Search Results (17,161)

Hypertension remains a major global health challenge, and pharmacological therapy is often constrained by tolerability issues. Adjunctive approaches targeting the nitric oxide synthase and soluble guanylate cyclase–cyclic guanosine monophosphate (sGC–cGMP) pathway may offer additional benefits. This study investigated the efficacy and safety of a nutraceutical formulation combining grape pomace extract (Taurisolo^®) and L-arginine in patients with grade 1 and grade 2 hypertension. The formulation was designed to enhance nitric oxide (NO) bioavailability and support sGC–cGMP signaling. Taurisolo^®, a polyphenol-rich extract, is known for its antioxidant and endothelial-protective properties, while L-arginine serves as the physiological substrate for endothelial NO synthase. Clinical outcomes included blood pressure changes, renal function parameters, and health-related quality of life assessed through the SF-12 questionnaire. Supplementation with Taurisolo^® plus L-arginine resulted in significant and sustained reductions in systolic and diastolic blood pressure, with renal function remaining stable throughout the study. Participants also reported meaningful improvements in perceived health, emotional well-being, vitality, and social functioning. The intervention was well tolerated, with no major adverse effects. These findings support the potential of Taurisolo^® combined with L-arginine as a safe and effective adjunctive strategy to conventional antihypertensive therapy, warranting further mechanistic investigation. Full article

In this study, the biological activities of extracts obtained from Ramaria obtusissima were optimized using response surface methodology (RSM) and artificial neural networks-genetic algorithm (ANN-GA) approaches, and the chemical and biological profiles of the obtained extracts were evaluated with a holistic approach. Antioxidant potential was determined using FRAP, DPPH, TAS, TOS, and OSI parameters. It was found that the extract optimized with ANN-GA had significantly higher FRAP (242 ± 3 mg Trolox equivalent/g), TAS (6.64 ± 0.04 mmol/L), and DPPH (154 ± 3 mg Trolox equivalent/g) values compared to the RSM extract, while its OSI value was lower. Anticholinesterase activities were evaluated using IC₅₀ values, and it was determined that the ANN-GA extract exhibited a stronger inhibitory effect on acetylcholinesterase (95 ± 2 µg/mL) and butyrylcholinesterase (125 ± 3 µg/mL) compared to the RSM extract. Antiproliferative effects were investigated in A549, MCF-7, and DU-145 cell lines, and a significant and dose-dependent suppression of cell proliferation was observed in all three cell lines, particularly at concentrations of 100 and 200 µg/mL. The chemical profile was determined using LC-MS/MS and GC-MS techniques. Higher levels of phenolic compounds such as gallic acid (6694.5 ± 4.9 mg/kg), caffeic acid (3374.8 ± 4.9 mg/kg), and quercetin (1563.1 ± 2.3 mg/kg) were found in the ANN-GA extract. GC-MS analyses showed that the ANN-GA extract has a richer lipophilic component profile in terms of biologically active fatty acids and ester derivatives. The findings reveal that AI-assisted optimization offers a powerful and effective approach to enhancing the biological efficacy of mushroom-derived natural products. Full article

Royal jelly, a nutrient-rich bee product characterized by high water content and active components, is particularly susceptible to quality deterioration during storage. While temperature effects have been extensively documented, the specific role of light exposure in quality degradation remains largely unexplored. despite its relevance during production, handling, transportation, and display. This study systematically investigated the volatile organic compounds (VOCs) of royal jelly under different storage conditions using gas chromatography-ion mobility spectrometry (GC-IMS) combined with fingerprint analysis. Results from dual-column validation demonstrated that even short-term light exposure at 25 °C induced pronounced alterations in VOC profiles, triggering the accumulation of off-flavor aldehydes (e.g., hexanal, nonanal) and ketones, along with 2-furfural generated via Maillard reaction. Concurrently, characteristic fresh-aroma esters and alcohols were significantly depleted. Multivariate statistical analysis confirmed light exposure as the predominant factor driving quality deterioration, with temperature variation under dark conditions producing comparatively minor effects within the same short timeframe. This work provides the first systematic evidence establishing insights into early volatile changes in royal jelly and identifies key VOC markers that offer valuable insights for optimizing storage strategies and developing rapid quality monitoring protocols. Full article

