GC-MS Fingerprinting Research Articles

Comprehensive characterization of volatile compounds in Iranian black teas using chemometric analysis of GC-MS fingerprints

Black tea, a widely popular non-alcoholic beverage, is renowned for its unique aroma and has attracted significant attention due to its complex composition. However, the chemical profile of Iranian tea remains largely unexplored. In this research, black tea samples from key tea cultivation regions in four geographical areas in northern Iran were firstly analyzed using headspace solid-phase microextraction followed by gas chromatography–mass spectrometry (HS-SPME-GC–MS) to separate, identify, and quantify their volatile organic compounds. Subsequently, employing a robust investigative strategy, we utilized for the first time the well-known multivariate curve resolution-alternating least square (MCR-ALS) method as a deconvolution technique to analyze the complex GC–MS peak clusters of tea samples. This approach effectively addressed challenges such as severe baseline drifts, overlapping peaks, and background noise, enabling the identification of minor components responsible for the distinct flavors and tastes across various samples. The MCR-ALS technique significantly improved the resolution of spectral and elution profiles, enabling both qualitative and semi-quantitative analysis of tea constituents. Qualitative analysis involved comparing resolved peak profiles to theoretical spectra, along with retention indices, while semi-quantification was conducted using the overall volume integration (OVI) approach for volatile compounds, providing a more accurate correlation between peak areas and concentrations. The application of chemometric tools in GC–MS analysis increased the number of recognized components in four tea samples, expanding from 54 to 256 components, all with concentrations exceeding 0.1 %. Among them, 32 volatile compounds were present in every tea sample. Hydrocarbons (including alkenes, alkanes, cycloalkanes, monoterpenes and sesquiterpenes), esters and alcohols were the three major chemical classes, comprising 78 % of the total relative content of volatile compounds. Analyzing black teas from four distinct regions revealed variations not only in their volatile components but also in their relative proportions. This integrated approach provides a comprehensive understanding of the volatile chemical profiles in Iranian black teas, enhances knowledge about their unique characteristics across diverse geographical origin, and lays the groundwork for quality improvement.

GC-MS Fingerprinting of Methanolic Extract of Moringa oleifera Stem, Leaf and Root

GC-MS Fingerprinting of Methanolic Extract of Moringa oleifera Stem, Leaf and Root

GC-MS fingerprinting, nutritional composition, in vitro pharmacological activities and molecular docking studies of Piper chaba from Uttarakhand region.

The online version contains supplementary material available at 10.1007/s13205-024-03996-7.

Ethnobotany and Phytochemical Composition of Alchornea Cordifolia in Abia State Nigeria

Alchornea cordifolia is one of the medicinal plants used in treatment of several medical challenges in Nigeria. The study was carried out to document the medicinal values of different parts of A. cordifolia in Ekebedi Oboro ancient Kingdom, and profile its bioactive constituents. The ethno-botanical survey involved oral interview of 81 randomly selected traditional medicine doctors and natives on medicinal values of A. cordifolia. The survey showed that A cordifolia is a major herb used in the treatment of various diseases in the ancient Kingdom and it is prescribed in different treatment regimens; coughs, bronchitis, post-partum bleeding, lacerations, wounds, vaginitis, diarrhea, sickle cell anaemia, venereal diseases including gonorhhea, and syphilis, and prostatitis. GC-MS fingerprinting was adopted to determine the bioactive constituents of methanol leaf extract of the plant. Phytochemical fingerprinting revealed the presence of several bioactive compounds as the major active ingredients of the plant. A total of 27 fatty acids and volatile hydrocarbons were detected in the leaf extract of the plant with 9, 12-Octadecadienoic acid (Z,Z)- methyl ester (18.42%), 9-Octadecanoic acid methyl ester (19.93%), Dodecanoic acid 1,2,3-propanetrieyl (15.87%) as most abundant ingredients. These phytochemical compounds of A. cordifolia possibly underscored its antimicrobial and therapeutic action and explains why natives use them in traditional medicine.

