Quantification of Lupeol as Secondary Metabolite by HPTLC Technique and Assessment of Antimicrobial Potential of Ethyl Acetate Extract of Betula alnoides Bark

The present work includes extraction of Betula alnoides bark using ethyl acetate as a solvent, preliminary phytochemical test, quantification of phytochemicals and quantification of lupeol in Betula alnoides by High Performance Thin Layer Chromatography (HPTLC) instrument along with the assessment of the antimicrobial potential of Ethyl Acetate Extract (EAE). The marc obtained after defatting of the coarsely powdered crude drug in Petroleum ether (60-80) was extracted using ethyl acetate. Afterward, preliminary phytochemical tests were done. For High Performance Thin Layer Chromatography (HPTLC), the solvent used was n-hexane: ethyl acetate (8:2 v/v) and scanning was performed at wavelength 254 nm. EAE was screened for antimicrobial potential. The extraction yield was 3.45% w/w. The result of the phytochemical analysis confirmed the presence of some important phytochemicals in EAE. A clear and resolved peak of lupeol was observed at Rf 0.61. The developed method was validated as per ICH guidelines. The concentration (%) of the marker compound (lupeol) was found to be 0.0168. Disk diffusion method using Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus subtilis as bacterial strains and Candida albicans, Aspergillus flavus and Epidermophyton floccosum as fungal strains against ciprofloxacin (for antibacterial activity) and fluconazole (for antifungal activity) as standard drugs was employed. The finding suggested that EAE possess significant antibacterial and antifungal activity when comparison was made with standard drugs. The proposed elucidated mechanism behind this action may be due to the presence of triterpenoids in EAE.

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PEMISAHAN DAN IDENTIFIKASI EKSTRAK KASAR SESKUITERPEN DAUN BUNGA MATAHARI (Helianyhus annuus L.) DENGAN KROMATOGRAFI LAPIS TIPIS

ALCHEMY ◽

10.18860/al.v0i0.2905 ◽

2013 ◽

Author(s):

Roihatul Muti'ah ◽

Elok Kamilah Hayati ◽

Yani Triastutik

Keyword(s):

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Ethyl Acetate Extract ◽

Hexane Extract ◽

Chemical Test ◽

Content Identification ◽

Rf Values ◽

Extract Fraction ◽

Layer Chromatography

The purpose of this research wasto separate and identify the leaf crude extracts esquiterpene of Sun flower (Helianthus annuus L.) using thin layer chromatography.Sun flower leaf maceration method performed with the solvent methanol. Then performed liquid extraction with ethyl acetate and n-hexane solvent. Ethyl acetate extract fraction and n-hexane extract fraction furth erphyto chemical test. After being test edphyto chemical with reagents, both extracts was followed by sesquiterpene content identification using thin layer chromatography (TLC) analytic.Phytochemical test result from ethyl acetate extract fraction was positive terpenoid, sesquiterpene and triterpene, while n-hexane extractfraction positive terpenoid, sesquiterpene andsteroid. All egedsesquiterpene with eluentdichloromethane: ethyl acetate (4,8:0,2) is shown with apurplestain. In the ethyl acetate extract fraction all egedsesquiterpene having Rf values of 0.89; 0.94, and 0.96. While n-hexane extract fraction, the resulting eluental legedsesquiterpene Rf 0.49; 0.8,and 0.99.

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Secondary Metabolites Production by Actinomycetes and their Antifungal Activity

KnE Life Sciences ◽

10.18502/kls.v3i4.713 ◽

2017 ◽

Vol 3 (4) ◽

pp. 256 ◽

Cited By ~ 2

Author(s):

Tirta Kumala Dewi ◽

Dwi Agustiani ◽

Sarjiya Antonius

Keyword(s):

Antifungal Activity ◽

Secondary Metabolites ◽

Thin Layer ◽

Fusarium Oxysporum ◽

Thin Layer Chromatography ◽

Pathogenic Fungi ◽

Diffusion Method ◽

Wide Range ◽

Layer Chromatography

Wilt desease of banana caused by Fusarium oxysporum f.sp. cubense (FOC) is one of the most destructive deseases of banana in the tropics. Actinomycetes are the most economically and biotechnologically valuable prokaryotes able to produce wide range of bioactive secondary metabolites. The aims of the present study are to isolate and screen the actinomycetes with high potential ability to produce secondary metabolites that have inhibitory activity against plant pathogenic fungi, Fusarium oxysporum f. sp. cubense. Two isolates from Lampung and Cianjur showed activity against fungi. The isolates designed as L.3.1 and CiIA5b. The metabolites from potent stain was produced by extraction of culture filtrate with ethyl acetate : methanol (4:1), it was tested for their antifungal activity by well diffusion method. Evidence for in vitro antibiosis of L.3.1 and CiIA5b isolates was demonstrated by the zone of fungal-growth inhibition. Production of secondary metabolites was analysis by thin layer chromatography (TLC) and bioautography assays. In this study, the metabolites from L.3.1 and CiIA5b have showed good antifungal activity.<div> Keywords: Actinomycetes; antifungal activity; bioautography; secondary metabolites; thin layer chromatography.</div>

