目的 建立超声辅助雄黄Realgar纳米化工艺，阐明纳米化改造在雄黄安全阈值内实现“减毒增效”的科学内涵。方法 采用星点设计-响应面法（central composite design-response surface methodology，CCD-RSM）优化雄黄锆球水飞参数，结合酸洗与超声剪切制备纳米化雄黄；通过激光粒度分析仪（laser particle size analyzer）、扫描电子显微镜（scanning electron microscope，SEM）、比表面积及孔隙度分析仪的Brunauer-Emmett-Teller（BET）分析法及液相色谱-原子荧光光谱联用仪（liquid chromatography-atomic fluorescence spectrometry，LC-AFS）对4种炮制品（干法粉碎雄黄、传统水飞雄黄、SDS水飞雄黄、纳米化雄黄）进行表征；利用线虫模型评价各炮制品毒性，小鼠创面模型评价其药效。结果 雄黄锆球水飞最佳工艺为研磨3.9 h、水飞3次、加锆球（直径分别为1、2、3、4 mm锆球质量比为1∶1∶2∶4）190 g；250 W功率下超声30 min可将水飞后雄黄样品粒径降至300 nm。BET分析结果显示，纳米化雄黄介孔发达；LC-AFS检测结果显示，与干法粉碎雄黄样品相比，3种炮制方法均可降低雄黄样品中As³⁺含量，纳米化雄黄样品略高但低于安全限度。线虫实验中，纳米化雄黄样品毒性最低（P＜0.001）；小鼠创面模型中，其创面愈合率最高，显著降低炎症因子并上调生长因子表达（P＜0.05）。结论 锆球水飞-超声辅助工艺可成功制备纳米化雄黄，在安全阈值内兼具减毒与增效双重优势，为雄黄炮制技术升级提供了理论依据。

Objective To establish an ultrasound-assisted nanonization process for realgar and elucidate the scientific connotation of achieving toxicity reduction and efficacy enhancement within safety thresholds through nanonization. Methods The parameters of zirconium ball water-grinding were optimized using central composite design-response surface methodology (CCD-RSM), and nano-realgar was prepared by combining acid washing with ultrasonic shearing. Four processed products (dry-ground Realgar, traditional water-floating Realgar, SDS-assisted water-floating Realgar and nano-Realgar) were characterized by laser particle size analyzer, scanning electron microscope (SEM), specific surface area and porosity analyzer based on the Brunauer-Emmett-Teller (BET) method, and liquid chromatography-atomic fluorescence spectrometry (LC-AFS). Toxicity was evaluated using a Caenorhabditis elegans model, and efficacy was assessed using a mouse wound model. Results The optimal process parameters of zirconium ball water-grinding were determined as milling for 3.9 h, three cycles of levigation, and the addition of 190 g of zirconia balls (the mass ratio of zirconium balls with diameters of 1, 2, 3, 4 mm is 1∶1∶2∶4); The particle size of Realgar sample can be reduced to 300 nm 30 min of sonication at 250 W. BET analysis showed that nano-realgar had well-developed mesopores. LC-AFS results indicated that, compared with the dry-milled product, all processing methods reduced the As³⁺ content, with nano-Realgar being slightly higher but still below the safety limit. In the C. elegans assay, nano-Realgar exhibited the lowest toxicity (P < 0.001). In the mouse wound model, it achieved the highest healing rate, significantly reducing inflammatory factors and upregulating growth factor expression (P < 0.05). Conclusion The zirconium ball water-grinding combined with ultrasound-assisted nanonization process successfully prepares nano-Realgar, which possesses dual advantages of toxicity reduction and efficacy enhancement within safety thresholds, providing a theoretical basis for upgrading the processing technology of Realgar.

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吉林省自然科学基金资助项目（YDZJ202501ZYTS271）；长春中医药大学大学生创新训练项目（S202510199014）