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Preliminary Phytochemical Screening, Pharmacognostical Analysis, and Identification of Bio-Active Compounds of Commelina Benghalensis Using Gc-Ms Technique

Mr. Dr. KL. Senthilkumar*1, Dr. P. D. Gokulan*2,Mr. S. Venkateshwaran*3, S.Arunadevi*4 , G. Ashwini*5 , M. Aslambasha*6

1 principal, sri vijay vidyalaya college of pharmacy, dharmapuri, tamilnadu, India

2 Head of the Department of Pharmaceutical Chemistry, Sri Vijay vidyalaya College of Pharmacy, Dharmapuri, Tamil nadu, India

3 Associate professor, Sri Vijay Vidyalaya College of Pharmacy, Dharmapuri, Tamil Nadu (India) 4,5,6 B. Pharm Students, Sri Vijay Vidyalaya College of Pharmacy, Dharmapuri, Tamil Nadu (India) ***

ABSTRACT- OBJECTIVE: The study deals with the preliminary phytochemical screening, pharmacological evaluation, and identification of bioactive compounds present in Commelina benghalensis using the GC-MS technique. The main objective of the work was to analyze the phytochemical constituent, conduct pharmacognostical studies, and identify biologically active compounds from the selected plant. METHOD: Fresh leaves of Commelina benghalensis were collected, shade-dried, and powdered into a coarse form. The powdered plant material was subjected to solvent extraction using ethanol and ethyl acetate. The obtained extract was then used for preliminary phytochemical screening to detect various secondary metabolites. Further analysis was carried out using GC-MS to identify the active phytochemical constituent present in the plant. RESULT: The results of the study emphasize the phytochemical and pharmacognostical importance of Commelina benghalensis and confirm the presence of several bioactive compounds identified through GC–MS analysis.

INTRODUCTION:

Traditional medicine plays a vital role in healthcare systems worldwide. Human survival would be challenging without the widespread use of plants and plant-derived products. Since ancient times, people from diverse civilizations have relied on plants for the treatment of numerous ailments. The World Health Organization (WHO) defines a medicinal plant as a herbaceousspeciesthatpossessesphytochemicalcompoundsresponsiblefortherapeuticactivities.Inrecentyears,therehas been growing interest among scientists and healthcare professionals in medicinal plants following the validation of their therapeuticbenefits.

Comedian beng halensis isa medicinal herb that belongs to thefamily Commelinaceae.It isa perennial speciesindigenous to thetropical regions ofAsia andAfrica and iscommonly referred toasBengal dayfloweror dewflower. Theplantisa robust, spreadingherbthatgrowsannuallyandreachesaheightofapproximately40cm.Itsrootsemergefromthebasalnodes.The species is readily recognized by its small, attractive bluish-violet flowers. The leaves are ovate to elliptical in shape, occasionally appearing slightly triangular, and are deep bright green in colour, measuring about 4–7 cm in length. The fruit capsulesarebroadlyovoidtooblong,approximately4–5mmlong,whiletheseedsareovoidinform.

Traditional uses: In India, Commelina benghalensis is traditionally used to treat ailments such as headache, constipation, leprosy, fever, snakebite, jaundice, mouth thrush, insanity, epilepsy, and psychosis. It is also known for its anti-inflammatory, demulcent, emollient, and depressant properties. In Lesotho, the plant is used to manage infertility in women. In China, it is usedasadiuretic.InPakistan,itisemployedinthetreatmentofleprosyandisalsousedasalaxativeandanti-inflammatory agent. In Bangladesh, the plant is used to treat insomnia, cataracts, night blindness, and other eye disorders, as well as suppurativesores,snakebites,inflammation,burns,conjunctivitis,headaches,toothaches,andeczema.

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

Taxonomical classification

Kingdom: Plantae

Subkingdom: Tracheobionta (Vascularplants)

Superdivision: Spermatophyta(Seedplants)

Division: Magnoliophyta (Angiosperms)

Class: Liliopsida (Monocotyledons)

Subclass: Commelinidae Order: Commelinales

Family: Commelinaceae

Genus: Commelina

Species: Commelina benghalensis L.

