooking. This is the first
report on the nutrient content of chekurmanis leaves at different stages of maturity, as
also on the bioaccessibility of minerals.
Keywords: Chekurmanis leaves, nutritional profile, stages of maturity, bioaccessibility
of minerals, alkaloid content
The Indian Journal of Nutrition and Dietetics
ISSN: 0022-3174; eISSN: 2348-621X
www.ijndonline.org
243
DOI:10.21048/ijnd.2017.54.3.15765
The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017
Introduction
Chekurmanis (Sauropus
androgynus) of the family Phyllanthaceae
martynov is a perennial shrub, growing
wildly in South and Southeast Asia. The
leaves of chekurmanis are commonly
consumed in Malaysia. Consumption of
chekurmanis, however, is not very
common in India, being restricted to
certain parts of Southern India1. The

exploited for use in the development of
nutritious food products, at a low cost,
after ascertaining its safety. As a result
of the high concentration of vitamin C
and E, chekurmanis may also have
antioxidant properties, which merits its
inclusion in health foods.
In the Southern parts of India, the
leaves of chekurmanis are usually
consumed either in the cooked form as
a vegetable, or in the form of a powder
incorporated with spice powder. In this
context, it would be interesting to see if
leaves of varying maturity differ in their
nutritive content, information not
available thus far. In addition, the effect
of heat processing on the alkaloid
content of chekurmanis also merits
examination. Thus the aim of the
present investigation was to determine
the nutritive content of chekurmanis
leaves at varying stages of maturity.
The influence of heat processing on the
total alkaloid content of chekurmanis
leaves was also studied. The basic data
on chekurmanis leaves generated in
this investigation will find application in
the development of nutritious food
products, which can be consumed
by people of all age groups and which
will help to overcome micronutrient
deficiencies.
Materials and Methods
Materials
Chekurmanis leaves at three
stages of maturity, viz. tender, partially
mature and fully mature, were collected
Kalpana Platel and Krishnapura Srinivasan
The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017
244
leaves of chekurmanis are highly
nutritious, being a very rich source of
β-carotene, vitamin E, vitamin C,
thiamine, riboflavin, calcium, iron, zinc
and protein. Thus, chekurmanis is an
unusual green leafy vegetable having
maximum number of nutrients being a
rich source of several vitamins,
minerals and protein1-5. Chekurmanis
leaves have been reported to possess
strong antioxidant properties, probably
owing to the vitamin C and E
contents3,4. In view of its rich nutrient
composition, this shrub can be explored
for the development of health beneficial
food products, which can also help
in the prevention of micronutrient
deficiencies. Chekurmanis leaf,
however, is reported to contain the
alkaloid papaverine in considerable
amounts5. Excessive consumption of
the leaves especially in raw form has
been reported to cause drowsiness and
respiratory disorders attributable to this
alkaloid. The effect of heat processing as
is involved in domestic cooking on the
papaverine content of chekurmanis
leaves merits to be examined.
In view of the widespread
micronutrient deficiencies in India, it
is desirable to explore natural sources of
micronutrients that can be easily grown
and used in households. Chekurmanis
is a perennial shrub which grows wildly
and which is a very rich source of
vitamins, minerals, as well as protein
as compared to other GLV. This easily
available plant can therefore be

