Journal of pharmacological and toxicological methods最新文献

{"title":"Characterization of reference vasoactive agents in a rodent safety pharmacology assay","authors":"Matt Skinner,&nbsp;Doris Anic,&nbsp;Pete Ceuppens","doi":"10.1016/j.vascn.2026.108474","DOIUrl":"10.1016/j.vascn.2026.108474","url":null,"abstract":"<div><div>Background: The effects of new drugs on arterial blood pressure are assessed in regulatory safety pharmacology studies in large animals. A HESI consortium is in the process of conducting studies, using a set of reference vasoactive drugs, to assess the ability of the standard dog telemetry assay to detect drug-induced changes in arterial blood pressure.</div><div>Objectives: The purpose of the current study was to assess the same set of reference drugs on hemodynamic parameters in conscious telemetered rats. The telemetered rat is the often the first assay used for assessing drug effects on arterial blood pressure and the translation of effects in this assay to large animal and to human are of interest.</div><div>Methods: Rats (<em>n</em> = 4–5) were implanted with telemetry transducers to record aortic blood pressure and ECG. Animals were dosed orally with either midodrine (0, 0.5, 1.5, 5 mg/kg), nifedipine (0, 0.3, 1, 3 mg/kg), hydralazine (0, 0.3, 1, 3 mg/kg), prazosin (0, 0.01, 0.03, 0.1 mg/kg) or milrinone (0, 0.3, 1 and 3 mg/kg). Telemetry data were collected from 1 h prior to dosing until 20 h post-dose. Satellite animal were also dosed with the same set of reference agents and blood samples were taken by capillary microsampling. Plasma was harvested and analyzed by LCMS for drug concentration determination.</div><div>Results: A dose-related decrease in systolic arterial blood pressure was observed after administration of the mid and high dose of nifedipine (−13 and −22 mmHg), hydralazine (−9 and -29 mmHg), prazosin (−12 and −18 mmHg) and milrinone (−20 and −30 mmHg). With the exception of hydralazine, a dose related-tachycardia accompanied these changes. Midodrine caused a dose-related increase in systolic arterial blood pressure (15 and 46 mmHg after the mid and high dose and a concomitant dose-related bradycardia was observed.</div><div>Conclusions: These data show that the rat telemetry assay is able to detect the expected effects of a set of reference vasoactive agents on arterial blood pressure. The results will be compared with emerging data using the same reference set of compounds in the dog and also with human data to assess the translation between species and the relevance of the rat as an early <em>in vivo</em> cardiovascular screen.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108474"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730676","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Reviving the organ bath: A classic approach to drug safety screening","authors":"Laura Coonen,&nbsp;Cristina Altrocchi,&nbsp;Mohamed Kreir,&nbsp;David Gallacher","doi":"10.1016/j.vascn.2026.108519","DOIUrl":"10.1016/j.vascn.2026.108519","url":null,"abstract":"<div><div>Drug-induced blood pressure changes are common side effects identified during drug development and post-marketing surveillance. These adverse events are concerning, especially in the case of hypertension, as it is associated with an elevated risk of cardiovascular events and mortality. The FDA recently emphasized the necessity to evaluate the potential of drugs to introduce such changes and with the growing awareness regarding animal welfare, the industry is encouraged to improve their safety screening portfolio. The organ bath was invented in 1904 and has been used for decades to evaluate the vasoactive properties of compounds. In the industry, the use of the organ bath is often limited to mechanistic studies and its value as a general tool for screening new drugs has not been thoroughly investigated. For this study, ten reference substances representing common pathways of drug-induced blood pressure changes were selected to be tested in the organ bath. The aorta, isolated from surplus Sprague-Dawley rats, were cleaned of fat tissue, cut into 1–3 mm rings and mounted in 5-mL organ baths. A standardized protocol was designed to optimize the experimental procedure, reducing testing time and data variability. The obtained data indicate that our set-up is robust across parameters such as tissue isolation time and ring position toward the heart, while other factors such as sedation method may impact the results. We show that the designed standardized protocol is suited to establish a dose-response curve for compounds with direct vasoactive effects (e.g., serotonin, gallopamil), while