Marker Compounds Research Articles

Phytoconstituents of Nymphaea rubra flowers and their anti-diabetic metabolic targets

Nymphaea rubra (N. rubra) flowers are prevalent in subtropical regions for both dietary and traditional medicinal purposes, attributing to their beneficial properties in supporting overall health. This study first time provides descriptions of the antidiabetic and dyslipidemic properties employing STZ induced high fat diet fed diabetic rats and inhibition of α-amylase enzyme activity first by in vitro analyses, followed by a confirmatory in silico study to create a stronger biochemical rationale. Furthermore, in 3 T3-L1 cells, this extract promoted the suppression of adipogenesis. GC–MS investigation of the ethyl acetate fraction of ethanolic extract of N. rubra flowers revealed the presence of marker compounds of N. rubra, Nuciferine, and Apomorphine, which were the focus of molecular docking studies. The acquired concentrations of Nuciferine (22.39%) and 10, 11-dimethoxy-Apomorphine (1.47%) were detected. Together with other alkaloids identified by GC–MS analysis from this extract, mechanistically suggested that it might be caused by the synergistic impact of these bioactive chemicals. Molecular docking has been done to check the binding affinities of various isolated phytochemicals with HPAA, the dose-response effect of 100 mg/kg and 250 mg/kg of flower extract after 30 days showed a significant effect on body weight, food, water intake, serum insulin, FBG, OGTT, lipid profile, glycated haemoglobin, liver and kidney function test. Kidney histopathology results show a significant effect. These findings offer a strong foundation for the potential application of the ethyl acetate fraction of ethanolic extract from Nymphaea rubra flowers and its bioactive constituent in an in vivo system for the treatment and control of diabetes and its associated condition dyslipidemia.

Risk of Enzyme- and Transporter-mediated Drug Interactions With Drugs Approved by the US Food and Drug Administration in 2022: A Detailed Analysis of In Vitro and Clinical Data Available in New Drug Application Reviews

PurposeThis analysis aimed to provide mechanistic understanding and clinical relevance of pharmacokinetic drug-drug interactions (DDIs) associated with drugs approved by the Food and Drug Administration in 2022. MethodsDrug metabolism, transport, and DDI data available in New Drug Applications (NDAs) of small molecular drugs approved (n = 22) was analyzed. The mechanism and clinical magnitude of these interactions were characterized based on in vitro, in silico, and clinical data. FindingsAs victims, 10 drugs were identified as clinical substrates. Of these, 7 drugs were substrates of CYP3A, including the sensitive substrates daridorexant and mitapivat. As perpetrators, 3 drugs (adagrasib, lenacapavir, and vonoprazan) were clinical inhibitors of CYP enzymes, and 2 drugs (mavacamten and mitapivat) showed induction. Regarding transporter data, abrocitinib and deucravacitinib were found to be substrates of OAT3 and P-gp/BCRP, respectively, and 4 drugs (abrocitinib, adagrasib, lenacapavir, and oteseconazole) were found to inhibit P-gp and/or BCRP. As expected, all clinical DDIs with AUC changes ≥ 2-fold triggered label recommendations. Over half of DDIs with an AUC change < 2 also had label recommendations, pertaining most often to the concomitant use of drugs with a narrow therapeutic index. Overall, CYP3A played a major role in the drug disposition of the drugs approved in 2022, mediating all strong drug interactions. ImplicationsThe mechanistic information obtained from studying these new therapeutics with marker compounds can be extrapolated to common concomitant medications sharing the same pharmacokinetic properties, enhancing the safe and effective administration of these products in situations of polytherapy.

Development of Simultaneous HPTLC Method and Validation for the Quality Assessment of Ayurvedic Formulation-Ayush Kvatha Churna by Using Marker Compound Rosmarinic Acid, Trans-Cinnamaldehyde and Piperine.

