The development of high-throughput sample pretreatment methods is essential for the rapid screening and accurate analysis of contaminants in food and environmental matrices. In this study， we established a novel high-throughput sample pretreatment approach for glyphosate using a 96-blade thin-film microextraction （TFME） device. The blade surface was coated with a functionalized metal-organic framework （MOF） as the extractive phase. Specifically， a fluorinated zirconium-based MOF （UiO-66-F₄） was synthesized via a hydrothermal method. Various characterization techniques， including scanning electron microscopy （SEM）， transmission electron microscopy （TEM）， Fourier-transform infrared spectroscopy （FT-IR）， X-ray diffraction （XRD）， and nitrogen adsorption-desorption isotherms， were employed to confirm the successful synthesis of UiO-66-F₄ crystals. The results demonstrated that the synthesized crystals possessed a high specific surface area （559.5 m²/g） and an irregularly stacked porous structure. The prepared UiO-66-F₄ was subsequently homogenized with a polyacrylonitrile （PAN） solution to form a slurry. This slurry was solidified onto 96 stainless-steel blades using an improved laboratory-developed spraying technique. As a result， an array of 96 UiO-66-F₄/PAN extraction coatings was obtained. Each coating in the array was fabricated with uniform dimensions of 2 cm in length， 0.2 cm in width， and approximately 100 μm in thickness. The relative standard deviations （RSDs， n=8） for the length， width， and thickness of the coatings measured were 0.29%， 0.48%， and 4.64%， respectively. The UiO-66-F₄ adsorbent offers multiple interaction forces with glyphosate， including metal coordination， hydrogen bonding， and electrostatic interactions. Therefore， the adsorption of glyphosate onto UiO-66-F₄ occurred rapidly， reaching equilibrium within 5 min. The pseudo-second-order kinetic model provides a superior fit for the adsorption kinetics behavior of glyphosate on the surface of UiO-66-F₄， suggesting that the adsorption process is predominantly controlled by chemisorption. Furthermore， the glyphosate adsorption isotherms indicate that its adsorption behavior closely conforms to the Langmuir isotherm model （R²>0.97）. This finding suggests that glyphosate is adsorbed as a monolayer on the adsorbent surface， giving rise to a maximum adsorption capacity of 239.4 mg/g. The developed TFME method efficiently enriches glyphosate from aqueous solutions at pH 5–7 within 30 min and allows for analyte desorption using an aqueous solution containing 5% （V/V） triethylamine. The UiO-66-F₄/PAN coatings displayed high repeatability and reproducibility， as evidenced by the RSD values below 2.3% （n=5）. In addition， the coatings exhibited remarkable robustness， maintaining a recovery of 98.2% after nine consecutive TFME processing cycles. Furthermore， coupling this TFME method with ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） enabled high-throughput analysis of glyphosate. The method demonstrated excellent linearity over a range of 0.5–500 ng/mL （R²=0.999 9）. The limits of detection （LOD） and quantification （LOQ） were 0.12 ng/mL and 0.5 ng/mL， respectively. Intra-day （n=3） and inter-day （n=5） precisions， expressed as RSDs， were 3.6% and 1.6%， respectively. The method was successfully applied to real-world environmental water and fruit samples， detecting glyphosate residues of （1.7±0.7） ng/g in orange peel and （1.2±0.5） ng/mL in irrigation river water. Spiked recoveries in real samples ranged from 88.1% to 107.1%， with precisions （RSDs， n=3） below 8.1%. This method is accurate， reliable， and reproducible， enabling parallel processing of 96 samples， which are well-suited for rapid screening and precise quantification of glyphosate contamination in large-scale sample analyses.