Phytochemical Investigation of Alysicarpus vaginalis by Gas Chromatography-mass Spectrometry and High-performance Thin-layer Chromatography

Introduction: Alysicarpus vaginalis belongs to the Fabaceae family, commonly known as buffalo clover and one-leaf clover. Alysicarpus vaginalis is medicinally used in disease-related kidney, diuretics, leprosy, pulmonary problems, jaundice, skin problems, respiratory difficulties, and as a hepatoprotective. Alysicarpus vaginalis is a medicinally important plant according to our review of literature on pharmacological activities, modern pharmacological activities, ethnomedicinal surveys, and traditional systems. However, very limited research related to phytochemical investigation by chromatographic techniques was studied. Methods: So, to investigate the phytochemicals of Alysicarpus vaginalis methanol extract by High-performance thin-layer chromatography and gas chromatography-mass spectrometry. Result: Phytochemical screening revealed the presence of steroids, flavonoids, polyphenols, terpenoids, carbohydrates, alkaloids, and tannins. GC-MS revealed, the presence of six out of nine phytochemicals found medicinally important. 4-O-Methylmannose, Undecane, Neophytadiene, 2-amino-5-[(2-carboxy) vinyl]- Imidazole, Lup-20(29)-en-3-one and stigmasterol are some of the important phytochemical useful as anticancer or as anti-inflammatory. A solvent system was developed by thin-layer chromatography, followed by the HPTLC analysis using the mobile phase toluene: methanol: ethyl acetate (7:3:1). High-performance thin-layer chromatography fingerprinting detected total of 8 peaks with Rf value of 0.007, 0.089, 0.142, 0.211, 0.468, 0.585, 0.772, and 0885 at 254nm and total of 9 peaks with Rf value of 0.007, 0.089, 0.140, 0.208, 0.468, 0.661, 0.772, 0887, and 0.940 at 366nm, respectively. Conclusion: Chemoprofiling by sophisticated spectral techniques such as GC-MS and HPTLC fingerprinting can serve as an important tool for quality control of assuring purity, safety, quality, and efficacy of herbal formulation, and herbal extracts from Alysicarpus vaginalis.

Anti-diabetic effect of <i>Eucalyptus camaldulensis</i> (Red gum) leaf-supplemented diet in streptozotocin-induced diabetic rats

Purpose: To investigate the potential anti-diabetic effect of supplementing a diet with Eucalyptus camaldulensis leaves in streptozotocin (STZ) induced Wistar rats. 
 Methods: Methanolic extract of E. camaldulensis was screened using gas chromatography-mass spectrometric analysis. After that, eighteen animals were separated into three groups: Group A served as control group. Group B, the diabetic control group, was induced with STZ, and Group C was induced with STZ and fed a 10 % E. camaldulensis leaf-supplemented diet for 14 days. Thereafter, rats were sacrificed, and fasting blood was collected for serum glucose, enzyme and protein activity tests. Organs were excised for biochemical and histological analysis. 
 Results: The GC-MS fingerprint identified 17 constituents, with 2-hydroxy carbazole being the most abundant. Rats on the E. camaldulensis leaf-based diet for 14 days exhibited a significant decrease (p < 0.05) in serum glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST), amylase, albumin, total bilirubin, urea, and creatinine concentrations compared to diabetic controls. Insulin levels significantly increased (p < 0.05). Additionally, the E. camaldulensis diet led to a significant decrease in serum lipid profile levels but increased high-density lipoprotein cholesterol (HDL-C). Superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities increased significantly, while malondialdehyde levels decreased significantly (p < 0.05). Histopathology revealed positive effects on hepatocytes, acini, pancreatic duodenal lymphoid activation, islets of Langerhans resurgence, and normal kidneys. 
 Conclusion: Eucalyptus camaldulensis leaf-supplemented diet demonstrates anti-diabetic, hypolipidemic and antioxidant enzyme activities. Carbazole identified by GC-MS may be the potential anti-diabetic and hypolipidemic agent in E. camaldulensis leaf-based diet.