from a local farm house. The source of
the leaves was maintained constant
throughout the study. The leaves were
considered to be tender before budding,
partially mature when they just started
to flower and fully mature when the
fruits (approx. 10 mm in diameter)
appeared. The leaves were washed to
remove adhering contaminants and
patted dry between filter paper sheets.
Fresh leaves were used for the analysis
of vitamins, while the leaves were dried
in a hot air oven for mineral analysis.
All analyses were carried out in four
replicates.
Total zinc, iron and calcium
The leaves were dried in a hot air
oven, powdered and ashed in a muffle
furnace at 550 ºC for 10 h and the ash
was dissolved in conc. HCl (Sp.Gr.1.18).
Zinc and iron contents were determined
by atomic absorption spectrometry
(Shimadzu AAF-6701, Shimadzu
Corporation, Kyoto, Japan). In the case
of calcium, lanthanum chloride
was added to the mineral solution
(final concentration 1%) to avoid
interference from phosphate.
Calibration of measurements was
performed using commercial standards.
All measurements were carried out
using standard flame operating
conditions, as recommended by the
manufacturer. The reproducibility
values were within 2.0% for both zinc
and iron.
Bioaccessibility of zinc and iron
Bioaccessibility of zinc and iron
from various food grain samples
was determined by an in vitro method
described by Luten et al 6, involving
simulated gastrointestinal digestion
with suitable modifications. The leaf
samples were finely ground in a
stainless steel blender. The ground
samples were subjected to simulated
gastric digestion by incubation with
pepsin (pH 2.0) at 37°C for 2 h. Titratable
acidity was measured in an aliquot
of the gastric digest, by adjusting the
pH to 7.5 with 0.2M sodium hydroxide in
the presence of pancreatin–bile extract
mixture (1 L of 0.1M sodium bicar-
bonate containing 4 g pancreatin + 25 g
bile extract). Titratable acidity was
defined as the amount of 0.2M sodium
hydroxide required to attain a pH of 7.5.
To simulate intestinal digestion,
segments of dialysis tubing (molecular
mass cut off 10 kDa) containing 25 ml
aliquots of sodium bicarbonate solution,
being equivalent in moles to the sodium
hydroxide needed to neutralize the
gastric digest (titratable acidity)
determined as above, were placed in
Erlenmeyer flasks containing the
gastric digest and incubated at 37 ºC
with shaking for 30 min or longer until
the pH of the digest reached 5.0. Five mL
of the pancreatin–bile extract mixture
were then added and incubation was
continued for 2 h or longer until the pH
of the digest reached 7.0. At the end of
simulated gastro-intestinal digestion,
Nutritional Profile of Chekurmanis (Sauropus androgynus), A Less Explored Green Leafy Vegetable
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The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017

zinc and iron present in the dialyzate
were analyzed by atomic absorption
spectrometry. The dialyzable portion of
the total minerals present in the sample
(expressed as per cent) represented the
bioaccessible minerals. Bioaccessibility
(%) was calculated as
Bioaccessibility (%) = 100 · Y/Z,
where Y is the element content of
the bioaccessible fraction (mg mineral
element/100 g leaves) and Z is the total
zinc or iron content (mg mineral
element/100 g leaves).
Vitamins
Thiamine was estimated by
fluorometry as described by the AOAC7.
The vitamin was extracted from the
leaves by acid hydrolysis, followed by
treatment with the enzyme taka-
diastase at 50 °C for 3 h. Thiamine was
then separated from other bio molecules
by column chromatography using
BioRex-70 and eluted using boiling acid-
KCl. Quantification of the vitamin was
done by oxidizing thiamine to
thiochrome using alkaline potassium
ferricyanide. The thiochrome was
extracted into iso-butanol, which was
used for measurement of fluorescence.
Estimation of riboflavin was done
after acid hydrolysis of flavin
mononucleotide and flavin adenine
dinucleotide to free riboflavin, removal
of protein by alkali hydrolysis and
stabilization of the vitamin in the
extract by glacial acetic acid8.
The vitamin was then quantified
fluorimetrically.
The procedure for estimation of
niacin involved extraction of the
vitamin from the leaves by calcium
hydroxide and removal of excess
calcium by oxalic acid9. Niacin was
measured by HPLC using C18 column
and a mobile phase consisting of
23% ACN + 0.1% H3PO4 + 0.1% SDS in
Milli-Q water9.
β-carotene was estimated by HPLC
as described by earlier10. The
provitamin was extracted from the
samples initially with a mixture of
acetone:ethanol (1:1) and subsequently
with petroleum ether, followed by
saponification to remove chlorophyll,
with 30% methanolic potassium
hydroxide at room temperature for 3 h.
Following saponification, the alkali was
removed completely by repeated
washing and the solvent was evaporated
to dryness in a rotary evaporator. The
residue was redissolved in petroleum
ether and stored in the cold pending
analysis. Prior to analysis, the
petroleum ether was evaporated under
nitrogen and the residue was dissolved
in the mobile phase used for HPLC
determination. Quantification of
β-carotene was carried out by reverse
phase HPLC (Shimadzu LC 10 AVP,
Shimadzu Corporation, Kyoto, Japan),
system consisting of a photo diode array
detector (SPD-M 20A), system controller
(SCL-10AVP), solvent delivery system D
The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017
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Kalpana Platel and Krishnapura Srinivasan

