摘要
目的：探讨中国药典2010年版盐酸苄丝肼有关物质检查方法的合理性,并对方法作出改进。方法：采用UPLC-MS联用技术对未知杂质进行鉴定,色谱柱为ACQUITY UPLC BEHC18柱（2.1mm×50mm,1.7μm）,流动相为三氟醋酸-甲醇-水（1∶20∶1000）,流速0.5mL·min^-1,采用正离子扫描（ESI^＋）方式,毛细管电压3.0kV,锥孔电压20V,源温120℃,脱溶剂气温度400℃,脱溶剂气流量600L·h^-1,锥孔气流量50L·h^-1。采用改进后的HPLC法检测该杂质,使用Phenomenex Luna C8色谱柱（4.6mm×250mm,5μm）,流动相为A（2.2g庚烷磺酸钠和6.8g磷酸二氢钾溶于900mL水,加入50mL甲醇,用磷酸调至pH3.5）-B（2.2g庚烷磺酸钠和6.8g磷酸二氢钾溶于500mL水,用磷酸调至pH35,加入500mL甲醇）,梯度洗脱（0～15min,0%B→100%B;15～25min,100%B;25～30min,100%B→0%B;30～35min,0%B）,流速1.3mL·min^-1,检测波长210nm,柱温30℃。结果：中国药典2010年版方法不能检出主要杂质三羟苄基苄丝肼,本文建立的HPLC法能够有效检出该杂质。结论：本文方法专属性强,灵敏度高,实验结果准确可靠,可作为测定盐酸苄丝肼有关物质的检查方法,为新版中国药典盐酸苄丝肼有关物质检查方法的完善提供参考。
Objective： To discuss and improve the determination method of related substances of benserazide hydrochloride in Chinese Pharmacopoeia 2010.Methods： An UPLC- MS system of an ACQUITY Waters UPLC coupled with a Micromass Quattro Premier XE ESI source mass spectrometer was used for the identification of benserazide impurity B.The BEH C18column（ 2.1mm × 50 mm,1.μm） was operated with a mobile phase consisting of trifluoroacetic acid- methanol- water（ 1∶ 20∶ 1000） at a flow rate of 0.5mL·min^-1.The analysis was performed with electric spray interface under positive ion mode,the capillary voltage was 3.0k V,the taper hole voltage was 20V,the ion source temperature 120℃,the temperature of the desolventizing agent 400℃,the desolventizing gas of N2at a flow rate of 600L·h^-1,the taper hole gas of He at a flow rate of 50L·h^-1.Benserazide impurity was detected by the improved method under the following chromatographic conditions.The Phenomenex Luna C8column（ 4.6mm × 250mm,5μm） was used at 30 ℃.The mobile phase composition included phase A（ 2.2g of sodium heptanesulfonate and 6.8g of potassium dihydrogen phosphate dissolved in 900mL of water,added with 50mL of methanol and adjusted to p H3.5 with phosphoric acid）,B（ 2.2g of sodium heptanesulfonate and 6.8g of potassium dihydrogen phosphate dissolved in 500mL of water,adjusted to p H 3.5with phosphoric acid and added with 500mL of methanol）,using a gradient elution（ 0- 5 min,0% B→100% B; 15- 25min,100% B; 25- 30 min,100% B→0% B; 30- 35 min,0% B） with a flow rate of 1.3mL·min^-1,and 210nm as the detection wavelength.Results： DL-2- amino- N＇,N＇- bis（2,3,4- trihydroxybenzyl） hydracrylohydrazide which was the main impurity of benserazide hydrochloride could not be detected by the determination method in Chinese Pharmacopoeia 2010.The HPLC method established in this paper could detect the above mentioned impurity effectively.Conclusion： The new method is accurate,sensitive and reliable.It can be used for the related substances determination in benserazide hydrochloride.It provided a useful reference for improving the related substances method of benserazide hydrochloride in the new edition of Chinese Pharmacopoeia.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2014年第11期2035-2040,共6页
Chinese Journal of Pharmaceutical Analysis