others require a customized approach (e.g., bosentan). This study demonstrates that the organ bath experiment is reliable to effectively reveal the direct vasoactive effects of different classes of drugs. Moreover, isolating multiple rings from one animal can significantly reduce and refine invasive <em>in vivo</em> studies. However, it is important to note that this standardized experiment does not encompass all pathways involved in drug-induced blood pressure changes such as those that are centrally mediated or related to chronic drug exposure. Therefore, a comprehensive and integrative safety screening strategy to investigate blood pressure changes should include a variety of complementary assays.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108519"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730630","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrated implanted telemetry and hemodynamic measurement on a general toxicology study for cynomolgus monkeys demonstrated minimal impacts of toxicology evaluation related to surgical instrumentation and supported use on chronic toxicology studies","authors":"Jui Tung R. Liu,&nbsp;Delilah Moreno,&nbsp;Kevin Norton,&nbsp;Stephen D. Tichenor","doi":"10.1016/j.vascn.2026.108479","DOIUrl":"10.1016/j.vascn.2026.108479","url":null,"abstract":"<div><div>Inclusion of safety pharmacology endpoints into repeated dose toxicology studies to reduce animal use has been highly recommended according to IHC M3 (R2) guideline. Implanted telemetry technology provides a high-quality electrocardiogram and hemodynamic measurement that allows an inclusion of cardiovascular assessment in a preclinical toxicology study. By integrating the implanted telemetry technology into a general toxicology study will not only reduce the animal usage but also provide a comparable analysis between safety pharmacology and toxicology parameters. However, the impact of surgical complication on study conducts, lifespan of the device, and toxicology data are not fully understood.</div><div>In this study, we reviewed integrated toxicology studies with implanted telemetry (with M‑11, L-11R, or L-21 devices) in Cynomolgus Monkeys conducted among Charles River Laboratory sites from 2020 to 2025 to investigate the impact of implanted instrumentation on a study over time. Data evaluated included animal exclusion rate post-surgery, abnormal clinical signs, clinical pathology parameters, and anatomic pathology evaluation. In addition, device functioning rate throughout the study duration was evaluated.</div><div>Analyses from studies with non-human primates (NHP) revealed that the implantation had minimal to no impacts on physiological endpoints, clinical pathology parameters, or histopathology parameters, with less than 2% of animals excluded from study and 0.9% of animals with abnormal clinical signs that were related to implantation. The major instrumentation-related pathology findings included neutrophilic inflammation/thrombus at the implant sites, blood pressure catheter tip, and fibroplasia in the gastrointestinal system close to the devices. With study duration up to 40 weeks, more than 90% of devices were functional, while the majority of the malfunctioning units were due to failed blood pressure measurement.</div><div>The data collected here supports the continued use of an implantable approach on a toxicology study and furthermore, emphasizes study power based on study design, which should be taken into consideration for chronic toxicology studies, when additional cardiovascular assessment is required in late-stage drug development.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"140 ","pages":"Article 108479"},"PeriodicalIF":2.1,"publicationDate":"2026-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148730645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Machine learning modeling of acute inhalation toxicity using an RFA-RFR framework, supported by explainable AI and SALI-based activity cliff analysis","authors":"Vijay H. Masand ,&nbsp;M.M. Rathore ,&nbsp;Jayant R. Bansod ,&nbsp;Meghshyam K. Patil ,&nbsp;Gaurav S. Masand ,&nbsp;Abdul Samad","doi":"10.1016/j.vascn.2026.108426","DOIUrl":"10.1016/j.vascn.2026.108426","url":null,"abstract":"<div><div>The present study develops machine learning models to predict the acute inhalation toxicity (pLC₅₀) of a diverse set of 552 molecules. Following dataset curation from the Integrated Chemical Environment (ICE) database, 3D molecular structures were generated and molecular descriptors (∼58,000) were calculated. After unsupervised pruning and redundancy reduction, 853 descriptors