目的: 基于气相色谱法,建立了对吸附破伤风疫苗中17种邻苯二甲酸酯类增塑剂含量测定方法。方法: 采用乙腈沉淀蛋白,氯化钠盐析分层,上清液转移、定容后经气相色谱分析,外标法定量。色谱参数:采用Agilent HP-5(30 m×0.32 mm×0.25 μm)毛细管柱;升温程序:初始温度为50 ℃,保持1 min,以20 ℃·min-1的速率升温至230 ℃,保持3 min,再以5℃·min-1的速率升温至300 ℃,保持5 min;色谱柱流量:1 mL·min-1;检测器温度:280 ℃;进样口温度:250 ℃;载气:氮气,线速度:19.148 cm·s-1;进样量:1 μL,分流模式:不分流,进样方式:自动进样。结果: 17种增塑剂在0.5~20 μg·mL-1范围内线性关系良好(r≥0.99)。在15、10、5 μg·mL-1 3个浓度水平下,加标回收率在83.6%~111.2%之间, RSD为0.39%~3.9%。采用该方法检测5批次吸附破伤风疫苗,药品中均未有增塑剂检出。结论: 该方法专属性好,准确度与精密度高,溶液稳定性好,方法耐用性高,能够用于吸附破伤风疫苗药品中增塑剂的含量测定。
Objective: To establish a GC-FID method to determine the content of 17 phthalic ester plasticizers in adsorbed tetanus vaccine. Methods: The protein was precipitated with acetonitrile, and the layers were separated by salting out with sodium chloride. The supernatant was transferred and analyzed by GC-FID with external standard method. Agilent HP-5 (30 m×0.32 mm×0.25 μm) capillary column was adopted. The temperature program: Initial temperature of 50 ℃ was maintained for 1 min, then was increased to 230 ℃ at the rate of 20 ℃·min-1 and was maintained for 3 min. The temperature was increased to 300 ℃ at a rate of 5 ℃·min-1 and maintained for 5 minutes. Column flow rate was 1 mL·min-1. The temperatures of detector and inlet were 280 ℃ and 250 ℃ respectively. The linear velocity of the carrier gas (nitrogen) was set to 19.148 cm·s-1. The injection volume was 1 μL and the split mode was splitless. The sampling mode: automatic injection. Results: The 17 plasticizers showed good linear relationship in the range of 0.5-20 μg·mL-1 with a correlation coefficient more than 0.99. At three different concentrations (15, 10, 5 μg·mL-1) of spiked samples, the recovery rates of 17 plasticizers ranged from 83.6% to 111.2%, with RSD between 0.39% and 3.9%. No plasticizers were detected in 5 batches of adsorbed tetanus vaccine by this method. Conclusion: The method has good specificity, high accuracy and precision, good solution stability, and high durability. It can be used to determine contents of plasticizers in adsorbed tetanus vaccine drugs.
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