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ORIGINAL ARTICLE
Year : 2021  |  Volume : 15  |  Issue : 1  |  Page : 61-67

Nutritional, anti-nutritional evaluation, and element analysis of Trichosanthes lobata Roxb. (Cucurbitaceae) through inductively coupled plasma-mass spectrometry technique


Department of Botany, Bharathiar University, Coimbatore, Tamil Nadu, India

Date of Submission15-Oct-2020
Date of Decision01-Dec-2020
Date of Acceptance07-Dec-2020
Date of Web Publication26-Mar-2021

Correspondence Address:
Aghil Soorya Aravindakshan
Department of Botany, Bharathiar University, Coimbatore - 641 046, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/joa.joa_239_20

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  Abstract 


Background: The search for a drug from plant source starts with its investigations on primary and preliminary compositions that further lead the research in its right path. Objective: The objective is to investigate the organoleptic, proximate composition and to quantify the mineral, nutritional, and anti-nutritional values of Trichosanthes lobata Roxb. Materials and Methods: Organoleptic, macroscopic, microscopic, physicochemical, proximate, nutritional, anti-nutritional compositions of T. lobata leaves and stem were evaluated using standard methods. Further mineral composition of the species was analyzed using inductively coupled plasma-mass spectrometry technique. Results: The organoleptic, macroscopic, microscopic, physicochemical, and proximate analysis revealed the good quality, purity, and active principles in the T. lobata. All the evaluations concluded that the leaf of the species possess better compositions that the stem. The fair quantity of carbohydrate (27.32 mg GU eq./g), starch (32.44 mg GU eq./g), protein (52.49 mg bovine serum albumin eq./g), amino acid (33.77 mg LE eq./g), Vitamin C (12.68 mg ascorbic acid equivalent eq./g), and Vitamin E (8.21 mg α-TOP eq./g) were seen in leaves. Similarly, minerals such as magnesium, potassium, calcium, sodium, lithium, iron, zinc, aluminum, copper, nickel, and chromium were found in notable quantities in leaves. The anti nutritional analysis of trypsin inhibitors (0.013 TIU/mg), oxalate (0.002%), cyanogen (below detection level (BDL)), and phytic acid (BDL) indicates the low toxicity profile of the species. All the results satisfy the requirements and receive the toxicity boundaries set by the World Health Organization and EFSA. Conclusion: T. lobata leaves possess notable nutritional values.

Keywords: Inductively coupled plasma-mass spectrometry, leaves, nutritional, physicochemical, Trichosanthes lobata


How to cite this article:
Aravindakshan AS, Thangavel S. Nutritional, anti-nutritional evaluation, and element analysis of Trichosanthes lobata Roxb. (Cucurbitaceae) through inductively coupled plasma-mass spectrometry technique. J Ayurveda 2021;15:61-7

How to cite this URL:
Aravindakshan AS, Thangavel S. Nutritional, anti-nutritional evaluation, and element analysis of Trichosanthes lobata Roxb. (Cucurbitaceae) through inductively coupled plasma-mass spectrometry technique. J Ayurveda [serial online] 2021 [cited 2021 Apr 13];15:61-7. Available from: http://www.journayu.in/text.asp?2021/15/1/61/311915




  Introduction Top


Trichosanthes lobata exhibits active therapeutics against headache, fever, skin allergy, inflammation, ulcer, malaria, etc.[1] The species reveals hepatoprotective property[2] and diabetics, Malaria, etc., as folklore uses.[3] It is an ingredient in the Siddha drug called “Punarnavasavam,” polyherbal formulation of 21 plant ingredients.[4] Being elements, minerals are not biochemically produced by living organisms/humans. Plants get minerals from soil. Majority of the minerals in human diets are through feeding plants, animals, or through drinking water.[5] The study aims the analysis of its organoleptic, macroscopic, microscopic, proximate composition and quantifying the mineral, nutritional and anti-nutritional contents.


  Materials and Methods Top


Collection, identification and preparation of samples

Fresh, healthy and matured leaves and stem of T. lobata Roxb. Were collected from Vattavada Panchayat (10°10′38.7″N 77°15′33.″E), Koviloor Post, Idukki District, Kerala state of India. The authenticity of the selected plant species is confirmed from the Botanical Survey of India, Southern Circle, Coimbatore (Vide No: BSI/SRC/5/23/2016/Tech./213). The leaves and stem were cleaned and dried in shade and ground well to a fine powder and stored in separate air-tight containers.

Organoleptic characters

Organoleptic evaluations of plant powder such as color, size, odor, and taste parameters of T. lobata stem and leaves as per Trease and Evans.[6]

Physicochemical analysis

The leaves and stem of the plant were shade dried and powdered using a mechanical grinder for powder analysis. The physicochemical characteristics were determined as per the World Health Organization (WHO).[7] The behavior of the powder to different reagents such as sodium hydroxide, hydrochloric acid, nitric acid, and sulfuric acid were analyzed and tested the staining of leaves powder in visible light and day light.

