Thin-layer Chromatography Assay Research Articles

Unveiling the Potential of Secang (Caesalpinia sappan L.) as a Novel Tanning Agent: A Promising Alternative for the Leather Industry

The exploration of vegetable tanning's natural resources is fascinating as an alternative to basic chromium sulfate for its natural abundance and considerable environmental impact. In this work, an attempt has been made to extract vegetable tannin from Caesalpinia sappan L. bark using water as a solvent with different levels of temperatures and incubation times. The highest yield efficiency was observed at 80 °C for 4 hours. The presence of tannin content and polyphenolic compounds were screened by the phytochemical analysis using Iron (III) chloride (FeCl3), sodium hydroxide (NaOH), and ensured by the thin layer chromatography (TLC), and the Fourier transform infrared (FTIR). The extracts were then determined by percentage of extract yield, phytochemical, tannin type, moisture, total solids, and tannin characteristics. The findings indicate that the extract of Secang contains 4.98±0.28% total moisture, 95.02 ± 0.28% total solids, and 72.12 ± 2.81% total soluble solids, 52.28 ± 1.79% tannins, and 19.84 ± 1.41% non-tannin. Moreover, Fourier transform infrared (FTIR) analysis revealed a clear functional group within Caesalpinia sappan L., as evidenced by the type of tannin that is condensed. The TLC assay exhibited a considerable intensity under UV lights. It included several shades of light blue and blue, which could indicate the presence of secondary metabolites and tannin. Compared to mimosa, as the commercial standard for tanning agents in the leather industry, Caesalpinia sappan L. presents promise as an eco-friendly substitute for basic chromium sulfate and a new alternative source of vegetable tannin for the leather industry.

Exploring the Therapeutic Potential of Ougeinia oojeinensis: A Comprehensive Analysis of Phytochemicals and Pharmacological Activities

Ougeinia oojeinensis, a plant indigenous to South Asia, is gaining recognition for its potential therapeutic benefits, particularly in cancer treatment. This study offers a thorough investigation into the phytochemical composition and pharmacological activities of O. oojeinensis, focusing on its promising role in cancer therapy. Through various analytical techniques, including solvent extraction, thin-layer chromatography, and biological assays, we elucidate the presence of bioactive compounds and their antioxidant and anticancer properties. Our findings underscore the significance of O. oojeinensis as a potential source of novel therapeutic agents for cancer treatment.

Assessment of total aflatoxin and ochratoxin A in poultry feed ingredients by thin-layer chromatography and enzyme-linked immunosorbent assay.

This study was conducted to evaluate the prevalence of total aflatoxin (AF) and ochratoxin A (OTA) in poultry feed ingredients under different environmental conditions during the summer and winter seasons, while the hygiene quality of the feed ingredient was assessed through viable fungal count (VFC). A total of 288 poultry feed ingredients (n = 96 each) samples were collected from different poultry shops, which were initially analyzed for the presence of AF and OTA through thin layer chromatography (TLC) and then confirmed the contamination concentration through the enzyme-linked immunosorbent assay method. The results of the current study confirmed the incidence of contamination with AF and OTA by TLC and ELISA methods. The contamination level of AF ranged from 26.09 to 50.56 (mean = 41.22 ± 9.45) μg/kg, whereas the contamination level of OTA ranged from 50.13 to 6.21 (mean 42.60 ± 6.21) μg/kg. The contamination level of AF was found to be above the permissible level set by the Food and Drug Administration (20 μg/kg), whereas the contamination level of OTA was below the permissible limits. Moreover, the VFC values were also below the recommended level. The results showed that the association between AF, OTA, and moisture content was significant (p < 0.05). Mycotoxin contamination was significantly (p < 0.05) highest in the winter season. These findings suggested that continuous monitoring regimes might prevent mycotoxin contamination in poultry feed ingredients.

A Boron-Dependent Antibiotic Derived from a Calcium-Dependent Antibiotic.

The prevalence of drug-resistant bacterial pathogens foreshadows a healthcare crisis. Calcium-dependent antibiotics (CDAs) are promising candidates to combat infectious diseases as many of them show modes of action (MOA) orthogonal to widespread resistance mechanisms. The calcium dependence is nonetheless one of the hurdles toward realizing their full potential. Using laspartomycin C (LspC) as a model, we explored the possibility of reducing, or even eliminating, its calcium dependence. We report herein a synthetic LspC analogue (B1) whose activity no longer depends on calcium and is instead induced by phenylboronic acid (PBA). In LspC, Asp1 and Asp7 coordinate to calcium to anchor it in the active conformation; these residues are replaced by serine in B1 and condense with PBA to form a boronic ester with the same anchoring effect. Using thin-layer chromatography, MS, NMR, and complementation assays, we demonstrate that B1 inhibits bacterial growth via the same MOA as LspC, i.e., sequestering the cell wall biosynthetic intermediate undecaprenyl phosphate. B1 is as potent and effective as LspC against several Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus. Our success in converting a CDA to a boron-dependent antibiotic opens a new avenue in the design and functional control of drug molecules.