Imaging metabolic mechanisms and the binding behavior of nutrients/transporters of edible Matricaria flowers VOCs

To promote the consumption of flowers and to utilize the nutritional value of proteins, the efficacy of the beneficial components of flowers has been intensively studied. Anthemis nobilis was used as the study object, and all its volatile components (VOCs) were fingerprinted using headspace solid-phase micro-extraction gas-mass spectrometry (HS-SPME/GC–MS). GC–MS fingerprints of five parts of Anthemis nobilis were established using three proteins, bovine lactoferrin (BLF), bovine lactoglobulin (β-Lg), and human serum albumin (HSA), as nutrient transporters. The interactions between the volatile components from different parts of the mother chrysanthemum plant and the nutrient/transport proteins were investigated. The results of fingerprinting showed that the flavor components were dominated by alkenes. In addition, this study revealed that among the three nutrient transporters, the strongest binding to the adsorbed volatile components was HSA, followed by BLF, and β-Lg was second. In addition, a characteristic molecule, camphene, was screened. Integrated molecular simulation using fluorescence spectroscopy was used to validate the results of the interaction of the nutrient/transport proteins systems with characteristic molecule. The properties of the characteristic molecules such as absorption, distribution, metabolism, excretion and toxicity in vivo were analyzed using ADMET to provide a theoretical basis for the preparation of flower-flavored dairy products.

GC-MS fingerprinting and antibacterial activity of in vitro raised plant parts of Solanum khasianum Clarke

GC-MS fingerprinting and antibacterial activity of in vitro raised plant parts of Solanum khasianum Clarke

GC-MS and HPTLC Analysis of Ocimum basilicum Ethanolic extract obtained from five locations of Tamilnadu

Background/objective: The purpose of this study is to analysis the bioactive compounds present in the medicinal plant Ocimum basilicum (whole plant) which was obtained from the five locations of Tamil Nadu. Methods: The whole plant of Ocimum basilicum was shade dried and it was extracted with ethanol. GC-MS and HPTLC fingerprints profiles were performed in the extract to find out the phytochemical constituents and different bioactive compounds with the help of retention time and peak value of the respective samples. Results: Forty bioactive compounds with their peaks and their low and high molecular weight were identified with the help of GC-MS analysis for all the five samples. HPTLC method was performed with the solvent system (i.e) Toluene: Ethyl acetate: Formic acid (6:5:1) and observed under UV-Vis 254nm and 366nmutilizing CAMAG REPROSTAR3 and flavonoid was quantified. Conclusion: The present study result of whole plant Ocimum basilicum ethanolic extract indicated the presence of various secondary metabolites and it is expected to exhibit healing properties.

Identification of secondary metabolites, antioxidant activity, and GC-MS analysis of <i>Launaea procumbens</i> (Roxb.) Amin (Angiosperm: Asteraceae): A neglected plant with therapeutic value

Although <i>Launaea procumbens</i> (Roxb.) Amin is not currently used by pharmacists; it has great potential for future medicinal use. According to both Ayurvedic texts and modern research, the leaves of Launaea procumbens have galactagogue, diuretic, antifungal, anorexic, anti-arthritic, and hepatoprotective properties. In this communication, various sections of <i>L. procumbens</i>, a member of the Asteraceae (Compositae) family is characterized pharmacognostically. Extracts were tested qualitatively for various constituents, revealing the presence of lignans, cardiac glycosides, alkaloids, phenolic compounds, tannins, steroids, and flavonoids. The leaves and stems were extracted using four different solvents: methanol, chloroform, distilled water, and petroleum ether, with methanol and chloroform extracts showing more significant results than petroleum ether and distilled water extract. These observations will aid in identifying and standardizing the drug in its crude form and distinguishing it from adulteration. This work aims to develop standardization parameters for <i>L. procumbens</i> leaves and stems by evaluating its pharmacognostics and conducting preliminary phytochemical screening. Additionally, GC-MS fingerprinting of the leaves revealed the presence of nine different chemical constituents, which could be useful for differentiating <i>L. procumbens</i> from other species of the genus <i>Launaea</i> in the herbal industry.