Botanical Name:CommelinabenghalensisLinn.

Common Name:BengalDayflower

VernacularName:

Tamil: Kana Vazhai,Aduthinna Palai

Telugu: Konda Gogu, Bodda Gaddi

Malayalam: Kakkapoo, Vazha Poo

Kannada: Kadu Kanagale, Bodda Soppu

Hindi: Kankaua, Benghal Dayflower

Bengali: Kanshira, Kana Shak

Marathi: Kena

English: Bengal Dayflower, Tropical Spiderwort

QUALITATIVE PHYTOCHEMICAL ANALYSIS

Phytochemicalsarenatural chemical substancesfoundinplantsthatarebeneficial.Chloroformsolventextractobtainedfrom theSynedrellanodiflorawassubjectedtovariousqualitativetestsfortheidentificationofvariousplantconstituentspresentin thespecies.

Fig. 1: Commelina benghalensis

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

1. Test for Alkaloids Dragen dorff’s test

To1mlofextract,add1mlofDragendorff’sreagent(potassiumbismuthiodidesolution).Anorange-redprecipitateindicates thepresenceofalkaloids.

Mayer’s test

To 1 ml of extract, add 1 ml of Mayer’s reagent (potassium mercuric iodide solution). A whitish yellow or cream-colored precipitateindicatesthepresenceofalkaloids.

2. Test for Glycosides

Test solution:Itwaspreparedbydissolvingthesampleinalcohol.

Killer Killani Test

Totestthesolution,afewdropsofferricchloridesolutionandconcentratedsulphuricacid wereadded;theformationoftwo layersoccurred,alowerlayerofreddish-browncolorandanupperlayerofbluish-greencolorsimultaneously.

3. Test for Triterpenoids

Test solution:Itwaspreparedbydissolvingtheextractinchloroform.

Salkowski test

Afewdropsofconc.sulphuricacidwereaddedtothetestsolutionandallowedtostandforsometime.Thelackofformationof yellowcolorinthelowercolorindicatestheabsenceoftriterpenoids.

4. Test for carbohydrate Molisch test

To 2-3 ml of the test solution, add a few drops of Molisch reagent solution and shake. Concentrated sulfuric acid was added fromthesideofthetesttube.Avioletringwasformedatthejunctionoftwolipids.

5. Test for Proteins

Test solution:Itwaspreparedbydissolvingextractinwaterandmakingaqueousextract.

Biuret test

Toabout3mloftheextract,40%sodiumhydroxidesolutionandafewdropsof1%coppersulfatesolutionwereadded.White producesbluecolor.

6. Test for Anthraquinone

Borntrager’s Test

Add 10 ml of 10% ammonia solution with few ml of Aq., shake with ml of benzene and filter (shake vigorously for 30 mins) appearsasapink,violet,orredcolorsolution,itshowsthepresenceofanthraquinones.

7. Test For Flavonoids

Test solution:

Toasmallamountofextract,anequalamountof2MHClwasaddedandheatedforabout30-40mins.Theextractwascooled down and again extracted with ethyl acetate, which was further concentrated to dryness and ready to be used as the test sample.

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

Lead acetate test

To a small quantity of extract, lead acetate was added. Formation of yellow-colored precipitate shows the presence of flavonoids.

8. Test for Quinone

Conc. HCL test

Plantextractwithconc.HClappearsagreencolor;itshowsthepresenceofquinones.

8. Test for Tannins

Test solution:Thetestsolutionispreparedbydissolvingtheextractinwaterandalcohol.

Ferric chloride solution test

To 1 ml of the extract, ferric chloride was added. Formation of a dark blue or greenish-black color shown in the presence indicatesthetannins.

Lead acetate test:

Afewmlof10%leadacetatewasaddedtothetestsolution.Thepresenceofthewhiteprecipitateindicatesthetannins.