Quantification of Pomalidomide Using Conventional and Eco-Friendly Stability-Indicating HPTLC Assays: A Contrast of Validation Parameters.

High-performance thin-layer chromatographic (HPTLC) assays for pomalidomide (PMD) measurement are lacking in the published database. Furthermore, eco-friendly stability-indicating analytical assays for PMD measurement are also lacking in the published database. In order to detect PMD in commercial products more accurately and sustainably than the conventional normal-phase HPTLC (NP-HPTLC) assay, an effort was made to design and verify a sensitive and eco-friendly reversed-phase HPTLC (RP-HPTLC) assay. The silica gel 60 NP-18F254S and 60 RP-18F254S plates were used as the stationary phases for NP-HPTLC and RP-HPTLC methods, respectively. The solvent system for NP-HPTLC was chloroform-methanol (90:10 v/v). However, the solvent system for RP-HPTLC was ethanol-water (75:25 v/v). The greenness scores for both assays were measured by AGREE approach. PMD measurement was performed for both assays at 372 nm. In the 50-600 and 20-1000 ng/band ranges, the NP-HPTLC and RP-HPTLC methods were linear for PMD measurement. The RP-HPTLC assay was superior to the NP-HPTLC method for measuring PMD in terms of sensitivity, accuracy, precision, and robustness. The ability of both methods to identify PMD in the presence of its degradation products suggests that both methods have stability-indicating features. When employing the NP-HPTLC and RP-HPTLC assays, respectively, the assay for PMD in commercial capsules was 88.68 and 98.83%. The AGREE scores for NP-HPTLC and RP-HPTLC assays were calculated to be 0.44 and 0.82, respectively, suggesting an outstanding greenness characteristic of the RP-HPTLC method than the NP-HPTLC method. The RP-HPTLC method was found to be superior to the NP-HPTLC method based on these findings. Therefore, the RP-HPTLC method could be successfully applied for the determination of PMD in pharmaceutical products.

A Metasurface Plasmonic Analysis Platform Combined with Gold Nanoparticles for Ultrasensitive Quantitative Detection of Small Molecules.

Food safety related to drug residues in food has become a widespread public concern. Small-molecule drug residue analysis often relies on mass spectrometry, thin-layer chromatography, or enzyme-linked immunosorbent assays (ELISA). Some of these techniques have limited sensitivity and accuracy, while others are time-consuming, costly, and rely on specialized equipment that requires skilled operation. Therefore, the development of a sensitive, fast, and easy-to-operate biosensor could provide an accessible alternative to conventional small-molecule analysis. Here, we developed a nanocup array-enhanced metasurface plasmon resonance (MetaSPR) chip coupled with gold nanoparticles (AuNPs) (MSPRAN) to detect small molecules. As sulfamethazine drug residues in poultry eggs may cause health issues, we selected this as a model to evaluate the feasibility of using MSPRAN for small-molecule detection. The MSPRAN biosensor employed competitive immunoassay technology for sulfamethazine detection. The limit of detection was calculated as 73 pg/mL, with sensitivity approximately twice that of previously reported detection methods. Additionally, the recovery rate of the biosensor, tested in egg samples, was similar to that measured using ELISA. Overall, this newly developed MSPRAN biosensor platform for small-molecule detection provides fast and reliable results, facile operation, and is relatively cost-effective for application in food safety testing, environmental monitoring, or clinical diagnostics.

