Biomedical Chromatography

Kadry, Kholoud Kh.; Moussa, Ashaimaa Y.; Youssef, Fadia S.; Afifi, Manal S.

doi: 10.1002/bmc.70480pmid: N/A

Ceiba insignis and Ceiba speciosa are utilized in traditional medicine for treating inflammatory diseases. The analysis of their extracts by LC‐ESI‐MS/MS revealed the presence of flavonoids like kaempferol hexoside and apigenin‐O‐deoxyhexosyl hexoside and fatty acids, namely, sterculic acid, α‐linolenic acid, and dihydrosterculic acid. C. insignis extract decreased 5‐lipoxygenase and tumor necrosis factor alpha (TNF‐α) gene expression by 0.2908‐fold and 0.3276‐fold, whereas C. speciosa extract reduced their expression by 0.4392‐fold and 0.2663‐fold, respectively. In response to lipopolysaccharide stimulation, both extracts had anti‐inflammatory effects like those of the control medication celecoxib, which decreased 5‐LOX and TNF‐α expression by 0.3554‐fold and 0.2263‐fold, respectively. Molecular docking highlighted that dihydrosterculic acid showed the tightest fit within 5‐LOX center revealing binding energy (∆G = −35.09 kcal/mol). Consequently, this study highlighted the relevance of the flowers of both Ceiba species in treating inflammatory disorders that in turn confirmed the traditional medicinal uses of these plants.

Zhang, Ze‐Chao; Ye, Xue‐Jin; Shang, Chang‐Jie; Chen, Yu; Guo, Lin; Li, Yi‐Wen; Zhu, Min; Tang, Bo‐Tao; Liang, Tai‐Sheng

doi: 10.1002/bmc.70455pmid: N/A

To investigate the molecular mechanism of Qianlong Shutong formula (QLSTF) in treating benign prostatic hyperplasia (BPH) through the regulation of metabolism and protein function. An integrated metabolomics and proteomics analysis was conducted using a BPH animal model. The animals were divided into a control group, a model group, and a QLSTF intervention group. Following drug intervention, metabolite extraction and protein sample processing were performed. Metabolomics results indicated the presence of metabolic disorders in the model group, which were significantly ameliorated by QLSTF through the regulation of relevant metabolic pathways. Quantitative proteomic analysis identified multiple differentially expressed proteins. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Clusters of Orthologous Groups (COG) enrichment analyses revealed that these proteins are involved in biological processes such as cell proliferation, apoptosis, and inflammatory responses. Correlation analysis demonstrated a synergistic relationship between metabolite levels and protein expression. Integrated analyses revealed metabolic disorders and differential protein expression, providing multiomics evidence for QLSTF's mechanism. QLSTF alleviates BPH by modulating metabolic pathways and protein networks, providing a molecular basis for TCM in BPH treatment and suggesting future research.

Sharma, Akshita; Katna, Sapna; Banshtu, Tanuja; Devi, Nisha; Kumar, Arvind; Singh, Shubhra; Sharma, Sakshi; Sharma, Isha; Tomar, Ankur; Thakur, Jyoti

doi: 10.1002/bmc.70471pmid: N/A

This study compares four different analytical methods for residue extraction to establish an efficient and cost‐effective sample analysis method for green chilli matrix after spiking. Modified Dutch mini‐Luke method, ethyl acetate extraction method and two modified QuEChERS methods with different amounts of sorbent were assessed for the detection of insecticide residue in green chilli fruits using liquid chromatography tandem mass spectrometry (LC–MS/MS). Chilli matrix was spiked with cyantraniliprole, lufenuron, spidoxamat and spinetoram and its metabolites (N‐demethyl and N‐formyl) at different fortification levels from 0.010 to 0.100 mg/kg (replication [n] = 5) to evaluate the efficiency of the aforementioned analytical method. Modified QuEChERS method with a higher amount of sorbent was found to be the most suitable of all methods (with recovery 91.6%–109.8% [ANOVA; p < 0.001]), whereas the modified QuEChERS with a lower amount of sorbent showed inconsistent results due to matrix interference. The rest of the tested methods were unable to give acceptable recoveries (17.23%–67.00%) (ANOVA; p < 0.001). Among these analytical methods, the modified QuEChERS analytical method was found to be the best suited for the extraction of the aforementioned insecticides and metabolite residues from a problematic complex matrix like green chilli showcasing acceptable method validation guidelines as per SANTE.

