Biochemical and Nutritional Assessment of Rhynchosia hirta (Andr.) Meikle and Verdc (Papilionaceae)
K. Sri Rama Murthy
Vivek Babu Kandimalla
Seeds of Rhynchosia hirta (Andr.) Meikle and Verdc. locally known as Adavi Kandulu in Telugu, used as food by local tribal in the Udayagiri forest of Nellore district of Andhra Pradesh of southern Peninsular India. The mature seed samples were analysed for proximate composition, total (true) seed protein fractions, amino acid composition, fatty acids profiles, mineral and antinutritional factors. The investigated seed samples contained higher amount of crude protein, crude fat, ash and nitrogen free extractives constitute 19.05, 8.73, 4.48 and 48.78 g/100 seed powder, respectively. The calorific value of seed material was 1690.9 kJ/100 g Dm. The essential amino acids, isoleucine, tyrosine and phenylalanine, were present in relatively large quantities. The fatty acid profiles revealed that the seed lipids contained higher concentrations of palmitic and linoleic acids. The seeds are rich in sodium, Phosphorous, calcium, zinc, manganese and iron. Anti-nutritional factors, such as total free phenols, (2.25%) tannins (4.75%), L-DOPA (0.65%), hydrogen cyanide (0.0654%) and phytic acid (1.32%) are, present in variable quantities. This paper presents the analytical data on the chemical composition and nutritional potential of Rhynchosia hirta seeds with regard to its suitability as a regular component in human diet.
Knowledge of the chemical composition of foods is key recommendations of foods
for providing a balanced nutritional diet. A table of food composition is used
at the macro-level in planning food demand and supply and at the micro level
in developing prescribed diet as well as in determining and correcting the nutritional
values of a given diet (Southgate, 1974). Inadequate availability
and consumption of protein foods in India due to both population exploration
and urbanization, will remain if efforts are not made toward finding alternate
and cheaper sources of proteins. In spite of an urgent need to meet the nutritional
requirements of the ever increasing populations, the available cheap protein
resources have remained relatively unexplored (Murthy et
al., 2003; Thangadurai et al., 2006). With
increasing in new food sources the seeds of wild plants, including the tribal
pulses, are receiving more attention because they are well adapted to adverse
environmental conditions, highly resistant to disease and pests and exhibit
good nutritional qualities (Maikhuri et al., 1991).
Some of the wild nuts and seeds are commonly used as proteinaceous foods in
different parts of the world (Amubode and Fetuga, 1983).
There are some 28 wild legumes commonly consumed by different tribal sects of
India (Arora et al., 1980; Murthy
and Pullaiah, 2005). However, most of the Indian legumes remain uninvestigated
biochemically and nutritionally. The tribal communities living in the forests
of Eastern Ghats in the vegetation of tropical moist deciduous and semi-evergreen
forests, collect the seeds of wild legumes genetic resource, randomly in the
vicinity of the forests, soak in water and consume the seed meal after boiling
and decanting for four to twelve times. This is tempted us to study the biochemical
composition of the seeds to know its potential use in human nutrition.
MATERIALS AND METHODS
The seeds of Rhynchosia hitra were collected from the moist deciduous
of Udayagiri forest of Nellore district, Andhra Pradesh, during April 2004 (early
summer) near the vicinity of tribal hamlets and were used for analysis. The
accessions were botanically identified by using the botanical keys of (Pullaiah
and Murthy, 2001) and deposited in the Herbarium of the Department of Botany,
Andhra Loyola College Herbarium. The moisture content was determined by drying
50 transversely cut seeds in an oven at 80°C for 24 h and is expressed on
a percentage basis. The oven dried and air-dried seeds were powdered separately
in a Kemi Mill (Scientific equipment works, Kerala), for 60-mesh size. The fine
powder obtained was used for further analysis. The total carbohydrate content
was estimated (Conrad and Palmer, 1976). The crude protein
content was calculated by multiplying the factor of 6.25 times percent
Kjeldahl nitrogen flowing Humphries (1956) method. The
crude fibre content was determined according to the methods described by Eggum
and Beame (1983). The contents of nitrogen free extractives (NFEs), crude
fat and ash were estimated by AOAC (1970) methods. The
energy content was determined by multiplying the percentage of crude protein,
crude fat and nitrogen free extractives by factors of 4, 9 and 4, respectively
(Osborne and Voogt, 1978).
The total true proteins were extracted by the method of Rajaram and Janardhanan
(1990). The extracted protein were purified by precipitation with cold 20% TCA
and determined by the method of Lowry et al. (1951).