To assess the therapeutic relevance of BRIP1 in gastric cancer (GC), we examine its functional role in conferring resistance to anoikis within GC cells and elucidate the oncogenic signaling pathways modulated by BRIP1. By integrating the Cancer Genome Atlas (TCGA) and Gene Set Enrichment Analysis (GSEA) databases with Least Absolute Shrinkage and Selection Operator (LASSO) regression, a novel risk score to stratify GC patients based on prognosis was generated, and a significantly differentially expressed gene, BRIP1, was validated through reverse transcription quantitative polymerase chain reaction (RT-qPCR). Protein expression associated with apoptosis, cell cycle, and epithelial-mesenchymal transformation (EMT) was quantified via RT-qPCR and Western blot (WB). 5-Ethynyl-2′-deoxyuridine (EdU) and cell counting kit-8 (CCK-8) assays were conducted to quantify proliferative activity. The protein level in axillary tumor tissues of nude mice was detected by immunohistochemistry (IHC). We established an eight-gene anoikis-related prognostic risk assessment model (DUSP1, VCAN, P3H2, TXNIP, BRIP1, FGD6, GPX3, and RLN2) for GC. Multivariate Cox regression confirmed the risk score as an independent prognostic factor. Among these genes, BRIP1 showed significant differential expression between tumor and normal tissues, as well as normal gastric mucosal epithelial cells and GC cells. Mechanistically, BRIP1 conferred anoikis resistance to GC cells by suppressing the generation of reactive oxygen species (ROS). We found that the PI3K inhibitor LY294002 counteracted BRIP1-driven oncogenic effects, which was evidenced by restored expression of key regulators governing apoptosis, cell cycle progression, and EMT, in addition to suppressed proliferation in GC cells. BRIP1 is postulated to function upstream of the PI3K/Akt signaling cascade. This study establishes a risk scoring model and identifies BRIP1 as a potential prognostic marker for GC. Full article

Background. NOTCH receptors play a pivotal role in carcinogenesis. Upon ligand binding, a cascade of proteolytic cleavages mediated by ADAM proteases and the γ-secretase complex activates the receptor, ultimately releasing the NOTCH intracellular domain (NICD). NICD translocates to the nucleus, where it regulates gene expression. This review mainly aims to evaluate γ-secretase inhibitors (GSIs) as anticancer agents in preclinical and clinical settings, with a focus on their ability to block tumor progression, target cancer stem cells, and overcome resistance to standard therapies. Methods. A systematic search was conducted in the ISI Web of Science, PubMed, and Scopus databases, following PRISMA guidelines. The review included preclinical in vitro and in vivo studies, as well as clinical trials, investigating GSIs, either as monotherapy or in combination with other treatments, in TNBC, metastatic melanoma, PDAC, gastric cancer, and NSCLC. Exclusion criteria included duplicates, non-English articles, studies published before 2010, studies on non-cancer conditions, research unrelated to NOTCH signaling, and studies outside the selected cancer types. Overall, 69 articles were included and categorized into the five types of cancer analyzed (20 on NSCLC, 22 on TNBC, 11 on metastatic melanoma, 7 on GC, and 9 on PDAC). Of these, 60 studies corresponded to preclinical research in the types of cancer, and 9 studies corresponded to clinical trials in the types of cancer except for GC. Two independent authors screened and extracted relevant data, with disagreements resolved by the corresponding author. Findings were synthesized qualitatively across cancer types under study. Results. This review summarizes therapeutic advances involving GSIs in cancers driven by oncogenic NOTCH signaling, based on the 69 articles included. Preclinical studies show that GSIs synergize with chemotherapy and radiotherapy, particularly in NSCLC, melanoma, and TNBC, and block EMT, overcome therapeutic resistance, and improve prognosis. Commonly used GSIs include DAPT and RO4929097, which enhance the efficacy of agents, such as gemcitabine (PDAC), paclitaxel, osimertinib, erlotinib, and crizotinib (NSCLC), and 5-FU (gastric cancer, TNBC). Promising strategies include combining GSIs with SAHA, ATRA, CB-103, and other NOTCH signaling targeting molecules, either alone or with chemo- and radiotherapy. Clinical trials with GSIs, however, remain limited. RO4929097 is the most extensively tested GSI in clinical settings. PDAC trials combining GSIs with gemcitabine showed no benefit; melanoma trials yielded modest outcomes; and TNBC trials demonstrated partial responses to GSIs but overall low efficacy and significant adverse events. Discussion and Conclusions. Despite encouraging preclinical evidence, clinical trials with GSIs have underperformed, largely due to tumor heterogeneity, dosing limitations, and the non-selective nature of γ-secretase inhibition. Other NOTCH inhibitors, such as DLL4 antibodies, also resulted in partial responses and secondary effects. Future strategies should prioritize receptor-specific NOTCH inhibitors, patient stratification based on NOTCH pathway activation, and optimized combination regimens. Emerging approaches include integrating immunotherapy with advanced technologies such as CRISPR, CAR-T cells, and bispecific antibodies, as well as targeted delivery systems to enhance efficacy and reduce toxicity. Additional research directions include addressing the tumor microenvironment and EMT-driven resistance, elucidating the mechanisms of immune evasion, and inhibiting tumor angiogenesis. Finally, leveraging artificial intelligence and big-data-driven personalized medicine, including sex-specific considerations, will be essential for improving patient outcomes. Full article