The disinfection of water is a fundamental and indispensable process in modern public health infrastructure. It plays a pivotal role in safeguarding drinking water safety by inactivating pathogenic microorganisms such as bacteria and viruses， thereby preventing waterborne disease outbreaks. However， a significant and well-documented drawback of chemical disinfection， particularly with chlorine-based agents， is the formation of disinfection byproducts （DBPs）. These DBPs are generated through complex reactions between disinfectants and natural organic matter present in source water. Among the vast array of identified DBPs， halogenated compounds are of particular concern due to their enhanced toxicity and persistence. 2，5-Dichloro-1，4-benzoquinone （2，5-DCBQ） is a prominent representative of these toxic halogenated DBPs. Toxicological studies have conclusively demonstrated that 2，5-DCBQ can induce pronounced oxidative stress in mammalian cells. This leads to a cascade of detrimental effects including a marked increase in intracellular reactive oxygen species. Subsequently， the detrimental effects may cause DNA single-strand breaks， oxidative base damage， and chromosomal aberrations. Consequently， developing effective technologies for the targeted removal of 2，5-DCBQ from aqueous environments is of paramount importance for mitigating its associated health risks. Current removal strategies， including adsorption using solid-phase materials， often suffer from several limitations. These include reliance on a single interaction mechanism， modest adsorption capacities， or slow kinetics. This hinders their practical application for efficient 2，5-DCBQ control. To address these challenges， this study presents the design， synthesis， and application of a novel porous organic adsorbent. Termed Tp-Azo， this adsorbent is specifically engineered for the rapid and efficient removal of 2，5-DCBQ. Tp-Azo was synthesized via a condensation reaction and possesses an exceptionally high specific surface area of 824.5 m²/g. It incorporates a high density of electron-rich azo （-N=N-） functional groups within its framework. These groups act as strong electron donors， enabling potent electron donor-acceptor interactions with the electron-deficient quinone ring of 2，5-DCBQ. Furthermore， the predominantly aromatic backbone of Tp-Azo facilitates strong π-π stacking interactions with the aromatic structure of 2，5-DCBQ. The inherent hydrophobicity of Tp-Azo promotes additional adsorption via hydrophobic effects. Critically， the material possesses an average pore diameter of approximately 2.0 nm. It is substantially larger than the estimated molecular kinetic diameter of 2，5-DCBQ. This suitable pore size ensures minimal diffusion resistance， allowing 2，5-DCBQ to rapidly access the internal adsorption sites.The adsorption performance of Tp-Azo was systematically evaluated under various conditions. Under pH 2-3， the adsorption efficiency was reduced. This is attributed to the protonation of 2，5-DCBQ， which diminishes its hydrophobicity. The shielding effect of high H⁺ concentration can also weaken electrostatic interactions. In contrast， within pH 4-6， Tp-Azo exhibited consistently high removal rates. Therefore， pH 5 was selected as the optimal condition for subsequent experiments to simulate typical water treatment scenarios. The adsorption kinetics were remarkably fast within the first 3 min and complete equilibrium was reached within 20 min. The kinetics were best described by the pseudo-second-order model， indicating that the adsorption process is primarily governed by chemisorption. Isotherm studies revealed that the adsorption capacity increased with the initial 2，5-DCBQ concentration. They were accurately modeled by the Langmuir isotherm， suggesting a homogeneous adsorption surface and monolayer coverage. The calculated maximum adsorption capacity was significantly higher than that reported for many conventional adsorbents.Swimming pool water and tap water were spiked with 2，5-DCBQ at 5 mg/L and 50 mg/L. Impressively， Tp-Azo achieved removal efficiencies of 96.2% and 96.6% at 5 mg/L， 87.9% and 88.7% at 50 mg/L， respectively. Given that realistic environmental concentrations of halogenated DBPs like 2，5-DCBQ are typically in the μg/L range， this suggests that Tp-Azo is highly capable of achieving virtually complete removal in real-world applications. The study provides a novel and highly effective material enabling targeted removal of halogenated DBPs. It also offers a strategic design principle for developing advanced adsorbents for DBPs in water treatment.

