Phytochemical Screening & Evaluation of Antimicrobial Activity of Ethanolic Extract of Flower and Leaves of Lantana Camara L.

Medicinal plants have been used to relieve illness for over 60,000 years ago, maybe even older. Scripts about medicinal plants from ancient civilizations like Egypt and China show people always sought to use nature to cure their diseases. According to WHO, most of the population worldwide (80%) utilize herbal substances to cure diseases. Plants contain phytochemicals that are helpful in the defense against herbivores. These phytochemicals have medicinal properties that are used by humans¹. Lantana camara (Umuhengeri), an ornamental weed garden plant that belongs to the Verbenaceae family, is a low erect, rugged hairy, evergreen shrub native to tropical America. It is considered as a valuable plant for traditional healers worldwide (Manish et al., 2011). L. camara extract has been shown to exhibit antimicrobial, insecticidal, and nematocidal activities. Besides, it exhibits immunosuppressive and antitumor activities². Throughout history, bacterial infections have plagued men. Antibiotics have changed the world in the past 70 years, saving and improving lives and making them the cornerstones of modern health systems. The therapeutic capacity for treating patients with bacterial infections is being depleted by antimicrobial resistance. Physicians encounter infections that are susceptible to few or even none of the available antibiotics³. Drug-resistant pathogens have made widespread infections more challenging to treat or even incurable in some cases, with catastrophic implications for patients. This was due to antibiotic misuse, incorrect dosage, etc⁴. In time, as many bacteria are developing antibiotic resistance to conventional antibiotics, natural products from plants are as alternative new antibiotics to cure diseases⁵. Plant-based medicines, which were previously only available in the form of crude medicines (teas, poultices, powders), are now used to develop new drug discoveries. Since plants contain various bioactive compounds such as alkaloids, flavonoids, and saponins, they can serve as antimicrobial agents⁶. Bacteria like P. aeruginosa, S. pneumonia, S. aureus, S. typhi and E. coli are the most problematic in Rwanda⁷. The development of antimicrobial resistance in patients to available antimicrobials is high and worries some. It is necessary to search for new antimicrobial agents. In search of a new alternative antibiotic drug that may help control drug-resistant pathogenic bacteria, this research focused on the effect of L. camara extracts on clinically isolated bacteria (S. typhi, S. aureus, S. pneumonia, E. coli

MATERIAL & METHODS

Collection, Identification & Authentication of Plant material

The flower and leaves of Lantana camara collected from a local market of Lucknow, U.P India. The plant materials, flowers & leaves of Lantana camara linn identified and authentication by Prof. N. K Dubey, scientist, plant diversity, Systemic and Herbarium division, Lucknow B.H.U (Banaras Hindu University) the plant specimens were authenticated in Herbarium with Ref. no. of Lat/CamL /2025/01.

Preparation of extract

The flower and leaves are dried in shade at room temperature. The dried flower and leaves of Lanata camara were pulverized into coarse powder and sieved through number 23 and stored in to a container⁸. Conventional hot Soxhlet extraction method was used for extraction of flower 200 gm of   powder flower were extract with 500 ml of ethanol at the temperature range 60-70°C successively 3-4 day the isolated extract was filtered and the solvent was removed using a vacuum rotary evaporator after a complete extraction, & the obtained residue was kept in a desiccators⁹. The percentage yield of extract was found to be 54.40 gm.

RESULT & DISCUSSION

Phytochemical Analysis of Extracts

The ethanolic extract of Leaves & flower of Lantana camara subjected for preliminary phytochemical analysis for detection of various phytoconstituent like Alkaloids, Glycoside, Tannins, Triterpenoids, Carbohydrates & proteins¹⁰. The detail result was given in table 1.

Table 1: Phytochemical screening of flower and leaves extract of L. camara extract

Fig. 1: Phytochemical screening of flower extracts (a) and leaves extract (b) of L. camara linn.

Fig. 2: U. V visible spectroscopy of L. camara linn. flower extract

Fig. 3: U. V visible spectroscopy of L. camara linn. leaves extract

Functional group analysis by IR Spectroscopy

FTIR Spectra of L. camara linn flower extract:

The FTIR spectra of ethanolic extract of L. camara linn flower analyses by FTIR (PerkinElmer spectrum version) spectrophotometer, CSIR-CDRI, Lucknow (U.P) India.

Fig. 4: FTIR Spectrum peaks of L. camara linn flower extract

Table 2 FTIR Spectrum peaks of L. camara linn flower extract

The FTIR spectra of ethanolic extract of L. camara linn. leaves linn analyse by FTIR (PerkinElmer spectrum version) spectrophotometer, CSIR-CDRI, Lucknow (U.P) India.