9. Test for Cholesterol:

Add2mlofchloroformwith10dropsofaceticanhydrideandadd2-3dropsofconc.H₂SO₄. Appearsared-rosecolorshows theabsenceofcholesterol.

10. Test for Diterpenes

Plant extract is dissolved in distilled water, and adding 3-4 drops of copper acetate solution appears emerald green, which showsthepresenceofditerpenes.

Test for Terpenoids : Add2mlofchloroformwith5mlofplantextractandadd3mlofconc.H₂SO₄(boiledinawaterbath). Donotappearinagrey-coloredsolutionintheabsenceofterpenoids.

11. Test for Saponins

Test solutions: It waspreparedbydissolvingextractinwaterandmakinganaqueoussolutionextract.

Foam Test

Thedrugextractwasvigorouslyshakenwithwater.Persistentfoamformationindicatestheabsenceofsaponins.

12. Test for Fat Solubility test

Fatsareinsolubleinwaterandethanol.Butsolubleinchloroform,benzene,andether.PresenceofFat.

13. Test for Sterols Libermann-Burchard Test

1gmofthetestsubstancewasdissolvedinafewdropsofchloroform,3mlofaceticanhydrideand3mlofglacialaceticacid wereadded,itwaswarmedandcooledunderthetap,anddropsofconc.H₂SO₄wereaddedalongthesideofthetesttube.The appearanceofabluish-greencolorshowstheabsenceofsterols.

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

QUALITATIVE

PHYTOCHEMICAL ANALYSIS

S.NO PHYTOCHEMICAL

1. Alkaloid

2. Glycosides

3. Triterpenoids

4. Carbohydrates

5. Proteins

6. Anthraquinones

7. Flavanoids

8. Quinone

9. Tannins

10. Cholesterol

11. Diterpenes

12. Terpenoids

13. Saponins

14. Fat

15. Sterols

PHARMACOGNOSTICAL STUDIES

METHODOLOGY

1. Microscopy (SOP No. PCOG-005): The sample was preserved in fixative FAA for more than 48 h. The preserved specimenswerecutintothintransversesectionsusingasharpblade,andthesectionswerestainedwith0.8%safranin and 0.5% Astra blue. Transverse sections were photographed using an Axiolab 5 trinocular microscope attached to a ZeissAxiocam208colordigitalcameraunderbrightfieldlight.Magnificationswereindicatedbyascalebar.

RESULT Microscopy Leaf sheath

TheTSoftheleafsheathisnearlycircular-shapedinoutlinewithanoutersingle-layeredepidermiscoveredbycuticle and bears simple covering trichomes; a hypodermal layer made up of aerenchyma cells can be seen beneath the epidermis, followed by parenchymatous ground tissue; vascular bundles are arranged in a ring from one end to the other; bundles are collateral and covered by parenchymatous bundles; xylem and phloem are formed of normal vascular elements; some of the parenchyma cells contain micro sphenoidal, acicular, rod-shaped, and prismatic crystals;andfewcellscontainanycellcontents(Fig.1).

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

TS of leaf shealth under polarized field

AC-aerenchyma; BS-bundlesheath; E-epidermis; Mcr-microsphenoidalcrystals; pa-parenchyma; pcr-prismaticcrystals; Phphloem; Rcr-rod-shapedcrystals; T-trichome; V-vessel; VB-vascularbundle

LEAF

TSofleafshowslowerconvexandupperdepressedmidribsurfacewithlaterallaminarextensions 9fig:2)

MIDRIB

TSofthemidribshowsupperandlowerepidermisformedofoval-shapedcellsbycuticleandbearssimplecoveringtrichomes; a patch of collenchymatous hypodermis is followed by a few layers of parenchyma cells; ground tissue is aeren chymatous, embeddedwith3vascularbundlesarrangedinahalfringinwhich thecenterbundleiscomparativelylargerthanthelateral bundle; the vascularbundle iscollateral andcovered by parenchymatous bundle sheathcells; xylem is arrangedtowards the uppersideandphloemtowardsthelower;xylemandphloemareformedofnormalvascularelements.(fig.2)