Tungapati, Sadakar; Nuvuluri, Srinivasu; Ramakrishna, M.

doi: 10.1002/bmc.70425pmid: N/A

Mirabegron, a Beta‐3 adrenoceptor agonist drug, and Solifenacin succinate, an antimuscarinics drug, are used to treat overactive bladder in combination. Till today, no bioanalytical method has been reported to estimate Mirabegron and Solifenacin succinate in type of matrix. Mirabegron and Solifenacin succinate were measured in rat plasma using Darifenacin as the internal standard, a newly developed and validated bioanalytical LC–MS/MS technique. The analytes were separated using a Luna C18 column and a mobile phase combination of acetonitrile: 0.1% formic acid in HPLC water (50:50 v/v). The flow rate was maintained at an isocratic level of 1.0 mL/min, and the runtime was about 5 min. Mirabegron (m/z 397.51 to 83.78) and Solifenacin succinate (m/z 481.56 to 148.79) were determined by using the mass spectra with positive mode of multiple reaction monitoring. The strategy was approved in accordance with USFDA regulations. Accuracy with mean % recovery of 96.25% to 98.45% and 96.46% to 98.05% and strong linearity with r2 value of 0.9998 in the ranges of Mirabegron (2.5–100 ng/mL) and Solifenacin succinate (0.5–20 ng/mL) are the outcomes. All other parameters fall within the approved range. Indicators of medication effectiveness and safety, pharmacokinetic parameters, may be determined using the proposed technique.

Fan, Jiaqi; Lin, Guimei; Li, Hongye; Yao, Yi; Zhao, Wenyuan; Bi, Qiuru

doi: 10.1002/bmc.70458pmid: N/A

The challenge of combating brain aging is significant due to its intricate pathogenesis. Polygalae radix (PT), a well‐known herbal remedy derived from the dried root of Polygala tenuifolia Willd., serves as a traditional Chinese medicine and is also utilized in health foods. The primary processed products of PT are PT processed with licorice (PT + L) and PT processed with honey (PT + ER). Both PT and its processed products exhibit anti‐brain aging properties, but their mechanisms remain unclear. This study investigated the brain‐penetrating components and mechanisms of PT, PT + L, and PT + ER using UPLC‐Q‐TOF‐MS, network pharmacology, molecular docking, and in vivo assays. Thirteen brain‐penetrating components were identified, including tenuifolin, 3,4,5‐trimethoxycinnamic acid, chlorogenic acid, liquiritigenin, and caffeic acid. Core targets (BDNF, Mfn1, Mfn2, Drp1, and Fis1) interacted with these components. In vivo, PT and its processed products improved memory, reduced hippocampal damage, regulated the HPA axis, and enhanced antioxidant capacity by modulating proteins involved in mitochondrial dynamics and BDNF. Processed products showed superior efficacy: PT + ER prominently regulated the HPA axis, while PT + L significantly upregulated BDNF. This study clarifies the material basis and multitarget mechanisms of PT and its processed variants, confirming traditional processing benefits and providing experimental evidence for clinical use in age‐related neurodegenerative disorders.

Qu, Zhizi; Kong, Xiaoni; Xv, Jiaxin; Cai, Meichao; Zhou, Honglei

doi: 10.1002/bmc.70464pmid: N/A

Hyperuricemia is a common metabolic disorder associated with gout, renal dysfunction, and cardiovascular complications. Although Astragalus membranaceus has long been used in traditional medicine, the influence of gut microbiota biotransformation on its potential urate‐lowering activity remains unclear. In this study, ethanol and water extracts of Astragalus membranaceus were subjected to in vitro fecal microbiota biotransformation, and their xanthine oxidase (XO) inhibitory activities before and after biotransformation were compared. The transformed ethanol extract was further fractionated by solvent partitioning, and the resulting fractions were characterized by UPLC‐MS/MS. The results showed that biotransformation altered the XO inhibitory activity of both extracts, with a more evident increase in the ethanol extract than in the water extract. Among the solvent fractions, the ethyl acetate fraction exhibited the strongest inhibitory activity. Chemical profiling tentatively characterized 62 compounds and revealed marked compositional changes after biotransformation, particularly a reduction in glycosidic constituents together with enrichment of corresponding aglycone‐related compounds. These findings suggest that gut microbiota biotransformation may be an important factor influencing the potential urate‐lowering activity of Astragalus membranaceus and provide a basis for further identification of its bioactive constituents.