Fig. 5: FTIR Spectrum of L. camara linn leaves extract

Table 3 FTIR Spectrum peaks of L. camara linn leaves extract

LC-MS (liquid chromatography & mass spectroscopy) Lantana camara linn flower extract

The LC-MS (liquid chromatography & mass spectroscopy) spectra of L. camara flower linn analyse by LC-MS, CSIR-CDRI, Lucknow (U.P) India.

Fig 6: LC-MS of the L. camara linn flower extract (F1)

LC-MS (liquid chromatography & mass spectroscopy) Lantana camara linn leaves extract

The LC-MS (liquid chromatography & mass spectroscopy) spectra of L. camara leaves linn analyse by LC-MS, CSIR-CDRI, Lucknow (U.P) India.

Fig. 7: LC-MS of the L. camara linn leaves extract (F2)

LC-MS (liquid chromatography & mass spectroscopy) Lantana camara linn flower and leaves extract

An incredibly sensitive and specific analytical method, liquid chromatography mass spectrometry (LC-MS/MS) can accurately identify the substances in your sample and their concentrations.

Table 4: LC-MS of the of L. camara linn leaves & flower extract

HPLC study of the Lantana camara flower extract

The HPLC method of the Lantana Camara flower extract analyzed by HPLC at CSIR-CDRI, Lucknow (U.P) India.

Fig. 8: HPLC chromatogram of the ethanolic extract of Lantana camara linn flower

HPLC study of the Lantana camara leaves extract

The HP-LC method of the Lantana Camaraleaves extract analyzed by HPLC at CSIR-CDRI, Lucknow (U.P) India.

Fig. 9: HPLC of the ethanolic extract of Lantana camara linn leaves

In phytochemical and analytical chemistry, HPLC is a chromatographic technique that can separate a mixture of compounds and is used to identify, quantify, and purify the individual components of the mixture. It is a flexible, reliable, and popular method for the isolation of natural products¹¹. This method is currently becoming more and more well-liked among other analytical techniques as the main choice for fingerprinting studies for herbal plant quality control.

Evaluation of Antimicrobial Activity

The Lantana camara linn. flowers & leaves were evaluated for antimicrobial activity against bacterial strains by and cup plate method¹². The microbes were procured from the Institute of Microbial Technology (IMTECH), Chandigarh, India. Ofloxacin   were used as standard.

Cup plate method

Test organism:  Microorganism staphylococcus aureus (MTCC1430), and candida albicans (MTCC 227) were procured from the institute of microbial technology (MTECH) Chandigarh India.

Preparation of inoculums: the bacterial cultures were kept in refrigerator with nutritional agar slats, and they were transferred every month. A loopful of organisms from 24- hour-old cultures of nutrient agar slants growth at 37^oC were inoculated in to the sterile saline to create the inoculum for the test organisms¹³. To achieve a culture density of roughly 107 CFU/ml, one millilitre using sterile saline.

 Preparation of test sample & standard drugs:

To prepare the 1000µg/ ml stock solution of the lantana camara leaves and flower extract, 10 mg of the synthesized chemical was dissolved in DMSO. After transferring 1 ml of the stock solution in to the volumetric flask, the same solvent was used to further dilute in up to 10 ml. Likewise, various concentration were generated, including 20,40, 60, 80, 100, μg/ml¹⁴. Fluconazole and norfloxacin were utilized and usual.

Table 5: Growth media, incubation temperature and pH for microbial strains.

Determination of zone of inhibition of the lantana camara leaves and flowers extract using cup plate method

The cup plate method was determined to the minimum zone of inhibition of lantana camara leaves and flower extract against the microbial stains using nutrition agar and sabouraud dextrose agar media¹⁵. A measured volume of the microbial inoculums was added to the sterilized agar media, which had been cooled to between 40 and 50 degrees Celsius, and it was thoroughly spun under the laminar air flow bench in an aseptic setting.  After sterilizing the petri plates, 25ml of inoculum was aseptically add and set aside to solidify¹⁶. Using sterile steel cork borer on solidified media, the cavities were created. The, using different micropipettes, the test chemical and references solution was added to each cavity. To allow for diffusion, inoculated plates were left at room temperature for two hours before being incubated for seventy two hours at 37^oC. After measuring the zone of inhibition, the MIC value was determined¹⁷.

Table 6: Zone of inhibition (diameter) of Lantana camara leaves and flower extract against bacterial trains by cup plate method