液质联用色谱测定聚丙烯酰胺水凝胶中丙烯酰胺含量

Determination of acrylamide in polyacrylamide hydrogel by high performance liquid chromatography-mass spectroscopy

目的探讨聚丙烯酰胺水凝胶中丙烯酰胺含量的可靠测定方法。方法采用液质联用色谱（HPLC—MS／MS）和同位素稀释定量技术，以选择反应监测（SRM）方式测定医用聚丙烯酰胺样本及20例聚丙烯酰胺临床样本中的丙烯酰胺含量。以丙烯酰胺标准品为对照，样本经水溶液振荡提取，硅藻土柱净化。色谱条件：MG120C18色谱柱（5μm粒径，150mm×4．6mm），柱温均26℃；流动相为甲醇：0．1％甲酸=1：9，流速为0．6mL/Mmin；进样量为10μL。结果丙烯酰胺分离良好。峰面积与浓度呈良好的线性关系，线性范围可达3000μg,／L。加标水平在50斗g／L时，回收率为103．1％，相对标准差（RSD）为6．20％。医用聚丙烯酰胺样本中丙烯酰胺含量为3．9×10^-9-3．1×10^-8g／L。20例聚丙烯酰胺临床样本中丙烯酰胺含量为（14．39±6．40）×10^-9g/L。结论该方法样本处理简单，准确度高，灵敏度高，适合聚丙烯酰胺水凝胶中丙烯酰胺含量的测定。

Objective To establish a method for determination of acrylamide in polyacrylamide hydrogel by high performance liquid chromatography-mass spectroscopy （HPLC-MS/MS）. Methods The standard sample was used as control. After adding 13 C3 labeled acrylamide , the sample was extracted by water and then cleaned up by Extrelut TM 20. Column：MGl20 C18 （5 μm , 150 mm ×4.6 mm）. Temperature： 26℃. Mobil phase：methanol： 0.1% formic acid = 1： 9. Flow rate： 6 mL/min. Sample quantity ：10 μL. Finally the polyaerylamide hydrogel sample and 20 clinical extract sample were analyzed by HPLC-MS/MS and isotope dilution quantify technique in selected reaction monitoring （SRM） mode. Results Acrylamide was perfectly separated from polyacrylamide hydrogel. The concentration of acrylamide in polyacrylamide hydrogel was in range of 3.9 ×10^-9- 3.1 ×10^-8 g/L and （ 14.39 ± 6.40 ） ×10^-9 g/L in 20 clinical samples. The peak area was favorable linear and the range was up to 3 000 μg/L. The recovery rate was 103.1% with relative standard deviation （RSD） 6.20% , when the mark level was 50 μg/L. Conclusions The results showed that the method is rapid , accurate ,sensitive , and suitable for the determination of acrylamide in polyacrylamide hydrogel.