Department of Chemistry and Biochemistry, South Dakota State University, Avera Health and Science Center 131, Box 2202, Brookings, SD, 57007, USA.
Anal Bioanal Chem. 2014 Jan;406(3):727-34. doi: 10.1007/s00216-013-7536-3. Epub 2013 Dec 12.
An analytical procedure for the simultaneous determination of cyanide and thiocyanate in swine plasma was developed and validated. Cyanide and thiocyanate were simultaneously analyzed by high-performance liquid chromatography tandem mass spectrometry in negative ionization mode after rapid and simple sample preparation. Isotopically labeled internal standards, Na(13)C(15)N and NaS(13)C(15)N, were mixed with swine plasma (spiked and nonspiked), proteins were precipitated with acetone, the samples were centrifuged, and the supernatant was removed and dried. The dried samples were reconstituted in 10 mM ammonium formate. Cyanide was reacted with naphthalene-2,3-dicarboxaldehyde and taurine to form N-substituted 1-cyano[f]benzoisoindole, while thiocyanate was chemically modified with monobromobimane to form an SCN-bimane product. The method produced dynamic ranges of 0.1-50 and 0.2-50 μM for cyanide and thiocyanate, respectively, with limits of detection of 10 nM for cyanide and 50 nM for thiocyanate. For quality control standards, the precision, as measured by percent relative standard deviation, was below 8 %, and the accuracy was within ±10 % of the nominal concentration. Following validation, the analytical procedure successfully detected cyanide and thiocyanate simultaneously from the plasma of cyanide-exposed swine.
建立并验证了一种用于同时测定猪血浆中氰化物和硫氰酸盐的分析方法。采用高效液相色谱-串联质谱法，在负离子模式下，通过快速简单的样品前处理，可同时分析氰化物和硫氰酸盐。将同位素标记的内标物 Na(13)C(15)N 和 NaS(13)C(15)N 与猪血浆（加标和未加标）混合，用丙酮沉淀蛋白质，离心后去除上清液并干燥。将干燥的样品用 10mM 甲酸铵重新溶解。氰化物与萘-2,3-二醛和牛磺酸反应，形成 N-取代的 1-氰基[f]苯并异吲哚，而硫氰酸盐则与单溴双马来酰亚胺发生化学修饰，形成 SCN-双马来酰亚胺产物。该方法对氰化物和硫氰酸盐的动态范围分别为 0.1-50 和 0.2-50μM，检测限分别为 10nM 和 50nM。对于质控标准，以相对标准偏差的百分比表示，精密度低于 8%，准确度在名义浓度的±10%范围内。经验证，该分析方法可成功地从暴露于氰化物的猪血浆中同时检测出氰化物和硫氰酸盐。
