Thin Layer Chromatography Analysis Research Articles

Forensic Panorama of Complex Suicide Case Study from India

Complex suicides involve two separate method of self inflicted harm to take ones own life. Toxicological analysis of these medico-legal cases are encountered by the forensic analyst for chemical examination to ascertain cause of death. Statistical data for complex suicides is at present largely unrecorded though National Crime Records Bureau records suggest 53.6% by hanging followed by 25.8% deaths by poisoning in India. Compared to death by hanging, fatal cases due to pesticide intake with hanging are barely seen. Here in, we are reporting a rare case where the deceased had used this complex combination of suicide to terminate his life. A complete forensic investigation, including the scene of crime and autopsy examination, was observed wherein 44-year-old man had hung himself at his residence. Chemical examination to ascertain presence of any poison in visceral tissues to rule out intoxication was conducted. The chemical tests, Thin Layer Chromatography analysis, ATRFTIR and GC-MS instrumental analysis established the presence of pesticides (Organophosphorus and Pyrethroid) in his visceral organs. The cause of death as stated by the medical expert was anti-mortem hanging, but after detection of massive amount of pesticides in visceral tissues and post mortem blood, suggests ingestion of pesticides as well as asphyxia that led to the death of the deceased. Thus, the method of death was more toward planned complex definite suicide which is alarming and needs severe attention from various sections of society.

Influence of in vitro culture age on morphology, antioxidant activities, reserpine production, and genetic fidelity in Indian snakeroot (Rauvolfia serpentina (L.) Benth. ex Kurz.)

The present study was carried out on Rauvolfia serpentina (L.) Benth. ex Kurz., for the first time, to assess the impact of in vitro culture age on morphology, antioxidant activities, and reserpine production. Fresh in vitro multiple shoot culture was established in Murashige and Skoog semisolid medium fortified with 1 mg/l N6-benzyladenine, while another pre-established in vitro multiple shoot culture was maintained in similar medium for two years. Comparisons were made among two-year-old in vitro culture, six-month-old in vitro culture and six-month-old ex vitro plantlet, wherein six-month-old ex vitro plantlet recorded significantly longer shoots than in vitro cultures. The internode lengths of six-month-old ex vitro plantlet, six-month-old in vitro and two-year-old in vitro cultures were significantly different from each other. The leaf area of six-month-old ex vitro plantlet was highest, followed by that of the six-month-old in vitro and two-year-old in vitro cultures, respectively. Reduction of superoxide dismutase activity was recorded in six-month-old in vitro culture than six-month-old ex vitro plantlet, but was found to be comparable to two-year-old in vitro culture. Marginal increase in peroxidase activity was detected in two-year-old in vitro culture when compared to six-month-old in vitro culture. High-performance thin-layer chromatography analysis detected least reserpine accumulation from six-month-old ex vitro plantlet, whereas, a surge in reserpine was recorded from both six-month-old in vitro and two-year-old in vitro cultures. Eventually, inter simple sequence repeats and start codon targeted polymorphism primers revealed a 6.79% of genetic variation in two-year-old in vitro culture from six-month-old ex vitro plantlet and six-month-old in vitro cultures. The study revealed that for morphological traits and antioxidant activities, a small-scale effect was imposed by tissue culture condition for at least up to two years. However, for enhanced (two-fold) accumulation of reserpine, long-term in vitro culture would be of commercial importance but in the cost of certain extent of genetic variation.

Phytochemical Constituent and Antioxidant Activity Evaluation of Red Seaweed Eucheuma sp.