Growing interest in natural therapies has increased the demand for essential oils; however, the complex interactions within their mixtures that dictate their final efficacy remain poorly understood. This study aimed to optimize a blend of ginger, cinnamon, tea tree, and geranium essential oils to develop an active ingredient, with synergistic multifunctional bioactivities, that was relevant to cutaneous healing. Initially, the composition and cytotoxicity for individual oils were determined; subsequently, a D-optimal mixture design was employed to evaluate three biological responses related to skin recovery: ultraviolet B radiation absorption, red blood cell lysis inhibition, and catalase enzyme activity. GC-FID analysis revealed the following major components (% w/w): cinnamon (cinnamaldehyde, 77.56%), ginger (α-zingiberene, 33.77%), geranium (citronellol, 33.6%), and tea tree (terpinen-4-ol, 38.38%). Dose–response data from essential oils tested against Detroit ATCC 551 skin fibroblasts revealed a clear cytotoxic hierarchy (IC₅₀ µg/mL): cinnamon (21.03) > ginger (25.3) > tea tree (41.67) > geranium (92.51). Cinnamaldehyde content was the primary contributor to photoprotective capacity, with a maximum sun protection factor (SPF) of 4.5. Inhibition against erythrocyte membrane lysis was not attributable to a single component; maximum protection (98.4%) was achieved through synergy between oxygenated monoterpenoids (geranium and tea tree), sesquiterpenes (ginger), and aromatic aldehydes (cinnamon). Highest catalase activity (160.86 kU/g Hb) was reached in mixtures with high cinnamaldehyde and eugenol contents, whereas an antagonistic effect was observed between tea tree and geranium oils. Finally, an optimal formulation (desirability = 0.927) was identified (% w/w): 31.7% ginger, 39.1% cinnamon, 14.5% tea tree, and 14.7% geranium. Experimental validation confirmed no significant difference compared with developed predictive models. This optimized mixture constitutes a bioactive natural component with potential for use in products aimed at promoting skin health, warranting further investigation into direct models of skin healing. Full article

Epilepsy remains one of the most prevalent neurological disorders, characterised by complex aetiology encompassing genetic, structural, metabolic, and inflammatory factors. Despite advances in neuroimaging and neurophysiological diagnostics, there is a persistent lack of sensitive and specific biomarkers to enable early diagnosis, risk stratification, and monitoring of therapeutic efficacy. Key epilepsy biomarkers include neurotransmitters, energy–related compounds, tryptophan pathway metabolites, and choline derivatives. Their determination employs liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS), high–performance liquid chromatography (HPLC) with electrochemical or fluorescence detection, gas chromatography with tandem mass spectrometry (GC–MS/MS), high–resolution mass spectrometry (HRMS), and proton nuclear magnetic resonance (¹H–NMR) spectroscopy, revealing metabolic disturbances in neurotransmission, energy metabolism, and oxidative stress associated with epileptogenesis. Among these techniques, LC–MS/MS currently provides the highest analytical sensitivity and specificity for quantifying low–abundance epilepsy–related metabolites, while HPLC with conventional detection remains a simpler and more cost–effective alternative for routine clinical laboratories. This review presents the current state of knowledge regarding chromatographic techniques applied to the analysis of mentioned metabolites, as well as therapeutic drug monitoring of antiepileptic drugs. Key sample preparation stages are also discussed. Various biological matrices–plasma, serum, urine, cerebrospinal fluid (CSF), dried blood spots (DBSs), and brain tissue—are evaluated. Novel approaches are also presented, including hair samples, microsampling techniques, and headspace analysis of volatile metabolites. Chromatographic techniques constitute the foundation of contemporary metabolomic research in epileptology, enabling biomarker identification and supporting personalised medicine. Further standardisation and translational validation remain necessary, as current evidence is insufficient for routine clinical implementation. Full article