To address the evolving demands for innovation and practical competencies in pharmaceutical education， and in alignment with the “student-centered， outcome-driven” pedagogical approach， our teaching team has undertaken a comprehensive reform of the Pharmaceutical Analysis curriculum. We designed and implemented an integrative， research-based laboratory experiment for undergraduate students. Centered on the design and application of functional materials， the experiment guides students through the independent synthesis of a halogen-bonded organic framework， designated as XOF-1， followed by systematic structural characterization， performance evaluation， and analytical application. The experiment is grounded in real-world environmental analysis challenges， requiring students to employ the synthesized material as a dispersive solid-phase extraction （DSPE） adsorbent for the selective enrichment of typical brominated flame retardants， including tetrabromobisphenol A （TBBPA）， tetrachlorobisphenol A （TCBPA）， and hexabromobenzene （HBB）， and to achieve the removal of TBBPA from actual water samples. The instructional process fully encompasses the workflow from material preparation and structural characterization to method development and performance assessment， enabling students to experience a complete research pathway from material design to practical application. This approach immerses them in the scientific inquiry cycle of theory-design-experiment-analysis. Through this experiment， students gain solid proficiency in synthesizing and characterizing functional materials and applying them to sample pretreatment. By engaging in experimental design， condition optimization， and data interpretation， they develop critical thinking， problem-solving skills， and capacity for independent inquiry. The exercise fosters deep integration of theoretical knowledge and practical skills， stimulates research interest， strengthens innovative and applied mindsets， and cultivates a sense of professional responsibility in environmental and public health. Ultimately， it achieves a cohesive blend of knowledge acquisition， competency development， and value cultivation， fulfilling the overarching goal of holistic education through experimental learning.

Synthetic phenolic antioxidants （SPAs） are a major class of chemical additives extensively employed in plastics， rubber， and food packaging to prevent polymer degradation and extend product lifespans. In recent years， the environmental and human health risks of SPAs have received great concern due to their widespread occurrence in various environmental media. Existing reviews primarily focus on environmental detection， human exposure， toxicity， and risk assessment of SPAs with limited discussion on recent advances in analytical methodologies. This review presents a systematic synthesis of research advances over the past five years， especially the state of the art analytical methodologies. This review systematically analyzes their occurrence in water， sludge， sediments， soils， dust， and biota tissues， revealing a clear pollution transition from conventional SPAs to emerging high-molecular-weight substitutes. Toxicological evidence demonstrates that SPAs exert adverse effects on rats， mice， and aquatic organisms， including genotoxicity， immunotoxicity， endocrine disruption， reproductive toxicity， etc. Notably， certain transformation products （TPs） of butylated hydroxytoluene （BHT） exhibit enhanced toxicity relative to their parent compounds. Methodological advances in sample preparation and analysis techniques of SPAs in different environmental matrices are reviewed for the first time. Recent applications of atmospheric pressure chemical ionization coupled with liquid chromatography-tandem mass spectrometry （APCI-LC-MS/MS）， as well as high-resolution mass spectrometry （HRMS） provide powerful approaches for target/non-target analysis of unknown SPA derivatives and TPs. Several critical research gaps have finally been identified. Environmental analytical methods of SPAs and their TPs face significant challenges due to their diverse physicochemical properties and chemical reactivity， particularly their susceptibility to oxidation. Another key issue is that current pretreatment approaches for SPAs in tissue samples overlook bound-SPAs， thereby underestimating their real body burdens. Moreover， existing studies mainly focus on traditional SPAs， while lacking systematic assessment of environmental fate and long-term ecological risks posed by emerging SPAs and potentially more toxic TPs. Additionally， ecotoxicological data of SPAs for terrestrial organisms are scarce compared to aquatic organisms and mammals， limiting comprehensive ecological risk assessment. Future research should prioritize： （1） developing oxidant-resistant pretreatment methodologies coupled with HRMS to overcome technical challenges in accurate quantification of trace SPAs and their bound species； （2） strengthening non-target screening and identification of emerging SPAs and TPs； （3） investigating long-term effects of SPAs in soil-plant-microorganism systems. This review advances our understanding of SPAs by bridging the knowledge gaps and providing updated insights into current research. It will help establish the scientific basis for comprehensive ecological risk assessment of SPAs.

