注射用阿莫西林钠克拉维酸钾卤化丁基胶塞中10种抗氧剂的提取和迁移研究

Extraction and Migration of Ten Kinds of Antioxidants in Halogenated Butyl Rubber Stopper of Amoxicillin Sodium Clavulanate Potassium for Injection

目的建立高效液相色谱法(HPLC)同时测定24批次注射用阿莫西林钠克拉维酸钾包装用卤化丁基胶塞中10种常见抗氧剂BHT-COOH、BHT-CHO、2,4-DTB、BHT-Q、BHT、抗氧剂3114、抗氧剂1010、抗氧剂330、抗氧剂1076、抗氧剂168的提取情况,并用超高效液相色谱/静电场轨道阱高分辨质谱(UHPLC/Orbitrap HRMS)法检测现有24批次卤化丁基胶塞及43批次注射用阿莫西林钠克拉维酸钾样品中抗氧剂迁移情况以补充验证。方法HPLC,SHIMADZU Shim-pack GIST C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相:水(A)-甲醇(B)-乙腈(C)梯度洗脱,检测波长:220 nm,流速1 mL·min^(-1),柱温25℃;UPLC/Orbitrap HRMS法,Thermo Hypersil GOLD色谱柱(2.1 mm×100 mm,1.9μm),流动相:水(A)-甲醇(B),梯度洗脱,检测波长:220 nm,流速1 mL·min^(-1),柱温25℃;离子源:电喷雾离子源(ESI),正负离子电离(ESI^(+)、ESI^(-))模式同用;质量数扫描范围m/z 150~1500;进样量:1μL。结果此新建HPLC方法液相条件下各待测成分分离度良好,分离度均大于2.0。精密度、重复性、回收率、稳定性符合规定,线性范围均在1.02~233.20μg·mL^(-1)(r≥0.9998)之间;10种抗氧剂的检测下限为0.06~3.90μg·mL^(-1),定量下限为0.18~11.72μg·mL^(-1)。UHPLC/Orbitrap HRMS法补充检验以上抗氧剂在胶塞及药品中的迁移量,胶塞中除抗氧剂BHT-CHO、BHT-Q和抗氧剂168外另7种抗氧剂均有检出,药品中有9批被检出抗氧剂BHT,4批被检出抗氧剂330。结论该HPLC联用UHPLC/Orbitrap HRMS验证方法可用于注射用阿莫西林钠克拉维酸钾与其包装用卤化丁基胶塞中以上10种抗氧剂的提取检测及迁移研究。

OBJECTIVE To determine the extraction of 10 kinds of antioxidants,BHT-COOH,BHT-CHO,2,4-DTB,BHT-Q,BHT,antioxidant 3114,antioxidant 1010,antioxidant 330,antioxidant 1076,and antioxidant 168,in 24 batches of halogenated butyl rubber stopper for packaging of amoxicillin sodium clavulanate potassium for injection by HPLC method,and detect the migration of the above antioxidants in 24 batches of halogenated butyl rubber stopper and 43 batches of amoxicillin sodium clavulanate potassium for injection by UPLC/Orbitrap HRMS as additional verification.METHODS HPLC method was established to determine the above components.The determination was performed by using SHIMADZU Shim-pack GIST C_(18) column(4.6 mm×250 mm,5μm)with mobile phase consisting of water(A)-methanol(B)-acetonitrile(C)gradient elution at the flow rate of 1 mL·min^(-1).The detection wavelength was set at 220 nm,and the column temperature was maintained at 25℃.UPLC/Orbitrap HRMS method consisted of Thermo Hypersil GOLD column(2.1 mm×100 mm,1.9μm)with mobile phase consisting of water(A)-methanol(B)gradient elution at the flow rate of 1 mL·min^(-1),220 nm detection wavelength,25℃column temperature,and an ion source in the electrospray ion source(ESI).Positive and negative ion ionization(ESI^(+),ESI^(-))mode was used for detection,the mass number scanning range was m/z 150-1500,and the injection volume was 1μL.RESULTS Under the established liquid phase conditions,the resolution of each component was good,and the resolution was greater than 2.0.The precision,repeatability,recovery rate,and stability were in line with the relevant requirements.The linear range was 1.02-233.20μg·mL^(-1)(r≥0.9998).The detection limits of the 10 antioxidants were 0.06-3.90μg·mL^(-1).The limits of quantification were 0.18-11.72μg·mL^(-1).The migration of the above antioxidants in the rubber stopper and the drug was verified by UHPLC/Orbitrap HRMS method.Seven kinds of antioxidants were detected in all halogenated butyl rubber stoppers,respectively,in addition to antioxidant BHT-CHO,BHT-Q and antioxidant 168.Antioxidant BHT was detected in 9 batches of the drug,and antioxidant 330 was detected in 4 batches.CONCLUSION This method can be used to study the extraction and migration of the above 10 kinds of antioxidants in halogenated butyl rubber stopper for packaging of amoxicillin sodium clavulanate potassium for injection.