Oxidative stress is a condition in which there is an imbalance between production of free radicals and protective response via antioxidant system. There are endogenous and exogenous antioxidants, however as age increases, there is a reduction in endogenous antioxidant, thus the search for potential exogenous antioxidants which could be derived from natural resources are needed. Indonesia is a megabiodiversity country which has more than 30,000 species of plants and animals. Red seaweed Eucheuma sp. is one of marine macroalgae species which shows potent biological activities. This study aims to determine the phytochemical constituent and to evaluate the antioxidant activity of red algae Eucheuma sp. Seaweed Eucheuma sp. obtained from Lombok, Nusa Tenggara Barat, Indonesia, were extracted by maceration process using three solvents, n-hexane, ethyl acetate, and ethanol, sequentially. Each extract was analyzed for its phytochemical constituents by phytochemistry screening, thin layer chromatography, total phenolic content, total flavonoid content, and total triterpenoid content. Evaluation of antioxidant activity for ethyl acetate extract and ethanol extract were done using DPPH method. Phytochemical analysis of Eucheuma sp. shows positive result for steroid and triterpenoid. Thin layer chromatography analysis of the Eucheuma sp. extracts showed total of 11 phytochemical constituents. Quantitative analysis revealed that the highest value in ethyl acetate extract, with total phenolic content of 29.57 mg gallic acid equivalent/g extract, total flavonoid content of 0.54 mg quercetin equivalent/g extract, and total triterpenoid content of 1.08 mg ursolic acid equivalent/g extract. Moreover, ethylacetate extract of Eucheuma sp. demonstrated an active antioxidant activity against DPPH free radical with IC50 value of 27.96 µg/mL. Thus, ethylacetate extract of Eucheuma sp. derived from Lombok, Indonesia, has a potential to be developed as a natural antioxidant.

UV Spectrophotometric Quantification of Sudan-IV Dye in Palm Oil from Major Markets of Benin Metropolis

The deficit in the supply of palm oil in Nigeria over the years has been met in part through smuggling of the product from different sources into the Nigerian market by marketers of which some of the products have been reported to be adulterated with azo dyes. The aim of this study was to detect and quantify sudan dyes in adulterated palm oil from the open market using simple and inexpensive procedures such as analytical thin layer chromatography (TLC) and Ultra violet spectrophotometric methods respectively. Seven palm oil samples from the open markets within Benin metropolis labelled as (AUC, AUC2, IK1, IK5, OLK1, ADIT, and OB) were screened for the presence of Sudan-III and Sudan-IV dyes. Four of the samples (IK1, IK5, OLK1, ADIT) were confirmed to contain Sudan-IV dye with quantities of 22.5mg/L, 21.7mg/L, 29.5mg/L, 23.8mg/L respectively. This experiment shows that the use of easy and cheap methods such as TLC and UV-spectrophotometry can be used to detect and quantify Sudan dyes in adulterated palm oil.

Comparison of the two extraction methods of fruit pulp of Aragvadha (Cassia fistula Linn.) by HPTLC

Background: The existence of significant bioactive compounds in herbs makes them pharmacologically treasured. Aragvadha (Cassia fistula Linn.), generally known as 'golden shower tree' is popular for its medicinal properties in Ayurveda. The fruit pulp of it owns medicinal values. Each plant has certain bioactive important compounds. For the extraction of different vital compounds, there is a need of specific method of extraction, which are known as ‘panchavidha kashaya kalpana’ (five varieties of extraction) in Ayurveda. These are - Svarasa (expressed juice), Kalka (paste), Kvatha (decoction), Hima (cold infusion) and Phanta (hot infusion). From centuries, Ayurvedic physicians are using extraction method, called as 'phanta' (hot infusion) for the fruit pulp of Aragvadha as a conventional technique; Ayurvedic scriptures also recommend the 'kvatha' (decoction) extraction process. To assess the superiority of these two extraction procedures, the present work used high-performance thin layer chromatography (HPTLC) analysis of Aragvadha (Cassia fistula Linn.) fruit pulp methanolic extract for phytochemical profiling. Results: The HPTLC analysis of Aragvadha (Cassia fistula Linn.) fruit pulp methanolic extract was carried out using winCATS Planar Chromatography Manager system, and the outcomes, which were obtained as chromatograms (scanned at the wavelength of 254 nm and 366 nm) representing multiple peaks. The plant's phytochemical profile of both samples were established, and tables indicating the total count of peaks with the height, area, percent area of peaks, and Rf values. Conclusion: The study revealed that the 'kadha' (decoction) of Aragvadha (Cassia fistula Linn.) fruit pulp methanolic extract contains a rich variety of phytochemicals as compare to 'phanta' (hot infusion) which might be accountable for its therapeutic value and thus justifies superiority of the 'kadha' (decoction) extraction method.

