pp. 42-49

Lokesh Kumar Soni1, Pradeep Parasher2  and M.P. Dobhal*3

1 Natural Products Laboratory, Centre of Advanced Studies, Department of Chemistry, University of Rajasthan, Jaipur, India.

2  Department of Chemistry, Govt. P.G. College, Jhalawar, Rajasthan, India.

3 Suresh Gyan Vihar University,Jaipur-302017, Rajasthan, India.

*Corresponding author Email: mpdobhal@yahoo.co.in, lsoni492@gmail.com

ABSTRACT

Natural products are the basis  of  synthetic  and traditional herbal medicine.  Phytochemicals  are compounds that present in plants.  There are hundreds of significant drugs and biologically active compounds developed from  the   traditional   medicinal   plants.   The   present   study   was   aimed   to   investigate   the phytochemical screening of Prosopis cineraria’ bark, Curcuma amada’ rhizomes, and Citrullus colocynthis’s fruits hull. The result of the qualitative phytochemical constituents of  these  plants  extracts showed presence of all the tested phytochemicals alkaloids, terpenoids,  flavonoids,  carbohydrates  and saponins. Thus, the results  suggests,  the  beneficial  role  of  their  plant  parts  and  supports  their  traditional uses  and  proved  to  be  useful  for pharmacological properties.  The   phytochemical   analysis   of   the plants is very significant commercially and has  great  interest  in  pharmaceutical  companies  for  the production of the new drugs for curing of various  diseases.

Keywords:   Phytochemicals,    medicinal    plants,    Prosopis   cineraria,   Curcuma    amada,    Citrullus colocynth.

INTRODUCTION

The medicinal plants are useful for  healing  as well as for curing of human diseases due to the presence of bioactive  phytochemicals (Nostro et al. 2000). Phytochemicals  are  naturally  occurring in the medicinal  plants, leaves, vegetables and roots     that    have     defense      mechanism   in pl a nt s a nd a r e  a l s o  r e s pons i bl e  f or th e tr e a tm e n t of various diseases due to presence  of  metabolites..  Chlorophyll,  proteins and common sugars are primary metabolites terpenoid,   alkaloids     and    phenolic  compounds are known as secondary metabolites  (Krishnaiah et al. 2007). Prosopis cineraria (L.) belongs  to the cosmopolitan  genus Prosopis, subfamily Mimosaceae, tribe Leguminosae/ Fabaceae (Rasanen & Lindstrom 2003) and it has common names such as Janti and Chonksa (Delhi), Jhind, Jhand and Jand (Punjab and Haryana), Banni (Karnataka), Sumri (Gujarat), Kandi (Sindh) and Khejri  (Sanskrit).  It   is   conjointly   referred   to as the  ‘wonder  tree’  and  therefore  as  the ‘king   of  desert’(Bari   et   al.   2007;   Gupta   & Prakash   1975;   Kaul  1967;   Burdak  1982).   Its flower is powdered and mixed with  sugar  and used throughout maternity as safeguard against miscarriage. The bark of Prosopis cineraria  is dry, acrid,  bitter  with  a  taste;  cooling anthelmintic, tonic, cures infectious disease, dysentery, bronchitis, asthma, leucoderma, piles, tremors of the muscles (Kirtikar &  Basu 1984). The bark is employed in rheumatism, cough and colds, diarrhea, worm infestations etc.(Sharma 1993). The bark of the plant offers immediate relief against snake or a scorpion bite (Chopra et al. 1956).

