A sensitive and selective ultra-performance liquid chromatography-tandem mass spectrometry cubed (UPLC-MS 3 ) assay was established for detection of fipronil in zebrafish tissue samples. Following protein precipitation pretreatment, the concentrations of fipronil in various zebrafish tissue samples were quantified by the developed UPLC-MS 3 assay. Fipronil was monitored by MS 3 transition ( m/z 435.0 → 398.9 → 330.0) with a ± 1.0 Da isolation window for secondary product ions. Chromatographic separation was achieved with 0.1 % formic acid-water ( v /v) as the aqueous phase and acetonitrile as the organic phase. Gradient elution with C 18 column was employed for separation of fipronil and probenecid (internal standard, IS). Method validation demonstrated excellent linearity over the range of 0.1–10 ng/mL (r 2 > 0.995), with the accuracies ranging from −10.33 % to 3.17 % and precisions between 3.25 % and 11.34 %. Consistent extraction recoveries (89.92–113.49 %) and acceptable matrix effects (92.54–110.27 %) were observed across all tested matrices. The implementation of MS 3 scanning significantly enhanced specificity by eliminating endogenous interference compared to conventional MRM detection. The successful application of this method in zebrafish tissue distribution studies confirmed its reliability for environmental toxicology research. This MS 3 -based approach provides a robust analytical platform for investigating biodistribution of fipronil in aquatic models.
Di Lu,Xinyue Zheng,Hongyu Xue,Jiansong You,Lei Yin,Meiyun Shi
The aim of this work was to develop a fast, simple, and reliable UPLC-MS 3 method for the sensitive detection of acetochlor in biological samples. In MS 3 mode, the ion transition m/z 270.1 → 224.1→148.1 was chosen for quantification with butachlor as the internal standard. In the UPLC system, separation was performed on a UPLC column (2.1 × 50 mm ID, 1.7 μm) with 0.1 % FA in water and acetonitrile as mobile phases . After simple protein precipitation via acetonitrile, the method was well validated with good linearity (0.5–20 ng/mL, r > 0.995), accuracy (−3.70 %-2.98 %), and precision (<15 %). The selectivity and sensitivity were improved obviously in MS 3 mode than that in MRM mode. The developed UPLC-MS 3 method was successfully applied to the cellular pharmacokinetics study of acetochlor in MCF-7 cells.
Yufeng Guo,Chunpeng Feng,Lai Mengchuan,Liu Meichen,Yalin Xi,Lei Yin,Meiyun Shi
Azoxystrobin is a widely used strobilurin fungicide. Its environmental persistence and potential toxicity to aquatic organisms demand accurate trace‑level quantification in biological tissues. However, sensitive and specific determination in small tissue samples using conventional methods remains challenging. In this study, a novel high‑sensitivity analytical method based on ultra‑performance liquid chromatography coupled with triple‑stage mass spectrometry (UPLC‑MS³) was established for the trace determination of azoxystrobin in zebrafish liver tissues. Following protein precipitation extraction, chromatographic separation was performed on a C₁₈ column using a gradient of 0.1% aqueous formic acid and acetonitrile. Detection relied on an optimized MS³ transition (m/z 404.0 → 371.9 → 344.2). The method exhibited excellent linearity (r > 0.9984) from 0.1 to 20 ng/mL, with accuracy between -3.33% and 3.67% and precision (CV) within 5.57-10.19%. Consistent recoveries (94.93-106.64%) and minimal matrix effects (99.45-104.79%) were achieved across all tissue matrices. Compared to conventional MRM, MS³ scanning significantly enhanced specificity by reducing endogenous interference. The validated approach was successfully applied to tissue distribution studies in zebrafish, confirming its reliability for environmental toxicology research and providing a robust platform for investigating fungicide biodistribution in aquatic organisms.
危险品化学品经营许可证（带存储）