Chemical index components and quality control of Traditional Chinese Medicine: “Never change a winning team”? -A case study of volatile oil from Bupleuri radix

Chemical index components, especially those defined as quality control (QC) markers through spectrum-effect relationship approach, are commonly suggested and adopted as indicator for quality control of Traditional Chinese Medicines (TCMs). However, are chemical index components and quality control of TCMs “never change a winning team”? In this study, under the ponderation of the applicability of QC markers strategy, spectrum-effect relationship and OPLS-DA between GC×GC-MS fingerprint and inhibitory effect on the expression of extracellular secretory TNF-α of volatile oil from Bupleuri radix (BVO) was studied with the purpose of discovery of QC markers and establish a bioactive compounds-based QC method. 290 compounds of BVO were identified by GC×GC-MS. Besides, BVO had significant inhibitory effects on the expression of extracellular secretory TNF-α in a dose-dependent manner. The potency of different batches of BVOs could be distinguished with this bioassay-based method, which has been validated in terms of intermediate precision, repeatability, linearity, range and credibility tests. The QC markers of BVO were investigated by Spearman’s correlation test and OPLS-DA. It is regrettable that there were no ideal QC markers of BVO could be found. In conclusion, quality control method relayed on chemical QC markers is not feasible for TCMs with complex composition but lack of ingredients that dominate in content, just like BVO. Alternatively, a bioassay-based method established in our study is suitable for quality control of BVO.

Discovery of quality markers in the rhizome of Atractylodes chinensis using GC–MS fingerprint and network pharmacology

BackgroundThe rhizome of Atractylodes chinensis (RAC), also known as “Cangzhu”, is a well-known Chinese herbal medicine (CHMs). Due to the scarcity of wild resources, cultivated RAC and counterfeit products derived from the rhizome of Atractylodes japonica (RAJ) are also prevalent on the market. Diverse origins lead to varying quality. At present, the Chinese Pharmacopoeia (2020 edition) uses atractylodin as the only quantitative index component for RAC, and there is a lack of research on quality markers (Q-markers) that are closely related to efficacy. PurposeThis work aims to discover the Q-markers of RAC based on chemical and pharmacological strategies. MethodsThe chemical composition of RAC was analyzed and identified through gas chromatography-mass spectrometry (GC–MS). The ″Chromatographic Fingerprint Similarity Evaluation System for Traditional Chinese Medicine″ was utilized to assess the similarity of the samples and identify the characteristic peaks. Network pharmacology was employed to screen targets and pathways for characteristic components of the RAC, construct “active ingredients–targets–pathways” network diagrams, and predict the Q-markers. The Q-markers for RAC were identified through an integrated analysis of the chemical components, network pharmacology, pharmacological activities and clinical applications. In addition, a preliminary evaluation of 28 batches of “Cangzhu” from different origins were conducted by GC–MS and chemical pattern recognition. ResultsA total of 16 specific components of RAC were identified by GC–MS fingerprint. The network pharmacology study found that these 16 components primarily function through 29 core targets and 23 pathways. Three compounds, hinesol, atractylon, and β-eudesmol were identified as the potential Q-markers for RAC. The evaluation of 28 batches of “Cangzhu” showed that these three Q-markers could effectively distinguish RAC from RAJ, and RAJ could not be used as a substitute for RAC. There was no significant difference between cultivated RAC and wild RAC, and the quality of cultivated RAC was more uniform than wild RAC. ConclusionThis study identified and verified three potential Q-markers for RAC by using a comprehensive strategy combing of “GC–MS fingerprinting – network pharmacology – chemical pattern recognition”. It provides a basis for the establishment of a more reasonable RAC quality evaluation system. Furthermore, it provides a reference for the discovery of Q-markers in other CHMs.