Ayurveda emphasizes the propagation of nature in maintaining health. In the present scenario, we have seen the faith of people in herbal drugs during the Covid 19 outbreak. The raises in the number of peoples have been using herbal drugs to boost immunity against infectious diseases shows the popularity of this ancient system of medicine. The standardization of Ayush Kvatha Churna (AKC), work set out to establish a straightforward, accurate and sensitive HPTLC method for the identification and quantification of marker compounds. The Rosmarinic acid, trans-Cinnamaldehyde and Piperine were used for the estimation of markers in Ayush Kvatha Churna by using HPTLC with a solvent system, consisting of Toluene: Ethyl acetate: Ethyl alcohol: Formic acid (5.6:2.4:2: 0.3v/v/v/v). The Rf value 0.33 for Rosmarinic Acid, 0.69 for Piperine and 0.77 for trans-Cinnamaldehyde was observed and it is exactly complying with the corresponding bands in Ayush Kvatha Churna. The technique has been effectively verified and validated, enabling it to be used for the standardization or quantitative analysis of Rosmarinic acid, trans-Cinnamaldehyde and piperine in Ayush Kvatha Churna.

Ultra-high performance liquid chromatography-tandem mass spectrometry assay for simultaneous determination of 22 marker compounds in traditional herbal medicine Ojeoksan

Traditional herbal medicines (THMs) have long been in use worldwide and are considered safe for use as tonics or complementary treatments for many diseases. Advanced quality control methods for THMs are required in the regulatory framework of modern medicines. In this study, an ultra-high performance liquid chromatography-tandem mass spectrometry assay was established for the simultaneous determination of 22 marker compounds in Ojeoksan (OJS), which is composed of 15 herbal substances. All marker compounds were analyzed within 20 min and successfully identified via scheduled multiple reaction monitoring. The method validation revealed excellent performance characteristics of the method such as specificity, linearity, sensitivity, precision, and accuracy, demonstrating its suitability for intended use. The developed method was applied to samples of commercial OJS tablet and soft-extract dosage forms. The 14 marker compounds corresponding to 12 component herbal substances were determined in the samples; ephedirine, albiflorin, paeoniflorin, ferulic acid, hesperidine, neohesperidin, cinnamic acid, platycodin D, 6-gingerol, atractylenolide III, glycyrrhizin, honokiol, decursin, and magnolol. A fast and easy assay method with sufficient discrimination power was established. As a novel assay, this method can contribute to the quality control of OJS products.

In-silico evaluation of Oroxylum indicum vent compounds in the plausible treatment and prevention of nasopharyngeal cancer

BackgroundShyonaka (Oroxylum indicum Vent) is widely used in Ayurveda and in ethnomedical practice for the treatment of inflammation, pain, diarrhea, non-healing ulcers, and cancer. Owing to the high prevalence of Epstein-Barr virus (EBV) infection in Nasopharyngeal carcinoma (NPC) patients, simultaneous targeting of proteins involved in both EBV replication and NPC proliferation might help to manage the disease effectively. ObjectivesThis study is designed to identify potential dual targeting inhibitors from Oroxylum indicum having the potential to inhibit both EBV and NPC. This study also attempted quantitative analysis of Shyonaka Bark Decoction (SBD) to confirm the presence of Baicalein and Chrysin which are predominant marker compounds of Shyonaka. MethodologyThe HPLC analysis of stem bark and root bark of Oroxylum indicum was done to estimate the presence of marker compounds Baicalein and Chrysalin. The in-silico analysis included ADMET analysis followed by molecular docking of known compounds from Oroxylum indicum (retrieved from IMPPAT database) onto the target proteins of EBV (BHRF1, NEC1, dUTPase, Uracil DNA glycosylase) and NPC (COX-2, EGFR, and MDM2) using DOCK6 tool. Further validations were done using the molecular dynamics simulations of top screened molecules onto the selected target proteins using AMBER20 package and their corresponding MMGBSA binding free-energy values were calculated. ResultsThe molecular docking revealed that the key molecules from the plant, scutellarein 7-rutinoside (S7R), scutellarin (SCU) and 6-hydroxyluteolin, Baicalein and 5,7-Dihydroxy-2-phenyl-6-[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromen-4-one (57D) are effectively intervening with the target proteins of EBV, one of the key causative factors of NPC and the NPC specific targets which have the potential to reduce tumor size and other consequences of NPC. The molecular dynamics simulations of S7R, Baicalein and 57D, Baicalein with MDM-2 protein and dUTPase protein, respectively, showed stable interactions between them which were further assessed by the binding energy calculations. ConclusionOverall, the in-silico evaluation of these phytochemicals with target proteins indicates their potential to inhibit both EBV and NPC which needs further in-vitro and in-vivo validations.