Bioactivity assays and phytochemical analysis upon Achillea filipendulina, focusing on xanthine oxidase inhibitory and antimalarial properties

BackgroundAchillea filipendulina (L.) is an important medicinal plant and an Asian species of the Asteraceae family. A. filipendulina that grows in Central and Southwest Asia. It was chosen for study with the aim of screening biologically active compounds in the essential oil of the aerial parts and the antioxidant, antimalarial and xanthine oxidase inhibitory activities of its extract. Essential oil was extracted by using Clevenger apparatus. Extraction was done by Soxhlet apparatus with solvents of n-hexane, DCM (dichloromethane) and MeOH (methanol), and then, fractionation of extracts was accomplished through various chromatographic techniques including VLC (vacuum liquid chromatography) approach and SPE (solid phase extraction) technique.ResultsApproximately 79% of A. filipendulina essential oil components were recognized by the gas chromatography mass spectrometry (GC–MS), of which monoterpenes were 63.2% the most abundant components, except for terpenes, other compounds such as fatty acids, phenols, aromatic compounds and ketones were identified, as well antimalarial activity test was done by β-hematin formation assay and xanthine oxidase inhibitory activity was accomplished by Ransod kit, then antioxidant activity was performed by DPPH (2,2-diphenyl 1-picrylhydrzyl) free radical scavenging technique. Preliminary phytochemical screening of MeOH and DCM extracts was done by TLC (thin layer chromatography) and total phenol assay. Monoterpenoids were the main component of the aerial part’s essential oil. Among different extracts, MeOH extract had significant antioxidant and xanthine oxidase inhibitory activities with RC50 (mg/mL) = 0.0707 and IC50 = 12.8741 µg/mL, respectively. Moreover DCM extract had significant antimalarial activities with IC50 (mg/mL) = 0.2511. Among the fractions, 40% and 60% Sep-Pak fraction of MeOH extract had stronger xanthine oxidase inhibitory and antioxidant activities. The 40% fraction of MeOH extract showed the strongest antioxidant and xanthine oxidase inhibitory activities, which might be related to the presence of phenolic compounds. The 80% VLC fraction of DCM extract also indicated the strong antimalarial activities.ConclusionAs this plant revealed a high content of phenols along with high antioxidant, antimalarial and xanthine oxidase inhibitory activities, it could have great medicinal value. Our results suggested the possible use of A. filipendulina in development of highly potent new pure medicinal compounds.

Tamarindus indica Extract as a Promising Antimicrobial and Antivirulence Therapy

The worldwide crises from multi-drug-resistant (MDR) bacterial infections are pushing us to search for new alternative therapies. The renewed interest in medicinal plants has gained the attention of our research group. Tamarindus indica L. (T. indica) is one of the traditional medicines used for a wide range of diseases. Therefore, we evaluated the antimicrobial activities of ethanolic extract of T. indica. The inhibitions zones, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and fractional inhibitor concentration indices (FICI) against Gram+ve and -ve pathogens were detected. The bioactive compounds from T. indica extract were identified by mass spectroscopy, thin-layer chromatography, and bio-autographic assay. We performed scanning electron microscopy (SEM) and molecular docking studies to confirm possible mechanisms of actions and antivirulence activities, respectively. We found more promising antimicrobial activities against MDR pathogens with MIC and MBC values for Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa), i.e., (0.78, 3.12 mg/mL) and (1.56, 3.12 mg/mL), respectively. The antimicrobial activities of this extract were attributed to its capability to impair cell membrane permeability, inducing bacterial cell lysis, which was confirmed by the morphological changes observed under SEM. The synergistic interactions between this extract and commonly used antibiotics were confirmed (FICI values < 0.5). The bioactive compounds of this extract were bis (2-ethylhexyl)phthalate, phenol, 2,4-bis(1,1-dimethylethyl), 1,2-benzenedicarboxylic acid, and bis(8-methylnonyl) ester. Additionally, this extract showed antivirulence activities, especially against the S. aureus protease and P. aeruginosa elastase. In conclusion, we hope that pharmaceutical companies can utilize our findings to produce a new formulation of T. indica ethanolic extract with other antibiotics.

Synthesis of biotinylated-LPG as a chemical biology tool enabling discovery of ALCAT1 modulators

As a mitochondrial signature phospholipid, cardiolipin (CL) is required for membrane structure, respiration, dynamics, fragmentation, and mitophagy. Alteration of CL by reactive oxygen species (ROS) can cause mitochondrial dysfunction, which is implicated in the pathogenesis of many diseases. The enzyme ALCAT1 (acyl-CoA: lysocardiolipin acyltransferase-1) facilitates the conversion of CL by incorporating polyunsaturated fatty acids into lysocardiolipin. Accumulating evidence suggests that overexpression of ALCAT1 is involved in pathological cardiolipin remodeling and mitochondrial bioenergetics. Few ALCAT1 modulators are reported in the literature, and the enzymatic activity was tested via a low-throughput TLC (thin layer chromatography) assay. To identify small molecule ALCAT1 inhibitors, a robust assay was needed to enable a full deck high throughput screen. Scintillation proximity assay (SPA) was the method of choice because it permits the rapid and sensitive measurement of a broad range of biological processes in a homogeneous system. A biotinylated ALCAT1 substrate was required as a chemical biology tool in developing SPA. Among a panel of phospholipids, lysophosphatidyl glycerol (LPG) was identified as the best substrate for ALCAT1. Herein we report the synthesis of biotinylated-LPG analogs with varied linker lengths and their activity towards ALCAT1.

