Designing Safer Solvents to Replace Methylene Chloride for Liquid Chromatography Applications Using Thin-Layer Chromatography as a Screening Tool

Methylene chloride, commonly known as dichloromethane (DCM), is a widely used chemical for chromatography separation within the polymer, chemical, and pharmaceutical industries. With the ability to effectively solvate heterocyclic compounds, and properties including a low boiling point, high density, and low cost, DCM has become the solvent of choice for many different applications. However, DCM has high neurotoxicity and is carcinogenic, with exposure linked to damage to the brain and the central nervous system, even at low exposure levels. This research focuses on sustainability and works towards finding safer alternative solvents to replace DCM in pharmaceutical manufacturing. The research was conducted with three active pharmaceutical ingredients (API) widely used in the pharmaceutical industry: acetaminophen, aspirin, and ibuprofen. Thin-layer chromatography (TLC) was used to investigate if an alternative solvent or solvent blend could show comparable separation performance to DCM. The use of the Hansen Solubility Parameter (HSP) theory and solubility testing allowed for the identification of potential alternative solvents or solvent blends to replace DCM. HSP values for the three APIs were experimentally determined and used to identify safer solvents and blends that could potentially replace DCM. Safer solvents or binary solvent blends were down-selected based on their dissolution power, safety, and price. The down-selected solvents (e.g., ethyl acetate) and solvent blends were further evaluated using three chemical hazard classification approaches to find the best fitting nonhazardous replacement to DCM. Several safer solvent blends (e.g., mixtures composed of methyl acetate and ethyl acetate) with adequate TLC performance were identified. Results from this study are expected to provide guidance for identifying and evaluating safer solvents to separate APIs using chromatography.

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Direct Fluorescent Detection of Organothiophosphorus Pesticides and Some of Their Sulfur-containing Breakdown Products After Thin Layer Chromatography

Journal of AOAC INTERNATIONAL ◽

10.1093/jaoac/50.5.1088 ◽

1967 ◽

Vol 50 (5) ◽

pp. 1088-1098

Author(s):

Mohamed Tawfik H Ragab

Keyword(s):

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Ultraviolet Light ◽

Congo Red ◽

Fluorescent Detection ◽

Bromine Vapor ◽

Layer Chromatography ◽

Sulfur Containing ◽

Dark Blue

Abstract A rapid, simple, convenient, and widely applicable method for the direct fluorescent detection of organothiophosphorus pesticides and some postulated breakdown products of these compounds is presented. The coinpounds were spotted on thin layer chromatographic sheets, developed in ethyl acetate :nhexane, and made visible by exposure to bromine vapor followed by spraying with ferric chloride and 2-(o-hydroxy phenyl) benzoxazole. Of the 47 compounds tested, 32 compounds produced fluorescent blue spots vinder longwave ultraviolet light; these consisted of 25 organothiophosphorus pesticides, 5 sulfur-containing breakdown products, and phosphoric and hypophosphorous acids. A superimposed Congo red spray destroyed the fluorescence and resulted in dark blue spots against a red backgrovind. The sensitivity of this method is in the range of 0.2 to 5.0 μg, depending on the specific compound.

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PHARMACOGNOSTIC STANDARDIZATION, PRELIMINARY PHYTOCHEMICAL SCREENING AND TLC FINGERPRINTING OF THE BARK OF CASCABELA THEVETIA L.

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2019v11i7.29218 ◽

2019 ◽

pp. 125-130

Author(s):

Neelutpal Gogoi ◽

Biman Bhuyan ◽

Trinayan Deka

Keyword(s):

