顶空气相色谱法测定A群脑膜炎球菌结合疫苗原液中乙腈及三乙胺的残留量  

Determination of residual acetonitrile and triethylamine contents in bulk of group A meningococcal conjugate vaccine by headspace gas chromatography

作　　者：赵荣丽[1] 刘峰 薛千成 潘海龙[1] 吕雪艳[1] 熊慧玲[1] 

机构地区：[1]成都生物制品研究所有限责任公司,四川成都610023 [2]四川省食品药品研究院化学室,四川成都610000

出　　处：《中国生物制品学杂志》2016年第10期1109-1112,1120,共5页Chinese Journal of Biologicals

摘　　要：目的建立A群脑膜炎球菌结合疫苗原液中乙腈及三乙胺残留量的顶空气相色谱测定法,并进行验证。方法采用标准溶液加入法,以10%无水碳酸钠溶液为溶剂制备混合对照品及供试品溶液,采用顶空气相色谱法进行检测,色谱条件为：采用Agilent DB-1毛细管柱（30 m×0.53 mm×5.0μm）;柱流速：4 ml/min;载气：氮气,分流比：10：1;程序升温：柱温起始温度为40℃,保持5 min,以10℃/min的升温速率升至200℃,保持2 min;进样口温度：200℃;FID检测器温度：250℃;顶空瓶平衡温度：80℃,平衡时间30 min。对方法的系统适应性、线性范围、重复性、检测限和定量限、准确性进行验证。采用建立的方法检测4批A群脑膜炎球菌结合疫苗原液的乙腈及三乙胺残留量。结果各待测组分理论塔板数不低于5 000,分离度不低于1.5;乙腈和三乙胺的线性方程分别为y=3 730.2 x＋1.210 6和y=64 699 x＋11.772,r均为0.999 9,线性范围分别为2.002~100.1μg/ml和1.508~75.40μg/ml;同一混合对照品溶液重复检测5次乙腈和三乙胺残留量的RSD值分别为1.34%和3.10%;乙腈和三乙胺检测限分别为0.020 02和0.007 54μg/ml,定量限分别为0.066 72和0.030 16μg/ml;样品高、中、低3个浓度的乙腈和三乙胺平均加样回收率分别为102.60%和101.92%,RSD分别为1.76%和4.12%。4批A群脑膜炎球菌结合疫苗原液中均未检出乙腈及三乙胺。结论该方法具有良好的重复性、准确性,可用于A群脑膜炎球菌结合疫苗原液中乙腈及三乙胺残留量的检测。Objective To develop and verify a headspace gas charomatography（GC）method for determination of residual acetonitrile and triethylamine contents in bulk of group A meningococcal conjugate vaccine. Methods The residual acetonitrile and triethylamine were determined by headspace gas chromatography（GC）, while standard addition method was used for quantification. The 10% anhydrous sodium carbonate solution was added as a solvent of sample, while the equilibrium zone temperature was 80 ℃, and the time for equilibrium was 30 min. The test was performed on aqgilent DB-1 capillary column（30 m × 0. 53 mm × 5. 0 μm）with a FID detector. Nitrogen was used as the carrier gas, and the split ratio was 10 ∶ 1. The programmed column temperature was set as follows： maintained at 40 ℃ for 5 min as initial temperature, and then raised to 200 ℃ at a rate of 10 ℃ / min and maintained for 2 min; the inlet temperature was 200 ℃ while the FID detector temperature was 250 ℃. The method was verified for systemic suitability, linear range, reproducibility,detection limit and quantitative limit, by which the residual acetonitrile and triethylamine contents in four batches of bulk of group A meningococcal conjugate vaccine were determined. Results The theoretical plate numbers of various compositions to be tested were not less than 5 000, while the separation degrees were not less than 1. 5. The linear equations of acetonitrile and triethylamine were y = 3 730. 2 x ＋ 1. 210 6 and y = 64 699 x ＋ 11. 772, with r values of both0. 999 9, while the linear ranges were 2. 002 ~ 100. 1 and 1. 508 ~ 75. 40 μg / ml, respectively. The RSDs of five test results of residual acetonitrile and triethylamine contents in the same pooled reference solution were 1. 34% and 3. 10%respectively. The detection limits of acetonitrile and triethylamine were 0. 020 02 and 0. 007 54 μg / ml, while the quantitative limits were 0. 066 72 and 0. 030 16 μg / ml, respectively. The mean recoveries of acetonitrile and triethylamine in sampl

关 键 词：顶空气相色谱法 A群脑膜炎球菌结合疫苗 乙腈 三乙胺 残留量 

分 类 号：Q657.72[生物学—生物物理学] R392-33[医药卫生—免疫学]