247
The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017
precipitation with Dragendorff’s
reagent, involving formation of yellow
bismuth complex in nitric acid medium
with thiourea developed by Narasimhan
and Shanta12.
Results and Discussion
Proximate composition of chekurmanis leaves
The moisture, protein and dietary
fibre content of chekurmanis leaves are
presented in Table I. There was no
significant difference in the moisture
content of the leaves at different stages
of maturity, the moisture content
ranging from 76 to 77.5%. Protein
content of chekurmanis leaves was
4.95, 5.11 and 4.87/100 g in tender,
partially mature and fully mature leaves
on fresh weight basis. As in the case of
protein and moisture, dietary fibre
content of the leaves was similar in the
tender, partially mature and fully
mature leaves. The total dietary fibre
content was 7.74, 8.46 and 8.40% at the
three stages of maturity, respectively.
More than 80% of the total dietary fibre
was in the form of insoluble fibre.
pump (LC-10AT VP) and manual sample
injector. Chromatographic separation
was accomplished using SGE 250 . 4.6 mm
ODS C18 column (SS Exil, Dandenong,
Victoria, Australia). β-Carotene
containing sample was eluted with an
isocratic mobile phase which consisted
of a mixture of 65% (v/v) acetonitrile,
15% (v/v) methylene chloride and 20%
(v/v) methanol containing 1.3 mm
ammonium acetate at a flow rate of
1 mL/min. Adequate precautions were
taken to minimize the exposure of
samples to light and air and thus
prevent oxidative destruction of
β-carotene.
α-tocopherol in chekurmanis
leaves was determined by HPLC method
described by Zaspel and Csallany11
using ODS column (C-18) and an UV-
visible detector (295nm) and a solvent
system acetonitrile - methanol (1:1).
Total alkaloids
Total alkaloids in chekurmanis
leaves was determined by the
spectrophotometric method after
TABLE I
Moisture, Protein and Dietary Fibre Content of Chekurmanis Leaves at Different Stages of Maturity
Moisture Protein Dietary fibre (%)
Stage of maturity (%) (g/100 g) Insoluble Soluble Total
Tender 77.5 ± 1.0 4.95 ± 0.27 6.39 ± 0.034 1.35 ± 0.038 7.74 ± 0.097
Partially mature 76.0 ± 0.1 5.11 ± 0.53 7.03 ± 0.17 1.44 ± 0.024 8.47 ± 0.18
Fully mature 76.8 ± 0.7 4.87 ± 0.02 6.89 ± 0.17 1.51 ± 0.042 8.40 ± 0.22
Values are mean ± SEM of four replicate analyses; Values are on fresh weight basis
Nutritional Profile of Chekurmanis (Sauropus androgynus), A Less Explored Green Leafy Vegetable

The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017
248
The protein content of chekurmanis
leaves examined is significantly higher
than that reported in literature, where
it ranged from 3 to 5 g/100 g, while
the moisture content is somewhat
comparable to reported values1-3, 13.
Mineral content of chekurmanis leaves
Iron content of chekurmanis
leaves was 0.88, 0.83 and 0.94 mg/100 g
in the tender, partially mature and fully
mature leaves on fresh weight basis,
while zinc content in the same ranged
from 1.26 to 1.48 mg/100 g (Table II).
Thus, fully mature leaves had
marginally higher content of iron and
zinc. The calcium content of the
fully mature leaves, however, was
significantly higher than tender leaves
(596 versus 301 mg/100 g). Calcium
content of the partially mature leaves
was somewhat similar to that of tender
leaves.
The calcium content of tender and
partially mature chekurmanis leaves
on fresh weight basis in the present
study corresponds to that reported by
Sheela et al 2, where it was 313 mg/100 g.
The calcium content of chekurmanis
leaves is similar to that of other green
leafy vegetables such as amaranth
(397 mg/100 g), drumstick leaves
(440 mg/100 g) and fenugreek leaves
(397 mg/100 g)13. The iron content
observed in the study (3.47-4.05 mg/100 g)
is lower than the values reported by
Sheela et al (10.1 mg/100 g)2 and the
Indian Council of Medical Research
(ICMR) (28.0 mg/100 g)12. The reason for
this difference could be the different
analytical methods adopted in these
studies. Amaranth leaves have similar
iron content (3.5 mg/100 g), while the
same is lower in drumstick, fenugreek
and spinach leaves13.
The bioavailability of minerals is a
matter of concern especially in plant
foods, which contain high amounts of
inhibitors of mineral absorption. In this
context, mere determination of the total
mineral content would not be adequate.
The bioaccessibility of iron and zinc
from chekurmanis leaves at different
stages of maturity was determined by a
TABLE II
Mineral Content of Chekurmanis Leaves at Different Stages of Maturity
Stage of maturity Iron (mg/100 g) Zinc (mg/100 g) Calcium (mg/100 g)
Tender 0.88 ± 0.08 0.28 ± 0.002 300.6 ± 43.4
Partially mature 0.83 ± 0.16 0.29 ± 0.005 378.7 ± 50.0
Fully mature 0.94 ± 0.05 0.34 ± 0.002 595.8 ± 55.0
Values are mean ± SEM of four replicate analyses; Values are expressed on fresh weight basis
Kalpana Platel and Krishnapura Srinivasan

