论文部分内容阅读
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 establish a method for the determination of ethanol residues in blood products by headspace gas chromatography (GC) and verify the method and preliminary application. Method Column: Rtx-1301 (30 m × 0.53 mm × 3 μm); gasification chamber temperature: 220 ℃; column temperature: 90 ℃; detector temperature: 250 ℃; carrier gas was nitrogen; ; Hydrogen flow rate: 40 ml / min; split ratio: 2: 1; headspace at 85 ° C for 30 min, run time: 5 min. The linear range of the standard curve was determined and the method was validated for repeatability, stability, accuracy, limit of detection (LOD) and limit of quantitation (LOQ). The established method was used to determine the ethanol residues in 14 batches of human serum albumin and human immunoglobulin (p H 4). Results The method showed good peak shape, single peak and no other interferences. The linear range of the standard curve was 1.064-21.280 μg / ml; the relative standard deviation (RSD) of 6 replicates of the same human serum albumin was 2.34%; the sample of human serum albumin was placed at 4 ° C The RSDs of detection results of ethanol residues were 2.64% at 0, 4, 8, 12, 16 and 20 h. The average spiked recoveries and RSDs were 99.58 ± 1.19% and 1.40%, respectively. The LOD and LOQ were 0.266 and 0.532 μg / ml. The ethanol residues of 14 batches of samples were the highest for human serum albumin # 3 (0.004 4%), which were much lower than the limit (0.025%) of the semi-finished products of the Chinese Pharmacopoeia (2010 edition). Conclusion Headspace gas chromatography is simple, reproducible, stable and accurate. It can be used for the determination of ethanol residues in blood products.