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Determination of Cimetidine, Famotidine, and Ranitidine Hydrochloride in the Presence of Their Sulfoxide Derivatives in Pure and Dosage Forms by High-Performance Thin-Layer Chromatography and Scanning Densitometry

Journal of AOAC International ◽

10.1093/jaoac/85.5.1015 ◽

2002 ◽

Vol 85 (5) ◽

pp. 1015-1020 ◽

Cited By ~ 9

Author(s):

Khadiga M Kelani ◽

Azza M Aziz ◽

Maha A Hegazy ◽

Laila Abdel Fattah

Keyword(s):

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Mobile Phase ◽

High Performance ◽

Accurate Method ◽

Dosage Forms ◽

Ranitidine Hydrochloride ◽

Layer Chromatography

Abstract A selective, precise, and accurate method was developed for the determination of cimetidine (C), famotidine (F), and ranitidine hydrochloride (R·HCI)in the presence of their sulfoxide derivatives. The method involves quantitative densitometric evaluation of mixtures of the drugs and their derivatives after separation by high-performance thin-layer chromatography on silica gel plates (10 × 20 cm) with ethyl acetate–isopropanol–20% ammonia (9 + 5 + 4, v/v) as the mobile phase for both C and F and ethyl acetate–methanol–20% ammonia (10 + 2 + 2, v/v) as the mobile phase for R·HCI; Rf values for C, F, and R·HCI and their corresponding derivatives were 0.85 and 0.59, 0.73 and 0.41, and 0.56 and 0.33, respectively. Developing time was approximately 20 min. For densitometric evaluation, peak areas were recorded at 218, 265, and 313 nm for C, F, and R·HCI, respectively. The relationship between concentration and the corresponding peak area was plotted for the ranges of 5–50 μg/spot for C and 2–20 μg/spot for F and R·HCl. Mean recoveries were 100.39 ± 1.33, 99.77 ± 1.30, and 100.09 ± 0.69% for C, F, and R·HCI, respectively. The proposed method was used successfully for stability testing of the pure drugs in the presence of up to 90% of their degradates, in bulk powder and dosage forms. The results obtained were analyzed statistically and compared with those obtained by the official methods.

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Antioxidant Activity of Daemonorops draco Resin

Jurnal Kimia Sains dan Aplikasi ◽

10.14710/jksa.22.5.179-183 ◽

2019 ◽

Vol 22 (5) ◽

pp. 179-183 ◽

Cited By ~ 2

Author(s):

Sri Purwanti ◽

Wulan Tri Wahyuni ◽

Irmanida Batubara

Keyword(s):

Antioxidant Activity ◽

Infrared Spectroscopy ◽

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Column Chromatography ◽

Ethyl Acetate Extract ◽

Preparative Thin Layer Chromatography ◽

Layer Chromatography

Jernang resin is secretion of jernang rattan (Daemonorops draco, Arecaceae family) fruits which is endemic in Southeast Asia. This resin has various biological activities and empirically used as wound healing, headache medicines, and fever remedies by Anak Dalam ethnic group from Jambi. This study was performed to evaluate the antioxidant activity of nonpolar fraction of D. draco resin which collected from Jambi Province, Sumatera, Indonesia. Resin was extracted with n-hexane, ethyl acetate, and methanol respectively. The antioxidant properties of the extracts were then evaluated using 1,1-diphenyl-2picryl-hidrazyl radical scavenging assay. The most active extract was further fractionated using n-hexane and methanol and separated using column chromatography and preparative thin layer chromatography. Separation of the extract was conducted through antioxidant assay-guided fractionation. Characterization of the active fraction was carried out by infrared spectroscopy. The result shows that ethyl acetate extract provides higher antioxidant activity (IC50 = 27.61 µg/mL) compare to methanol and n-hexane extracts. N-hexane fraction of ethyl acetate extract used for further separation using column and preparative thin layer chromatography due to its antioxidant activity. Separation using column chromatography resulting in 9 fractions (F.1-9). Fraction F.5 provide high antioxidant activity (IC50 = 17.27 µg/mL) and further separated using preparative thin layer chromatography resulting two fractions with lower antioxidant activity F.5.1 (IC50 = 85.18 µg/mL) and F.5.2 (IC50 = 34.94 µg/mL). Characterization of fraction F.5.2 using infrared spectroscopy showed that component in fraction F.5.2 contains NH-substituted benzene.