Molecular docking of gingerol and shogaol for immunomodulatory effect in lupus disease

Background: One of the autoimmune diseases associated with high mortality is lupus or Systemic Lupus Erythematosus (SLE). In addition to symptomatic therapy, the treatment management of this condition includes immunomodulatory therapy. Various studies have been carried out on immunomodulators from natural products. Ginger rhizome (Zingiber officinale Roxb.) is a plant with potential immunomodulatory activity. Shogaol, which gives a spicy taste, is a metabolite of gingerol, a marker compound in ginger. Both of these compounds become important components of pharmacological activity. Objective: This study aimed to determine the in silico immunomodulatory activity of gingerol and shogaol compounds contained in ginger against S100A9, CTLA-4, SRSF1, JAK3, and MYD88 receptors. Method: In silico, a test was carried out using Pymol and PyRx applications, and receptors were involved in developing the immune system and SLE disease. Result: Docking results showed negative binding affinity and an RMSD of 2˚Angstroms. The shogaol, gingerol, and tofacitinib had several amino acid residues in common. Conclusion: In-silico analysis suggests that shogaol and gingerol could modulate the immune response against lupus. The resulting protein residues were similar between shogaol, gingerol, and the control, supporting their potential for immunomodulatory activity against lupus disease.

Chemical Relationship among Genetically Authenticated Medicinal Species of Genus Angelica.

The genus Angelica comprises various species utilized for diverse medicinal purposes, with differences attributed to the varying levels or types of inherent chemical components in each species. This study employed DNA barcode analysis and HPLC analysis to genetically authenticate and chemically classify eight medicinal Angelica species (n = 106) as well as two non-medicinal species (n = 14) that have been misused. Nucleotide sequence analysis of the nuclear internal transcribed spacer (ITS) region revealed differences ranging from 11 to 117 bp, while psbA-trnH showed variances of 3 to 95 bp, respectively. Phylogenetic analysis grouped all samples except Angelica sinensis into the same cluster, with some counterfeits forming separate clusters. Verification using the NCBI database confirmed the feasibility of species identification. For chemical identification, a robust quantitative HPLC analysis method was developed for 46 marker compounds. Subsequently, two A. reflexa-specific and seven A. biserrata-specific marker compounds were identified, alongside non-specific markers. Moreover, chemometric clustering analysis reflecting differences in chemical content between species revealed that most samples formed distinct clusters according to the plant species. However, some samples formed mixed clusters containing different species. These findings offer crucial insights for the standardization and quality control of medicinal Angelica species.

Nanostructured lipid carriers loaded with essential oils: a strategy against SARS-CoV-2

This work aimed to investigate the effectiveness of Lippia sidoides and Syzygium aromaticum essential oils (EOs) encapsulated in nanostructured lipid carriers (NLCs) as SARS-CoV-2 inhibitors through virucidal activity assessment. We developed anionic and cationic NLCs loaded with the EOs and assessed their physicochemical properties and SARS-CoV-2 virucidal activity, focusing on the effects of EO type and the NLCs composition. The NLCs exhibited particle sizes of 141.30 to 160.53 nm for anionic and 109.30 to 138.60 nm for cationic types, with PDIs between 0.16 and 0.25. High zeta potentials (>29.0 in modulus) indicated stable formulations. The NLCs effectively encapsulated the EOs, achieving encapsulation efficiencies between 84.6 to 100% w/w of marker compound. The EOs-loaded NLCs reduced the SARS-CoV-2 virion count, exceeding 2 logs over the control. NLCs loaded with Lippia sidoides and Syzygium aromaticum EOs represent an innovative strategy for combating SARS-CoV-2.