The seed protein fractions, albumins and globulins were extracted following
the method of Murray (1979), from the remaining pellet;
the prolamine protein fraction was extracted by treating it with 80% ethanol
(1:5 w/v) overnight. After centrifugation at 20,000 x g for 20 min the supernatant
containing prolamine was air dried and dissolved in 0.1 N NaOH. The remaining
pellet was extracted with 0.4 N NaOH (1:10 w/v) overnight and centrifuged at
20,000 x g for 20 min. The supernatant thus obtained was assumed to be the glutelin
protein. The purified total seed proteins were acid hydrolyzed with 6 N HCl
at 100°C for 24 h in vaccuo. After evaporation, the dried residue
were dissolved in citrate buffer (pH 2.2), known aliquots were analyzer in LKB-Biochrome
Automated Amino acid Analyzer Model 4151-Alpha Plus. For the determination of
cystine, samples were oxidized with formic acid and hydrogen peroxide. Methionine
amino acids recovered are presented as mg/100 g proteins. The contents of different
amino acids recovered were presented as mg g-1 protein. The essential
amino acids were scored and compared with FAO, WHO, UNO (1985)
The total lipids from the seed fluor were extracted (Folch
et al., 1957) using chloroform and methanol mixtures in the ratio
of 2:1 (V/V). Methyl esters were prepared from the total lipids by the method
of Metcalfe et al. (1966). Fatty acid analysis
was prepared (Mohan and Janardhanan, 1993) by gas chromatography
(Shimadzu Model RIA) using an instrument equipped with a flame ionization detector
and a glass column (2 mx3 mm) packed with 1% diethylene glycol succinate on
chromosorb W (Silanised 80/100 mesh). The carrier gas was nitrogen, at flow
rate of 32 mL mim-1. The column temperature was 190°C. Peaks
were identified by comparison with authentic standards, quantified by peak area
integration and relative weight percentage of pack fatty acid was determined
from integrated peak areas.
The macrominerals and trace elements were estimated (Issac
and Johanson, 1975; Meines et al., 1976)
in Perkin Elmer Model 5000 Atomic Absorption spectrophotometer. Dry ashing procedures
were used for the preparation of mineral solutions. The samples were ignited
at 450°C for 12 h in a muffle furnace and dissolved in 3 N HNO3.
For correction of error for the determination of calcium and magnesium, a 1%
lanthanum solution was added to the sample. Phosphorus was measured by calorimetric
means (Virmani and Narula, 1995).
Anti-nutritional factors like total free Phenols, tannins L - DOPA (3,4-dihydrioxy
phenylalanine), hydrogen cyanide and phytic acid were qualitative. The concentration
of total free phenols was determined using the method of Mole
and Waterman (1987). Tannins were captured and determined in a polyamide
chromatography column following the method described by Burns
(1971). L-DOPA content was determined by Brain method
(1976). Hydrogen cyanide was estimated by extraction with 0.1 M orthophosphoric
acid. After extraction, sample was neutralized and estimated with chloramines,
sample was neutralized and estimated with chloramines T and barbituric acid
reagent (Cooke and Madugwu, 1978; Nambisan
and Sunderasen, 1984). The colorimetric technique of Wheeler
and Ferrel (1971) as modified by Reddy et al.
(1978) was used to estimate phytic acid.
RESULTS AND DISCUSSION
The proximate composition (Table 1) shows that Rhynchosia
hitra seed meal contained high amount crude protein of 19.05%, crude fat
of 8.73% than in other commonly consumed legumes Cicer arietinum (Srivastava
and Ali, 2004), Vigna umbellata (Rajaram and Janardhanan, 1990),
Canavalia virosa (Thangadurai et al., 2001a)
and Tamarindus indica (Pugalenthi et al.,
2004). The remarkably high level of protein in the wild legume under study
underscores their importance as source of this vital nutrient. The food energy
value of the seed was 1690.9 kJ due to the protein, lipid and NFEs rich nature.
Albumins and globulins (5.12 and 6.75 g/100 g, respectively) (Table
2) constitute the major bulk of the seed proteins as in many other legumes
and percentage distribution of both proteins are more or less equal to that
of Vigna sesquipedalis (Rajaram and Janardhanan, 1990), Vigna trilobata
(Sidduraju et al., 1992) Phaseolus lunatus
(Vijayakumari et al., 1993) and Abrus
precatorius (Mohan and Janardhanan 1995; Pugalenthi
et al., 2004). The data on fatty acid composition of the seed lipids
(Table 3) indicated that palmitic, linoleic, oleic and stearic
acids are the predominant fatty acids. The occurrence of unsaturated fatty acids
which account for more than 60% of the seed lipids were comparable with some
other wild legumes (Vidivel and Janardhanan, 2004).