Antihyperlipidemic and Hepatoprotective Effect of Zingiber Cassumunar Rhizome Extract in High-Fat Diet-Induced Hyperlipidemic Rats: The Role of Antioxidant Activity

Hyperlipidemia and oxidative stress are major risk factors for the onset of cardiovascular diseases, and the oxidative stress caused by high level of lipids can cause liver damage. Zingiber cassumunar has been reported to contain a high antioxidant content that may provide therapeutic advantages. The present study was to evaluate the antihyperlipidemic and hepatoprotective effects of Z. cassumunar rhizome extract (ZCRE) in high-fat diet (HFD)-induced hyperlipidemicrats model and investigate the mechanism through its effect on the endogenous antioxidant enzymes. In this study, the rhizomes of Z. cassumunar was extracted using ethanol 96% (v/v) and evaporated to get the concentrated Z. cassumunar rhizome extract (ZCRE). Thin layer chromatography (TLC)-densitometry was performed to determine the curcumin content in the extract. High fat diet-induced hyperlipidemia model was used to evaluate the anti-hyperlipidemic and hepatoprotectiveactivities of ZCRE in rats. Male Wistar rats were randomly divided into five groups: normal control; High fat diet-induced hyperlipidemic rats (HFD); High Fat Diet and 100 mg/kgBW of ZCRE (HFD + 100 mg/kgBW); High Fat Diet and 200 mg/kgBW of ZCRE (HFD + 200 mg/kgBW); and High Fat Diet and 400 mg/kgBW of ZCRE (HFD + 400 mg/kgBW). The antihyperlipidemic and hepatoprotective potential of ZCRE were assessed through a series of analyses of body weight, blood biochemical parameters, which include total cholesterol (TC), triglycerides (TG), the serum glutamic-oxaloacetic transaminase (SGOT) and serum glutamic-pyruvic transaminase (SGPT). The antioxidant activity of catalase (CAT) and glutation peroxidase (GSH-Px) were assessed on the liver homogenate. Data of the study were presented as mean ± SD and analyzed by using one way analysis of variance (ANOVA) followed by Least Significant Difference (LSD) test for multiple comparisons. The TLC analysis showed that ZCRE contained a significant amount of Curcumin. In addition, the study has also shown that ZRCE was able to significantly lower the levels of total cholesterol, triglyceride, SGPT, and SGOT as compared to hyperlipidemic rats (p <0.05). Concomitantly, the activity of CAT and GSH-Px was found significantly increased (p <0.05) when compared to hyperlipidemic control, with the dose of 400 mg/kg BW being the most effective. This study showed the significant antihyperlipidemic and hepatoprotective effects of ZCRE in HFD-induced hyperlipidemic rats, which mechanism might possibly connect to the increased antioxidant enzyme activities.

Chitosan-Based Nanogels Designed for Betanin-Rich Beetroot Extract Transport: Physicochemical and Biological Aspects.

Nanotechnology has emerged as a possible solution to improve phytochemicals' limitations. The objective of the present study was to encapsulate beetroot extract (BR Ext) within a chitosan (CS)-based nanogel (NG) designed via ionic crosslinking with tripolyphosphate (TPP) for betanin (Bet) delivery, mainly in the ophthalmic environment. BR Ext is rich in betanin (Bet) according to thin layer chromatography (TLC), UV-visible spectroscopy, and HPLC analysis. NG presented a monodisperse profile with a size of 166 ± 6 nm and low polydispersity (0.30 ± 0.03). ζ potential (ζ-Pot) of +28 ± 1 is indicative of a colloidally stable system. BR Ext encapsulation efficiency (EE) was 45 ± 3%. TEM, with the respective 3D-surface plots and AFM, showed spherical-elliptical-shaped NG. The BR Ext release profile was biphasic with a burst release followed by slow and sustained phase over 12 h. Mucoadhesion assay demonstrated interactions between NG with mucin. Moreover, NG provided photoprotection and pH stability to BR Ext. FRAP and ABTS assays confirmed that BR Ext maintained antioxidant activity into NG. Furthermore, in vitro assays using human retinal cells displayed absence of cytotoxicity as well as an efficient protection against injury agents (LPS and H2O2). NGs are a promising platform for BR Ext encapsulation, exerting controlled release for ophthalmological use.