Proximate analysis

The proximate composition of the plant T. lobata leaf and stem powder includes estimation of foreign matter, ash, soluble solids, moisture, crude fat, and fiber contents as per Raghuramulu et al.[8] The determination of all parameters was performed in triplets.

Nutritional component estimation

Nutritional estimation of T. lobata includes determination of total carbohydrate, total free amino acids, and starch as per the methods of Sadasivam and Manickam[9] and the results were expressed in mg GU equivalents/g sample, mg LE equivalents/g sample, and mg GU equivalents/g sample respectively. The estimation of Vitamin E and Vitamin C was done according to Achikanu et al.[10] and total proteins by Lowry et al.[11] and the results were expressed in α-tocopherol equivalents, mg ascorbic acid equivalent equivalents/g sample and mg bovine serum albumin equivalents/g sample, respectively. All the estimations were done in triplets.

Anti-nutritional component estimation

Determination of cyanide

For cyanide determination, alkaline picrate method was adopted.[12] 5 g of the T. lobata plant powder was dissolved in corked conical flask with 50 ml distilled water and macerated for 12 h. After this, the extract was filtered and used for the determination of cyanide. One ml of the filtrate is mixed with 4 ml of alkaline picrate and incubated for 5 min in the water bath and absorbance was read at 490 nm. A cyanide standard curve was used for the extrapolated cyanide content.[13]

Determination of oxalate

Acid digestion method was used to determine the oxalate contents of T. lobata. The sample was treated with sulfuric acid and filtered using Whatman No. 1 filter paper. The filtrate was titrated hot (80°C–90°C) aligned with 0.1M potassium permanganate solution to a dim pink color that remains only for 30 s.[13]

Determination of phytate

The method used in order to determine the phytic acid contents of T. lobata was according to Wheeler and Ferrel.[14] In order to calculate the ferric ion contents a standard curve of various FE (NO3)3 concentrations were plotted alongside the successive readings on spectrophotometer. Phytate phosphorus was calculated from the concentrations of ferric ion.

Determination of trypsin inhibitors

0.5 g of the plant powder was macerated with distilled water for 12 h., filtered and evaporated to dryness on boiling water bath. Different concentrations of the extract were made up to 2 ml using distilled water and 2 ml of trypsin solution is added, agitated, and heater using water bath at 37°C. Five ml of heated benzoyl-Dl-arginine-p-nitroanilide hydrochloride solution is added and replaced in water bath. After 10 min, acetic acid is used to stop the reaction and the solution was filtered and absorbance is taken at 410 nm against blank.

Inductively coupled plasma – mass spectrometry

Two hundred mg of powdered sample (leaf/stem/flower) is heated with 10 ml tri-acid in a 100 ml conical flask at 60°C–80°C in fume hood using heating block or mantle until all liquid got evaporated from the flask. Then, the sample flask is washed with double distilled water and made up to 100 ml in a standard flask. From this, 10 ml of the sample is filtered using Whatman No. 1 filter paper and subjected for quantitative analysis of 18 elements, namely, sodium (Na), potassium (K), calcium (Ca), lithium (Li), beryllium (Be), magnesium (Mg), aluminum (Al), chromium (Cr), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), arsenic (As), selenium (Se), molybdenum (Mo), cadmium (Cd), and lead (Pb) through inductively coupled plasma-mass spectrometry (ICP-MS). The apparatus run details are depicted in [Table 1].
Table 1: Apparatus details used for the analysis

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Statistical analysis

All the result values were expressed as mean ± standard deviation according to its relevance.


  Results and Discussion Top


The basics of Ayurvedic medicines are the concept of universal interconnectedness, prakriti (the body's constitution) and doshas (life forces). In Ayurveda, by eliminating impurities, increasing resistance to diseases, reducing symptoms and worry and increasing harmony in life are the goals of a treatment aid individual. Unfortunately, there remain a lack of scientific proof of various concepts of Ayurveda and these blessings from our ancients are rambling. Hence, the evidence-supportive research is highly recommended for the global recognition and acceptance of Ayurveda and its concepts.

