注射用酒石酸吉他霉素胶塞中挥发性物质迁移的GC-MS/HPLC测定及与溶液澄清度相关性分析

Determination of volatile substances and migration in kitasamycin tartrate for injection by GC-MS/HPLC and analysis of the correlation between the volatile substances and the clarity of solution

目的:建立GC-MS法测定注射用酒石酸吉他霉素胶塞中二硫化碳和挥发性物质的迁移;建立HPLC法测定注射用酒石酸吉他霉素中硫化剂和抗氧剂(BHT)的迁移;并对该类挥发性物质与溶液澄清度的相关性进行分析研究。方法:GC-MS法,采用毛细管色谱柱DB-5MS(30.0 m×0.25 mm,0.25μm),载气为He,流速1.0 mL·min-1,进样口温度200℃,分流比10∶1,程序升温(起始柱温50℃保持10 min,以10℃·min-1升至170℃,维持20 min),顶空温度121℃,平衡30 min,离子源温度230℃,接口温度250℃,质量分辨率1 amu,二硫化碳采用sim模式,其他挥发物采用全扫描方式。HPLC法,采用Luna C18色谱柱(250 mm×4.6 mm,5μm),以含0.05%三氟乙酸的乙腈-含0.05%三氟乙酸的水(90∶10)为流动相,柱温35℃,检测波长为280 nm,流速1.0 mL·min-1,进样量10μL。结果:GC-MS测得胶塞中二硫化碳和挥发性物质随加速时间的推移向样品渗透并迁移至样品中,引起溶液澄清度变差;HPLC法测得硫和BHT的质量浓度均在0.1~100μg·mL-1范围内与峰面积呈良好的线性关系(r均为1.000);该方法硫和BHT的检测下限均为0.015μg·m L-1,定量下限均为0.03μg·mL-1;通过添加标准回收实验,硫的的回收率为96.9%(RSD=5%,n=9),抗氧剂(BHT)的回收率为99.8%(RSD=5%,n=9)。结论:GC-MS/HPLC法可作为注射用酒石酸吉他霉素胶塞的质量控制方法。

Objective:To establish a GC-MS method and an HPLC method for determination of the migration of carbon disulfide and volatile substances and the migration of sulfidizer and antioxidant(BHT)in kitasamycin tartrate for injection,respectively and to analyze the correlation between the volatile substances of rubber stopper and the clarity of solution.Methods:GC-MS method was used with capillary column DB-5 MS(30.0 m×0.25 mm,0.25 μm),carrier gas was He at the flow rate of 1.0 mL·min-1,the temperature of injection port was200 ℃,split ratio was 10∶1,programmed temperature:initial column temperature was 50 ℃ and kept for 10 min,and with a temperature rising rate at 10 ℃·min-1 to 170 ℃ and kept for 20 min,the headspace temperature was 120 ℃,equilibrium for 30 min,the ion source temperature was 230 ℃ and the interface temperature was 250 ℃,the mass resolution was 1 amu,the carbon disulfide mode was sim,and the other volatile substances were using full scan.A Luna C18 column(250 mm×4.6 mm,5 μm)was used for the HPLC analysis.The mobile phase consisted of acetonitrile containing 0.05% trifluoroacetic acid and water containing 0.05% trifluoroacetic acid(90∶10)at the flow rate of 1.0 mL·min-1.The column temperature was 35 ℃,the detection wavelength was 280 nm and the injection volume was 10 μL.Results The results determined by GC-MS showed that carbon disulfide and volatile substances in rubber stopper would permeate into the sample and migrate to the sample with acceleration time,and which effecting the clarity of solution.A good linearity was observed over the range of 0.1-100 μg·mL-1(r=1.000).The detection limits of both sulfur and antioxidant(BHT)were 0.015 μg·mL-1,and the quantitative limits were 0.03 μg·m L-1.The recovery rate of sulfur was 96.9%(n=9,RSD=5%)and that of antioxidant(BHT)was 99.8%(n=9,RSD=5%).Conclusion:The GC-MS/HPLC method can be used for the quality control of rubber stopper used for kitasamycin tartrate for injection.