Antimalarial drugs quality monitoring in some provinces of Viet Nam

This study was conducted between January and August, 2016 in some malarial prevalence areas of Vietnam including Dak Nong and Gia Lai provinces. Total of 114 anti-malarial medicine samples (of which 55 available antimalarials and 59 selected antibiotics for malaria treatment) were collected from public and private health facilities. All of the collected samples were screen analized by GPHF-minilab kits at National Institute of Malariology, Parasitology and Entomology (NIMPE) according to GPHF monographs for specifications of visual and physical inspections, disintergration tests and thin layer chromatography assays. None of them was suspicious as substandard quality drugs. Fifteen randomly chosen samples (13.16% of total samples) were sent to National Institute of Drug Quality Control (NIDQC) for comfirmatory tests according to United States Pharmacopoeia or Vietnamese Pharmacopoeia criteria. The results showed that: three of them (03/114, 2.63%) were found as substandard drugs. All three artesunate 50 mg tablet samples (had the same lot 012013, expiry date in March, 2016, registration number VD-13186-10, collected from private pharmacies in Dak Nong and Gia Lai) did not pass purity tests. All three substandard drug samples were antimalarials with proportion of 2.63% of total samples and 5.45% of altimalarials.

Phytochemical and Comparative Assessment of Bark and Root Extracts of Ixora barbata Roxb. ex. Sm. and Ixora coccinea L. Using High-Performance Thin Layer Chromatography Assay

The present study deals with the extraction and quantification of phytochemicals, which play a key role in the pharmaceutical world. In this article, we focused on the study of plant secondary metabolites and analyzed ethanolic and methanolic extracts of bark and roots of two species of Ixora using the highperformance thin layer chromatography technique. Alcoholic extracts of bark and root of both the plants revealed the presence of alkaloids, terpenoids, glycosides, phenols, saponins, sterols, tannins, etc., but both extracts were devoid of flavonoids and coumarins. Analysis of alkaloid content revealed the highest amount of total alkaloid content in the methanolic extract of roots of Ixora coccinea (146 mg/ gm) followed by the methanolic extract of bark of Ixora coccinea (138 mg/g). The lowest total alkaloid content was observed in the methanolic extract of bark of Ixora barbata (64 mg/g). Both ethanolic, as well as methanolic extracts of Ixora coccinea, showed higher amounts of alkaloid content when compared to Ixora barbata in similar solvents. As per high-performance thin layer chromatography analysis, the highest amount of camptothecin was recorded in ethanolic extract of root of Ixora coccinea (117 μg/ml) followed by ethanolic extract of bark of Ixora barbata (64.43 μg/ml). The lowest amount of camptothecin was observed in methanolic extract of bark of Ixora coccinea (12.64 μg/ml). The results obtained from the present high-performance thin layer chromatography analysis also revealed that the above-mentioned selected plants are a source of a number of other phytoconstituents. This is the first reported study on the high-performance thin layer chromatography analysis of Ixora barbata. The study includes quantification of camptothecin compound in both methanolic and ethanolic extracts of Ixora barbata for which there are no previously published reports.

Effective In Vitro Control of Two Phytopathogens of Agricultural Interest Using Cell-Free Extracts of Pseudomonas fluorescens and Chitosan.

A biofungicide is a natural product that can be derived from various sources such as, among others, microorganisms, higher plants, animal products, phytochemicals, semiochemicals, and antagonist microorganisms. One of the most important approaches for the production of biofungicides is the combination of biocontrol agents. This study showed the inhibition growth of Alternaria alternata and Fusarium solani treated with cell-free extracts of P. fluorescens. Using thin-layer chromatography and plate assays it was also demonstrated that the cell-free extracts of P. fluorescens contained siderophores and derivates of 4-diacetylphloroglucinol and phenazine. Moreover, the combination of cell-free extracts of P. fluorescens and chitosan [50–1.5% (v/v)] had a synergistic effect since they notably inhibited the mycelial growth of A. altenata and F. solani. Various morphological alterations to the mycelia and conidia of the treated fungi as a result of this combination were also observed. The present study could be a starting point to control other fungal phytopathogens using different cell-free extracts and chitosan as biocontrol agents.

