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.
[1] 张虹. 邻苯二甲酸二(2-乙基)己酯和偏苯三酸三辛酯在临床不同药液中溶出量的比较[J].色谱, 2015, 33(5):522
ZHANG H. Comparison of the release behaviors of di(2-ethylhexyl) phthalate and tri(2-ethylhexyl)trimellitate from the polyvinyl-chloride infusion set into pharmaceutical solutions[J].Chin J Chromatogr, 2015, 33(5):522
[2] 王婷, 高星梅, 郭会彩, 等. 增塑剂DEHP神经毒性作用及机制的研究进展[J].生态毒理学报, 2022, 17(2):33
WANG T, GAO XM, GUO HC, et al. Research progress on neurotoxic effects and mechanism of plasticizer DEHP[J].Asian J Ecotoxicol, 2022, 17(2):33
[3] 胡希俅. DEHP增塑剂应用于聚氯乙烯医疗器械中的安全性分析[J].塑料助剂, 2021, 2: 59
HU XQ. Safety analysis of DEHP plasticizer used in PVC medical devices[J].Plastics Addit, 2021, 2: 59
[4] PASQUALE V, DOMENICA V, EMILIO R, et al. Phthalates: european regulation, chemistry pharmaco kinetic and related toxicity[J].Envir Toxicol Pharmacol, 2013, 36(1):88
[5] 刘晓毅, 蒋可新, 石维妮. 国内外食品接触材料中邻苯二甲酸酯类塑化剂迁移限量对比分析[J].食品工业科技, 2011, 32(10):397
LIU XY, JIANG KX, SHI WN. Comparative analysis of specific migration limits of phthalate plasticizer in food contact materials[J].Sci Technol Food Ind, 2011, 32(10):397
[6] 黄婵媛, 蔡玮红, 莫锡乾. 邻苯二甲酸酯类的特性及在食品中的限量分析[J].包装与食品机械, 2014,32(2):66
HUANG CY, CAI WH, MO XQ. The toxicity and limited provisions of phthalate esters in food[J].Pack Food Machin, 2014, 32(2):66
[7] 化学药品注射剂与药用玻璃包装容器相容性研究技术指导原则(试行)[S].2015: 1
Technical Guidelines for Compatibility Study between Chemical Injection and Medicinal Glass Packaging Container (Trial)[S].2015: 1
[8] 化学药品注射剂与塑料包装材料相容性研究技术指导原则(试行)[S].2012: 1
Technical Guidelines for Compatibility Study between Chemical Injection and Plastic Packaging Materials (Trial) [S].2012: 1
[9] 化学药品与弹性体密封件相容性研究技术指导原则(试行)[S].2018: 1
Technical Guidelines for Compatibility Research between Chemicals and Elastomer Seals (Trial)[S].2018: 1
[10] FDA: Container Closure Systems for Packaging Human Drugs and Biologics[S].1999: 1
[11] EMEA: Guidance on Plastic Immediate Packaging Materials[S].2005: 1
[12] 张广湘. 重组人促红素注射液与预灌封注射器相容性研究[D].广州: 华南理工大学, 2016
ZHANG GX. Compatibility Studies on Recombinant Human Erythropoietin Injection with Prefillable Syringes[D].Guangzhou: South China University of Technology, 2016
[13] NATALIA N, KATELL F, KONI G, et al. Contamination of vegetable oils marketed in Italy by phthalic acid esters[J].Food Control, 2011, 22(2):209
[14] 张璇, 陈大舟, 汤桦, 等. 顶空固相微萃取/气相色谱-质谱法测定液态化妆品中8种增塑剂[J].分析测试学报, 2012, 31(3):317
ZHANG X, CHEN DZ, TANG H, et al. Determination of 8 phthalate esters in liquid cosmetics using solid-phase microextraction/gas chromatography-mass spectrometry[J].J Instrum Anal, 2012, 31(3):317
[15] SALAZAR-BELTRÁN D, HINOJOSA-REYES L, RUIZ-RUIZ E, et al. Determination of phthalates in bottled water by automated on-line solid phase extraction coupled to liquid chromatography with uv detection[J].Talanta, 2017, 168: 291
[16] 赵世文, 林佶, 孙灿, 等. 液相色谱-质谱/质谱双柱法直接测定白酒中15种邻苯二甲酸酯[J].中国卫生检验杂志, 2013, 23(16):3192
ZHAO SW, LIN Z, SUN C, et al. Determination of 15 phthalates in liquor by double-column liquid chromatography tandem mass spectrometry[J].Chin J Health Lab Technol, 2013, 23(16):3192
[17] MARÍA CBA, NEREA OL, PILAR BB. Solid phase extraction using molecular imprinted polymers for phthalate determination in water and wines samples by HPLC-ESI-MS[J].J Microchem, 2017(132):233
[18] 马康, 汤福寿, 何雅娟,等. 食品包装材料中13种增塑剂的毛细管气相色谱法测定[J].分析测试学报, 2011, 30(3):284
MA K, TANG FS, HE YJ, et al. Determination of thirteen plasticizers in food plastic packaging materials by capillary column gas chromatography[J].J Instrum Anal, 2011, 30(3):284
[19] 杨瑛, 樊文娟. GC-FID法测定塑料包材中16种邻苯二甲酸酯类化合物[J].食品科学, 2013, 34(20):250-254
YANG Y, FAN WJ. Determination of 16 kinds of phthalate esters in plastic food packaging materials by GC-FID[J].Food Science, 2013, 34(20):250