simulated gastrointestinal procedure
with equilibrium dialysis. It was
surprising to note that there was a
decrease in bioaccessibility of iron as
the leaves matured, with a drastic
reduction in the fully mature leaves.
Bioaccessibility of iron in the tender
and partially mature leaves was 15.2
and 13.4%, respectively, while it was
2.72% in the fully mature leaves
(Table III). The same trend was
observed in zinc bioaccessibility, with
the reduction being prominent in the
partially mature stage itself. Per cent
zinc bioaccessibility in tender, partially
mature and fully mature leaves were
9.44, 2.19 and 2.93, respectively. The
reason for this drastic reduction in
mineral bioaccessibility in mature
leaves is not clear, since there were no
significant differences in either the
total content of these minerals, or in the
content of dietary fibre, a known
inhibitor of mineral absorption, in the
leaves of different maturity. One
probable reason could be the
significantly higher content of the
alkaloid in the fully mature leaves.
Although not specifically examined in
the investigation, it could be possible
that the alkaloid is an inhibitor of
mineral availability.
Vitamin content of chekurmanis leaves
The concentrations of vitamins B1,
B2 and B3 in the leaves at different
stages of maturity are presented in
Table IV. There was a significant
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The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017
TABLE III
Bioaccessible Iron and Zinc from Chekurmanis Leaves at Different Stages of Maturity
Stage of maturity % Bioaccessible iron % Bioaccessible zinc
Tender 15.20 ± 0.8 9.44 ± 0.55
Partially mature 13.40 ± 0.7 2.19 ± 0.09
Fully mature 2.72 ± 0.27 2.93 ± 0.24
Values are mean ± SEM of four replicate analyses
TABLE IV
B-complex Vitamin Content of Chekurmanis Leaves at Different Stages of Maturity
Stage of maturity Thiamine (mg/100 g) Riboflavin (mg/100 g) Niacin (mg/100 g)
Tender 0.15 ± 0.01 0.23 ± 0.003 74.2 ± 2.80
Partially mature 0.20 ± 0.03 0.20 ± 0.002 69.4 ± 0.40
Fully mature 0.26 ± 0.01 0.21 ± 0.002 47.4 ± 1.60
Values are expressed on fresh weight basis; Values are mean ± SEM of four replicate analyses
Nutritional Profile of Chekurmanis (Sauropus androgynus), A Less Explored Green Leafy Vegetable

