Comprehensive phytochemical profiling of Phyllanthus emblica L. flowers on UHPLC/MS quadrupole time of flight, HPTLC, HPLC, and NMR analytical platforms reveals functional metabolites with potent anti-inflammatory effects in human (THP-1) macrophages.

INTRODUCTION: Phyllanthus emblica L., renowned for its pharmacological benefits found in its fruits and leaves, has received considerable attention. However, there is a notable lack of research on its flowers, specifically on metabolite profiling and pharmacological activity. OBJECTIVE: The present study aims to delineate the phytochemical constituents of hydromethanolic extract of P. emblica flowers by ultra-high-performance liquid chromatography coupled with quadrupole time of flight mass spectrometry (UHPLC-QToF-MS), high-performance thin layer chromatography (HPTLC), high-performance liquid chromatography (HPLC), infrared and nuclear magnetic resonance spectroscopic methods and subsequent evaluation of its anti-inflammatory potential. MATERIALS AND METHODS: The identification and characterization of phytochemicals in P. emblica flowers was performed by UHPLC/MS-QToF in both positive and negative ionization modes. Additionally, marker compounds present in flower extract were analyzed using HPTLC, HPLC, FT-IR, and NMR methods. The anti-inflammatory potential was evaluated in lipopolysaccharide-stimulated THP-1 macrophages by evaluating inflammatory biomarkers. RESULTS: UHPLC/MS-QToF analysis facilitated the identification of 51 compounds from P. emblica flowers including gallic acid derivatives, flavonoid glycosides, and tannins based on their fragmentation patterns and previous literature reports. Notably, the study also identified spermidine compounds for the first time in this species. Optimization of HPTLC and HPLC methods marked the presence of corilagin as major compound followed by FT-IR and NMR spectral methods. Moreover, treatment with hydromethanolic extract of P. emblica flowers resulted in decreased levels of proinflammatory cytokines, TNF-α, IL-1ß, and IL-6, alongside modulation of nuclear factor-κB activity in lipopolysaccharide-induced THP-1 macrophages. CONCLUSION: Chromatographic techniques in conjunction with spectral methods found robust prevalence in the identification of signature phytometabolites present in P. emblica flowers, which sets the basis for its anti-inflammatory potentials. The studies established a foundation for further exploration of potential applications of P. emblica flowers across various domains.

Exploring the potential of Mustard (Brassica spp.) seeds through 'Kolhu' traditional method of extraction and novel identification of an anti-cancer dipeptide, Aurantiamide acetate (Asperglaucide) on ultra-performance liquid chromatography-mass spectrometry coupled with quadrupole time-of-flight (UPLC/MS-QToF) analytical platform.

Mustard (Brassica spp.) is one of the world's oldest condiments in the food basket, which holds a significant place in the global culinary landscape due to historical prominence and perceived health benefits. This study explores the extraction of oils from Mustard seeds by employing traditional 'Kolhu' method, modern supercritical fluid, and solvent extraction techniques. This study, for the first-time, identified Aurantiamide acetate, a potent anti-cancer dipeptide in Mustard seeds using ultra-performance liquid chromatography-mass spectrometry coupled with quadrupole time-of-flight (UPLC/MS-QToF) analytical platform. The analytical methodology was meticulously validated encompassing optimal parameters such as limit of detection, limit of quantification, precision, accuracy, linearity and robustness, within the range. Interestingly, 'Kolhu' method of oil extraction exhibited better yield of Aurantiamide acetate, suggesting superior efficiency of traditional methods. This study accentuates the importance of classical extraction methods, used traditionally, and emphasizes that naturally occurring substances indeed could be harnessed for better health.

Dissecting the natural phytochemical diversity of carrot roots with its colour using high performance liquid chromatography and UV-Visible spectrophotometry.

The research provides insights into the phytoconstituents of black, orange and red carrots (Daucus carota subsp. Sativus (Hoffm.) Schübl. & G. Martens), a highly nutritious food crop widely appreciated across age groups. Recognising carrots as a repository of health-promoting compounds, our study employs UV-Visible spectrophotometric and HPLC methods to discern significant variations in bioactive components among carrot varieties. Black carrots emerge as potent contenders, displaying the highest levels of total phenolics (2660 ± 2.29 mg GAE/100 g F W.), total flavonoids (831 ± 1.74 mg QE/100 g F W.), proanthocyanins (10910 ± 1.11 mg CE/100 g F W.), and tannins (713 ± 0.84 mg/100 g F W.). Red carrots, conversely, showcase higher anthocyanin content (6870 ± 1.85 mg CyGE/100 g F W.) by UV-Vis spectrophotometry. Additionally, orange carrots exhibit heightened ß-carotene levels, confirmed at 0.03 µg/mg through HPLC. HPLC analysis unveils substantial chlorogenic acid variability (1.29 µg/mg) in black carrots, accompanied by the discovery of unique compounds such as cryptochlorogenic acid (0.05 µg/mg), caffeic acid (0.01 µg/mg), ferulic acid (0.11 µg/mg), methyl caffeate (0.01 µg/mg), and quercetin (0.02 µg/mg), marking the first detection of methyl caffeate in black carrots. The analytical methodology was meticulously validated encompassing optimal parameters such as linearity, precision, limit of detection, limit of quantification, accuracy, and robustness, within the range. In conclusion, our study underscores the health benefits of black carrots due to their rich polyphenolic content and endorses orange carrots for elevated ß-carotene levels. These findings contribute to a deeper understanding of the diverse phytoconstituents in carrots, aid in informed dietary choices for improved health.