Chemical profile of common varieties of Allium cepa L. sold in FCT-Abuja, Nigeria

Onions (Allium cepa L.) belong to the family of vegetables and herbs with strong antioxidants properties that combats the formation of free radicals and other microbial based disease conditions. The study aimed at investigating chemical profile of common varieties of Allium cepa L. sold in FCT-Abuja, Nigeria. The samples were randomly purchased at point of sales, processed and analyzed for Phytochemical, antioxidant, mineral elements, microbial activities and GC-MS fingerprints using standard methods. Results indicated presence of alkaloid, flavonoids and tannin; antioxidant inhibitory concentrations (IC50) range of 1.496 - 2.961 mg/mL and presence of Ca, Mg, Cu, Cr and Fe were observed. The GC-MS fingerprints identified 17 compounds with retention times ranging from 3.09 to 9.77 minutes. The presence of flavonoids, alkaloids and tannins corroborated report from the earlier studies of white onions. Inhibitory concentrations of the antioxidants obtained are lower than values reported from previous similar studies. The levels of the mineral contents of all the samples are within the safe limits in terms of mineral overloads and contamination, while the GC-MS fingerprints indicated nine (9), eight (8) and seven (7) compounds in the red, white and brown varieties respectively. The allyl group from organosulfur compounds is responsible for the antimicrobial activities. All the varieties of Allium cepa L analyzed contained alkaloid, flavonoids, free from heavy metals contamination and also had potent antimicrobial activities due to the presence of the 17 compounds revealed by the GC-MS fingerprints which was not previously reported. The findings of the present study has therefore contributed to additional knowledge on the existing database on Nigerian cultivars of Allium cepa .

HPTLC and GC–MS finger-printing of two potential multifunctional siddha tailams: Mathan and maha megarajanga tailam

The Siddha system of medicine is an ancient medical lineage that is practiced primarily in the southern part of India. Siddha system of medicine has been in practice for thousands of years with documented evidence dating back to the 6th century BCE. According to siddha system of medicine’s basic fundamental principle, the human body is made up of 96thathuvam(primary components), which encompass physical, physiological, psychological, and intellectual aspects. Medicine (marunthu) is classified as a wide range of internal and external medicines. The major components of its medical formulations include plant parts, minerals and animal products. Various methods were carried out for the purification process to eliminate the toxins.Choornam, Guligai, Tailam, Parpam, Chendooram, Kattu, Pasai and Poochuare the most common medicines used in Siddha system of medicine for the treatment of various diseases. The pathophysiological classification of diseases is elaborated in detail in the classical Siddha literature. Siddha system of medicine plays an important role in protecting people from diseases such as COVID-19 by providing immune-protecting and immune-boosting medicines in today's world.Mathan tailam and maha megarajanga tailam are the two unique preparations used widely for various skin diseases including chronic wounds and burns. Scientific validation of both medicines will help in understanding their effectiveness against a typical wound condition. In the present study physio-chemical and phytochemical, HPTLC, and GC–MS analyses were carried out and discussed in detail on the multifunctional properties exhibited in the patient communities.

GC–MS Fingerprinting and Antibacterial Activity of Solanum khasianum Stem and Fruit Extracts

GC–MS Fingerprinting and Antibacterial Activity of Solanum khasianum Stem and Fruit Extracts

Cuscuta reflexa Possess Potent Inhibitory Activity Against Human Malaria Parasite: An In Vitro and In Vivo Study