The level of 43.76 and 21.75 g/100 g of palmitic and linoleic acids, respectively
(Table 3), were more than the cultivated legumes of Vigna
(Viswanathan et al., 2001).
|| Data on proximate composition of Rhynchosia hirta*
|*Mean of triplicate determinations expressed on dry weight
basis (except moisture)
|| Data on protein fractions of seeds of Rhynchosia hirta*
|*Mean of triplicate determinations expressed on dry weight
|| Data on fatty acid composition of seeds of Rhynchosia
|* Mean of triplicate determinations expressed on dry weight
||Amino acid composition of acid hydrolysed purified total seed
proteins of seeds of Rhynchosia hirta (mg/100 g crude protein)
The data on amino acids profile of the purified seed proteins revealed that
the essential amino acids, cystine, methionine and tryptophan are the conspicuous
limiting amino acids. Whereas, the other essential amino acids leucine, lysine,
isoleucine, valine, threionine and histidine are present in higher concentrations
(91, 55, 48, 30, 50 and 26 mg/100 g crude protein, respectively) when compared
with FAO/WHO/UNO (1985) provisional pattern adequate for
human maintenance and normal growth (Table 4). Minerals are
rich (Table 5). The seeds were found to be a potential source
of minerals such as calcium, potassium, magnesium, manganese and copper than
in the legumes of Phaseolus lunatus, Leucaena leucocephala and
Lathyrus sativus (Duke, 1981) and in comparison
with recommended dietary allowance value (NRC/NAS, 1989).
Phosphorus content is more than in the legumes of Abrus precatrius and
Cassia obtusifolia (Mohan and Janardhanan 1995),
Tamarindus indica, Erythrina indica and Sesbania bispinosa
(Pugalenthi et al., 2004).
The anti-nutritional factors of seed flour are present variably (Table
6). The seed contains relatively hig her amount of 2.25 and 1.32 g of total
free phenols and phytic acids, respectively, than the commonly cultivated legumes
as observed earlier (Bressani et al., 1983; Khan
et al., 1979; Rajaram and Janardhanan, 1992;
Rodrigues and Thorne, 1991). The contents of tannin
and non-protein amino acids L-DOPA are found to be very low (4.75 and 0.65 g,
respectively) when compared with other species in Vigna (Rajaram
and Janardhanan, 1990; Siddhuraju et al., 1992).
A part from these anti-nutritional factors, the presence of negligible amount
of hydrogen cyanide (0.0654 g/100 g seed flour) was also noticeable. The phytohaemagglutinating
activity of albumins and globulins are similar showing with out any specificity
against human ABO system as observed earlier (Siddhuraju
et al., 1992).
The conventional method of repeated soaking and boiling of seeds in water followed
by decanting five to six times, before, consumption is being practiced by the
local tribes to eliminate most of the antinutritional factors. All the antinutritional
factors reported except L-DOPA are heat labile. Hence they can be removed wet
or dry thermal treatments (Geervani and Theophilus, 1981).
In an earlier study, it has been demonstrated that the L-DOPA contents can also
be significantly reduced by repeated soaking and boiling of the seeds in water
under optimum heat conditions to realize the maximum nutritional advantages
(Thangadurai et al., 2001b).
|| Data on selected mineral composition of Rhynchosia hirta
|* Mean of triplicate determinations expressed on dry weight
|| Data on antinutritional factors present in the seed flour
of Rhynchosia hirta*
|* Mean of triplicate determinations expressed on dry weight
Therefore, the presence of these antinutritional factors may not be a limiting
factor in the utilization of these seeds for food and other purpose.
In view of the above facts, Rhynchosia hirta may further be exploited in breeding programs and popularized for mass cultivation and consumption in third world countries such as India to alleviate hunger and poverty. More efforts are needed to encourage the increased production and consumption of processed indigenous legumes, especially the Rhynchosia hirta as they seem to be economical sources of vegetable protein. As its domestication for commercial exploitation is considered in a number of biogeographical regions, such nutritional information is also very crucial to overcome the food crisis of ever expanding worlds population.
KSM is grateful to the Department of Science and Technology, New Delhi for the award of SERC-Young Scientist research grant No. SR/FT/L-16/2003.
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