remained, which were further refined to 47 descriptors. Final feature selection was carried out exclusively on the training set using the Red Fox Algorithm (RFA). Random Forest Regressor (RFR) models were then developed and interpreted using Shapley additive explanations (SHAP) values. The resulting RFA–RFR model demonstrated robust statistical performance (R²_tr = 0.907, R²_L10%O = 0.902, and Q₂F₃ = 0.887). Model robustness was confirmed through leave-10%-out cross-validation and external set validation method. Importantly, benchmarking against previous QSAR models for inhalation toxicity shows improved predictive accuracy and broader chemical applicability. Mechanistic interpretation revealed that toxicity is strongly influenced by molecular weight, hydrogen bond acceptor count, lipophilic-polar balance, and sp³-hybrid carbon with oxygen arrangements. This study highlights the value of integrating feature optimization with interpretable machine learning approaches for deriving predictive and mechanistically meaningful models of inhalation toxicity.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"139 ","pages":"Article 108426"},"PeriodicalIF":1.8,"publicationDate":"2026-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148007746","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Study on pharmacokinetic behaviors of colchicine sustained-release microspheres in beagle dogs and its relevance to toxicity mitigation via LC-MS/MS","authors":"Chuyao Yang ,&nbsp;Jingtao Wang ,&nbsp;Qiannan Zhang ,&nbsp;Yuli Wang ,&nbsp;Lingchao Wang ,&nbsp;Wenpeng Zhang ,&nbsp;Jinghua Xu ,&nbsp;Xiaomei Zhuang","doi":"10.1016/j.vascn.2026.108418","DOIUrl":"10.1016/j.vascn.2026.108418","url":null,"abstract":"<div><h3>Objective</h3><div>This study aimed to develop a simple and highly sensitive LC-MS/MS (Liquid Chromatography-Tandem Mass Spectrometry) method for quantifying colchicine in beagle plasma and use it to investigate the pharmacokinetic (PK) characteristics of colchicine sustained-release microspheres (SRMs) in beagles.</div></div><div><h3>Methods</h3><div>Plasma proteins were precipitated using methanol. Following centrifugation, the supernatant was vacuum-concentrated and reconstituted in 0.1% formic acid aqueous solution. Separation was performed on a Kromasil 100–5-C8 column (2.1 × 50 mm), with detection carried out via positive ion electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode. A total of 12 beagles (9 males, 3 females) were enrolled in two crossover pharmacokinetic studies with washout periods of 3 weeks, respectively. Single-dose studies (6 males, 4-treatment, 4-period) included intravenous colchicine API (1 μg/kg), oral SRMs (0.25/0.5 mg) and commercial tablets (0.5 mg); multiple-dose studies (3 males/3 females, 2-treatment, 2-period) involved daily oral gavage of 0.5 mg SRMs or tablets for 5 consecutive days.</div></div><div><h3>Results</h3><div>The method exhibited good linearity (r² &gt; 0.990, 0.025–50 ng/mL), with qualified precision, accuracy, extraction recovery, matrix effects and stability. Colchicine had moderate clearance (CL, 0.98 ± 0.18 L/h/kg) and extensive volume of distribution (V_z, 1.27 ± 0.24 L/kg). Compared with isodose tablets, single-dose SRMs had lower Cₘₐₓ (8.27 ± 2.64 vs 2.81 ± 0.24 ng/mL) and prolonged Tₘₐₓ [1.0 (0.88–1.25) vs 2.0 (2.0–2.5) h]. Both gropus of multiple-dose PK showed no accumulation (accumulation factor, <em>R</em> &lt; 1.2); colchicine SRMs exhibited a lower fluctuation index (FI, a key PK parameter calculated as the ratio C_max/C_min after repeated dosing) (30.04 ± 8.99 vs 50.07 ± 9.37, <em>P</em> &lt; 0.01).</div></div><div><h3>Conclusion</h3><div>The developed LC-MS/MS method meets the requirements for colchicine SRMs pharmacokinetic studies in beagles. Compared with conventional tablets, colchicine SRMs offer improved safety and efficacy.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"138 ","pages":"Article 108418"},"PeriodicalIF":1.