NADES extraction, UHPLC-ELSD-based quantification, and network pharmacology-guided target identification of fourteen specialised metabolites from Trillium govanianum Wall. ex D.Don.

Trillium govanianum Wall. ex D.Don is a folk medicinal herb rich in structurally diverse steroidal saponins. The annual demand for this herb in India is about 200-500 metric tons, highlighting the need for a thorough quality assessment. The objective of this study is to develop an easy and reliable ultrahigh-performance liquid chromatography-evaporative light scattering detector (UHPLC-ELSD)-based quality assessment method with 14 specialised metabolites of T. govanianum and identify the potential targets of this herb using network pharmacology. A UHPLC-ELSD method was developed and validated with 14 markers of T. govanianum. The developed method and natural deep eutectic solvent (NADES)-assisted extraction were utilised for the recovery enhancement study of targeted specialised metabolites from rhizome samples (collected from five geographically distinct areas). In addition, the network pharmacology approach was performed for these 14 markers to predict the plausible biological targets of T. govanianum. The developed method showed good linearity (r2: 0.940-0.998), limit of detection (LOD) (2.4-9.0μg), limit of quantification (LOQ) (7.92-29.7μg), precision (intra-day relative standard deviations [RSDs] 0.77%-1.96% and inter-day RSDs 2.19-4.97%), and accuracy (83.24%-118.90%). NADES sample TG-1* showed the highest recovery (yield: 167.66 ± 4.39 mg/g of dry weight) of total saponin content (TSC) as compared to its hydroethanolic extract (yield: 103.95 ± 5.36 mg/g of dry weight). Sample TG-1* was the most favourable (yield: 167.66 ± 4.39 mg/g) in terms of TSC as compared to other analysed samples (32.68 ± 1.04-88.22 ± 6.79 mg/g). Govanoside D (yield: 3.43-28.06 mg/g), 22β-hydroxyprotodioscin (yield: 3.22-114.79 mg/g), and dioscin (yield: 1.07-20.82 mg/g) were quantified as the major metabolites. Furthermore, network pharmacology analysis of targeted 14 markers indicated that these molecules could be possible therapeutic agents for managing neuralgia, diabetes mellitus, and hyperalgesia. The current study represents the first report for the simultaneous quantification and a network pharmacology-based analysis of 14 chemical marker compounds isolated from T. govanianum.

Identification of Codonopsis Radix from different sources by integrating high‐performance liquid chromatography fingerprinting with chemometric approaches and Lobetyolin determination

AbstractCodonopsis Radix (CR) is often used as both a medicine and a food ingredient. However, identifying CR can be difficult due to various factors such as variety, sulfur fumigation, and storage time. Consequently, a comprehensive strategy based on fingerprinting and chemometric approaches was developed to distinguish different CRs. First, the fingerprints of CR were acquired by high‐performance liquid chromatography (HPLC). Secondly, various chemical pattern recognition methods were employed to identify CR. Additionally, 14 and 16 major compounds were identified as marker compounds to differentiate between different varieties and sulfur‐fumigated samples, respectively. Furthermore, the amount of Lobetyolin present in different samples was determined, showing that the content of Lobetyolin varied significantly across the samples. Codonopsis pilosula Nannf. var. modesta (Nannf.) L.T.Shen had the highest content of Lobetyolin, followed by Codonopsis tangshen Oliv., and Codonopsis pilosula (Franch.) Nannf. In addition, the content of Lobetyolin was highest when it was non‐sulfur‐fumigated and stored for 0 years. Conclusively, HPLC fingerprint in conjunction with chemical pattern recognition and component content determination, can be employed to differentiate different varieties and sulfur‐fumigated CR. Additionally, it is a reliable, comprehensive, simple, and rapid method for the identification of CR.