Curcuma amada Roxb. is commonly known as mango ginger. It is a perennial, rhizomatous , aromatic herb belonging to the family Zingiberaceae. Additional health benefits of C. amada rhizome  reported  were  biliousness, itching, skin diseases, asthma and  inflammation due to injuries.  According  to the Unani systems of medicine, it is a  diuretic,   emollient, expectorant, antipyretic and appetizer. The ability of C. amada rhizome’s  against  inflammation  in the mouth and ear, gleet,  ulcers  on  the  male sex organs,  scabies,  lumbago  and  stomatitis have  been  reported        (Kirtikar  and  Basu 1984; Warrier et al. 1994; Hussain et al. 1992). Mango ginger has a typical unusual  flavour  of raw unripe mango. Therefore, it is used as  a basic ingredient in pickles, candies, sauces and curries. (Elsayed 2016; Shankaracharya 1982). Citrullus colocynthis is a perennial herbaceous vine, belongs to the family Cucurbitaceae (Dane and  Liu  2007).  The  Citrullus  colocynthis  is well known for its  therapeutic  activity  in folklore. The fruits and pulp  of  this  plant are. well known natural cathartics  since  ancient times.  The  fruit is pungent, cooling      purgative, anthelmintic,     antipyretic     carminative.     It    is beneficial   in   treatment    of    tumor, leucoderma, ulcer, asthma, bronchitis, urinary troubles, enlargement of spleen, tuberculosis, dyspepsia, constipation, anemia’s and throat diseases. The leaves of this herb are used to treat asthma and jaundice, whereas the roots are used traditionally for the treatment for amenorrhea, breast infla mmation, arthralgias, seizures, tuberculosis, syphilis, and parasitic infections and ophthalmic diseases. (Blaskovich et al. 2003). C. colocynthis is used as an antidiabetic agent in developing  countries  (Errajraji  et  al.  2010; Ziyyat  et  al.   1997).   In  the  present  study three different  medicinal  plants  each  belonging to different families were evaluated for the presence of various class of  phytochemicals.

MATERIAL AND METHODS

Plant Material

The fresh rhizomes of Curcuma amada were procured from local market , Jaipur, Rajasthan. Bark of Prosopis cineraria and  fruits  of Citrullus colocynthis were collected locally and authenticated by Prof. N.J. Sarana Department of Botany, University of Rajasthan,  Jaipur, Rajasthan,  India.

Preparation of crude plant extract

Shade dried plant materials (100 gm) were powdered  and  extracted  with  ethanol separately.  Each extraction  step was completed in 48 hours. The extracts were filtered hot and solvent was removed under reduced pressure. The concentrated extracts were dried under vaccuo.

PHYTOCHEMICAL ANALYSIS

The test samples (plant extracts) were subjected to phytochemical analysis in order to find out the presence of various class of compounds. The colour  reactions  were  employed  for   the detection  of  alkaloids,  tannins,  cardiac glycosides, saponins, flavonoids  and  terpenoids etc. (Uthayarasa et al. 2010; Trease and Evans 1989; Harborne 1973;  Sofowara  1993).

TEST FOR ALKALOIDS

Ethanolic extracts were dissolved individually in dilute hydrochloric acid in te s t  tu b e s  a n d filtered. Wagner’s test: Filtrates were treated with Wagner’s reagent (iodine in potassium iodide). A brown/reddish precipitate formed indicates the presence  of alkaloids.

Mayer’s test: Filtrates were treated with Mayer’s reagent (potassium mercuric iodide). Formation of a yellow precipitate indicates the presence  of alkaloids.

Drage ndroff  test:   Filtrates   were   treated with Dragendorff’s  reagent  (solution  of potassium bismuth iodide). Formation of a red precipitate  indicates the presence of alkaloids.

Hager’s test: Filtrates were treated with Hager’s reagent (saturated picric acid solution). Formation of yellow  colour  indicates   the presence  of alkaloids.

TEST FOR TANNINS

Lead test: Extracts were dissolved in distilled water in separate test tubes and  filtered.  The filtrate was treated with 3-4 drops of ferric chloride solution, development of blue or green coloration  indicated the presence of tannins.

TEST FOR FLAVONOIDS

Alkaline reagent test: Extracts were treated with  a  few  drops  of  sodium  hydroxide  solution. Formation of intense yellow colour, which becomes  colorless  on  the  addition  of   dilute acid,  indicates  the  presence  of flavonoids.

Lead acetate  test: Extracts  were  treated with a few drops  of  lead acetate  solution.  Formation of a yellow color precipitate  indicates  the presence  of flavonoids.

TEST FOR SAPONIN

Foam test: Extracts  were  dissolved  with distilled water and shaken vigorously till a stable persistent froth was obtained. The  froth  was mixed with 3 drops of  olive oil and shaken vigorously  and  then observed for emulsion.

TEST FOR CARBOHYDRATES

Extracts were dissolved individually in  5  ml distilled water and filtered. The filtrates  were used to test the presence of carbohydrates.

Molisch’s test: Filtrates were treated with 2 drops of alcoholic α-Naphthol solution in a  test tube. A violet ring observed at  the  junction indicates  the presence  of carbohydrates.

Benedict’s  test: Filtrates  were  treated  with Benedict’s reagent (complex mixture of sodium carbonate, sodium citrate and copper  (II)  sulfate pentahydrate ) and heated gently. Orange red precipitate indicates the presence of reducing sugars.