Substance P Serum Degradation in Complex Regional Pain Syndrome – Another Piece of the Puzzle?

In a previous study, we demonstrated that the serum peptidase system might be less efficient in complex regional pain syndrome (CRPS). Since the neuropeptide substanc P (SP) contributes to inflammation in CRPS, we now investigated the metabolism of SP in CRPS specifically. An SP metabolism assay was performed in 24 CRPS patients, which constitute a subgroup of our previous investigation on BK degradation. In addition, we included 26 healthy controls (24 newly recruited plus 2 from our previous investigation), and 13 patients after limb trauma, who did not fulfil the CRPS diagnostic criteria (trauma controls, TC) were included. We adapted a thin layer chromatography assay (TLC) to quantify SP disappearance after incubation with 7.5 µL of serum. These results were compared with bradykinin (BK) metabolization to BK1-8 and BK1-5 fragments from our previous study. In addition, TC were clinically and quantitative sensory testing (QST) phenotyped; the phenotyping of CRPS patients was retrieved from our existing database. SP metabolism was less efficient in CRPS and TC patient serum vs human control (HC) serum (P < .03) suggesting reduced activity of the neutral endopeptidase (NEP) and/or the angiotensin converting enzyme (ACE). Together with the decreased occurrence of BK1-5 fragment in CRPS and TC, this suggests a reduced activation of the angiotensin converting enzyme (ACE). There was no clear clinical phenotype related to impaired SP degradation; duration of disease and gender were also not associated. Most importantly, results in TC did not differ from CRPS. Collectively, our current and previous experimental results suggest that limb trauma reduces serum peptidase metabolism of SP ex vivo, specifically serum ACE activity. However, this finding is not CRPS-specific and seems to be rather a long-term consequence of the trauma itself. PerspectiveThe experimental data from this study further support the hypothesis that impaired metabolism of inflammatory peptides potentially contribute to the development of posttraumatic pain in CRPS or limb trauma patients.

Development and validation of a high-performance thin-layer chromatography assay for the analysis of tacrolimus ointments

Development and validation of a high-performance thin-layer chromatography assay for the analysis of tacrolimus ointments

Development of a specific and sensitive high-performance thin-layer chromatography assay method for the determination of linagliptin in tablet dosage form

Development of a specific and sensitive high-performance thin-layer chromatography assay method for the determination of linagliptin in tablet dosage form

The Effect of Freeze-Drying Procedures on Frangipani Sap Chloroform Extract Quality

The water content in the frangipani sap enormously influences the product of the extraction process of natural materials. Before extraction, water content on the herbal material based sap is reduced by using the drying process. This study aimed to analyze the impact of water content reduction through freeze and non-freeze dry in the quality chloroform extract of frangipani sap. This research was designed into two groups to reduce frangipani sap water content through freeze-drying and without freeze-drying. Both groups were extracted by maceration method and chloroform solvents. The result of extraction was seen the physical quality and phytochemical screening through the Thin Layer Chromatography method (flavonoids, anthraquinone, polyphenols, alkaloids, terpenoids, and triterpenoids). Furthermore, the color intensity assay is produced from the Thin Layer Chromatography assay results and then carried out statistical tests. The results of this study are that without the freeze-drying group there is a fungus in the storage period. Both groups do not contain active ingredients flavonoids, anthraquinone, and polyphenols but contain alkaloids, terpenoids, and triterpenoids. The results of the color intensity test on the active ingredients contained showed that the freeze-drying group was higher and there were statistically significant differences in values. The results of this study indicated that chloroform extract of frangipani extract by freeze-drying showed higher quality than non-freeze drying. In the freeze-drying group, it can be found that there were no fungi and had the right color intensity. However, this study requires further study to know the compound properties quantitatively.