India with its varied climatic conditions is a home for many fascinating medicinal plants, typically the Western Ghats. The genus Trichosanthes is with commonly cultivated species. Although it is native to Southern and Eastern Asia, Australia, an Island of Western Pacific, a wide variety of its species occur in India. The genus shows the presence of various active constituents such as flavonoids, phenolics, carotenoids, saponins, triterpenoids, and cucurbitacins. It also exhibits active therapeutics against headache, fever, malaria, bronchitis, abdominal tumor, boils, acute colic, bilious, diarrhea, skin allergy, hematuria, alopecia, cholesterol, wounds, inflammation, diabetes, and HIV. Trichosanthes species is used as vermifuge, purgative, abortifacient, laxative, anti-bacterial, anti-spasmodic, anti-ulcer, anti-herpetic, anti-oxidant, cardioprotective, hemagglutinant, cathartic, larvicidal, hepato-protective, emetic, anthelmintic, etc., Novel compounds such as glucosidal, 4'-methylene-dioxyisoflavone-7-O-beta-D-(2''-O-p-coumaroylgluco-pyranoside), etc., been characterized from the species.[2]

The genus shows the presence of various active constituents such as flavonoids, phenolics, carotenoids, saponins, triterpenoids, and cucurbitacins. It also exhibits active therapeutics against headache, fever, malaria, bronchitis, abdominal tumors, boils, acute colic, bilious, diarrhea, skin allergy, hematuria, alopecia, cholesterol, wounds, inflammation, diabetes, HIV, etc., Trichosanthes species is used as vermifuge, purgative, abortifacient, laxative, anti-bacterial, anti-spasmodic, anti-ulcer, anti-herpetic, anti-oxidant, cardioprotective, hemagglutinant, cathartic, larvicidal, hepatoprotective, emetic, and anthelmintic. Novel compounds such as glucosidal, 4'-methylene-dioxyisoflavone-7-O-beta-D-(2''-O-p-coumaroylgluco-pyranoside), etc. been characterized from the species.[2]

Organoleptic, macroscopic, and microscopic characters

Herbal formulation for various diseases includes the use of fresh and dried plant parts. It is to be ensured the quality of the plant samples which is much essential for the proper identification and contribute immensely to its safety and efficacy.[15] Macroscopic, microscopic and organoleptic character analysis serves as an important tool for the selected species of Trichosanthes [Table 2]. The color, odor, and taste indicated the purity and presence of compounds with therapeutic importance. The absence of sand, silica, insect infestation, and rodent contamination were noted. The macroscopic characters such as the size, shape, texture, and venation displayed a good growth and metabolism of the plant. It also supports the identification of the species. The taste, odor, texture, and color of the leaves and stem indicated they may possess therapeutic importance with the presence of secondary metabolites. Similarly, the microscopic evaluation displayed the presence of calcium oxalate crystals, oils, and different types of trichomes. All depicted results indicate the good quality, maturity, and medicinal significance of T. lobata.
Table 2: Macroscopic, organoleptic, and microscopic characteristics of leaf and stem of Trichosanthes lobata

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Physicochemical analysis

The result of physicochemical analysis includes the nature of the plant powder treated with various acids and reagents under visible light and day light are presented in [Table 3]. The color emitted by the powders in day light and visible light was particular to each of them. Varied color indicated the presence of series of active principles in them. The study may be helpful in identification and authentication of the medicinal plant and also to analyses the quality, safety, and standardization of active phytoconstituents for its safe use.
Table 3: Physicochemical analysis of leaf and stem of Trichosanthes lobata

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Proximate analysis

Among the leaves and stem of T. lobata analyzed for their proximate parameters and are shown in [Table 4]. The leaves possessed the predominant contents than stem. Although the moisture content, acid soluble ash, and crude fiber were found to be more in stem than the leaves, the other parameters such as the ash, water soluble ash sulfated ash, ether soluble extractive yield, alcohol soluble extractive yield, water soluble extractive yield, total soluble solids, and crude fat were found to be more in the leaves than the stem. Both the leaves and stem were free of foreign matters.
Table 4: Proximate analysis of leaf and stem of Trichosanthes lobata

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Since the ash content is the refection of the minerals in the sample, the species contained good mineral content. The plants are good source of crude fiber when consumed as its adequate intake can lower the serum cholesterol levels, hypertension, constipation, heart diseases, breast cancer, and diabetes.[16]

Nutritional and anti-nutritional estimation

The leaves and stem parts of T. lobata were analyzed for their nutritional and anti-nutritional values and the results are revealed in [Table 5]. The leaves of the species possessed comparatively greater contents of nutritional values of carbohydrate, starch, protein, amino acids, Vitamin C, and vitamin E than the stem. The results support that the species is a good source of nutrition since it meets the recommended dietary allowance values.[17]
Table 5: Nutritional and anti-nutritional estimation of leaf and stem of Trichosanthes lobata

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Vitamin C, an effective antioxidant, facilitates the reduction of folic acid intermediates, the transport and uptake of nonheme iron at the mucosa and the synthesis of cortisol. Its deficiency includes fragility to blood capillaries, scurvy, and gum decay. Vitamin E is a powerful antioxidant which helps to protect cells from damage and normal function of red blood cell (RBC) and muscles.[18] The anti-nutritional quantity revealed the absence of toxic substances.