Thin Layer ◽

Ethyl Acetate ◽

Petroleum Ether ◽

Thin Layer Chromatography ◽

Water Soluble ◽

Phytochemical Analysis ◽

Alcoholic Extract ◽

Phytochemical Screening ◽

Cork Cell ◽

Layer Chromatography

Objectives: In this study, systematic pharmacognostic study and preliminary phytochemical screening of the bark of Cascabela thevetia L. were carried out. Methods: The selected plant part was collected, processed and stored in an airtight container. From the bark different pharmacognostic studies like macroscopic and microscopic evaluation, physicochemical parameters, fluorescence analysis were done. Powdered bark was successively extracted by petroleum ether, chloroform, ethyl acetate, and methanol using a Soxhlet apparatus and finally macerated with the hydro-alcoholic solvent system (30:70). The preliminary phytochemical analysis and thin layer chromatography of the extracts were done to find the nature and number of the different phytoconstituents present. Results: Transverse microscopy reveals the presence of crystal oxalate, cork cell, starch granules, vascular bundle, phloem fiber, parenchyma cells, and collenchyma cells. Powder microscopy also showed the presence of cork cell, fiber and calcium oxalate crystal. Results obtained in different physicochemical analysis like total ash, acid insoluble ash, water soluble ash, alcohol-soluble extractive, water-soluble extractive, and moisture content were 8.67%, 0.83%, 5.33%, 4.53%, 12.27%, and 7.83% respectively. Phytochemical analysis showed the presence of alkaloid, flavonoid, triterpenoid, phytosterol, tannin, saponin, anthraquinone, carbohydrate and fatty acid in the different extracts. TLC (Thin Layer Chromatography) study revealed 4 spots in petroleum ether, chloroform, ethyl acetate, and methanol extracts and 3 spots in the Hydro-alcoholic extract with different solvent systems. Conclusion: The results obtained from the study will provide a reliable basis for identification, purity, and quality of the plant.

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Phytochemical Investigation of the Aerial Part of Iraqi Convolvulus arvensis

Iraqi Journal of Pharmaceutical Sciences ( P-ISSN 1683 - 3597 E-ISSN 2521 - 3512) ◽

10.31351/vol29iss2pp62-69 ◽

2020 ◽

Vol 29 (2) ◽

pp. 62-69

Author(s):

Mustafa H. Alwan ◽

Maha N. Hamad

Keyword(s):

Liquid Chromatography ◽

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Mass Spectroscopy ◽

Morning Glory ◽

Convolvulus Arvensis ◽

Aerial Parts ◽

Phytochemical Investigation ◽

Layer Chromatography

Convolvulus arvensis is a species of bindweed that is rhizomatous and is in the morning glory family (Convolvulaceae) native to Europe and Asia. The plant is naturally grown in Iraq. The plant was reported to be used in traditional medicine from as early as 1730s. The Aerial parts of Convolvulus arvensis were macerated in 80% ethanol for 6 days. The concentrated extract was partitioned with n-hexane, chloroform, ethyl acetate- and n-butanol successively. The n-hexane and ethyl acetate, fractions were examined for the presence of phytochemicals by thin layer chromatography and high performance liquid chromatography and its steroid and flavonoid contents were investigated. Stigmasterol was isolated from n-hexane fraction and identified by liquid chromatography/mass spectroscopy. Rutin was isolated from the ethyl acetate fraction and identified by liquid chromatography/mass spectroscopy. The aim is to examine the phytochemical constituents of the aerial parts of Convolvulus arvensis, literature survey available so far revealed that there were no studies about the phytochemical investigation for Convolvulus arvensis in Iraq. Different chromatographic techniques like Thin Layer Chromatography and mass spectroscopy were used and the presence of Stigmasterol and Rutin in aerial parts of Convolvulus arvensis was indicated.

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Identifikasi Senyawa Metabolit Sekunder pada Durian (Durio zibethinus Murr)

Jurnal Ilmiah Biologi UMA (JIBIOMA) ◽

10.31289/jibioma.v1i1.138 ◽

2019 ◽

Vol 1 (1) ◽

pp. 1-6

Author(s):

Aji Suteja ◽

Emmy Harso Kardhinata ◽

Rosliana Lubis

Keyword(s):

Thin Layer ◽

Ethyl Acetate ◽

Experimental Method ◽

Thin Layer Chromatography ◽

Secondary Metabolite ◽

Phytochemical Screening ◽

Screening Analysis ◽

Durio Zibethinus ◽

Rf Values ◽

Layer Chromatography

This study aims to determine how to identify the types of secondary metabolite compounds contained in durian leaves. The study was conducted by an experimental method by identifying secondary metabolite compounds on durian leaves using phytochemical screening. The sample criteria used were the leaf buds and all the leaf strands (except the leaf bones). The results of the study showed that of the three types of durian leaves namely copper, baskets and Sp A. There are several types of secondary metabolite compounds including alkaloids, steroids, and terpenoids. Phytochemical screening analysis was carried out using thin layer chromatography by showing RF values on three types of durian leaves using methanol and ethyl acetate solvents in a ratio of 3: 1. RF value on copper durian is 0.97, bakul durian is 0.95 and Sp durian. A 0.94.

