超高效液相色谱-四级杆-飞行时间质谱法测定珠芽瓜叶乌头中毒患者血液、尿液中乌头生物碱及其代谢产物
目的 建立超高效液相色谱-四级杆-飞行时间质谱法(ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry, UPLC/Q-TOF-MS)定性定量检测珠芽瓜叶乌头中毒患者血液、尿液中乌头生物碱及其相关代谢产物。方法 血样经离心后, 取上层血清稀释后过0.22 ?m微孔滤膜;尿样经稀释后过0.22 ?m微孔滤膜。食物经前处理后, 过Oasis MCX 60 mg/3 mL 固相萃取小柱, 洗脱、吹干、定容, 过0.22 ?m微孔滤膜。草乌块根经匀浆、超声、离心后, 取上清液过0.22 ?m微孔滤膜。所有样品溶液经Aglient SB-C18色谱柱分离后用UPLC-Q-TOF进行测定。结果 14种乌头类生物碱及其代谢产物在1.0~100 ng/mL的浓度范围内线性关系良好, 相关系数均大于0.99, 检出限在0.1~5.0 ng/mL之间, 样品加标回收率为78.9%~116.9%, 相对标准偏差为0.5%~4.1%(n=6)。2位患者血液中检出苯甲酰乌头原碱、滇乌头碱和草乌甲素,其中苯甲酰乌头原碱未达到定量限, 滇乌头碱含量分别为0.18和0.27 mg/L,草乌甲素含量分别为0.25和0.62 mg/L; 2位患者尿液中检出苯甲酰乌头原碱、滇乌头碱、草乌甲素, 苯甲酰乌头原碱含量分别为0.063和0.055 mg/L, 滇乌头碱含量分别为1.66和3.79 mg/L草乌甲素含量分别为2.31和4.89 mg/L, 其中一份尿样检出印乌头碱,含量为0.159 mg/L。结论 此方法操作简便、准确度好、能够快速检测检材中乌头类生物碱。珠芽瓜叶乌头含有较高的滇乌头碱和草乌甲素, 尿液中乌头生物碱及代谢产物高于血液中产物含量, 更适合作为检验材料。
Objective To establish a method for qualitative and quantitative detection of aconitum alkaloids and their metabolites in urine and blood of patients with Aconitum hemsleyanum var. hsiae poisoning by ultra performance liquid chromatography coupled with quadrupole time of flight mass spectrometry(UPLC/Q-TOF-MS). Methods After centrifugation of the blood sample, the upper serum was diluted and passed through a 0.22 ?m microporous membrane. The urine sample was diluted and filtered with 0.22 ?m microporous membrane. The food samples were eluted, dried and fixed volume on a solid-phase extraction column of Oasis MCX 60 mg/3 mL after pretreatment and passed through a 0.22 ?m microporous membrane. Aconite roots were homogenized, sonicated, and centrifuged, then the supernatant was filtered with a 0.22 ?m microporous membrane . All the sample solutions were separated on an Aglient SB-C18 column and determined by UPLC-Q-TOF. Results The linear relationship of 14 aconitoid alkaloids and their metabolites were good in the concentration range of 1.0-100 ng/mL, with all correlation coefficients greater than 0.99, and the detection limit was 0.1-5.0 ng/mL. The recoveries of standard addition were 78.9%-116.9% with the relative standard deviation 0.5%-4.1% (n=6). Benzoylaconitine, yunaconitine and bulleyaconitine A were detected in the blood of 2 patients, among which the content of benzoylaconitine did not reach the quantitative limit, yunaconitine content was 0.18 and 0.27 mg/L, and bulleyaconitine A content was 0.25 and 0.62 mg/L respectively. Benzoylaconitine, yunaconitine and bulleyaconitine A were detected in the urine of 2 patients. The content of benzoylaconitine was 0.063 and 0.055 mg/L, the content of yunaconitine was 1.66 and 3.79 mg/L, and the content of bulleyaconitine A was 2.31 and 4.89 mg/L respectively. Indaconitone was found in one urine sample, with the content of 0.159 mg/L. Conclusion This method is simple, accurate and can be used to detect aconitine alkaloids in samples. Aconitum hemsleyanum var. hsiae contains extremely high bulleyaconitine A and yunaconitine, and urine contains a higher content of aconitine alkaloid and its metabolites than those in the blood, which is more suitable to be used as the test material.
标题：超高效液相色谱-四级杆-飞行时间质谱法测定珠芽瓜叶乌头中毒患者血液、尿液中乌头生物碱及其代谢产物
英文标题：Determination of aconitum alkaloids and its metabolites in blood and urine of patient with Aconitum hemsleyanum var. hsiae poisoning by ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry
作者：
张志清 云南省疾病预防控制中心
伍福仙 云南省疾病预防控制中心
林佶 云南省疾病预防控制中心
中文关键词:珠芽瓜叶乌头,草乌甲素,滇乌头碱,超高效液相色谱-四级杆-飞行时间质谱法,血液,尿液,
英文关键词:Aconitum hemsleyanum var. hsiae,bulleyaconitine A,yunaconitine,ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry,blood,urine,
发表日期：2020-03-26
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