Identification of volatile organic compound of oil palm plants infected with Ganoderma sp.

The success of controlling basal stem rot (BSR) disease caused by Ganoderma sp. is mostly determined by the early detection of the fungi. Hence, when the signs of infection begin to appear, plants are generally difficult to rescue since the pathogen infection has spread widely to all parts of the plant. Therefore, early detection through the application of biosensors for Ganoderma sp. infection is absolutely necessary. This study aim was to identify biomarker compounds of Ganoderma sp. infection in oil palm plants with GC-MS for volatile compounds, and LC-MS for non-volatile compounds. The results showed that Ganoderma sp. mycelium produced pyrimidinamine compounds. Meanwhile in early infected nursery plants, were found benzo[h]quinoline, hexaoxa-7,9,11-trisilaheptad, tris-(trimethyl-silyl ester and methyl-tris(trimethyl-siloxy)-silane). Whereas in Bekri, Rejosari, and Adolina plantation, the compound methyl-tris(trimethyl-siloxy)-silane was also found in healthy mature plants. GC-MS test results showed that for early, moderate and severe plants produced several benzene derivative compounds such as ethylbenzene, xylene, and benzaldehyde. These compounds were assumed to be resulted from the breakdown of the lignin structure which build plant cell walls, and have potency to be used as marker compounds for early infected Ganoderma sp. detection. The result of the produced gas quantification concluded that in the produced oil palm plants released less CO2 compared to healthy plants. On the other hand, the NH3 produced was higher than the healthy plants. Meanwhile, two non-volatile compounds were found that they were only produced by infected trunk and root tissue, namely pseudobrucine and picrasidine.

Evaluation of antiplasmodial potential of Cissampelos pareira L.: Characterization and quantification of isoquinoline alkaloids responsible for the biological activity

Globally, based on traditional knowledge, different parts of Cissampelos pareira plant are being used individually or in combination in various forms to manage malaria. However, the scientific investigation for validating the most effective part of this plant against malaria parasite has not been done. Thus, the study aims to explore the antiplasmodial potential of different parts of the plant against malaria parasite by bio-assay guided isolation of compounds from different extracts and fractions. Antiplasmodial activities of the whole plant parent extract and its fractions along with decoctions (roots, stem, leaves and whole plant) and isolated molecules were evaluated against chloroquine-sensitive Pf3D7 and chloroquine-resistant PfINDO strains of Plasmodium falciparum. A UPLC-DAD based method was developed to quantify the marker compounds in all samples by utilizing isolated molecules as analytical standards. The chloroform fraction of the whole plant was found to be the most potent with IC50 (µg/mL) of 0.79 (Pf3D7) and 2.26 (PfINDO) followed by root decoction with IC50 (µg/mL) 10.22 (Pf3D7) and 7.7 (PfINDO). Further, phytochemical analysis of active chloroform fraction led to the isolation of two potent bisbenzylisoquinoline compounds, namely O,O-dimethylcurine (1) [IC50 (µM) 1.46 (Pf3D7) and 0.51 (PfINDO)], and curine (2) [IC50 (µM) 1.46 (Pf3D7) and 0.51 (PfINDO)]. Analysis of developed UPLC-DAD method showed the highest quantities of curine (2) (∼107 mg/g) and O,O-dimethylcurine (1) (∼15 mg/g) in chloroform fraction which might be responsible for its potent activity. This study showed that the root decoction was more effective than decoctions of each of the other parts of the plant and the whole plant hydroalcoholic extract.

Identification of Bis(methylsulfanyl)methane and Furan-2(5H)-one as Volatile Marker Compounds for the Differentiation of the White Truffle Species Tuber magnatum and Tuber borchii.