Drug resistance to practicallyallantimalarialdrugs in use necessitate the development of new chemotherapeutics against malaria. In this aspect, traditionally used plants with folklore reputation are the pillar for drug discovery. Cuscuta reflexa being traditionally used in the treatment of malaria in Odisha, India we aimed to experimentally validate its antimalarial potential. Different solvent extracts of C. reflexa or column fractions from a promising solvent extract were evaluated for in vitro anti-plasmodial activity against Plasmodium falciparum strain Pf3D7. Potent fractions were further evaluated for inhibition of parasite growth against different drug resistant strains. Safety of these fractions was determined by in vitro cyto-toxicity, and therapeutic effectiveness was evaluated by suppression of parasitemia and improvement in survival of experimental mice. Besides, their immunomodulatory effect was investigated in Pf-antigen stimulated RAW cells. GCMS fingerprints of active fractions was determined. Column separation of methanol extract which showed the highest in vitro antiplasmodial activity (IC50 = 14.48μg/ml) resulted in eleven fractions, three of which (F2, F3, and F4) had anti-plasmodial IC50 ranging from ≤ 10 to 2.2μg/ml against various P. falciparum strains with no demonstration of in vitro cytotoxicity. F4 displayed the highest in vivo parasite suppression, and had a mean survival time similar to artesunate (19.3 vs. 20.6days). These fractions significantly modulated expression of inflammatory cytokines in Pf-antigen stimulated RAW cells. The findings of the study confirm the antimalarial potential of C. reflexa. Exploration of phyto-molecules in GCMS fingerprints of active fractions is warranted for possible identification of lead anti-malarial phyto-drugs.

GC-MS fingerprinting and in silico molecular interaction of antioxidant agents from A. malabarica as tyrosinase inhibitors

Free radicals are generated by the chemical reaction of oxidation and they set off chain reactions that damage cells. In this study, the antioxidant activity of Anisomeles malabarica methanolic leaves extract were examined by DPPH and ABTS radical scavenging assays (in vitro). Molecular docking (in silico) using SeeSAR-9.2 was performed to find out the possible antioxidant phytoconstituents that inhibit the tyrosinase receptor (3NM8) from A. malabarica. Physicochemical and ADMET properties of phytocompounds were evaluated using StarDrop software. About 6 phytoconstituents were identified from A. malabarica methanolic leaves extract by using GC-MS analysis. DPPH and ABTS free radical scavenging activities of leaves extract reveal dose dependent and IC50 values of 81.28 ± 0.02 and 81.49 ± 0.15 μg/ml and the standard L-ascorbic acid showed 52.19 ± 0.05 and 66.46 ± 0.23 μg/ml respectively. Molecular docking studies report that 2,5-ditert-butylphenol and phytol had stronger binding efficiency on tyrosinase enzyme. Physicochemical and ADMET analysis of phytocompounds showed that 2,5-ditert-butylphenol had higher druglikeness properties.

Simultaneous component analysis as an interesting preliminary data analysis method in GC-MS – An example of headspace screening of Polish grasses

Abstract70 species of grasses family (Poaceae), coming from genera: Agrostis, Alopecurus, Anthoxanthum, Apera, Arrhenatherum, Avena, Brachypodium, Briza, Bromus, Calamagrostis, Corynephorus, Cynosurus, Dactylis, Danthonia, Deschampsia, Digitaria, Echinochloa, Elymus, Eragrostis, Festuca, Glyceria, Helictotrichon, Hierochloe, Holcus, Hordeum, Koeleria, Leymus, Lolium, Milium, Molinia, Nardus, Panicum, Phalaris, Phleum, Phragmites, Poa, Saccharum and Setaria, collected mostly from natural stands in Poland during 2020 season, were subjected to GC-MS fingerprinting of headspace volatile fraction above dried material. Obtained mass spectrometry data were analyzed by means of principal component analysis (PCA) and hierarchical cluster analysis (HCA). Five species: Glyceria maxima (Hartm.) Holmb., Lolium multiflorum Lam., Hordeum jubatum L., Bromus tectorum L. and Bromus secalinus L. were identified as outliers, which is consistent with our earlier analysis by thin layer chromatography. These species deserve further look and their outliance is orthogonal to coumarin content, which was independently observed for odorant species of grasses.