Investigation of Chemical Components of Hexane Extract from the Stem of Arcangelisia flava via Phytochemical Test, GC-MS Analysis, FTIR Fingerprinting, and In Vitro Antioxidant Activity

Arcangelisia flava, also known as yellow root, is a woody vine indigenous to the Philippines. It has been established that it possesses medicinal properties, and several of its bioactive compounds have been identified. However, most of its possible bioactive nonpolar chemical compounds remain unidentified. In this study, the stem of A. flava was investigated for its chemical components, FTIR fingerprinting, and antioxidant activity. The stem of A. flava was chopped, ground, and then extracted with hexane per gram of A. flava. Thin-layer chromatography (TLC), phytochemical screening, GC-MS and FTIR analyses, cupric-reducing antioxidant capacity (CUPRAC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays were performed to evaluate the chemical components of A. flava hexane extract. TLC and phytochemical screening revealed the presence of some organic acids, triterpenes, sterols, essential oils, phenols, and flavonoids. Further characterization using GC-MS analysis revealed 64 chemical components classified into hydrocarbon, ketone, alcohol, aromatic compounds, aldehyde, ester, organosilicon, and other chemicals. The most abundant compound was hexacosane with a relative abundance of 43.17%. This was followed by tetracosane (8.85%), o-xylene (8.09%), and hexatriacontane (5.72%). The FTIR spectrum also revealed the presence of hydrocarbons such as alkanes and alkenes, confirming the 60% abundance of such class of compounds as revealed by GC-MS. Meanwhile, based on the CUPRAC and DPPH assay conducted, the hexane extract from the stem of A. flava had low antioxidant capacity. Based on the literature search, this is the first time the chemical composition of A. flava hexane extract has been evaluated and its chemical compounds identified. The findings presented in this paper should be taken into account for any future research on the species’ therapeutic properties.

Neuroprotective potential of Marsilea quadrifolia Linn against monosodium glutamate-induced excitotoxicity in rats.

Background: Excitotoxicity is a condition in which neurons are damaged/injured by the over-activation of glutamate receptors. Excitotoxins play a crucial part in the progression of several neurological diseases. Marsilea quadrifolia Linn (M. quadrifolia) is a very popular aquatic medicinal plant that has been utilised for a variety of therapeutic benefits since ancient times. Its chemical composition is diverse and includes phenolic compounds, tannins, saponins, flavonoids, steroids, terpenoids, alkaloids, carbohydrates and several others that possess antioxidant properties. Objective: The objective of the present study was to investigate the neuroprotective potential of M. quadrifolia against monosodium glutamate (MSG)-induced excitotoxicity in rats. Methods: A high-performance thin-layer chromatography (HPTLC) analysis of chloroform extract of M. quadrifolia (CEMQ) was conducted to identify the major constituents. Further, the in silico docking analysis was carried out on selected ligands. To confirm CEMQ's neuroprotective effects, the locomotor activity, non-spatial memory, and learning were assessed. Results and discussion: The present study confirmed that CMEQ contains quercetin and its derivatives in large. The in-silico findings indicated that quercetin has a better binding affinity (-7.9kcal/mol) towards the protein target 5EWJ. Animals treated with MSG had 1) a greater reduction in the locomotor score and impairment in memory and learning 2) a greater increase in the blood levels of calcium and sodium and 3) neuronal disorganization, along with cerebral edema and neuronal degeneration in the brain tissues as compared to normal control animals. The changes were however, significantly improved in animals which received standard drug memantine (20mg/kg) and CEMQ (200 and 400mg/kg) as compared to the negative control. It is plausible that the changes seen with CEMQ may be attributed to the N-methyl-D-aspartate (NMDA) antagonistic properties. Conclusion: Overall, this study indicated that M. quadrifolia ameliorated MSG-induced neurotoxicity. Future investigations are required to explore the neuroprotective mechanism of M. quadrifolia and its active constituents, which will provide exciting insights in the therapeutic management of neurological disorders.

Equilibrium Thermodynamics of Macropa Complexes with Selected Metal Isotopes of Radiopharmaceutical Interest.