Taro Ice Cream: Addition of Colocasia esculenta Stem to Improve Antioxidant Activity in Ice Cream

Colocasia esculenta (C. esculenta) is a widely cultivated plant for consumption and traditional medicine. Recent evidences showed lack studies about the stem of C. esculenta as medicinal agent. Besides, ice cream is a food, which contains milk that play beneficial role as antioxidant. Addition of C. esculenta in ice cream may improve antioxidant activity by its phytochemical compounds. Thus, the aim of this nutritional food study is to determine the antioxidant activity, phytochemical compounds and nutritional minerals. The methods used were done by thin layer chromatography (TLC) test and antioxidant assays characterized using 2,2-diphenyl-1-picrylhydrazyl (DPPH). The nutritional minerals was also measured on the study. The results showed the extract had antioxidant activity that measured at 675.283pg/ml. The stem extract of C. esculenta positively contained Flavonoid, Terpenoid, Saponin, and Steroid based on TLC assays. Based on nutritional minerals test, C. esculenta contained vitamin C, vitamin E, Potassium, and % Calcium. Flavonoids, Terpenoid, Saponin, and steroid had been proved as potent treatment for oxidative stress. Vitamin C and E also had evidence as antioxidant properties. Activity of antioxidant enzymes were improved in Potassium and Calcium treatment studies. Therefore, the addition of C. esculenta in ice cream could improve the activity of antioxidant.

ANTIFUNGAL ACTIVITY OF ACTINOBACTERIA WITH A POTENTIAL TO INHIBIT RICE BLAST FUNGUS MAGNAPORTHE ORYZAE (ANAMORPH PYRICULARIA ORYZAE)

Objective: The aims of the present study were to screen the actinobacteria with high potential ability to produce secondary metabolites that have inhibitory activity against plant pathogenic fungi, Magnaporthe oryzae. Production of secondary metabolites was analysis by thin-layer chromatography and bioautography assay.&#x0D; Methods: Screening and selection of potential Streptomyces sp. morphological, cultural, physiological, and biochemical characterization of the screened isolate was carried out. Antifungal compound was confirmed by bioautography assay.&#x0D; Results: Bioautography method use in this study was found to be antifungal fraction from the crude extract. Antifungal secondary metabolites can be readily located on the plates by observing clear zones where active compounds inhibit fungal growth.&#x0D; Conclusion: The bioautography assay shows that this isolates can produce antifungal compound. Therefore, this isolate proves to be a promising microbe which can be further studied for its applications a biocontrol agent against rice blast fungi.

Assessment of Diadenylate Cyclase and c-di-AMP-phosphodiesterase Activities Using Thin-layer and Ion Exchange Chromatography.

All living cells use cyclic nucleotides as second messengers for signal sensing and transduction. Cyclic di-3',5'-adenosine monophosphate (c-di-AMP) is primarily involved in the control of bacterial and euryarcheal osmoadaptation and is produced by diadenylate cyclases from two molecules of ATP. Specific phosphodiesterases hydrolyze c-di-AMP to the linear phosphoadenylate adenosine 5'-pApA or to AMP. Different methods including high-performance liquid chromatography (HPLC), thin-layer chromatography (TLC) and ion exchange chromatography (IEX) can be used to determine activities of c-di-AMP-synthesizing and degrading enzymes. Here, we describe in detail the TLC and IEX methods adapted for characterization of the diadenylate cyclase DisA and the phosphodiesterase AtaC from Streptomyces venezuelae. TLC allows quick and easy separation of radioactive-labeled substrates and products, while IEX avoids utilization of potentially hazardous radioactive substrates and can be used as a good substitute if an HPLC system is not available. Unlike in TLC assays, samples cannot be analyzed in parallel by using the IEX assay, thus it is more time consuming.

Isolation and Identification of Terpenoid Compound from Vetiver Grass-Root (Vetiveria zizanioides Stapf) as a Repellent against Termite (Cyrptotermes sp.) through Bioactivity Assay

This study reported the isolated compound from vetiver grass-root (Vetiveria zizanioides Stapf) have bioactivity as a repellant against termites. The compounds were extracted by n-hexane solvent using the soxhletation method. The compounds were tested through bioactivity assay using wood pieces added isolated compound as a sample on the final assay, acetone as a blank, and wood pieces without treatment as a control. The result showed that the isolated compound was acquired by separation and purification in the form of colorless oil 0.0225 g. Of the thin-layer chromatography (TLC) assay using eluents of n-Hexane eluent 100%, n-Hexane : DCM 95%, and n-Hexane : Acetone 70%, were obtained a red-purple spot and Rf values which are 0.27, 0.59, and 0.91, respectively. 2-methyl butane-1-amine (C5H13N) was supposed by the structure elucidation of the isolated compound. The molecular weight was 87.1 g/mol as a precursor terpenoid based on the spectrum examination of FTIR and GC-MS. The bioactivity showed that an isolated compound has acted as a repellent against termite (Cryptotermes sp.) with % attractive (0.025 %) and attractiveness index (AI) (-0.039). A negative value of AI indicated that the compound is potential as a repellent against termites.