Chromatography remains a cornerstone technique in analyzing chiral compounds. The development and selection of chiral stationary phases （CSPs） are central to achieving high-resolution enantiomeric separation. This study presents a novel regeneration and functionalization strategy for the preparation of high-performance superficially porous particle （SPP）-CSP， offering a promising alternative to conventional fully porous particle （FPP）-CSP. First， bonded phases on the surface of spent chromatographic column silica packing materials were removed using ultraviolet light （λ=254 nm） and hydrogen peroxide solution （3%， mass fraction）， thereby obtaining sub-3 μm regenerated SPP. The regenerated SPP were then amino-functionalized. Subsequently， under mild reaction conditions， carbonyldiimidazole-activated （2-hydroxyethyl）-β-cyclodextrin （HE-β-CD） was bonded onto the surface of the amino-functionalized SPP to prepare the SPP-CSP. For comparative evaluation， a CSP based on 3 μm commercial FPPs was prepared. Additionally， a commercial （2-hydroxypropyl）-β-cyclodextrin （Hp-β-CD） chiral column with identical particle size and structure was selected as a reference. This commercial column was used to investigate the selectivity differences among different cyclodextrin derivatives. The enantioseparation and overall chromatographic performance of the CSPs were systematically evaluated under both supercritical fluid chromatography （SFC） and reversed-phase liquid chromatography （RPLC） modes. The evaluation was performed using ten chiral molecular probes： zopiclone， equol， 2-phenylpropionic acid， dobutamine， flutriafol， miconazole， folic acid， catechin， valacyclovir， and chlorphenamine. Results revealed that， under identical chromatographic conditions， the SPP-CSP consistently outperformed the FPP-CSP in terms of enantiomeric resolution， peak sharpness， theoretical plate number， and retention time efficiency. More importantly， the FPP-CSP exhibited a significant loss in resolution when chromatographic conditions were altered to yield equivalent analysis times. In contrast， the SPP-CSP showed only minor adverse effects on resolution when the flow rate was increased to shorten analysis time， highlighting its inherent suitability for ultrafast separations. Moreover， SFC and RPLC modes displayed complementary chiral recognition ranges. Within the overlapping separation scope of both modes， SFC mode achieved higher enantiomeric resolution and superior chromatographic performance. Notably， under RPLC mode， catechin showed high enantiomeric resolution on both CSPs， underscoring system-dependent selectivity differences. Multiple factors—including mobile phase composition （particularly the additive type）， and the stereochemical structure of the analytes—were found to significantly influence enantioseparation outcomes. Under SFC mode， none of the analytes could be eluted when acetonitrile was used as the mobile phase modifier， whereas alcohols effectively enabled elution， with weaker hydrophobicity correlating to stronger elution strength. Among the alcohols tested， methanol offered distinct advantages over isopropanol by maintaining lower backpressure and superior chromatographic performance without compromising resolution. Under RPLC mode， although isopropanol as a mobile phase component provided higher enantiomeric resolution， it resulted in relatively poorer chromatographic performance and higher column pressure drop. Using acetonitrile offered limited flexibility in adjusting elution strength. In both chromatographic modes， the influence trends of mobile phase additives on enantioseparation performance and overall column performance were generally consistent. Mobile phases with acidic additives were more suitable for analyzing acidic analytes such as 2-phenylpropionic acid and folic acid. Mobile phases with ammonium formate were better suited for analyzing basic analytes like chlorphenamine and flutriafol， as well as analytes with pK_a values near 7， such as zopiclone. In contrast， NH₃ as an additive yielded unsatisfactory results. Compared with the commercial Hp-β-CD chiral column of the same particle size and structure， the SPP-CSP exhibited different selectivity， which varied with the chromatographic mode. Under RPLC mode， the SPP-CSP showed higher resolution for zopiclone， dobutamine， and folic acid. Under SFC mode， it demonstrated higher resolution for equol， folic acid， and valacyclovir. Despite these advances， the study has certain limitations. To enhance compatibility with mass spectrometric detection， normal-phase mode was not investigated. Additives commonly used in chiral compound analysis， such as triethylamine and ammonium trifluoroacetate， were also not employed. Polar organic solvent mode was not investigated either. Additionally， peak symmetry and theoretical plate numbers on the SPP-CSP were generally inferior to those observed on the commercial Hp-β-CD chiral column across both chromatographic modes.