To pursue the design of in vivo stable chelating systems for radiometals, a concise and straightforward method toolbox was developed combining NMR, isothermal titration calorimetry (ITC), and europium time-resolved laser-induced fluorescence spectroscopy (Eu-TRLFS). For this purpose, the macropa chelator was chosen, and Lu3+, La3+, Pb2+, Ra2+, and Ba2+ were chosen as radiopharmaceutically relevant metal ions. They differ in charge (2+ and 3+) and coordination properties (main group vs lanthanides). 1H NMR was used to determine four pKa values (±0.15; carboxylate functions, 2.40 and 3.13; amino functions, 6.80 and 7.73). Eu-TRLFS was used to validate the exclusive existence of the 1:1 Mn+/ligand complex in the chosen pH range at tracer level concentrations. ITC measurements were accomplished to determine the resulting stability constants of the desired complexes, with log K values ranging from 18.5 for the Pb-mcp complex to 7.3 for the Lu-mcp complex. Density-functional-theory-calculated structures nicely mirror the complexes' order of stabilities by bonding features. Radiolabeling with macropa using ligand concentrations from 10-3 to 10-6 M was accomplished by pointing out the complex formation and stability (212Pb > 133La > 131Ba ≈ 224Ra > 177Lu) by means of normal-phase thin-layer chromatography analyses.

Chemical analysis of endophytic fungi isolated from mangrove trees in Playa San Pedro Nature Reserve, Buenaventura, Valle del Cauca, Colombia

Endophytic fungi are well known for their association with a wide variety of plant species, likewise, mangrove plants are well known for harboring a vast variety of fungi with a valuable diversity of bioactive compounds originating from the secondary metabolism that is synthesized in part as a response to the chemical defense against microorganisms, hostile environments, and antagonistic insects. The objective of the present study was to analyze the chemical composition of endophytic fungi isolated from mangrove trees in Buenaventura, Colombia. Analyses of DNA sequences from the internal transcribed spacer ribosomal nuclear region (ITS) were conducted to determine the fungi’s identity. The results revealed 17 isolates, belonging to eight fungal families. All isolates were subjected to thin-layer chromatography analysis, observing different phytochemical nuclei eluted in the system (7: 3 hexane: acetone), of these, 23 compounds were recognized using gas chromatography coupled to mass spectrometry; cytotoxicity tests were carried out in human foreskin fibroblast cell line, which did not show a trend in cell viability. The selected endophytic fungi derived from mangrove trees reveal the presence of different chemical compounds, representing an alternative resource of great interest in bioprospecting and bioremediation.

Chitosanase Production from the Liquid Fermentation of Squid Pens Waste by Paenibacillus elgii.

Chitosanases play a significant part in the hydrolysis of chitosan to form chitooligosaccharides (COS) that possess diverse biological activities. This study aimed to enhance the productivity of Paenibacillus elgii TKU051 chitosanase by fermentation from chitinous fishery wastes. The ideal parameters for achieving maximum chitosanase activity were determined: a squid pens powder amount of 5.278% (w/v), an initial pH value of 8.93, an incubation temperature of 38 °C, and an incubation duration of 5.73 days. The resulting chitosanase activity of the culture medium was 2.023 U/mL. A chitosanase with a molecular weight of 25 kDa was isolated from the culture medium of P. elgii TKU051 and was biochemically characterized. Liquid chromatography with tandem mass spectrometry analysis revealed that P. elgii TKU051 chitosanase exhibited a maximum amino acid identity of 43% with a chitosanase of Bacillus circulans belonging to the glycoside hydrolase (GH) family 46. P. elgii TKU051 chitosanase demonstrated optimal activity at pH 5.5 while displaying remarkable stability within the pH range of 5.0 to 9.0. The enzyme displayed maximum efficiency at 60 °C and demonstrated considerable stability at temperatures ≤40 °C. The presence of Mn2+ positively affected the activity of the enzyme, while the presence of Cu2+ had a negative effect. Thin-layer chromatography analysis demonstrated that P. elgii TKU051 chitosanase exhibited an endo-type cleavage pattern and hydrolyzed chitosan with 98% degree of deacetylation to yield (GlcN)2 and (GlcN)3. The enzymatic properties of P. elgii TKU051 chitosanase render it a promising candidate for application in the production of COS.

Characterization and antibacterial activity of alkaloids and polyphenols extracts from Haplophyllum tuberculatum (forssk.)

