目的:建立GC-MS法测定注射用头孢呋辛钠瓶胶塞中有机物的迁移;建立HPLC法测定注射用头孢呋辛钠中硫化剂和抗氧剂的迁移;对胶塞中有机浸出物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Rtx-5MS(30.0 m×0.25 mm,0.25 μm),载气为He,流速1.8 mL·min-1,进样口200 ℃,分流比10:1,柱温为程序升温(起始温度40 ℃保持15 min,以10 ℃·min-1升至100 ℃,维持2 min,25.0 ℃·min-1至230 ℃,保持10.0 min),顶空温度120 ℃,平衡30 min,接口温度250 ℃,扫描范围m/z 29~600,采用全扫描方式。HPLC法,采用Agilent SB C18色谱柱(250 mm×4.6 mm,5 μm),流动相A为含0.05%三氟乙酸的水,流动相B为含0.05%三氟乙酸的乙腈,梯度洗脱,流速1.0 mL·min-1,柱温35 ℃,检测波长为220 nm,进样量10 μL。结果:GC-MS测得胶塞中抗氧剂BHT(264)、硅油等有机物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法测得硫化剂(二硫化碳、硫)和抗氧剂(168、264、330、1076、1010)质量浓度均在0.5~100 μg·mL-1范围内与峰面积呈良好的线性关系(r均＞0.990);该方法硫化剂和抗氧剂的检测限均约为0.03 μg·mL-1,定量限均约为0.1 μg·mL-1;通过添加标准回收试验,抗氧剂(168、264、330、1076、1010)和硫化剂(二硫化碳、硫)的平均回收率(低、中、高浓度水平)均在90.0%~110.0%,对应的RSD均<5%(n=9);稳定性试验表明抗氧剂和硫化剂对照溶液在24 h内稳定。试验结果表明注射用头孢呋辛钠样品溶液中检出了抗氧剂264、1010和硫化剂,含量分别为3.83~14.06、2.02~3.60和2.25~3.21 μg·mL-1;且相关性分析表面抗氧剂264、1010和硫化剂发生迁移是导致注射用头孢呋辛钠澄清度不合格的重要因素。结论:GC-MS法结合HPLC法可作为注射用头孢呋辛钠瓶胶塞中浸出物的质量控制方法。
Objective: To establish a GC-MS method for determination of the migration of organic substances in the rubber stopper of cefuroxime sodium injection bottle, establish an HPLC method for determination of the migration of vulcanizing agent and antioxidant in cefuroxime sodium for injection respectively, and to analyze and study the correlation between the organic extract in rubber stopper and the clarity of solution. Methods: GC-MS method was used with capillary column Rtx-5MS(30.0 m×0.25 mm, 0.25 μm), the carrier gas was He at the flow rate of 1.8 mL·min-1, the temperature of injection port was 200 ℃, shunt ratio was 10:1, the column temperature was programmed temperature(initial column temperature was 40 ℃ and kept for 15 min, and with the speed of 10 ℃·min-1 upping to 100 ℃ and kept for 2 min, and with the speed of 25 ℃·min-1 upping to 230 ℃ and kept for 10 min), the headspace temperature was 120 ℃, the balance was 30 min, the interface temperature was 250 ℃, and the scanning range was from 29 to 600(m/z), and the full scanning mode was adopted. An Agilent SB C18 column(250 mm×4.6 mm, 5 μm) was used for the HPLC analysis. The mobile phase A consisted of water containing 0.05% trifluoroacetic and the mobile phase B consisted of acetonitrile containing 0.05% trifluoroacetic acid, the gradient elution was used and the flow rate was 1.0 mL·min-1. The column temperature was 35 ℃, the detection wavelength was 220 nm and the injection volume was 10 μL. Results: Antioxidant BHT(264), silicone oil and other organic compounds in rubber stopper permeated and migrated to the sample with acceleration time by GC-MS, resulting in deterioration of solution clarity. The mass concentration of vulcanizing agent (carbon disulfide, sulfur) and antioxidant (168, 264, 330, 1076, 1010) measured by HPLC had a good linearity with the peak area in the range of 0.5-100 μg·mL-1(r＞0.999). The detection limits of vulcanizing agent and antioxidant were about 0.03 μg·mL-1, and the quantitation limits were about 0.1 μg·mL-1. The average recoveries (low, medium and high concentration level) of antioxidants (168, 264, 330, 1076, 1010) and vulcanizing agents (carbon disulfide, sulfur) were between 90.0% and 110.0%, and the corresponding RSD were less than 5%(n=9) by adding standard recovery experiments. The stability test showed that the antioxidant and the vulcanizing agent solution had good stability and was stable within 24 h.The results showed that antioxidant 264, 1010 and vulcanizing agent were detected in the sample solution of cefuroxime sodium for injection. The content was 3.83-14.06 μg·mL-1, 2.02-3.60 μg·mL-1 and 2.25-3.21 μg·mL-1. The migration of surface antioxidant 264, 1010 and vulcanizing agent was an important factor that leads to the unqualified clarity of cefuroxime sodium for injection. Conclusion: The GC-MS combined with HPLC method can be used for the quality control of the extract in rubber stopper of cefuroxime sodium for injection.