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UJI TOKSISITAS DAN IDENTIFIKASI SENYAWA EKSTRAK ALGA MERAH (Eucheuma cottonii) TERHADAP LARVA UDANG Artemia salina LEACH

ALCHEMY ◽

10.18860/al.v0i0.2892 ◽

2013 ◽

Cited By ~ 1

Author(s):

Noer Mardlatillah Sharo ◽

Rachmawati Ningsih ◽

Ahmad Hanapi ◽

Ahmad Nasichuddin

Keyword(s):

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Red Algae ◽

Ethanol Extract ◽

Artemia Salina ◽

Probit Analysis ◽

Active Compounds ◽

Layer Chromatography ◽

Eucheuma Cottonii

Eucheuma cottonii is a multicellular algae (multicellular) that have bioactive compounds used in medicine, such as anticancer. The purpose of this research is to know the level of toxicity of the crude extract of ethanol and n-hexane red algae (Eucheuma cottonii) against larvae of shrimp A. salina Leach and to know the active compounds contained in crude extracts of ethanol and n-hexane red algae (Eucheuma cottonii).This research was conducted with the maceration extraction continuing by test toxicity against the shrimp larvae of A. salina Leach (BSLT). The result of the toxicity test then proceeds with the test reagents and phytochemicals with thin layer chromatography for positive test of compound phytochemicals. Thin-layer chromatography using eluen n-hexane: ethyl acetate (7: 3) to ethanol extract and n-hexane: ethyl acetate (17: 3) for n-hexane extract. Data on toxicity of A. salina Leach analyzed by probit analysis to find out the value of LC50 on each extract.The results of this research indicate that each of the rough red algae extract (Eucheuma cottonii) by using a solvent of ethanol and n-hexane have toxicity values (LC50) of shrimp larvae of A. salina Leach consecutive 58,0128 ppm and 61,7571 ppm. Active compounds content of Phytochemical test results i.e. triterpenoid (ethanol extract) and steroids (n-hexsane extract). The TLC triterpenoid obtained 7 stains, and the results obtained steroids 8 TLC stains.

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

Oriental Journal Of Chemistry ◽

10.13005/ojc/370223 ◽

2021 ◽

Vol 37 (2) ◽

pp. 426-432

Author(s):

Shahbaz Khan ◽

Harpreet Singh ◽

Arun K Mishra ◽

Najam Ali Khan

Keyword(s):

Antifungal Activity ◽

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

High Performance ◽

Ethyl Acetate Extract ◽

Diffusion Method ◽

Antimicrobial Potential ◽

Betula Alnoides ◽

Layer Chromatography

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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Identifikasi Kromatografi Lapis Tipis Sudamala (Artemisia vulgaris L.)

Majalah Kedokteran Gigi Indonesia ◽

10.22146/majkedgiind.9226 ◽

2015 ◽

Vol 20 (2) ◽

pp. 167 ◽

Cited By ~ 1

Author(s):

Ira Arundina ◽

Theresia Indah Budhy S. ◽

Muhammad Luthfi ◽

Retno Indrawati

Keyword(s):

Squamous Cell Carcinoma ◽

Oral Cavity ◽

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Artemisia Vulgaris ◽

Anti Cancer ◽

Vacuum Column ◽

Layer Chromatography ◽

Primary Risk Factor

Karsinoma sel skuamosa rongga mulut merupakan jenis kanker yang paling sering ditemukan di rongga mulut. Faktor risiko utama terjadi keganasan di rongga mulut meliputi riwayat serta kebiasaan mengkonsumsi tembakau dan atau alkohol. Tanaman sudamala (Artemisia vulgaris L.) sering digunakan di masyarakat sebagai anti tumor pada organ pencernaan termasuk di rongga mulut, namun belum ada penelitian tentang bahan aktif yang berperan sebagai anti kanker di rongga mulut. Banyak didapatkan spesies dari genus Artemisia, sedangkan yang banyak tumbuh di Indonesia adalah spesies Artemisia vulgaris L. Tujuan penelitian ini adalah untuk menjelaskan identifikasi menggunakan Kromatografi Lapis Tipis (KLT) dari sudamala. Penelitian meliputi ekstraksi sudamala, identifikasi ekstrak sudamala, fraksinasi sudamala menggunakan Kromatografi Kolom Vakum dan identifikasi dari fraksi sudamala menggunakan Kromatografi Lapis Tipis (KLT). Ekstrak heksan sudamala yang dilakukan fraksinasi menggunakan n-heksan: etil asetat menghasilkan 11 fraksi. Fraksi n-heksan: etil asetat (3:7,v/v) dari sudamala yang teridentifikasi menggunakan Kromatografi Lapis Tipis (KLT) mengandung terpenoid.Identification of Sudamala ( Artemisia vulgaris L.) Thin Layer Chromatography. Oral squamous cell carcinoma is the type of cancer which is most frequently found in oral cavity. The primary risk factor of malignancy in oral cavity includes the habit of consuming tobacco and or alcohol. The plant sudamala (Artemisia vulgaris L.) is often used in the community as anti-tumor in digestive organ, including in oral cavity. However, there have been no studies on active ingredients playing the role as anti-cancer in oral cavity. The species are mostly from the genus Artemisia, while those generally growing in Indonesia are the species Artemisia vulgaris L. The objective of this study is to explain the identification by TLC of sudamala. The study was sudamala extraction, identification of sudamala extract, sudamala fractionation with Vacuum Column Chromatography and identification of sudamala fractionation with TLC (Thin Layer Chromatography). The result shows that the fractionation using hexane extract resulted in 11 fractions of n-hexane: ethyl acetate. The conclusion of this study is that the fraction of n-hexane: ethyl acetate (3:7, v/v) sudamala (Artemisia vulgaris L.) identified contains terpenoids compounds.