The illegal addition of substances in ordinary food poses a serious threat to food safety. Therefore， the detection and identification of such additives have long been important research areas in the field of food safety. Especially， illegal additives are showing new trends now， such as emerging functional varieties， new structural derivatives， and clandestine addition practices. Meanwhile， along with the development of online shopping and logistics， new threats pose a substantial challenge to social governance and food safety. In recent years， there are frequent cases of people consuming wild mushrooms containing hallucinogenic tryptamine toxins out of curiosity， and the abuse of tryptamine psychoactive substances also occurs from time to time. Therefore， the development of a robust and sensitive analytical method is necessary for accurately identifying the risks associated with tryptamine psychoactive substances.To address this issue， a novel method based on ultra performance liquid chromatography-tandem mass spectrometry （UPLC-MS/MS） combined with an isotopic internal standard and solid-phase extraction purification technique was developed for the simultaneous determination of three tryptamines （psilocin， bufotenine and psilocybin） in ten types of ordinary foods. Several optimization procedures were undertaken， involving the chromatographic conditions， mass spectrometric parameters， and sample extraction and purification protocols. The food samples were extracted and centrifuged， followed by purification using a strong cation exchange solid-phase extraction column. The separation was carried out using a Capcell PAK ST （S-5） hydrophilic interaction chromatographic column. The multi-reaction monitoring （MRM） mode was employed for data acquisition in the positive ion mode， whereas isotopic internal standards were used to quantify the three tryptamines. It was verified that the calibration curves of psilocin， bufotenine， and psilocybin exhibited good linearities （R²>0.995） within the range of 0.025-10.0 ng/mL. The limits of detection （LODs） for psilocin， bufotenine and psilocybin were determined as 0.4， 0.5， and 0.6 µg/kg， respectively， while the limits of quantification （LOQs） were determined as 1.4， 1.7， and 1.9 µg/kg， respectively. The LODs and LOQs demonstrate that the method has the ability to detect minute quantities of tryptamines. The recoveries of the tryptamines were 94.3%-113.7% under three spiked levels， with relative standard deviations （RSDs） ranging from 0.4% to 4.9% （n=6）. The consistent and satisfactory recoveries of tryptamines indicate that this method meets the requirements to accurately quantify the target analytes even in a complex matrix. In summary， this newly developed method is rapid， sensitive， accurate， and compliant with relevant regulatory standards； hence， it is suitable for the determination of psilocin， bufotenine and psilocybin in food matrices. As part of the application of this method， a comprehensive investigation was conducted on ten types of actual food samples， including candies， beverages， wines， succade， coffee， health care products， condiments， cooked meat products， spices and hot pot sauce. As a result， out of 100 samples， 14 samples were found to contain bufotenine. Additionally， one sample had a bufotenine concentration of 7.6 mg/kg. By tracing back， it was discovered that the bufotenine in the positive samples was derived from Zanthoxylum armatum DC. or Zanthoxylum Bungeanum Maxim. Subsequently， 12 samples， containing Zanthoxylum armatum DC. and Zanthoxylum Bungeanum Maxim， were collected and processed using the same method， which revealed the bufotenine in all samples with the content ranging from 23.8 to 885.0 mg/kg. At the same time， it was found that the daughter ion scanning mass spectra of the Zanthoxylum Bungeanum Maxim sample were almost consistent with those of the bufotenine standard. Based on this， it can be firmly concluded that bufotenine is naturally present in Zanthoxylum armatum DC. and Zanthoxylum Bungeanum Maxim. In conclusion， the developed method and the discovery of the content of bufotenine in Zanthoxylum armatum DC. and Zanthoxylum Bungeanum Maxim provide a reference framework for future research in the areas of food safety risk assessment， the screening of illegal additives， and food product development.

