顶空气相色谱法测定血液制品中乙醇残留量  

Determination of residual alcohol content in blood products by headspace gas chromatography

作　　者：张晓杰 李荣贞 郑霞[1,3] 

机构地区：[1]山东泰邦生物制品有限公司,山东泰安271000 [2]山东省生物制品研究所,山东泰安271000 [3]泰安市妇幼保健院,山东泰安271000

出　　处：《中国生物制品学杂志》2016年第1期84-88,共5页Chinese Journal of Biologicals

摘　　要：目的建立顶空气相色谱法测定血液制品中乙醇残留量,并对方法进行验证及初步应用。方法色谱柱：Rtx-1301（30 m×0.53 mm×3μm）;气化室温度：220℃;柱温：90℃;检测器温度：250℃;载气为氮气,柱流速：2.5 ml/min;氢气流速：40 ml/min;分流比：2∶1;顶空85℃平衡30 min,运行时间：5 min。确定标准曲线的线性范围,并对方法进行重复性、稳定性、准确性、检出限（limit of detection,LOD）和定量限（limit of quantitation,LOQ）验证。应用建立的方法测定14批人血白蛋白、静注人免疫球蛋白（p H 4）样品的乙醇残留量。结果该方法色谱峰形良好,峰单一且无其他干扰。标准曲线的线性范围为1.064~21.280μg/ml;同一人血白蛋白样品重复检测6次乙醇残留量的相对标准偏差（relative standard deviation,RSD）为2.34%;人血白蛋白样品于4℃放置0、4、8、12、16、20 h,乙醇残留量检测结果的RSD为2.64%;样品的平均加标回收率及RSD分别为（99.58±1.19）%和1.40%;LOD和LOQ分别为0.266和0.532μg/ml。14批样品的乙醇残留量最高为3号人血白蛋白半成品（0.004 4%）,均远低于《中国药典》三部（2010版）的半成品限量要求（0.025%）。结论顶空气相色谱法操作简便,重复性、稳定性、准确性良好,可用于血液制品中乙醇残留量的检测。Objective To develop a method for determination of residual alcohol content in blood products by headspace gas chromatography. Methods A headspace capillary gas chromatography method was adopted using Rtx-1301（30 m ×0. 53 mm × 3 μm） capillary column under the following condition： gasification room temperature： 220 ℃; column temperature： 90 ℃; detector temperature： 250 ℃; carrier gas： nitrogen; column flow rate： 2. 5 ml / min; hydrogen flow rate：40 ml / min; split ratio： 2 ∶ 1. The heating temperature of the headspace oven was 85 ℃, the heating time was 30 min,and the runtime was 5 min. The method was determined for linear range, and verified for reproducibility, stability,accuracy, limit of detection（LOD） and limit of quantification（LOQ）. Fourteen batches of human albumin and human immunoglobulin for intravenous injection were determined for residual alcohol content by the developed method. Results The chromatographic peak in this method was in normal shape, and only a single peak appeared without other interference. The linear range of standard curve was 1. 064 ~ 21. 280 μg / ml. The relative standard deviation（RSD）of residual alcohol contents in one human albumin sample determined for 6 times was 2. 34%, while that of determination results of human albumin after storage at 4 ℃ for 0, 4, 8, 12, 16 and 20 h was 2. 64%. The mean and RSD of spike recovery rate were（99. 58 ± 1. 19）% and 1. 40% respectively. The LOD and LOQ of the method were 0. 266 and0. 532 μg / ml respectively. The residual alcohol content in 14 batches of samples was（0. 004%）at most, which was in sample 3（final bulk of human albumin）, and was remarkably lower than the requirement in Chinese Pharmacopoeia（Volume Ⅲ, 2010 edition）（0. 025%）. Conclusion The developed headspace gas chromatography method was simple,reproducible, stable and accurate, which might be used for the determination of residual alcohol content in blood products.

关 键 词：顶空气相色谱 血液制品 乙醇 残留 

分 类 号：O657.71[理学—分析化学] R392-33[理学—化学]