Some truffles are expensive and, therefore, are prone to food fraud. A particular problem is the differentiation of high-priced Tuber magnatum truffles from cheaper Tuber borchii truffles, both of which are white truffles with similar morphological characteristics. Using an untargeted approach, the volatiles isolated from samples of both species were screened for potential marker compounds by comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-TOFMS) and statistical analysis of the obtained semiquantitative data. Results suggested bis(methylsulfanyl)methane and furan-2(5H)-one as compounds characterizing T. magnatum and T. borchii, respectively. Exact quantitation of both volatiles by conventional one-dimensional gas chromatography-mass spectrometry in combination with stable isotopologues of the target compounds as internal standards confirmed both as marker compounds. The method is suitable to be used in the routine analysis for the objective species differentiation of T. magnatum and T. borchii.

Tracing of Gelsemium elegans by UPLC-Q-TOF MS fingerprint analysis technique

Fingerprints of 13 batches of Gelsemium elegans (G. elegans) were established by ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF MS). The study aimed to explore the compositional variations in G. elegans before and after high-temperature processing and to distinguish it from honeysuckle and Ficus hirta Vahl. Chemical constituents of 25 peaks in G. elegans were identified by precisely determining retention times, MS2 spectra fragments, and validation against relevant databases and reference materials (RM). The entire analysis process was completed within 15 min. Moreover, the similarities and significant differences of chemical components of G. elegans before and after processing were analyzed via principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLS-DA). The results showed that the contents of 10 components (kouminol, celastrol, sempervirine, 14,15-dihydroxygelsenicine, 4,20-dehydrogelsemicine, 4-O-caffeoylquinic acid, Nb-Methylgelsedilam, 3-hydroxykoumidine,14-acetoxygelsenicine, and humantenine) were related to the degree of processing. In addition, 15 alkaloids exhibited relative stability before and after processing, serving as distinctive marker compounds for traceability studies. This research provides a valuable basis for the source analysis and species traceability of toxin in forensic science.

SECONDARY METABOLITE PROFILE IN STEM AND ROOT OF KEJI PLANT (Staurogyne elongata (Blume) Kuntze)

Keji plant (Staurogyne elongata (Blume) Kuntze) is an herbal medicinal plant that is often found on the islands of Java to Sumatra. The leaves and roots of keji plants (S. elongata) are used by the community as diuretic drugs and increase blood pressure and are believed to treat bladder stones, kidney stones, and joint problems. However, research on secondary metabolites in the stems and roots of keji plants has never been done. The aims of the study were to identify the secondary metabolites content and marker compounds in the stems and roots of keji plants. The research stages were drying and pollination of samples, extraction, and analysis using Gas Chromatography-Mass Spectrometry (GC-MS). The results showed that the compounds in the stem were nine secondary metabolite compounds from the phenol, terpenoid, alcohol and ester group. Meanwhile, secondary metabolite compounds in the roots of keji plants were ten compounds from coumaran, phenols, and ester group. The marker compounds in the stem were phenol, 2,2'-methylenebis[6-(1,1-dimethylethyl)-4-methyl- while the marker compounds in the roots were octadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester.

Evaluation and method development for quantification of Piperine in Hutabhugadi Churna by RP- HPLC

Aim and Objective: The current work was aimed at preparing the Hutabhugadi Churna in the laboratory and evaluating the same including the method development for the estimation of a marker compound Piperine by using RP-HPLC. Methods: Prepared Hutabhugadi churna was subjected for macroscopic, physical, and chemical evaluation considering WHO guidelines. The methanolic extract was subjected for estimation of Piperine as marker using RP-HPLC. Results: The macroscopic characteristics like colour, odour and taste are recorded. The physical characteristics like loss on drying, ash value, extractive value, swelling index, foaming index, powder properties like angle of repose, bulk density, tapped density, compressibility index etc. were determined. Total phenolic content, total flavonoid content, preliminary phytochemical screening was also carried out. The results are compared with marketed formulation of Hutabhugadi churna. The retention time of the standard Piperine was found to be 5.517, while the Piperine in extracts of laboratory and marketed formulations was found to be was found to be 5.554 and 5.562 respectively. The concentration of Piperine in laboratory and marketed formulation was found to be 0.17 %w/w and 0.18 % w/w respectively. The method developed was also validated. Conclusion: The laboratory made Hutabhugadi churna and marketed formulation of Hutabhugadi churna was comparatively evaluated. The resulting data will be useful for the standardization of the Hutabhugadi churna, an Ayurvedic formulation.