Zhang, Minyue; Sun, Jiakang; Tang, Shuhan; Li, Xintong; Wang, Yige; Xu, Yaqi; Ma, Rong; Wang, Xuexi; Li, Xianna; Wang, Ying; Xu, Hongying; Zhang, Hailong; Wang, Zhigang

doi:

Yusuf, Abdulla; Yin, Yu; Zhao, Xi; Ma, Yuqian; Shen, Bingyue; Gao, Jie; Liu, Yi; Ren, Xiaoliang; Fu, Zhiming

doi: 10.1002/bmc.70462pmid: N/A

Aurantii Fructus (AF), the dried immature fruit of Citrus aurantium L., is a widely used qi‐regulating herb in clinical practice. Traditional medical texts describe AF as “the older, the better,” implying that aging moderates its medicinal properties and enhances efficacy, although the mechanisms remain unclear. Here, dynamic changes in volatile and nonvolatile components during AF aging were investigated using long‐term natural aging and accelerated aging models, involving raw AF and four processed forms. Compositional variations across aging stages were characterized by high‐performance liquid chromatography (HPLC), electronic nose analysis, and multivariate chemometrics. During long‐term aging, flavonoid glycosides such as naringin gradually decreased and stabilized, whereas small‐molecule flavonoids and polymethoxyflavonoids increased, likely due to microbial transformation. Compared with raw AF, processed products better preserved key active constituents during aging, indicating improved quality stability. Electronic nose results showed an overall decline in volatile components, particularly hydrocarbons such as D‐limonene, supporting the traditional concept that aging reduces “dryness.” However, high‐temperature processing accelerated volatile loss, and accelerated aging caused more drastic compositional changes, potentially compromising quality. Overall, this study clarifies the chemical basis of the “the older, the better” concept of AF and provides scientific support for quality evaluation and aging process optimization.

Song, Xiaoming; Zang, Liyan; Li, Jiamin; Liu, Zixuan; Sun, Jing

doi: 10.1002/bmc.70457pmid: N/A

Sinopodophyllum hexandrum, a Berberidaceae medicinal herb, contains lignans and flavonoids with anticancer, antibacterial, antiviral, and expectorant activities, and its antioxidant capacity underpins therapeutic effects. This study evaluated the quality and antioxidant capacity of S. hexandrum from 12 Qinghai Plateau origins, and analyzed their correlations with environmental factors. Cluster analysis classified the origins into three groups. Principal component analysis (PCA) showed that the first four principal components contributed a cumulative variance of 86.395%, with podophyllotoxin‐4‐O‐glucoside (PPG) as the main contributor. Orthogonal partial least squares‐discriminant analysis (OPLS‐DA) further confirmed PPG as a key origin discriminant (VIP > 1). Comprehensive quality evaluation recognized P3 (Huzhu), P2 (Ledu), and P11 (Nangqian 2) as high‐quality origins. While antioxidant capacity analysis showed strong activity in P10 (Nangqian 1) samples—with 4′‐demethylepipodophyllotoxin (4′‐DMEP) as the key related component. Redundancy analysis (RDA) and Mantel Test results showed that component contents (especially quercetin, PPG, and podophyllotoxin [PPT]) and antioxidant capacity of S. hexandrum correlated significantly with environmental factors. Notably, antioxidant capacity correlated significantly with B1 (precipitation supply capacity and temperature fluctuation stability), and quercetin was the most environment sensitive component. This study provides a scientific basis for resource protection, elite germplasm screening, and targeted cultivation of S. hexandrum.

Singh, Manju; Gaurav, ; Gaur, Praveen Kumar

doi: 10.1002/bmc.70475pmid: N/A

Quality standardization of Indian medicinal plants is crucial for confirming their safety, efficiency, and validity in pharmaceutical applications. The current investigation explores ethnopharmacological perspectives of Allium sativum and Syzygium jambolanum through phytochemical screening using HPTLC and LC–MS profiling, and systems‐based pharmacology investigation using network pharmacology and molecular docking studies. The results indicated that both plants are enriched in polyphenols, terpenoids, glycosides, etc., responsible for various biological activities. In vitro antioxidant activity demonstrated significant free‐radical neutralization potential. HPTLC showed major and minor metabolites at different Rf values, and LC–MS analysis showed key bioactive substances including ferulic acid, gallic acid, naringenin, catechin, quercetin, and myricetin. The Network pharmacology study identified interactions with key proteins, including IL‐1β, TLR4, IL6, NOS3, ABCA1, with prominent interaction of quercetin and kaempferol with the protein IL‐1β. The DisGeNET mapping findings point toward their participation in key pathophysiological conditions like inflammation, interstitial inflammation, myofibroblast activation causing tubulointerstitial fibrosis, and microbial infection. The study concludes that A. sativum and S. jambolanum actively play a significant role in inflammation and associated dysfunctions, hence providing novel plant‐based therapeutics as A. Sativum and S. jambolanum by highlighting the significance in traditional medicine as well as their potential relevance in contemporary pharmacology.