Tea is an important cash crop in China， and pesticide residues in tea products are among the main issues affecting tea exports. Glufosinate （GLU） is a widely used herbicide in tea gardens and its residues in tea samples have raised health concerns. However， there are few reports on analytical methods for the simultaneous determination of GLU and its metabolite residues in tea. Therefore， the development of suitable analytical methods to monitor the content of GLU and its metabolite residues in tea is essential to ensure compliance with the export quality standards of tea products. In this study， a sensitive analytical method was developed for the determination of GLU and its two main metabolites， N-acetyl-glufosinate （NAG） and 3-［hydroxy（methyl）phosphinoyl］propionic acid （MPP）， in tea. The method is based on ultra-high performance liquid chromatography-tandem mass spectrometry （UHPLC-MS/MS） coupled with precolumn derivatization. Tea samples （2.00 g） were extracted with 20 mL of ultrapure water by shaking， and followed by centrifugation. After freezing treatment， aliquots of the extract were centrifuged at 15 000 r/min. Then 200 μL of the supernatant was evaporated to complete dryness using nitrogen blowdown. The resulting residue was derivatized with triethyl orthoacetate （TEOA） at 80 ℃ for 1.5 h， in the presence of acetic acid as the catalyst. The derivatization mixture was evaporated to dryness and reconstituted with methanol. NAG was transformed into the same reaction product as GLU， and therefore quantified together with GLU. The derivatization products were separated on an Agilent ZORBAX Eclipse Plus C18 column （100 mm×2.1 mm， 1.8 μm）. The mobile phase consisted of an aqueous solution of 0.1% （V/V） formic acid （containing 5 mmol/L ammonium formate） as phase A and acetonitrile as phase B. The gradient elution was carried out as follows： 0–0.5 min， 90%A； 0.5–7.5 min， 90%A–80%A； 7.5–8.0 min， 80%A–10%A； 8.0–10.0 min， 10%A； 10.0–10.1 min， 10%A–90%A； 10.1–12.0 min， 90%A. The separated compounds were detected by tandem mass spectrometry with positive electrospray ionization （ESI⁺） in multiple reaction monitoring （MRM） mode， and stable isotope-labeled internal standards were used for quantitative analysis. Good linearity was obtained in the 0.5–20 μg/L range； the correlation coefficients （r²） were higher than 0.995， the limits of detection were 2.2–5.3 µg/kg and the limits of quantification were 25 µg/kg. For two representative matrices of green tea and black tea， the average recoveries of GLU and MPP spiked at four levels of 0.025， 0.05， 0.1， and 0.5 mg/kg were 83.9%–105.0%. The intra-day precisions （n=5） ranged from 1.9% to 6.9% and the inter-day precisions （n=10） ranged from 2.6% to 10.3%. Furthermore， the developed method was used to detect GLU and its metabolite residues in 34 tea samples， and the positive detection ratio was 26.5%. In summary， the proposed method is simple， reliable， practical， and suitable for determining GLU and its metabolite residues in various types of tea.

Non-steroidal anti-inflammatory drugs （NSAIDs） are often used to treat horses’ injuries and pain due to long-term training and competitions. In equestrian competitions， NSAIDs do not affect the upper limit of a horse’s athletic ability， but they can enable the horse to perform close to its maximum capacity by reducing inflammation and pain. This not only affects the competition results， but also fails to ensure the health and welfare of the horses. Therefore， it has become crucial to effectively monitor and control NSAIDs residues during equestrian competition. In this study， a rapid screening and determination method for 33 NSAIDs in horse urine was established using QuEChERS pretreatment combined with ultra performance liquid chromatography-tandem mass spectrometry （QuEChERS-UPLC-MS/MS）. The pretreatment process， chromatographic conditions， and mass spectrometric conditions were also optimized. In brief， the target analytes in 1 mL of horse urine samples were extracted with 5 mL of acetonitrile. During extraction， 100 mg of NaCl was added as a salting-out agent， and the mixture was shaken for 3 min. Subsequently， the extract was purified using a QuEChERS method with 300 mg of anhydrous MgSO₄， 40 mg of octadecylsilane-bonded silica （C₁₈）， and 40 mg of primary secondary amine （PSA） as sorbents. After centrifugation， the supernatant was dried under nitrogen and reconstituted. The 33 NSAIDs were separated on an Agilent Poroshell 120 EC-C₁₈ analytical chromatographic column （100 mm×3.0 mm，2.7 μm） with gradient elution using （A） 0.1% formic acid aqueous solution and （B） methanol solution as the mobile phases. The gradient elution program was as follows： 0–1 min， 30%B； 1–3 min， 30%B–60%B； 3–12 min， 60%B–85%B； 12–12.1 min， 85%B–95%B； 12.1–14 min， 95%B； 14–14.1 min， 95%B–30%B； 14.1–17 min， 30%B. Then， the target analytes were determined by UPLC-MS/MS in dynamic multiple reaction monitoring （dMRM） mode with positive/negative ion switching. Quantification was performed using the matrix-matched external standard method. The results showed that the method can rapidly and simultaneously determine the 33 NSAIDs in horse urine. The analytes had good linear relationships within their respective ranges， with correlation coefficients of >0.99. The limits of detection （LODs） and quantification （LOQs） were 0.1–1.0 μg/L and 0.9–5.4 μg/L， respectively. The recoveries of the 33 NSAIDs in horse urine were between 70.8% and 123.8% at three spiked levels of LOQ， 20 μg/L and 200 μg/L. The relative standard deviations of all targeted compounds ranged from 1.9% to 14.8%. The method was applied to 10 actual horse urine samples. The results revealed the presence of three NSAIDs， including ketorolac， lornoxicam and naproxen， with contents ranging from < LOQ to 6.87 μg/L. According to the Equine Prohibited Substances List published by the Federation Equestre Internationale （FEI）， ketorolac and naproxen are controlled medications. They are prohibited for use during competition period， as they may mask symptoms and aggravate clinical conditions. However， they are permitted for use during non-competition times. Lornoxicam is a banned substance and is strictly prohibited for use in horses at any time. The proposed method is simple， efficient and accurate， and is suitable for the simultaneous determination of the 33 NSAIDs residues in horse urine samples.