Ultrafine dark tea powder (UDTP) was prepared by superfine grinding and sieving through a 200-mesh sieve, and incorporated into cookies to improve their textural properties, sensory acceptability and flavor characteristics. Through single-factor experiments and orthogonal testing, the optimal formulation was determined. The quality of cookies was evaluated by texture analysis, sensory evaluation, electronic nose (E-nose), and gas chromatography-mass spectrometry (GC-MS). Results showed that cookies supplemented with 4 g UDTP per 80 g butter exhibited significantly lower hardness and comparable fracturability, along with higher sensory scores in texture, odor and taste compared to basic butter cookies. E-nose and GC-MS analyses revealed that UDTP enrichment promoted the formation of desirable volatile compounds, particularly aldehydes, ketones, and heterocyclic compounds, which contributed to floral, fruity, roasted nutty, and caramel aromas. This study demonstrates that UDTP can effectively improve both the textural and flavor properties of cookies, providing a viable approach for developing tea-fortified baked products with enriched sensory profiles. Full article

Photocatalytic processes have been studied as promising solutions to mitigate the impact of pollutants on aquatic environments. Here, the enhancement of photocatalytic performance and stability of titanate nanostructures (TNS), a well-established photocatalyst, were investigated through Sr modification. Structural characterization confirmed Sr in-corporation in the crystalline structure, mainly in the interlayers. The sample Sr(5%)TNS, synthesized with 5% (wt.), exhibited fine lamellar morphology, different from the elongated nanowires of pristine TNS. The photocatalytic performance of the Sr-modified sample was studied for the removal of a model pollutant, caffeine, under UV-Vis and visible irradiation. A clear enhancement in the caffeine removal rate was observed using Sr(5%)TNS as a photocatalyst, when compared with the pristine material. Further improvement in the photocatalytic performance was obtained by combining Sr(5%)TNS with graphitic-like carbon nitride (g-C₃N₄) as a novel composite film. This proved to be a promising strategy for enhancing both the visible-light photocatalytic efficiency and the stability of the films, while also facilitating their reuse. Various configurations of the hybrid system were tested, and the best results for caffeine degradation and catalyst robustness were achieved with a 4:1 ratio of Sr(5%)TNS to g-C₃N₄. Mechanisms for charge transfer in irradiated Sr(5%)TNS particles, and in Sr(5%)TNS/g-C₃N₄ composite films are proposed and discussed. Full article

In vitro fecal inoculation coupled with gas chromatography–mass spectrometry (GC-MS) has been used for evaluating fecal deodorants. However, high cost and complex data interpretation limit its routine application. An electronic nose (eNose) offers a rapid, cost-effective alternative. This study aimed to evaluate the eNose as a screening tool for fecal odor compared with solid-phase microextraction gas chromatography–mass spectrometry (SPME GC-MS) and to examine the in vitro effects of fecal deodorant supplements on fecal odor profiles. Feces from ten healthy cats were serially diluted (1:1 to 1:8) and analyzed using both instruments. Four dietary supplements—Yucca schidigera extract (YSE), Quillaja saponaria extract (QSE), fructooligosaccharides (FOS), and oat beta-glucans (OBG)—were tested at concentrations of 0.0, 0.2, 0.4, and 0.8 g/100 mL. The eNose showed comparable performance to GC-MS in discriminating among sample dilutions. In vitro fermentation showed that FOS and OBG significantly increased volatile fatty acid (VFA)-related sensor responses while signals linked to ammonia and sulfur compounds were reduced. QSE had minimal effect, whereas YSE produced moderate changes. The total sensor response intensities did not differ between treatments. These findings indicate that prebiotic supplements exert stronger effects than saponin-based supplements and highlight the potential of eNoses with in vitro fermentation for rapid screening of fecal deodorants. Full article