GCMS fingerprints and phenolic extracts of Allium sativum inhibit key enzymes associated with type 2 diabetes

ObjectivesInhibition of carbohydrate digestion enzymes (α-amylase and α-glucosidase) has been reported in studies as a therapeutic approach for the management or treatment of type 2 diabetes mellitus, owing to its potential to decrease postprandial hyperglycemia. The anti-diabetic potential of Allium sativum (also known as garlic) against diabetes mellitus has been established. Therefore, in this study, we assessed the antidiabetic potential of A. sativum using in vitro enzyme assays after which we explored computational modelling approach using the quantified GC-MS identities to unravel the key bioactive compounds responsible for the anti-diabetic potential. MethodsWe used in vitro enzyme inhibition assays (α-amylase and α-glucosidase) to evaluate antidiabetic potential and subsequently performed gas chromatography–mass spectroscopy (GC-MS) to identify and quantify the bioactive compounds of the plant extract. The identified bioactive compounds were subjected to in silico docking and pharmacokinetic assessment. ResultsA. sativum phenolic extract showed high dose-dependent inhibition of α-amylase and α-glucosidase (p < 0.05). Interestingly, the extract inhibited α-glucosidase with a half maximal inhibitory concentration of 53.75 μg/mL, a value higher than that obtained for the standard acarbose. Docking simulation revealed that morellinol and phentolamine were the best binders of α-glucosidase, with mean affinity values of −7.3 and −7.1 kcal/mol, respectively. These compounds had good affinity toward active site residues of the enzyme, and excellent drug-like and pharmacokinetic properties supporting clinical applications. ConclusionsOur research reveals the potential of A. sativum as a functional food for the management of type 2 diabetes, and suggests that morellinol and phentolamine may be the most active compounds responsible for this anti-diabetic prowess. Therefore these compounds require further clinical asessment to demonstrate their potential for drug development.

Standardization of the Leaves of Ajuga remota Benth

Herbal medicines are extensively utilized across the world. In Ethiopia, people use Ajuga remota Benth to treat high blood pressure, hyperlipidaemia, diabetes, malaria, fever, ascariasis, stomach pain, diarrhoea, evil eye, and retained placenta. However, work has not been done on standardizing the herb. Thus, the aim of this study was to standardize the leaves of A. remota. Fresh leaves, powdered leaves, 80% methanol, chloroform, and methanol extracts of leaves of A. remota were evaluated using macroscopic and microscopic characterization, fluorescence analysis, physicochemical characterization, phytochemical analysis and residual analysis. Macroscopically, fresh leaves of A. remota were 17 - 62 mm (length) × 7.5 - 23 mm (width); 1 - 2 mm thick and dark sea green coloured, soft, fibrous, had a very bitter taste and a fruity odour. Microscopic characterization showed turgid guard cells, anisocytic stomata, epidermal cells (stratum spinosum), parenchymatous (aerenchyma) cells, pointed trichomes, spiral tracheids, and xylem fibre in powdered leaves of A. remota. The powder of the plant gave characteristic colours in fluorescence analysis under visible and 366 nm UV light. The powder was also studied for swelling index (2.392), foaming index (100), bitterness value (8400), extractive value (aqueous: 34.33% and methanol: 21.16%), moisture content (6.43%), total ash value (27.10%), water-soluble ash (11.43%), acid-insoluble ash (7.38%), tannin content (0.016%), and powder fineness (moderately fine). TLC, LC-MS, GC-MS, and FT-IR fingerprinting were produced. Ajugarin I, dodecyl acrylate, and hexacosane were identified as analytical marker compounds. In the residual analysis of the leaves of A. remota, soil took the highest percent (0.83%) among foreign matters. Heavy metals (Cd, Co, Pb, Cr, Ni, Cu, Zn, Fe, and Mn) and aerobic bacterial count, mould, yeast, coliform, Escherichia coli, Salmonella and Shigella species were quantified. 228Ac (7 ± 1.8 Bq kg-1) and 40K (530 ± 34 Bq kg-1) radionuclides with a low dose were detected. The findings provide the standards for identification and authentication of the plant.