Acetohexamide （AH） and chlorpropamide （CP） can stimulate pancreatic secretion of insulin and regulate water-salt balance， which may indirectly lead to an increase in adrenaline and potentially enhance the athletic performance of horses. For the protection of horse welfare and the consideration of fair competition， AH and CP were listed as pancreatic stimulants by the Federation Equestre Internationale （FEI）. A method for the detection of pancreatic stimulants in equestrian sports needs to be established. Before analyzing these drugs from biological samples such as urine， it is necessary to perform pre-treatment to separate and concentrate the target analytes. However， there are several disadvantages to using traditional pre-treatment methods， including toxicity and inefficiency. A rapid and environmentally friendly pre-treatment method utilizing ionic liquid-based deep eutectic solvent vortex-assisted liquid-liquid microextraction （IL-DES-VALLME） was developed. Coupled with ultra-high performance liquid chromatography-tandem mass spectrometry （UHPLC-MS/MS）， the approach enabled efficient extraction and sensitive quantification of AH and CP from equine urine. N-Butyl-N-methylmorpholinium bis（（trifluoromethyl）sulfonyl）imide （EMMPNTF₂） was chosen as the hydrogen bond acceptor （HBA）， combined with pentafluorophenol （PFP） acting as the hydrogen bond donor （HBD）. A molar ratio of 1∶4 （HBA/HBD） was identified as the optimum composition of IL-DES. Differential scanning calorimetry （DSC） was utilized to measure the melting point of the synthesized IL-DES. Fourier-transform infrared （FT-IR） spectra and proton nuclear magnetic resonance （¹H NMR） spectra were employed to characterize the structure of the IL-DES and investigate the hydrogen bonding interactions between its components. Under the optimum conditions， only 100 μL of IL-DES， 1 mL of equine urine， and 6 min were required. An external standard calibration method was utilized for quantification， and a linear relationship was achieved with a mass concentration range of 0.2-20 000 ng/mL for AH and 0.5-20 000 ng/mL for CP. The coefficients of determination （r²） were all greater than 0.999. The limits of detection （LOD） of the method based on a signal-to-noise ratio of 3 were 0.1 ng/mL for AH and 0.2 ng/mL for CP. The limits of quantification （LOQ） of the method based on a signal-to-noise ratio of 10 were 0.2 ng/mL for AH and 0.5 ng/mL for CP. The absolute values of matrix effects were all less than 20% （-8.9% for AH， -11.2% for CP）. Method accuracy， assessed through recovery studies at multiple concentrations across the linear range， yielded values between 90.2% and 110.4%. Precision， expressed as relative standard deviation （RSD）， was excellent， with intra-day RSDs≤6.6% and inter-day RSDs≤5.8%. The RSDs for robustness were less than 5.7%， and the average carry-over ratios were in the range of 11.4%-17.3%， indicating good method robustness and acceptable carry-over levels for the quantitation of AH and CP in equine urine. The greenness was assessed by the AGREEprep metric （score： 0.63）. Compared with traditional extraction methods， the IL-DES-VALLME method has the advantages of rapidity， simplicity， efficiency， low toxicity， and low cost. Additionally， the application of the proposed approach to real equine urine samples proved its practicability. These advantages make the method very appealing for doping control in equestrian sports.