Inductively coupled plasma – mass spectrometry

Trace elements are those minerals included in diet in very minute quantities (<0.01%) of the mass of the organism. The theory of requirement of nutrients may vary from culture to culture for genetic, dietary, lifestyle, and geographic reasons. Essential trace elements play an important role as a cofactor for certain enzymes involved in metabolism, cell growth, and development, muscle and nerve function, normal cellular functioning, synthesis of some hormones and connective tissues and further in the metabolism of protein, carbohydrates, lipids, and energy. The ability of trace elements to function as substantial affecter in a variety of processes necessary for life such as regulating homeostasis and prevention of free radical damage, can provide an answer to the definite correlation between content of trace elements and many common diseases.[19]

One of the essential components for healthy living is micronutrients/trace elements with specific biochemical function in human body are zinc, manganese, molybdenum, iodine, selenium, sulfur, iron, chlorine, cobalt, copper, etc.,[20] The current study species T. lobata is being examined quantitatively for the presence of 18 elements through ICP-MS. Considering the plant parts, the leaf of the species possessed higher amount of potassium (236 mg/g) followed by calcium (64.2 mg/g), magnesium (37.18 mg/g), and sodium (10.73 mg/g). The stem of the species possessed higher quantity of magnesium (9856.77 mg/g) followed by potassium (236 mg/g), calcium (36.87 mg/g), and sodium (10.13 mg/g). Elements such as beryllium, arsenic, selenium, molybdenum, cadmium, and lead were absent or in negligible quantity that supports the nontoxic nature of the species [Table 6].
Table 6: Major and trace element profile of Trichosanthes lobata plant parts

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The elements possess various role in the human body: Sodium helps in extra cellular fluid, cellular homeostasis, electrolyte balance, blood pressure, excitability of muscle, and nerve cells and transport nutrients; potassium in water conduction, protein and glycogen synthesis, acid base balance and blood pressure; calcium is essential for formation, rigidity, strength and elasticity of skeleton, intercellular signaling and secretion of parathyroid; lithium has role to maintain mood stability; magnesium in energy production, oxidative phosphorylation, glycolysis, bone development, synthesis of DNA, RNA, antioxidant glutathione, muscle contraction and heart rhythm;[21] aluminum is used to treat fluorosis and reduce phosphorus absorption in uremic patients; chromium in glucose tolerant factor, enhance protein, carbohydrate and lipid metabolism, treat Dysthymia, Bipolar disorder, turner's syndrome, and Polycystic Ovary Syndrome.[22]

Manganese in mechanisms of antioxidant, bone health and development, wound healing and activator and constituent of enzymes; Iron carry oxygen in hemoglobin, cellular electron transport system, synthesis of steroid hormones and bile acids, detoxification in liver, neurotransmitter, immune function, wound healing. Nickel Stabilize RNA structure, prolactin production, iron absorption, adrenaline and glucose metabolism, hormones, lipid, cell membrane and production of RBC; copper as energy source, antioxidants, formation and regulation of hormones, collagen, RBC and iron absorption; zinc for healthy Immune system, cell division, against skin problem and sore throat, development of hair, tissue nail, skin and muscle, protein, collagen, sex; selenium in cellular functioning, antioxidant enzyme glutathione peroxidase and thioredoxin reductase, thyroid hormone metabolism, DNA synthesis, reproduction; molybdenum in the production of molybdoprotein, xanthine oxidase enzyme, remove sulfite, and toxins.[22]

With the above references, the study species possess elements and nutrition in considerable amount as per nutritional requirements and receives the toxicity boundaries set by the WHO,[7] European food safety authority.[23]


  Conclusion Top


Almost 90% of the ayurvedic formulations include plants since they possess vital performances on body either as food or drug. Those actions activate the plants to reverse the pathophysiological processes and to stabilize the doshas. The study species T. lobata was found to contain important constituents needed to combat diverse ailments in man. From the results, it is quite fascinating that the species has good carbohydrate, starch, protein, and fiber content that adds its nutritional benefits. In addition, the study has highlighted that the species delivering sufficient, safe and nutritious with minerals satisfying the daily required diet to the human population. Thus, the current research documents the various constituents of T. lobata and gives a nutritional and safety of the species to be used in the treatments through Ayurveda. Further research on the bioactive components, phyto-constituents can be derived after the identification, isolation, and characterization spectroscopically.

Financial support and sponsorship

This study was supported by Bharathiar University, Coimbatore, Tamil Nadu, India.

Conflicts of interest

There are no conflicts of interest.





 
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Wheeler EL, Ferrel RE. A method for phytic acid determination in wheat and wheat fractions. Cereal Chem 1971;48:312-20.  Back to cited text no. 14
    
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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]



 

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