In vitro studies were conducted in a series to investigate if the ruminal microbes are capable of degrading chlorpyrifos. This in vitro study presents the results from three experiments: Exp. I was conducted without feed, while Exp II and III were conducted with feed, either with or without methanol for dissolving chlorpyrifos, respectively. A basal diet comprising finger millet straw and concentrate was prepared. Incubation medium with feed but without chlorpyrifos served as the control. A total of six replicates each of control and chlorpyrifos spiked were used for the incubation. The pesticide concentration in the incubation medium before and after 24 h of incubation was analyzed using GC-MS/MS. The genomic DNA was isolated from the incubation fluid of the individual samples, and the shotgun metagenomic sequencing was performed. The clean reads were taxonomically classified using the Kraken2 database, and microbial classification at different taxonomic ranks was separated using Pavian v1.0. The microbial genes in the metagenome data were predicted and assigned functional roles using the MetaErg v1.2.3 pipeline. The assigned KEGG Orthology (KO), EC numbers (Enzyme Commission number), Gene Ontology (GO), and corresponding NCBI taxonomy information relevant to chlorpyrifos metabolism/degradation were retrieved. Results from the study revealed that the chlorpyrifos concentration was decreased from 5.78 to 1.64 ppm over 24 h of in vitro incubation with feed. Similar alpha and beta diversity indices between control and chlorpyrifos treatments revealed that the richness and the evenness of the microbial population were not affected by the presence of chlorpyrifos in the rumen fluid. There was no difference in the microbiota affiliated to the major phyla such as Bacteroidota, Fibrobacterota, Bacillota, and Pseudomonadota. The EC 3.1.8.1, EC 3.1.3.1, EC 1.14.13.-, and EC 1.1.1.- reported for chlorpyrifos degradation were not detected in the metagenome, and only EC 3.1.1.1 was identified, which demonstrated that degradation of chlorpyrifos was carried out by the affiliated enzyme carboxylesterase. The presence of GO:0004035, GO:0004364, GO:0019637, GO:0016791, and GO:0042178 in the metagenome strengthens that the chlorpyrifos degradation in the present study was primarily assigned to the rumen microbiota. This in vitro study provided insights into the rumen microbiota involved in the chlorpyrifos degradation and the initial clue that the rumen microbes are capable of degrading chlorpyrifos. Further, the animal studies in different species with the variable levels of chlorpyrifos are also warranted to confirm the efficacy of rumen microbes in mixed syntrophy and determine the threshold capabilities of the ruminal microbes. Full article

Biopolymers are increasingly explored as safer and more sustainable food packaging materials. This study evaluated the migration behavior of intentionally and non-intentionally added substances (IAS and NIAS), as well as the safety of gelatin and xanthan gum blends reinforced with microcrystalline cellulose, with and without oxygen plasma treatment, incorporating glycerol and limonene as plasticizers. Migration tests were conducted according to European Union (EU) Regulation No. 10/2011 using simulants of different polarities, and IAS/NIAS were analyzed by gas chromatography–mass spectrometry and ultra-high-pressure liquid chromatography–quadrupole time-of-flight mass spectrometry (GC–MS and UPLC-QTOF-MS). Films containing limonene were also evaluated for antioxidant activity. Results showed that plasticizer migration is strongly influenced by simulant polarity, glycerol predominantly migrated into hydrophilic media, whereas limonene and its derivatives exhibited higher migration in fatty simulants. Ethanol 95% acted as a conservative worst-case simulant, promoting extensive migration, while substantially lower migration levels were observed in isooctane and tenax plasma treatment resulted in modest changes in volatile compound migration, while significantly enhancing the antioxidant activity of limonene-containing films. Although overall migration levels were low under most of the tested conditions, NIAS formation, particularly from limonene degradation, highlights the need to account for chemical stability and simulant type when assessing bio-based films. Overall, the study demonstrates that film composition, surface modification, and simulant characteristics jointly influence migration behavior and functional performance under the evaluated conditions reinforcing the need to adapt current regulatory frameworks to the specific behavior of biopolymeric packaging materials. Full article