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147319055","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Streamlined dispersive solid phase extraction for determination of amitriptyline, cocaine, and benzoylecgonine in urine","authors":"Isabela Nucci Galetti ,&nbsp;Heloísa Peres Luz ,&nbsp;Bruna Espíndola da Silva ,&nbsp;Maitê Perin ,&nbsp;Camila Marchioni","doi":"10.1016/j.vascn.2026.108416","DOIUrl":"10.1016/j.vascn.2026.108416","url":null,"abstract":"<div><div>This study aimed to develop and validate an analytical method using dispersive solid phase extraction (d-SPE) with analysis by liquid chromatography coupled with diode array detector (HPLC-DAD). The developed technique showed detection limit of 100 ng/mL and 1300 ng/mL for cocaine, amitriptyline, and benzoylecgonine, respectively. The precision was less than 15% for 3 different analyte concentrations. The method shows no carryover, and stability was proven for cocaine and amitriptyline. When compared to other techniques, the current study presented a lower detection limit and used a smaller sample volume. The method was compared with the rapid immunoassay test on samples from patients with suspected intoxication, and was satisfactory, showing adequate sensitivity for the proposed application. The method using d-SPE/HPLC-DAD proved to be an appropriate technique to complement emergency toxicological analysis with a satisfactory total run time of 14 min for the detection of the 3 analytes of interest, which fits well into a fast-paced workflow.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"138 ","pages":"Article 108416"},"PeriodicalIF":1.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146055817","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analytical method development for detection and quantification of AMB-FUBINACA in mouse brain tissue using GC–MS/MS","authors":"Jihad Al-Qudsi ,&nbsp;Malak Alghamdi ,&nbsp;Abdulsallam Bakdash ,&nbsp;Elkhatim H. Abdelgadir ,&nbsp;Tulsidas R. Baggi ,&nbsp;Sachil Kumar","doi":"10.1016/j.vascn.2026.108420","DOIUrl":"10.1016/j.vascn.2026.108420","url":null,"abstract":"<div><h3>Background and objective</h3><div>Identifying and quantifying the indazole-derived synthetic cannabinoid, AMB-FUBINACA, in mouse brain tissue is indeed possible. However, evaluating the extraction efficiency, validating the GC–MS/MS method, and demonstrating its effectiveness in real samples require detailed experimental data and expertise in analytical forensic toxicology.</div></div><div><h3>Methodology</h3><div>Male Swiss albino mice (<em>n</em> = 18) were divided into three groups: group 1, control (without administration of AMB-FUBINACA); group 2, orally gavaged with (5 mg/kg) of AMB-FUBINACA; and group 3, orally gavaged with (50 mg/kg) of AMB-FUBINACA. After 24 h, mice were sacrificed and decapitated. The brain tissue was collected, homogenized, and extracted using liquid-liquid extraction (LLE) with ethyl acetate as the extraction solvent. Finally, an analytical method for the analysis of samples using GC/CI-MS/MS was developed and validated.</div></div><div><h3>Results</h3><div>The results demonstrated the high efficiency of the employed method in extraction, detection, and quantification processes. The limit of detection (LOD) and limit of quantification (LOQ) were 0.5 ng/g and 1 ng/g, respectively. Analytical linearity was observed across a wide concentration range (1–1000 ng/mL), achieving a strong correlation coefficient (r²) of 0.99. The study also noted an absence of matrix interference at the retention times of all tested ions. The %RSD for intra-day precision varied from 1.28% to 7.51%, with an accuracy of −10.75 to −0.423, while the %RSD for inter-day precision was 2.31% to 4.4%, with an accuracy of −8 to −1.9%. The LLE approach yielded a recovery rate of 76%.</div></div><div><h3>Conclusion</h3><div>The results suggested implementing this analytical method in the forensic and clinical toxicology laboratories to identify and quantify the drug (AMB-FUBINACA) in brain samples. This will assist clinicians and pathologists treating AMB-FUBINACA intoxications in describing their results in the scientific literature, particularly in post-mortem forensic toxicology samples and future medical and forensic investigations.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"138 ","pages":"Article 108420"},"PeriodicalIF":1.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147398983","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Special Issue of Journal of Pharmacological and Toxicological Methods: Cutting edge pharmacological and toxicological methods","authors":"Reza Tabrizchi,&nbsp;Noriko Daneshtalab","doi":"10.1016/j.vascn.2026.108413","DOIUrl":"10.1016/j.vascn.2026.108413","url":null,"abstract":"","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"138 ","pages":"Article 108413"},"PeriodicalIF":1.