[1] 刘又宁, 王睿, 邓宝军, 等. 头孢呋辛钠治疗细菌性感染临床评价[J]. 中国临床药理学杂志, 2000, 16(4): 251
LIU YN, WANG R,DENG BJ, et al. Clinical study of cefuroxime in the treatment of bacterial infection[J]. Chin J Clin Pharmacol, 2000, 16(4): 251
[2] 董秀风. 头孢呋辛与头孢呋辛酯治疗细菌感染性疾病及体外抗菌作用[J]. 青海医药杂志, 2008, 38(9): 41
DONG XF. Cefuroxime and cefuroxime axetil in the treatment of bacterial infectious diseases and their antibacterial effect in vitro[J]. Qinghai Med J, 2008, 38(9): 41
[3] 肖全英, 谌雪芳, 钟萍, 等. 注射用头孢曲松钠的胶塞相容性研究[J]. 广东药学院学报, 2004, 20(6): 610
XIAO QY, ZHAN XF, ZHONG P, et al. Study on compatibility of ceftriaxone sodium for injection[J]. Acad J Guangdong Coll Pharm, 2004, 20(6): 610
[4] 严幼达, 顾惠敏, 侯惠民. 胶塞对抗生素粉针剂质量的影响[J]. 中国医药工业杂志, 2003, 34(4): 190
YAN YD, GU HM, HOU HM. Elastomeric closures and quality of antibiotics for injection[J]. Chin J Pharm, 2003, 34(4): 190
[5] 徐俊, 蔡荣, 陆维怡. 胶塞可挥发性物质的迁移及其对头孢曲松钠的可能影响[J]. 药学实践杂志, 2010, 28(6): 448
XU J, CAO R, LU WY. The migration and possible influences of the volatile substances from rubber stoppers on ceftriaxone sodium[J]. J Pharm Pract, 2010, 28(6): 448
[6] 胡君, 王瑞红. 注射用头孢曲松钠的胶塞相容性研究[J]. 中国科技投资, 2016, 26: 382
HUN J, WANG RH. Study on compatibility of ceftriaxone sodium for injection[J]. China Vent Capit, 2016, 26: 382
[7] 赵霞.药用丁基胶塞与头孢曲松钠的相容性研究[D]. 北京: 中国协和医科大学, 2006
ZHAO X. Compatibility of Medical Butyl Rubber Plug with Ceftriaxone Sodium, Doctoral Dissertation[D]. Beijing: Peking Union Medical College, 2006
[8] 郭艳娟, 李伟曹, 杨倩, 等. 国产注射用头孢地嗪钠质量分析[J]. 中国抗生素杂志, 2018, 43(3): 291
GUO YJ, LI WC, YANG Q, et al. Quality assessment of cefodizime sodium for injection[J]. Chin J Antibiot, 2018, 43(3): 291
[9] 姚世杰, 樊云峰. 橡胶硫化剂进展[J]. 中国橡胶, 2003, 21(19): 23
YAO SJ, FAN YF. Development on rubber curing ingredients[J]. China Rubber, 2003, 21(19): 23
[10] 李樾, 杨会英, 王峰, 等. 药用卤化丁基橡胶塞中可提取硫的检测研究[J]. 中国药事, 2014, 28(11): 1257
LI Y, YANG HY, WANG F, et al. Study on detection of extractable sulphur from medicinal halogenated butyl rubber[J]. Chin Pharm Aff, 2014, 28(11): 1257
[11] 张磊, 黄葆华. 5%葡萄糖注射液中外源性抗氧剂的测定[J]. 安徽医药, 2011, 14(12): 400
ZHANG L, HUANG BH. Determination of antioxidant 1010 and antioxidant 168 contained in 5% glucose injection by HPLC[J]. Anhui Med Pharm J, 2011, 14(12): 400
[12] 赵霞, 胡昌勤, 金少鸿. 对考察药用丁基胶塞与药物相容性的思考与建议[J]. 中国药事, 2007, 21(10): 810
ZHAO X, HU CQ, JIN SH. Consideration and suggestion to estimate the compatibility between rubber closures and drugs[J]. Chin Pharm Aff, 2007, 21(10): 810
[13] 周劫, 殷果, 刘洪伟, 等. 药用丁基胶塞中BHT的GC-MS/MS法测定[J]. 中国医药工业杂志, 2013, 44(8): 801
ZHOU J, YIN G, LIU HW, et al. Determination of BHT in butyl rubber closures by GC-MS/MS [J]. Chin J Pharm, 2013, 44(8): 801
[14] 赵霞, 胡昌勤, 金少鸿. 药用丁基胶塞中易挥发性成分的成分分析[J]. 药物分析杂志, 2006, 26(3): 315
ZHAO X, HU CQ, JIN SH. The analysis of volatile substances of butyl rubber closures[J]. Chin J Pharm Anal, 2006, 26(3): 315
[15] 姜源, 程福军. 注射液用氯化丁基胶塞中的抗氧剂的测定和迁移测定[J]. 黑龙江医药, 2016, 29(3): 436
JIANG Y, CHENG FJ. Determination of the antioxidant concent and migration of the chlorinated butyl rubber plug with injection[J]. Heilongjiang Med J, 2016, 29(3): 436
[16] 陈壮生, 刘丽平. 注射用头孢呋辛钠粉针澄清度和颜色[J]. 广东药学院学报, 2010, 26(5): 459
CHEN ZS, LIU LP. Study on the improvement of clarity and color in cefuroxime sodium for injection[J]. Acad J Guangdong Pharm Coll, 2010, 26(5): 459
[17] 冯靖, 蔡心怡, 刘言. 药用丁基胶塞中抗氧剂-BHT和硫化剂-可提取留的测定[J]. 药物分析杂志, 2017, 37(4): 702
FENG J, CAI XY, LIU Y. Determination of antioxidant-BHT and vulcanizing agent extractable sulphur in medicinal butyl rubber[J]. Chin J Pharm Anal, 2017, 37(4): 702