Background: Chrysopogon zizanioides (L.) Roberty (vetiver) is a perennial medicinal grass with deep aromatic roots traditionally used for several ailments. Its root essential oil (CZEO) is rich in phytochemicals with documented antimicrobial, anti-inflammatory, and antioxidant activities. Although its anticancer potential remains underexplored, the complex phytochemical profile of CZEO positions it as a promising multi-target therapy, particularly for colorectal (CRC) and lung cancers where resistance and pathway redundancy often limit conventional treatments. Therefore, this study aimed to investigate the phytochemical composition and antiproliferative activity of CZEO from Qatar against colorectal (HCT-116) and lung (A549) cancer cells and to elucidate its molecular targets and mechanisms of action in CRC and lung cancer using network pharmacology and in silico approaches. Methods: CZEO was extracted by steam distillation and characterized using GC–MS. In vitro proliferation assays with HCT-116 colorectal and A549 lung cancer cells were conducted using the Alamar Blue assay. The ten most abundant phytochemicals identified by GC–MS were assessed for drug-likeness and ADMET properties and further analyzed through network pharmacology, molecular docking, and molecular dynamics (MD) simulations to elucidate the molecular targets and mechanisms underlying CZEO’s anticancer activity. Results: GC-MS profiling identified 40 compounds, predominantly sesquiterpenoids (93%), including khusimol, β-eudesmol, α-vetivone, and rosifoliol. CZEO inhibited cancer cell viability in a dose-dependent manner, with IC₅₀ values of 62.95 ± 2.19 µg/mL for HCT-116 and 167.82 ± 6.51 µg/mL for A549 cells, demonstrating greater potency against colorectal cancer. CZEO did not affect the growth of normal human neonatal fibroblasts (HDFn), suggesting potential selectivity for cancerous cells. ADMET predictions indicated favorable pharmacokinetics and low toxicity of CZEO’s top 10 abundant compounds (TACs). Network pharmacology revealed 373 and 394 overlapping gene targets between TACs and lung and colorectal cancer, respectively. The overlapping genes were used to construct a protein–protein interaction (PPI) network to identify hub genes. STAT3 and AKT1 consistently emerged as common top-scoring hub genes in both cancers. Molecular docking of TACs showed strong binding affinities of rosifoliol and α-vetivone to AKT1 (−6.20 and −5.93 kcal/mol, respectively) and STAT3 (−5.19 and −5.09 kcal/mol, respectively), surpassing reference inhibitors. MD simulations confirmed stable ligand–protein interactions and structural stabilization, particularly with α-vetivone. Conclusions: CZEO from Qatar exhibits potent antiproliferative activity against colorectal and lung cancer cells, supported by a sesquiterpenoid-rich phytochemical profile. Integrative computational analyses highlight AKT1 and STAT3 as key molecular targets, with rosifoliol and α-vetivone emerging as promising lead compounds. These findings support CZEO as a natural, multi-target anticancer agent, warranting further mechanistic and in vitro and in vivo validation. Full article

Slovenia preserves one autochthonous pig breed, the Krškopolje pig, whose meat has been reported to exhibit a favourable fatty acid profile compared with that of modern breeds. However, meat quality is not solely determined by genetics; the production system also influences it, as organic and conventional farming differ in feed composition, housing and outdoor access. This study aimed to compare the effects of pig breed (Krškopolje vs. modern) and production system (organic vs. conventional) on the fatty acid composition and volatile organic compound (VOC) profile of pork. Fatty acid composition was determined by GC-FID after methylation, and the VOCs profile was obtained using headspace solid-phase microextraction (HS-SPME) coupled with GC-MS. Results showed that Krškopolje meat had higher SFA and MUFA, while modern pig meat had higher PUFAs, particularly n-6, reflecting genetic and dietary influences. Modern breeds also showed greater fatty acid response to the rearing system than the Krškopolje breed. Several VOCs were unique to modern breed pigs, indicating breed-specific differences in lipid composition, amino acid metabolism, and microbial activity. Aldehydes were the dominant VOC class in both breeds, slightly higher in Krškopolje meat. OPLS-DA model revealed breed-related differences in VOCs, pinpointing compounds likely responsible for breed-specific aroma and flavour. Full article

(1) Background: Primary metabolism is essential for tea quality formation, however systematic analysis of tea primary metabolites remains limited; (2) Methods: An orthogonal design was used to optimize the pre-column derivatization reaction conditions for the tea matrix. Using the optimized method, gas chromatography–mass spectrometry (GC–MS) was employed to analyze Longjing green tea (LJGT) samples from different processing stages and regions; (3) Results: Optimal derivatization was achieved with 75 μL of methoxamine hydrochloride pyridine solution at 30 °C for 1.5 h. A total of 52 primary metabolites were identified in LJGT. Processing analysis showed that 8 metabolites associated with carbohydrate metabolism significantly decreased during spreading. Five reducing sugars significantly decreased, while sucrose, turanose, and quinic acid significantly increased due to the thermal action during pan-fixation. Additionally, 15 key differential compounds were identified among three regions (Xihu, Qiantang, and Yuezhou) of LJGT. Quantitative analysis revealed that shikimic and quinic acid contents were significantly higher in the Xihu region compared to other regions; (4) Conclusions: This study established a pre-column derivatization GC-MS method for primary metabolite profiling, elucidated metabolic regulation during LJGT processing, and identified differences in primary metabolite content among LJGT from different geographical origins. Full article