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145992458","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"QbD-based, greenness and whiteness-assessed LC–MS/MS method for simultaneous determination of sodium phenylbutyrate and Taurursodiol with pharmacokinetic application","authors":"Thirumalavasu Palla ,&nbsp;Venkata Kanaka Srivani Maddala ,&nbsp;Kumaraswamy Gandla","doi":"10.1016/j.vascn.2026.108417","DOIUrl":"10.1016/j.vascn.2026.108417","url":null,"abstract":"<div><div>Sodium phenylbutyrate (NaPB) and taurursodiol (TRS) are clinically important agents frequently investigated in combination for therapeutic applications. To facilitate pharmacokinetic studies, a sensitive, robust, and environmentally sustainable bioanalytical method for their simultaneous determination in plasma was developed. A Liquid Chromatography-Tandem Mass Spectrometry (LC–MS/MS) method was systematically optimized using a Box–Behnken design within the Design of Experiments framework. The influence of organic solvent proportion, flow rate, and mobile phase pH was evaluated, yielding optimal conditions of 40% (<em>v</em>/v) organic modifier, a flow rate of 1.0 mL/min, and mobile phase pH 2.6. Detection was achieved <em>via</em> electrospray ionization in positive selected reaction monitoring (SRM) mode with transitions <em>m</em>/<em>z</em> 187.2 → 144.7 for NaPB, m/z 500.7 → 218.9 for TRS, and m/z 516.5 → 440.0 for taurocholic acid (internal standard). Method validation, performed according to EC 2002/657/EC guidelines, confirmed excellent linearity (r² &gt; 0.999), accuracy, precision within 15% CV, and analyte stability under various storage and handling conditions. The method was successfully applied to a pharmacokinetic study in Wistar rats. Greenness and whiteness assessment further demonstrated strong environmental compatibility. Overall, the developed LC–MS/MS protocol is precise, reproducible, and sustainable, providing a valuable tool for preclinical pharmacokinetic investigations of NaPB and TRS.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"138 ","pages":"Article 108417"},"PeriodicalIF":1.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147278112","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid quantification of CSF total protein via smartphone digital colorimetry: Enabling early diagnosis of central nervous system diseases","authors":"Yonghua Zhao ,&nbsp;Dandan Liang ,&nbsp;Yufei Guo ,&nbsp;Ziqi Li ,&nbsp;Zehua Yang","doi":"10.1016/j.vascn.2026.108412","DOIUrl":"10.1016/j.vascn.2026.108412","url":null,"abstract":"<div><h3>Objective</h3><div>This study evaluated a smartphone-based system for measuring cerebrospinal fluid (CSF) total protein, verifying its compliance with clinical requirements and exploring its application in diagnosing central nervous system (CNS) diseases in remote/medically underserved areas.</div></div><div><h3>Methods</h3><div>The system analyzed CSF total protein standards to establish a standard curve (concentration vs. R, G, Y values). Per Clinical and Laboratory Standards Institute (CLSI) guidelines, key parameters (precision, detection capability, linear range [LR], clinical reportable range [CRR]) were assessed. Forty-two clinical samples from neurological patients were tested, with results compared to the Beckman Coulter AU5800 analyzer for consistency. Accuracy at three medical decision levels (based on CSF total protein reference ranges) was verified.</div></div><div><h3>Results</h3><div>The limit of blank (LOB), limit of detection (LOD), and limit of quantitation (LOQ) were 5.18 mg/dL (0.05 g/L), 11.78 mg/dL (0.12 g/L), and 12.59 mg/dL (0.13 g/L), respectively. The LR was 29.75–127.20 mg/dL (0.30–1.27 g/L), and the CRR was 12.59–508.80 mg/dL (0.13–5.09 g/L; via 1:3 dilution), both meeting clinical requirements. Total imprecision coefficients of variation (CVs) were 3.83% (high-concentration: 91.52 mg/dL [0.92 g/L]) and 3.59% (low-concentration: 31.32 mg/dL [0.31 g/L]), within acceptable ranges. The 42 clinical samples showed strong consistency with AU5800 results (R² = 0.991). In the evaluation of the smartphone-based CSF total protein detection system, although the 95% confidence intervals of expected biases at 45 mg/dL (0.45 g/L) and 60 mg/dL (0.60 g/L) exceed 1/4 total allowable error per CLSI EP9-A2, the deviations are clinically irrelevant—with the maximum bias (5.29 mg/dL [0.0529 g/L]) fails to alter the pathological threshold classification of CSF total protein (≥45 mg/dL [0.45 g/L]), thus confirming the acceptable bias of the system for auxiliary CSF protein detection.</div></div><div><h3>Conclusions</h3><div>The system meets clinical standards, is user-friendly and cost-effective, and has high potential for aiding CNS disorder diagnosis in resource-constrained areas.</div></div>","PeriodicalId":16767,"journal":{"name":"Journal of pharmacological and toxicological methods","volume":"138 ","pages":"Article 108412"},"PeriodicalIF":1.8,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145994665","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}