increase in the thiamine content of the
leaves when they were fully mature, the
per cent increase being 73. Such an
increase was not visible in partially
mature leaves. On the other hand,
there was a decrease in niacin content
of the leaves with increasing maturity,
the decrease being significant in the
fully mature leaves. While the tender
leaves contained 74.2 mg/100 g, the
niacin content of the partially mature
and fully mature leaves was 69.4 and
47.4 mg/100g respectively. This is
significantly higher than the content of
niacin in other green leafy vegetables12.
The riboflavin content of the leaves
ranged between 0.20 and 0.23 mg/100 g.
The concentration of these water-
soluble vitamins in chekurmanis
leaves is significantly higher than any
of the other commonly consumed green
leafy vegetables such as amaranth,
fenugreek leaves, drumstick leaves
and spinach.
β-Carotene content of
chekurmanis leaves increased
significantly as the leaves matured,
with the fully mature leaves containing
around 30% more β-carotene than the
tender leaves. While the β-carotene
content of tender leaves was
7403 µg/100 g, the same in partially
mature and fully mature leaves was
8600 and 9250 µg/100 g, respectively
(Table V). The β-carotene content of
chekurmanis leaves has been reported
to be 5700 µg/100 g, by the ICMR12.
Chekurmanis leaves were found to
be a very good source of vitamin E, the
fat-soluble vitamin with antioxidant
properties. Vitamin E content of the
tender and partially mature leaves was
around 17 mg/100 g, while there was a
slight decrease in the fully mature
leaves (15.6 mg/100 g). This is probably
the first study to report the vitamin E
content of this green leafy vegetable.
Vitamin C content of chekurmanis
leaves ranged from 282 to 314 mg per
100 g fresh leaves, thus registering only
a marginal increase in matured leaves.
The leaves of chekurmanis are
reported to contain the alkaloid
papaverine in considerable amounts5.
TABLE V
Content of ββ
ββ
β-carotene, Vitamin E and Vitamin C in Chekurmanis Leaves at Different Stages of Maturity
Stage of maturity β-Carotene (µg/100 g) Vitamin E (mg/100 g) Vitamin C (mg/100 g)
Tender 7403 ± 10.3 17.6 ± 0.15 282.1 ± 15.6
Partially mature 8600 ± 20.0 17.8 ± 0.90 319.2 ± 19.3
Fully mature 9250 ± 50.0 15.6 ± 0.05 314.5 ± 0.5
Values are expressed on fresh weight basis. Values are mean ± SEM of four replicate analyses
Kalpana Platel and Krishnapura Srinivasan
The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017
250

The present investigation determined
the alkaloid content in the leaves at
different stages of maturity. Alkaloid
content of the tender leaves was around
1400 mg/100 g, whereas the same in
the partially and fully mature leaves was
around 1700 mg/100 g (Table VI). Thus,
there was an 18% increase in the
alkaloid content as the leaves matured.
Excessive consumption of the raw
leaves is reported to cause drowsiness
and respiratory disorders, which is
attributed to the alkaloid content of the
leaves. In view of this, the effect of heat
treatment by pressure-cooking (as
encountered in our cooking practices)
on the alkaloid content was examined.
Pressure-cooking of the leaves of
chekurmanis significantly decreased
the alkaloid content, the decrease being
more than 50% in the tender and
partially mature leaves, while in the
fully mature leaves, the decrease was
86%.
Thus, chekurmanis leaves are a
rich storehouse of micronutrients
such as vitamins and minerals. The
free radical scavenging activity of
chekurmanis leaves, as reported by
Nahak and Sahu3, could be attributable
to their high content of the antioxidant
vitamins E and C. By virtue of this
rich concentration of micronutrients,
chekurmanis leaves are far superior to
other common green leafy vegetables
such as amaranth, drumstick leaves,
fenugreek leaves and spinach.
This is the first documentation on
the nutrient content of chekurmanis
leaves at different stages of maturity.
The tender leaves, the form in which
chekurmanis is commonly consumed,
seem to have a higher content of all the
nutrients. The bioaccessibility of
minerals such as iron and zinc is also
significantly higher in the tender
leaves. Although chekurmanis leaves
contain high amounts of an alkaloid
that is reported to cause respiratory
disorders, the present study has
demonstrated that the alkaloid can be
destroyed by domestic heat treatment
methods such as pressure-cooking.
Summary and Conclusion
Thus, chekurmanis leaves can be
exploited to derive the maximum
TABLE VI
Total Alkaloid Content of Chekurmanis Leaves at Different Stages of Maturity
Total alkaloids (mg/100 g)
Tender Partially mature Fully mature
Raw 1439.0 ± 135.0 1714.7 ± 115.5 1740.6 ± 156.8
Pressure-cooked 714.4 ± 61.6 756.0 ± 62.3 191.2 ± 6.0
Values are expressed on fresh weight basis; Values are mean ± SEM of four replicate analyses
Nutritional Profile of Chekurmanis (Sauropus androgynus), A Less Explored Green Leafy Vegetable
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The Indian Journal of Nutrition and Dietetics, Vol.54 (3), July - September 2017

amount of micronutrients. The
consumption of this green leafy
vegetable should be promoted, especially
among the vulnerable sections of the
population. This green leafy vegetable
merits use in supplementary feeding
programmes and midday meal
schemes. This could be a highly feasible
strategy to combat micronutrient
malnutrition in our country.
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