Haplophyllum tuberculatum is a medicinal plant belonging to the Rutaceae family, is renowned for its various therapeutic properties. This study aims to characterize the alkaloids and polyphenol extracts from this plant and assess their antibacterial activity. Herein, the extraction of polyphenols and alkaloids from this plant was performed by the maceration method. Folin Ciacalteu’s method was used to estimate the total phenolic content, and the qualitative characterization of the two extracts was performed by thin-layer chromatography. Whilst, the antibacterial activity of the two extracts was tested with the disk diffusion method on a solid medium and the minimal inhibitory concentration (MIC) of susceptible bacteria was determined using the agar dilution method. Our results indicate respective yields of 8.39% in polyphenols and 0.37% in alkaloid extracts, while the total phenolic content was estimated to be 74.45 mg GAE/g of dry matter in polyphenolic extract. Thin-layer chromatography analysis allowed choosing the system toluene-acetate-ethanol-concentrated ammonia (40:4:8:3, v/v) to separate H. tuberculatum alkaloids, and ethyl acetate-methanol-water (100:13.5:10, v/v/) for the separation of its polyphenols. The same analysis detected traces of quercitin, catechin, and rutin in the polyphenolic extract. Our findings demonstrated good antibacterial activity on Gram-positive strains such as Staphylococcus aureus ATCC 25923 and Bacillus subtilis ATCC 6633, and moderate activity on Pseudomonas aeruginosa ATCC 27953, with MICs ranging from 0.625 to 10 mg/mL for alkaloids and from 5 to 20 mg/mL for polyphenols.

Effects of extraction temperature on phytoconstituents of leaf extracts of Azadirachta indica and its antiparasitic efficacy against Argulus sp.

Aim: To understand the effect of extraction temperatures on the phytoconstituents of dried and fresh neem (Azadirachta indica) leaf extracts and its antiparasitic efficacy (AE %) against Argulus sp., an ectoparasite of fish. Methodology: Extracts of fresh and dried neem leaf were prepared using water at three different temperatures viz. room temperature (26 °C) for 24 hr, cold extraction (4 °C) for 24 hr and hot extraction (60°C) for 15 min, respectively. The preliminary qualitative and high-performance thin liquid chromatography analysis was conducted to estimate the phytoconstituents of each extract. The antiparasitic efficacy and 6 hr median effective concentration (EC50) of each extract was estimated against Argulus sp. Results: The qualitative test revealed all the estimated phytoconstituents in aqueous dried and fresh leaf extract prepared at 26 °C. Whereas the hot extract of dried leaf lacked alkaloids, and similarly among the fresh leaf extract, the absence of saponin in the cold, and hot extract lacked terpenoids and tannin. Further, thin layer chromatography analysis revealed more phytoconstituents in dried leaf than fresh neem leaf extracts at all extraction temperatures. Significantly higher 6 hr antiparasitic efficacy of 100 % and 61 % with an estimated median effective concentration (EC50) of 2.75 and 3.24 were found for dried and fresh neem leaf extracts, respectively, extracted at 26 °C in 3.5 g l-1 compared to cold and hot extracts. Interpretation: The present study recommends room temperature as the best condition for extracting active phytoconstituents from neem leaf extracts, additionally dried neem leaf extract could be a potent argulocidal agent in the aquaculture industry Key words: Antiparasitic efficacy, Argulus sp., Azadirachta indica, Extraction temperature, Phytoconstituents

Analysis of Methamphetamine Using Necrophagous Insects For Forensic Entomotoxicology Applications

The high use of methamphetamine (MA) has resulted in many cases of overdose leading to death, where bodies are generally found after several days which complicates the investigation so that a forensic entomotoxicological analysis using insects is carried out. The use of necrophagous insects was chosen because necrophagous insects such as maggots were at the crime scene. Maggot was then analyzed using Gas Chromatography-Mass Spectrometry (GCMS) with several stages. The preliminary test was carried out using the marquis reagent which changed the color of the sample to orange if positive for methamphetamine, then continued with TLC (Thin Layer Chromatography) analysis and obtained the Rf value close to the comparative Rf value, which means that maggot positively contains methamphetamine. Maggot was extracted by maceration and sonication methods using methanol:chloroform 1:3 solvent. Finally, a confirmatory test was carried out using a GCMS and it was found that methamphetamine had a retention time of 3.554 minutes and a peak of 58.1. The results are matched with NIST (National Institute of Standards and Technology) library data. It can be concluded that the use of maggot in analyzing methamphetamine using GCMS can be realized properly

In Vitro Biofilm-Mediated Biodegradation of Pesticides and Dye-Contaminated Effluents Using Bacterial Biofilms.