Fehling’s test: Filtrates were hydrolyzed with dilute HCl, neutralized with alkali and heated with Fehling’s A solution (aqueous  solution  of copper(II) sulfate) and Fehling’s B solutions (aqueous potassium sodium tartrate with sodium hydroxide). Formation of  a  red  precipitate indicates the presence of reducing sugars.

TEST FOR PHENOL

Ferric  chloride  test:  Extracts   were   treated with 3-4 drops of  ferric  chloride  solution. Formation of bluish black colour indicates the presence  of phenol.

TEST    FOR    PROTEIN     AND    AMINO ACID

Xanthoprote in test: the extracts were treated with a few drops of conc. Nitric acid. Formation of yellow colour indicates  the  presence  of proteins.

Ninhydrin Test: to the extract, 0.25% w/v Ninhydrin reagent was added and boiled  for  a few minutes.Formation of blue- purpule color indicates  the presence of amino acid.

Biuret test: the extracts were  treated  with sodium hydroxide solution , 1%  CuSO4  solution and warm the mixture for about 5 minutes. Bluish violet coloration indicates the  presence  of proteins.

TEST FOR GLYCOSIDES

Extracts were hydrolyzed with dilute  HCl,  and then subjected to test for glycosides.

Modified Borntrager’s test: Extracts were treated  with  ferric  chloride  solution   and immersed in boiling water  for  5  min.  The mixture was cooled and extracted with equal volume of benzene. The organic layer was separated  and   treated   with   ammonia   solution. A  rose   pink   color   observed   in   the ammoniacal layer which indicates  the  presence of anthranol glycosides

Test for cardiac glycosides : 5 ml of each extract was treated with 2 ml of  glacial acetic acid, 4-5 drops of ferric  chloride  solution and then 1ml of conc. sulphuric acid was added. A brown ring at the  junction of two  layers  indicated the  presence  of  cardenolides.

TEST FOR FIXED OILS AND FATS

Spot tes: A small quantity of the extract was pressed between two filter  papers.  Oil stain  on the paper indicated the presence of fixed oil. Detection of gum and mucilage: Extract was mixed with 10 ml distilled water and 25 ml of alcohol with constant stirring. White or cloudy precipitate indicated the presence of gum and mucilage.

TEST FOR PHYTO STEROLS/ TERPENOIDS

Liebermann Burchard’s test: Extracts were treated with  chloroform  and  filtered.  The filtrated  were  treated  with  a  few  drops  of acetic anhydride, boiled  and  cooled.  Conc. sulphuric acid was added.  The  formation  of brown ring at the junction of two liquids indicates the presence  of phytosterols.

Salkowaski’s  test:  Extracts  were   dissolved in chloroform and  concentrated  sulphuric  acid was added to it.  A  reddish  brown  discolouration at the interface showed  the  presence  of terpenoids .

Table 1: Phytochemical  constituents  of three medicinal  plants  studied.

RESULTS AND DISCUSSION

Phytochemical analysis of Prosopis cineraria, Curcuma amada and Citrullus colocynthis plant extracts showed the presence of various constituents which  are  known  to exhibit medicinal as well as physiological activities  (Sofowara 1993). Prosopis cineraria  gives positive result for all constituents where as Curcuma amada showed negative result  for  saponins  and  positive   for all other on the other hand Citrullus colocynthis gave positive  result for carbohydrates, alkaloids, flavonoids terpenoids etc.

The phytoconstituents  i.e.  alkaloids and  flavonoids  are  active   principles of plants. These active principles provides defensive mechanism of the plants against different pathogens (Hafiza, 2000). The terpenoids have significant pharmacological activities, such as anti-viral, anti-bacterial, anti- malarial,  anti-inflammatory,   inhibition of cholesterol synthesis and   anti- cancer activities (Mahato and Sen,1997). Saponins which are used to stop bleeding and in treating wounds and ulcers as it helps in red blood cell coagulation (Okwu and Josiah, 2006). Further studies will need to isolate and characterize the bioactive chemical entities of these plants.

CONCLUSION

presence   of   phytochemical    constituents in Prosopis cineraria’ bark, Curcuma amada’ rhizomes, and Citrullus colocynthis’s  fruits  hull  extract  supports its traditional uses and may be utilized for different biological activities

ACKNOWLEDGEMENT

The author (LKS) is grateful to UGC, New Delhi for granting the senior research fellowship.

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