In-depth phytochemical profiling of Roscoea purpurea (Kakoli): Comparative UHPLC-MS/QToF and GC-MS/MS analysis of supercritical CO2 fluid - and conventional solvent - based extractive processes.

The remarkable biodiversity of medicinal plants worldwide highlights their significance in traditional and alternative medicine. Astavarga, a group of eight medicinal herbs from the Himalayan region of India, including Roscoea purpurea (commonly known as Kakoli), is esteemed in Ayurveda for its health-promoting and rejuvenating properties. In this comprehensive study, we aimed to develop and optimise robust UHPLC-MS/QToF (Ultra-high-performance liquid chromatography-mass spectrometry coupled with quadrupole time of flight) and GC-MS/MS (Gas chromatography-tandem mass spectrometry) methods to identify the phytochemicals in R. purpurea root hydromethanolic extract and essential oil. We also conducted a comparative assessment of supercritical fluid extraction and conventional solvent extraction methods for the first time in R. purpurea root, highlighting their relevance to the medicinal field. Using the UHPLC/MS-QToF method, we identified a total of fifty-six phytometabolites, while sixteen volatile constituents were discerned within the essential oil of R. purpurea by GC-MS/MS method. Among the volatile constituents, ß-eudesmol (40.84 %), guaiac acetate (10.55 %), and γ-eudesmol (10.31 %) were emerged as the principal components. Our findings were further compared with the volatile constituents extracted via supercritical fluid extraction and conventional solvent extraction methods. Notably, our research unveiled the presence of a carotenoid metabolite, 15-methyl retinol, for the first time. Furthermore, our fatty acid analysis of the supercritical fluid extract revealed elevated levels of unsaturated fatty acids, particularly oleic and linoleic acids. The methods were validated in terms of system specificity also. The discovery of these well-recognised therapeutically active components in R. purpurea significantly enhances its potential, highlighting its unique profile among medicinal plants in the Himalayan region and its suitability for traditional Ayurveda.

Modulation of psoriatic-like skin inflammation by traditional Indian medicine Divya-Kayakalp-Vati and Oil through attenuation of pro-inflammatory cytokines.

Background and aim: Psoriasis (Ps) is a chronic skin inflammatory disorder, that progresses to scaly-red dermal plaque formations associated with inflammation. Divya-Kayakalp-Vati (DKV) and Divya-Kayakalp-Oil (DKO) are traditional Ayurveda herbo-mineral formulations, that are prescribed for the treatment of inflammatory dermal ailments. In the present study, we evaluated the efficacy of Divya-Kayakalp-Vati and Divya-Kayakalp-Oil (DKV-O) combined treatment in ameliorating Ps-like skin inflammation under in-vitro and in-vivo conditions. Experimental procedure: Efficacy of DKV-O was analyzed in λ-carrageenan-treated Wistar rats paw edema and 12-O-tetradecanoylphorbol 13-acetate (TPA)-treated CD-1 mouse ear edema models through physiological and histopathological analysis. Mode of action for the DKV-O was studied in LPS-stimulated THP-1 cells through pro-inflammatory cytokine analysis. Observed effects were correlated to the phytochemicals constituents of DKV-O analyzed using the HPLC method. Result and conclusion: DKV and DKO formulations were individually found to contain phytochemicals Gallic acid, Catechin, Berberine, Curcumin, Phenol and Benzoic acid. DKV-O treatment significantly reduced the paw volume and edema in Wistar rats stimulated through λ-carrageenan-treatment. Furthermore, DKV-O treatment significantly reduced the ear edema and enhanced biopsy weight, epidermal thickness, inflammatory lesions and influx of neutrophils stimulated by TPA-treatment in CD-1 mice. DKV alone ameliorated the LPS-stimulated release of Interleukin (IL)-6, IL-17A, IL-23, and Tumor Necrosis Factor-alpha cytokines in the THP-1 cells.Taken together, DKV-O showed good efficacy in ameliorating acute systemic inflammation stimulated by effectors such as, λ-carrageenan and TPA in animal models. Hence, Divya-Kayakalp-Vati and Divya-Kayakalp-Oil co-treatment can be further explored as an anti-inflammatory treatment against dermal diseases like psoriasis and atopic dermatitis.