Overuse of pesticides in agricultural soil and dye-polluted effluents severely contaminates the environment and is toxic to animals and humans making their removal from the environment essential. The present study aimed to assess the biodegradation of pesticides (cypermethrin (CYP) and imidacloprid (IMI)), and dyes (malachite green (MG) and Congo red (CR)) using biofilms of bacteria isolated from pesticide-contaminated soil and dye effluents. Biofilms of indigenous bacteria, i.e., Bacillus thuringiensis 2A (OP554568), Enterobacter hormaechei 4A (OP723332), Bacillus sp. 5A (OP586601), and Bacillus cereus 6B (OP586602) individually and in mixed culture were tested against CYP and IMI. Biofilms of indigenous bacteria i.e., Lysinibacillus sphaericus AF1 (OP589134), Bacillus sp. CF3 (OP589135) and Bacillus sp. DF4 (OP589136) individually and in mixed culture were tested for their ability to degrade dyes. The biofilm of a mixed culture of B. thuringiensis + Bacillus sp. (P7) showed 46.2% degradation of CYP compared to the biofilm of a mixed culture of B. thuringiensis + E. hormaechei + Bacillus sp. + B. cereus (P11), which showed significantly high degradation (70.0%) of IMI. Regarding dye biodegradation, a mixed culture biofilm of Bacillus sp. + Bacillus sp. (D6) showed 86.76% degradation of MG, which was significantly high compared to a mixed culture biofilm of L. sphaericus + Bacillus sp. (D4) that degraded only 30.78% of CR. UV-VIS spectroscopy revealed major peaks at 224 nm, 263 nm, 581 nm and 436 nm for CYP, IMI, MG and CR, respectively, which completely disappeared after treatment with bacterial biofilms. Fourier transform infrared (FTIR) analysis showed the appearance of new peaks in degraded metabolites and disappearance of a peak in the control spectrum after biofilm treatment. Thin layer chromatography (TLC) analysis also confirmed the degradation of CYP, IMI, MG and CR into several metabolites compared to the control. The present study demonstrates the biodegradation potential of biofilm-forming bacteria isolated from pesticide-polluted soil and dye effluents against pesticides and dyes. This is the first report demonstrating biofilm-mediated bio-degradation of CYP, IMI, MG and CR utilizing soil and effluent bacterial flora from Multan and Sheikhupura, Punjab, Pakistan.

Thin Layer Chromatographic Profiling, Antioxidant and Antidiabetic Activity of Selected Medicinal Plants

Diabetes mellitus is a disorder whose frequency is speedily increasing everywhere on the planet. According to the World HealthOrganization (WHO), a United Nations Agency, polygenic diseases are going to be the seventh leading reason for death in 2030. Several healthful plants like Citrullus lanatus, Trigonella foenum-graecum, and Murraya koenigii are used for the treatment of diabetes disorders in the Indian medicine system and different ancient systems of the world. These three plant leaves were collected from Vadodara and Anand, Gujarat. Thin layer chromatography profiling of all three plant leaves confirmed the presence of various common classes of phytochemicals such as rutin, gallic acid, tannic acid, and quercetin in both cities. Thin layer chromatographic analysis of a particular leaf revealed various retention factor values in the range of 0.01 to 0.97. Also, C. lanatus, T. foenum-graecum, and M. koenigii plant leaves confirmed the antioxidant and antidiabetic activity but C. lanatus from Anand city was found to be the best plant. The study will help in the future to identify this plant for further research in industries and pharmaceutical companies.

Evaluation of Marketed Rosemary Essential Oils (Rosmarinus officinalis L.) in Terms of European Pharmacopoeia 10.0 Criteria.