Drug abuse is a major global public health problem. Fentanyl analogs and amphetamine-type stimulants （ATS） are widely abused worldwide. Hair analysis can trace drug use history over months. This technique has unique advantages in forensic toxicology. However， its application is limited by a lack of matrix-certified reference materials （CRMs）. At present， the existing CRMs are unable to simultaneously cover both fentanyl analogs and amphetamine-type stimulants. This study established a hair sample preparation method based on a water‑dimethyl sulfoxide‑hydrochloric acid soaking approach， targeting 15 analytes （nine fentanyl analogs and six ATS）. The soaking time was systematically investigated. Drug incorporation into hair reached a maximum at 24 days. A slight decrease was observed at 25 days. This decrease may result from desorption and solvent-induced structural changes in keratin. Therefore， 24 days was selected as the optimal soaking time. Preparation experiments demonstrated that 14 of the 15 drugs were successfully embedded into the hair matrix. This study also applied a dual-platform quantitative strategy for certification. The two platforms were liquid chromatography-tandem mass spectrometry （LC-MS/MS） and gas chromatography-mass spectrometry （GC-MS）. Cross-validation between the two platforms improved the reliability of certified values. Sample pretreatment conditions were optimized using Box-Behnken response surface methodology. Four factors were investigated： extraction temperature， extraction time， liquid-to-solid ratio， and hydrochloric acid concentration. The effects of the four factors on extraction efficiency followed a decreasing order： extraction temperature>extraction time>liquid-to-solid ratio>hydrochloric acid concentration. The optimal extraction conditions were as follows： extraction temperature 40 ℃， extraction time 50 min， liquid-solid ratio 100∶1 （mL/g）， and hydrochloric acid concentration 0.01 mol/L. Method validation was performed according to international guidelines. For LC-MS/MS， the limits of detection （LODs） and quantification （LOQs） were 0.05 pg/mg and 0.25 pg/mg， respectively. For GC-MS， the LODs and LOQs were 0.02 ng/mg and 0.08 ng/mg， respectively. Good linearities were obtained for all 15 drugs. Correlation coefficients （r） were greater than 0.999 for both platforms. Good linearities were obtained for all 15 analytes on both platforms with correlation coefficients >0.999 over the respective concentration ranges Intra-day precisions （n=6） were evaluated as relative standard deviations （RSDs）. Inter-day precision （n=30 over 5 days） was also calculated. For 14 drugs （except amphetamine）， RSD values ranged from 0.2% to 9.4%. All RSDs were below 10%， indicating good repeatability and intermediate precision. Stability of hair extracts was assessed at 1， 24， 48， 72， and 96 h at room temperature. The extracts were stable for at least 96 h when stored at room temperature， with RSDs below 6.0% for LC-MS/MS and below 10% for GC-MS. Recovery tests were performed at three quality control levels （low， medium， high）. For LC-MS/MS， average recoveries ranged from 84.46% to 116.4% （n=3）. For GC-MS， average recoveries ranged from 85.50% to 116.4% （n=3）. All RSDs were below 15%. Matrix effects （MEs） were evaluated by comparing matrix-matched standards with solvent standards. ME values between 80% and 120% （n=3） were observed for both platforms. These results indicate negligible ion suppression or enhancement. Homogeneity of the prepared hair reference material was assessed. The results show that the uniformity of the hair samples is good. The contents of the 14 drugs in the prepared hair samples were determined. For LC-MS/MS， the values ranged from 1.012 ng/mg to 7.830 ng/mg. For GC-MS， the values ranged from 1.087 ng/mg to 7.712 ng/mg. Amphetamine was not detected by either platform， likely due to its high polarity and weak binding to keratin. Most drugs showed good agreement between the two platforms. Relative deviations were below 10% for most analytes. However， methamphetamine showed a deviation of -12%. This may be due to matrix effects in LC-MS/MS. The GC-MS results for methamphetamine were more robust. The dual-platform approach allowed cross-validation and enhanced the reliability of the certified values. In conclusion， a dual-platform method based on LC-MS/MS and GC-MS was successfully applied to certify a new hair matrix reference material. The cross-validation strategy enhances the accuracy and traceability of the certified values. This work provides a valuable tool for quality control in forensic toxicology laboratories. The prepared reference material and validated method will facilitate inter-laboratory comparisons. They will also support the monitoring of drug abuse in forensic and clinical settings.