Metabolite profiling of potential bioactive fractions from ethanol extract of Boehmeria nivea flowers by GC–MS/MS analysis

BackgroundRamie (Boehmeria nivea) has been reported to have a wide variety of secondary metabolite compounds in its vegetative organ that have distinct organic activities. Regarding the chemical composition of flowers, there have been no study considering their pharmacological effects. PurposeThis research aims to determine profile metabolites contained in B. nivea flowers employing a metabolomics approach. Study DesignThis research was designed to obtain a metabolite database from five dissolved fractions of ramie flower using a Gas Chromatography-Mass Spectrometry (GC–MS/MS) instrument. This study includes metabolite profiles from each fraction of the spectrum obtained, which are then investigated for potential bioactivity based on previously reported studies. Metabolite profiles were developed using Principal Component Analysis (PCA) to obtain information on clustering of the compounds and determination of marker compounds from each fraction analyzed. MethodsThe active compounds of the flowers were separated utilizing the maceration method with ethanol solvent and multilevel fractionation using n-hexane, dichloromethane, ethyl acetic acid derivation, butanol, and water employing a GC–MS/MS approach. Principal Component Analysis (PCA) was optimized with a total represented data diversity range of 60 % - 65 %. This allowed for the identification of characteristic compounds and similarities in compound substances accross the fractions. ResultGC–MS/MS interpretation showed the dominant compounds contained in each fraction of n-hexane, dichloromethane, n-butanol, ethyl acetate, and water are 9,12-Octadecadienoic acid, ethanol, 2-phenoxy-, butyl ester, and hexanedioic acid, bis(2-ethylhexyl) ester. PCA clustering showed combined similarity of the compound for ethyl acetate fraction with butanol, and also butanol with n-hexane. Meanwhile, the dichloromethane and water fractions have very small content similarities. The score plot indicated marker compounds of each fraction are: ethanol, 2-butoxy; hexane, 2,2,3-trimethyl-; phthalic acid, in (2-propylpentyl) ester; n-hexadecanoic acid, and Hexanedioic acid, bis(2- ethylhexyl) ester. ConclusionsThe abundance of compounds in the fractions shows potential for application as a source of new drug candidates. In addition, there are still many compounds that are unknown in structures or biological activities which could lead to further research to provide potential clues towards the bioprospecting of the plant. AbbreviationsCH2Cl2, dichloromethane; Dm1, Dimension 1; Dm2, Dimension 2; EtOAc, ethyl acetate; GC–MS/MS, Gas Chromatography–Mass Spectrometry; n-BuOH, n-butanol; PCA, Principal Component Analysis.

Development and Validation of a High-Performance Liquid Chromatography Method to Quantify Marker Compounds in Lysimachia vulgaris var. davurica and Its Effects in Diarrhea-Predominant Irritable Bowel Syndrome.

Irritable bowel syndrome (IBS), a common gastrointestinal disorder worldwide, is characterized by chronic abdominal pain, bloating, and disordered defecation. IBS is associated with several factors, including visceral hypersensitivity, gut motility, and gut-brain interaction disorders. Because currently available pharmacological treatments cannot adequately improve symptoms and may cause adverse effects, the use of herbal therapies for managing IBS is increasing. Lysimachia vulgaris var. davurica (LV) is a medicinal plant used in traditional medicine to treat diarrhea. However, information on whether LV can effectively improve diarrhea-predominant IBS (IBS-D) remains limited. In this study, using an experimental mouse model of IBS-D, we elucidated the effects of the LV extract. The methanol extract of LV decreased fecal pellet output in the restraint stress- or 5-hydroxytryptamine (5-HT)-induced IBS mouse model and inhibited 5-HT-mediated [Ca2+]i increase in a dose-dependent manner. Furthermore, we developed and validated a high-performance liquid chromatography method using two marker compounds, namely, chlorogenic acid and rutin, for quality control analysis. Our study results suggest the feasibility of the methanol extract of LV for developing therapeutic agents to treat IBS-D by acting as a 5-HT3 receptor antagonist.