Various pure rosemary essential oil containing commercial products are in demand for their health-promoting and cosmetic claims in Türkiye. Although they are natural and harmless, they should be in compliance with European Pharmacopoeia (EP) criteria. Therefore, in this study, 15 rosemary oil samples sold in pharmacies, herbal shops, and online platforms in Türkiye were investigated in terms of "Rosemary Oil" EP 10.0. monograph criteria. In the current study, we aimed to evaluate the current quality status of rosemary essential oils in the Turkish market. Appearance, fatty oils and resinified essential oils, relative density, refractive index, optical rotation, and acid value tests were performed according to EP 10.0 and compared with the given standards. In addition, thin layer chromatography (TLC) and gas chromatography-mass spectrometry (GC-MS) analysis were conducted on all samples for advanced understanding of their phytochemical profile and harmony with EP standards. Fifteen pure rosemary oil-containing products from the Turkish market were evaluated. All of the samples were licensed as cosmetic products in Türkiye via the Ministry of Health. 83.1 to 96.9% of the ingredients of all samples were determined via GC-MS analysis. Results demonstrated that none of the samples from the Turkish rosemary essential oil market fully complied with the EP rosemary oil monograph standards. Considering our data, it was revealed that enhanced regulations and auditing mechanisms are needed to improve the quality of products. When the difference between the sources of purchase is assessed, pharmacies are still better locations to obtain such products.

Analytical Profiling of Saffron (Crocus sativus) using 1H-NMR and FTIR based Metabolomics approach and UV-Vis, HPTLC and TLC Chromatography Fingerprinting

Crocus sativus L. commonly known as saffron or Kesar in India, is an important medicinal herb in Ayurveda and has been traditionally used for treatment of neurological disorders, for depression, anxiety and sleep disorders. It is uses as a coloring and flavouring agent in the preparation of various foods. Modern research has high lighted its beneficial effects in treatment of cardiovascular action, diabetic cataract, and as a potent antiinflammatory herb. Due to its high cost its quality control is of utmost importance to ensure its authenticity, purity and its medicinal properties. In the preset study the we have used Thin Layer Chromatography (TLC) analysis, High performance thin layer chromatography (HPTLC) fingerprint, UV-Vis spectrophotometric analysis, 1H nuclear magnetic resonance (1H NMR) and Fourier transform infrared spectroscopy (FTIR) based metabolomic study for quality control and characterization of saffron. The antibacterial and antifungal activity were evaluated using agar well-diffusion method in two pathogenic bacterial strains, Escherichia coli, Staphylococcus aureus and two pathogenic fungal strains, Candida albicans, and Aspergillus brasiliensis. The 1H NMR spectroscopy couples with FTIR analysis leads to identifcation of the secondary metabolites of saffron like crocetin, picrocrocin and safranal on basis of reported diagonostic signals and peaks. The antimicrobial activity showed moderate antibacterial and antifungal activity. The TLC and HPTLC profile reveals the characteristic fingerprint. Overall the present study showed that the 1H-NMR, FTIR based metabolomics approach and TLC and HPTLC metabolite profiling can be powerful strategy for maintaining the holistic quality of the saffron.

Comparative Study of Densitometry and Videodensitometry for Quantitating the Active Pharmaceutical Ingredients Using Thin Layer Chromatography – Systematic Review

Background: Chromatography is one of the analytical techniques widely used for the quality control process in the pharmaceutical industry. One of the analytical methods used in drug analysis is Thin Layer Chromatography (TLC). The analysis process of TLC can be performed using densitometry (scanner) or videodensitometry (videoscan). The principal analysis of densitometry (scanner) is based on the density measured from each spot on the TLC plate using a specific wavelength range, and videodensitometry (videoscan) is performed by taking pictures of the plate using a Visualizer at a specific wavelength. Objective: This review article discusses the application of densitometry and videodensitometry methods for quantitative analysis of pharmaceutical products. Methods: This study was conducted using a systematic review method using the PRISMA statement from January to April 2023. Four databases were searched: PubMed, ScienceDirect, Scopus, and Google Scholar with inclusion criteria: studies on thin layer chromatography analysis using densitometry and videodensitometry. Results: Based on the ten articles in this study, it is known that the active ingredient concentrations in pharmaceutical products can be determined using densitometry and videodensitometry. The statistical analysis results show no significant difference between the two methods' chemical concentrations of active ingredients in pharmaceutical products. Conclusion: TLC densitometry and videodensitometry is a valid methods analysis that can be used for quantitating the active pharmaceutical ingredient concentration in finished pharmaceutical products.