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Determination of Rosmarinic Acid and Caffeic Acid in Thymus daenensis and Thymus lancifolius Extracts by High-Performance Thin Layer Chromatography

Letters in Applied NanoBioScience ◽

10.33263/lianbs104.28042809 ◽

2021 ◽

Vol 10 (4) ◽

pp. 2804-2809

Keyword(s):

Thin Layer ◽

Formic Acid ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Caffeic Acid ◽

Rosmarinic Acid ◽

Mobile Phase ◽

High Performance ◽

Layer Chromatography

Thymus species belong to the Lamiaceae family, of which 18 species in the flora of Iran, 6 are endemic to Iran. In the current research, high-performance thin-layer chromatography (HPTLC) technique as an easy, fast, reproducible, and low-cost method was used for the determination of rosmarinic acid and caffeic acid in Thymus lancifolius (T. lancifolius) and two species of Thymus daenensis (T. daenensis) from Iran. Toluene-ethyl acetate-formic acid with a ratio of 67.72-22.90 and 9.38% was selected as the mobile phase of rosmarinic acid, and ethyl acetate-methanol-formic acid-water with a ratio of 85-8-2 and 5% was designated as the mobile phase of caffeic acid. The highest and lowest amount of rosmarinic acid was observed for T. daenensis 1 (10.54±0.12 mg/g) and T. lancifolius (0.46±0.01 mg/g), respectively. The amount of rosmarinic acid for T. daenensis 2 was obtained as 7.85±0.02 mg/g for each of the dried plants. In the following, HPTLC analysis of caffeic acid for T. daenensis 1, T. daenensis 2, and T. lancifolius was acquired amounts of 0.78±0.007, 0.13±0.007, and 0.26±0.007 mg/g for each of dried plants, respectively. Therefore, regarding the special effects of phenolic acids and properties of the Thymus genus, the acquired results are suitable for application in pathogenic research, infections, immunology diseases, and evaluation of the antioxidant activity.

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QUANTITATIVE PHYTOCHEMICAL SCREENING, THIN-LAYER CHROMATOGRAPHY ANALYSIS, HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY FINGERPRINTING, AND ANTIOXIDANT ACTIVITY OF LEAVES OF DIOSPYROS MONTANA (ROXB.)

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2019.v12i2.29738 ◽

2019 ◽

pp. 325-331

Author(s):

ABHIJEET V PURI

Keyword(s):

Antioxidant Activity ◽

Thin Layer ◽

Ethyl Acetate ◽

Petroleum Ether ◽

Thin Layer Chromatography ◽

High Performance ◽

Butylated Hydroxytoluene ◽

Antioxidant Potential ◽

Total Phenolic ◽

Layer Chromatography

Objective: The objective of this study was to investigate important phytochemical constituents and antioxidant potential of Diospyros montana Roxb. leaves belonging to the family Ebenaceae. Methods: Leaves were exhaustively extracted with ethanol and fractionated into petroleum ether, chloroform, and ethyl acetate extracts. The various fractions were further analyzed for phytochemical composition and concentration-dependent antioxidant activity using conventional methods and high-performance thin-layer chromatography (HPTLC) fingerprinting. Since leaves contained phenolic compounds, extracts were evaluated for total phenolic content, flavonoids contents, and in-vitro antioxidant activity. Antioxidant potential was assessed using parameters such as superoxide radical scavenging, nitric oxide inhibition, and β-carotene/linoleic acid antioxidant activity. Results: Primitive phytochemical investigation highlighted the presence of steroids, saponins, flavonoids, alkaloids, and tannins which were confirmed by TLC and HPTLC fingerprinting. The antioxidant activity of leaf extracts decreased in the following order ethyl acetate > ethanolic > chloroform > petroleum ether and it was comparable with standards such as ascorbic acid and butylated hydroxytoluene. Conclusion: The present study concludes that the ethanolic extract and fractions of D. montana (Roxb.) leaves have prominent antioxidant activity comparable to standards. Therefore, D. montana (Roxb.) leaves may be used as a probable source of natural antioxidants in the pharmaceutical industry.

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