Characterization of Flavor Profiles of Cigar Tobacco Leaves Grown in China via Headspace-Gas Chromatography-Ion Mobility Spectrometry Coupled with Multivariate Analysis and Sensory Evaluation.

Although cigar tobacco leaves (CTLs) have a high economic value, research regarding the flavor characteristics of CTLs is currently limited. A comprehensive study of the flavor characteristics of CTLs from different regions of China was conducted by identifying their volatile-flavor-containing compounds (VFCs) and flavors. The samples were analyzed via gas chromatography-ion mobility spectrometry (GC-IMS) and sensory evaluation. Results revealed considerable differences in the VFC contents of CTLs from different regions of China, suggesting that the VFLs of CTLs could be influenced by geographical origin. Mainly, phenols, pyrazines, and aldehydes were present in the CTLs from Sichuan. High contents of esters and pyrazines were present in the CTLs from Hubei, while esters were the major components of the CTLs from Hainan. Multivariate analysis results showed the effective differentiation of samples from different geographical origins based on the GC-IMS results. Sensory evaluation revealed that the flavors of CTLs from different geographical origins were different. 1,8-Pinene, 3-methyl-3-butene-1-ol, 2,3-dimethyl-5-ethylpyrazine, 4-methyl-3-penten-2-one, and (E)-2-pentenal might serve as geographical marker compounds, indicating the geographical origin of CTLs based on the results of GC-IMS and sensory evaluation. This study may be beneficial for the trade of CTLs and the development of cigar products.

Formulation and bioavailability studies on orodispersible tablets containing standardized extracts of Glycyrrhiza glabra, Curcuma longa and Piper nigrum

BackgroundGlycyrrhizin, Curcumin and Piperine indicated good antiasthamatic properties owing to their ability to inhibit airways constriction and anti-inflammatory properties. Orodispersible tablets have advantage of faster drug release and has patient compliance in asthmatic patients. Hypothesis / PurposeTo develop an orodispersible polyherbal tablets using the standardized extracts of Glycyrrhiza glabra, Curcuma longa and Piper nigrum with respect to glycyrrhizin, curcumin and piperine respectively and evaluate bioavailability of the markers from the formulation. MethodsAlcoholic extracts of G. glabra, C. longa, and P. nigrum were standardized using HPTLC for glycyrrhizin, curcumin, and piperine content, respectively. The orodispersible tablet formulation was created and standardized for curcumin (50 mg), glycyrrhizin (75 mg), and piperine (2.5 mg) per tablet, using the HPTLC method. The absolute Bioavailability studies were conducted for glycyrrhetinic acid and curcumin in rats by administering pure marker compounds intravenously (2 mg/kg b.w) and orally (10 mg/kg b.w.), followed by determining relative bioavailability through oral administration of the orodispersible tablet formulations. ResultsOrodispersible tablets could be successfully developed using the standardized extracts. The HPTLC bioanalytical method was developed and validated as per M 10 guideline and results were in accordance to specifications. The bioavailability studies in rats indicated absolute bioavailability of Glycyrrhetinic acid and Curcumin to be 20.35 and 2.57% respectively and relative bioavailabilities of orodispersible formulation F1 were found to be 75 and 38.67%. ConclusionA standardized orodispersible formulation can be a better option for asthma patients. The low absolute bioavailability of curcumin was significantly (P<0.05) increased in both the formulations F1 and F2. This may be attributed to the extracts containing saponin glycosides like Glycyrrhizin aiding into greater dissolution of curcumin. Formulation with piperine (F1) showed significant increase in bioavailability is due to inhibition of human P-glycoprotein, cytochrome P450 3A4 (CYP3A4) and increase secretion of bile acids.