高效液相色谱-电感耦合等离子体质谱法分析经牛黄解毒片暴露后大鼠血清中砷形态
Arsenic Species in Rat Serum after Oral Administration of Niuhuang Jiedu Tablet by HPLC-ICP-MS
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摘要: 采用高效液相色谱-电感耦合等离子体质谱联用技术测定经牛黄解毒片暴露后大鼠血清中的砷形态。以乙腈为提取剂，采用60 ℃水浴超声提取法对血清样品进行前处理，提取液以13 000 r/min离心10 min，上清液过0.45 μm滤膜。采用Dionex IonPac AS19分析柱(250 mm×4 mm×10 μm)，以20 mmol/L碳酸铵(pH 9.7)-3%甲醇溶液作为流动相，分析经牛黄解毒片暴露后大鼠血清中的砷形态。结果表明，AsB、DMA(Ⅴ)、As(Ⅲ)、MMA(Ⅴ)和As(Ⅴ)的检出限分别为0.05、0.05、0.08、0.10和0.10 μg/L；线性相关系数（R2）大于0.999；加标回收率在86.3%~109.2%之间，相对标准偏差RSD小于5%。通过对经牛黄解毒片暴露后大鼠血清中砷形态的分析发现，DMA(Ⅴ)和U1为主要的砷形态，另外含有少量的AsB和U2。该方法灵敏度高、提取效率好，实现了多种已知和未知砷形态的同时分离，可为研究牛黄解毒片在大鼠血液中的代谢提供技术支持。
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关键词:
- 高效液相色谱-电感耦合等离子体质谱（HPLC-ICP-MS） /
- 牛黄解毒片 /
- 血清 /
- 砷形态
Abstract: The preliminary study of arsenic species in rat serum after oral administration of Niuhuang Jiedu tablet was established by high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (HPLC-ICP-MS). With acetonitrile as extraction agent, water bath ultrasonic extraction at 60 ℃ was used as the pretreatment of serum samples. The extracting solution was separated by centrifugation at 13 000 r/min for 10 min, then filtered through 0.45 μm membrane. 20 mmol/L (NH4)2CO3 (pH 9.7) and 3% methanol were used as mobile phase, and Dionex IonPac AS19 (250 mm×4 mm×10 μm) was used as analytical column for analysis of arsenic species in rat serum after oral administration of Niuhuang Jiedu tablet. Experimental results show that the detection limits of AsB, DMA(Ⅴ), As(Ⅲ), MMA(Ⅴ) and As(Ⅴ) are 0.05, 0.05, 0.08, 0.10 and 0.10 μg/L, respectively. The linear coefficients (R2) are more than 0.999, the recoveries are between 86.3% and 109.2% with the relative standard deviation less than 5%. The preliminary study found that DMA(Ⅴ) and U1 are the main arsenic species in rat serum after oral administration of Niuhuang Jiedu tablet, and a small amount of AsB and U2 are also detected in rat serum. The method is sensitive and high extraction efficiency, which can realize the simultaneous separation of various known and unknown arsenic forms, and provide technical support for the metabolic study of arsenic in rat blood. -
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