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PROFIL KROMATOGRAFI LAPIS TIPIS MINYAK ATSIRI SEREH (Cymbopogon citratus)

Viva Medika: Jurnal Kesehatan, Kebidanan dan Keperawatan ◽

10.35960/vm.v10i3.431 ◽

2019 ◽

Vol 10 (3) ◽

pp. 72-75

Author(s):

Rani Prabandari

Keyword(s):

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Mobile Phase ◽

Distillation Method ◽

Cymbopogon Citratus ◽

Citronella Oil ◽

Lemongrass Oil ◽

Silent Phase ◽

Layer Chromatography

Abstract. Rhodinol has been isolated from citronella oil. Isolation of lemongrass oil is done by using the distillation method. Samples used were citronella harvested for 6 months and 9 months. Silent phase used TLC plate slika gel F254 with a size of 2 x 10 cm and the mobile phase used was Toluen: Ethyl Acetate (93: 7). Results of Thin Layer Chromatography HRF profile obtained for experiment I (citronella harvesting age 6 months) 32, 77, 87 and Experiment II (lemongrass harvest age 9 months) respectively 25, 73, 81.

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Preliminary Phytochemical Screening and Thin Layer Chromatography Analysis of Stem Bark Extracts of African Mistletoe Parasitic on Vitellaria paradoxa, Piliostigma thonningii and Combretum fragrans

Asian Journal of Applied Chemistry Research ◽

10.9734/ajacr/2019/v3i230087 ◽

2019 ◽

pp. 1-6

Author(s):

T. Agber Cyprian ◽

Shaakaa Sewuese ◽

Linus U. Akacha

Keyword(s):

Thin Layer ◽

Ethyl Acetate ◽

Thin Layer Chromatography ◽

Heart Diseases ◽

Stem Bark ◽

Phytochemical Screening ◽

Vitellaria Paradoxa ◽

Chromatography Analysis ◽

Plant Hosts ◽

Layer Chromatography

Aim: Mistletoes are highly utilized in traditional medicine to treat different kinds of diseases such as heart diseases, diabetes and malaria, among others. The chemistry of African mistletoe is not sufficiently documented. This paper is therefore, aimed at determining the phytochemicals present in the crude extracts of mistletoe parasitic on plants that are commonly seen as hosts. Study Design: This study was designed to compare the phytochemical profiles of mistletoe stem barks obtained from different plant hosts. Place and Duration of Study: Department of Chemistry, University of Agriculture, Makurdi, Benue State Nigeria, between August and September, 2018. Methodology: Powdered stem bark of mistletoe was extracted successively with hexane, ethyl acetate and methanol. Preliminary phytochemical screening was carried out on the extracts. Thin layer chromatography (TLC) was carried out on silica gel precoated plates in 9:1 (hexane/ethyl acetate), 1:1 (hexane/ethyl acetate), and 7:3 (ethyl acetate/methanol) mobile phases for hexane, ethyl acetate and methanol extracts respectively. Results: The study revealed the presence of secondary metabolites such as alkaloids, flavonoids, tannins/phenols, cardiac glycosides, steroids and triterpenoids. It was evident from TLC analysis that mistletoes from various plant hosts contain similar chemical profile. Conclusion: We therefore debunk the claim by some herbalists that medicinal values of mistletoes vary due to host plant. This is the first time a study of this kind is reported on mistletoe parasitic on Vitellaria paradoxa Pilostigma thonningii, Combretum fragrans.

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