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

LAMINA

TSoflaminashowstheupperandlowerepidermisconsistsofovaltobarrel-shapedcellscoveredbycuticleandbearssimple coveringtrichomes;mesophylltissueisdifferentiatedintouppersingle-layeredlong,columnarpalisadecellsfollowedby3to 4layersoflooselyarrangedspongyparenchymacellswithintercellularspaces;vascularbundlesarecollateralandcoveredby parenchymatous bundle sheath and some patches of pericyclic fibers at the lower side; xylem and phloem are formed of normalvascularelements;rod-shapedcrystalsarefounddistributedinsomeoftheepidermalcellsandmesophylltissues(Fig. 2).

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

GAS CHROMATOGRAPHY-MASS SPECTROMETRY

The present study focuses on the analysis of phytochemical constituents performed using gas chromatography-mass spectrometry.Intheethylacetateandethanolicextractof Commelina benghalensis, bioactivephytochemicalsweredetectedby the screening method of GC-MS. These extracts show relative concentrations of the bioactive phytoconstituents at different peakheights.Thelistofphytoconstituentswiththeirpercentageofpeakareaandretentiontime.

PROCEDURE

Sample preparation:

Weigh1gofhomogenizedsample(chloroformandethanol)intoa15mlplastictube.Add1mlofethylacetateandvortexfor5 minutes.

Thentransfertoa1mlvialforinjectionofµlofthesampleintoGC-MS

INSTRUMENT CONDITIONS:

GC–MSanalysis wasconductedusinga ShimadzuQP2020gaschromatograph–massspectrometerfittedwitha split/splitless injector and an electron impact (EI⁺) ionization source. Chromatographic separation was performed on an HP-5MS capillary column(30m×0.32mminternaldiameter,0.25µmfilmthickness).Asamplevolumeof1.0µLwasinjected,andtheinjector temperaturewasmaintainedat285°C.

High-purity helium (99.999%) served as the carrier gas at a constant flow rate of 1.0 mL/min. The oven temperature was initially set at 50°C and programmed to increase gradually to 290°C, where it was held for 10 minutes to ensure complete elution of analytes. The split valve was opened 3 minutes after injection to purge the injector. All injections were performed usinga10µLmicrosyringe.

Forsamplepreparation,1gofthehomogenizedplantmaterialwasweighedintoa15mLplasticcentrifugetube,followedby theadditionof5mLofethylacetate.Themixturewasvortexedfor5minutes,andtheresultingextractwastransferredintoa1 mLvial.Analiquotof1µLoftheextractwastheninjectedintotheGC–MSsystemforanalysis.

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

TABLE 1:Bioactivephytoconstituentsofethanolicextractof Commelina benghalensis

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

Volume: 13 Issue: 01 | Jan 2026 www.irjet.net p-ISSN: 2395-0072

TABLE 2:BioactivephytoconstituentsofEthylacetateextractof Commelina benghalensis

International Research Journal of Engineering and Technology (IRJET) e-ISSN: 2395-0056

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Conclusion:

Commelina benghalensis was extracted using different solvents, and the percentage yield obtained from each solvent was determined.Phytochemicalscreeningwascarriedouttoidentifythepresenceofbioactiveconstituentsintheselectedextracts. Thedetectionofthesephytochemicalsthroughpreliminaryanalysisindicatesthepotentialmedicinalvalueoftheplant.

The plant extract was further analyzed using gas chromatography–mass spectrometry (GC–MS) under standard operating conditions. Compounds were separated based on their retention times and detected using a flame ionization detector (FID). The resulting chromatogram displayed multiple peaks, each representing distinct bioactive compounds. Gas chromatography confirmed the chemical diversity of the extract by effectively separating volatile constituents. The identified compounds are likelycontributorstothetherapeuticpropertiesattributedto Commelina benghalensis

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