高效液相色谱-线性离子阱质谱鉴定瑞舒伐他汀中间体中相关杂质

目的：建立高效液相色谱-线性离子阱质谱法分离鉴定瑞舒伐他汀药物中间体N-[5-腈基-4-（4-氟苯基）-6异丙基嘧啶-2-]-N-甲基甲磺酰胺中的有关杂质，并探讨这些杂质的质谱裂解规律。方法：采用Cortecs C₁₈色谱柱（100 mm×2.1 mm，2.7 μm），以乙腈-0.1%甲酸溶液为流动相，进行梯度洗脱，流速为0.25 mL·min^-1，柱温35℃，检测波长为242 nm，电喷雾离子化正负2种离子模式扫描。结果：中间体与各已知杂质均分离良好，根据一级和多级质谱信息，共鉴定出7种杂质，包括对甲基苯磺酸、4-（4-氟苯基）-2-羟基-6-异丙基-5-氰基-嘧啶、4-（4-氟苯基）-2-氨基-6-异丙基-5-氰基-嘧啶、5-氰基-6-（4-氟苯基）-4-异丙基-2-N-甲磺酰胺嘧啶-1-N-氧化物、4-（4-氟苯基）-2-甲氨基-6-异丙基-5-氰基-嘧啶、中间体的氯代产物和5-甲酸内酯化产物。杂质类型主要为氧化、降解产物及合成副产物。结论：本法分析结果为有关杂质确证提供分析方法，为瑞舒伐他汀合成中的质量控制提供指导。

Objective: To develop a high performance liquid chromatography coupled with linear ion trap mass spectrometric method for the identification of impurities in rosuvastatin intermediate:N-[5-cyano-4-(4-fluorophenyl)-6-isopropylpyrimidin-2-yl]-N-methylmethane sulfonamide. The fragmentations were also discussed.Methods: The analysis was conducted on a Waters Cortecs C₁₈ column(100 mm×2.1 mm, 2.7 μm)with a mobile phase of acetonitrile-water(0.1% formate acid)by gradient elution at the flow rate of 0.25 mL·min^-1. The column temperature was 35℃ and the detection wavelength was 242 nm. MS was scanning in ESI source, positive and negative ions mode.Results: The impurities and intermediate were completely separated from each other; Seven impurities were identified by MS1 and MSⁿ fragments. The impurities were identified as 4-methylbenzenesulfonic acid, 4-(4-fluorophenyl)-2-hydroxy-6-isopropylpyrimidine-5-carbonitrile, 2-amino-4-(4-fluoro phenyl)-6-isopropylpyrimidine-5-carbonitrile, 5-cyano-6-(4-fluorophenyl)-4-isopropyl-2-(N-methylmethylsulfonamido)pyrimidine-1-oxide; N-[8-fluoro-4-isopropyl-5-oxo-5Hchromeno(4, 3-d)pyrimidin-2-yl]-N-methylmethanesulfonamide; 4-(4-fluorophenyl)-6-isopropyl-2-(methylamino)pyrimidine-5-carbonitrile and N-[4-(chloro-4-fluorophenyl)-5-cyano-6-isopropylpyrimidin-2-yl]-N-methylmethanesulfonamide. The impurities include oxidative products, degradation products and synthesis byproducts. The mass fragmentation patterns and structural assignments of these seven impurities were studied.Conclusion: The established method is suitable for the identification of related substances and provides a tool for the quality control in the synthesis of rosuvastatin.

[1] TADA H, KOBAYASHIJ, KAWASHIRI MA, et al. Changes in lipoprotein lipase and endothelial lipase mass in familial hypercholesterolemia during three-drug lipid-lowering combination therapy[J]. Lipids Health Dis, 2016, 15(1):66.
[2] CARTER NJ. Rosuvastatin:A review of its use in the prevention of cardiovascular disease in apparently healthy women or men with normal LDL-C levels and elevated hsCRP levels[J]. Am J Cardiovasc Drugs, 2010, 10(6):383
[3] 浙江大学.瑞舒伐他汀中间体的制备方法及中间体化合物:中国, 104059024[P]. 2014-9-24
[3] Zhejiang University. Improved Method for Preparation of Rosuvastatin Intermediate and Intermediate Compound:China, 104059024[P]. 2014-09-24
[4] XIONG F, WANG H, YAN L, et al. Stereocontrolled synthesis of rosuvastatin calcium via iodine chloride-induced intramolecular cyclization[J]. Org Biomol Chem, 2016, 14(4):1363
[5] KUMAR N, DEVINENI SR, GAJJALA PR, et al. Four processrelated potential new impurities in ticagrelor:identification, isolation, characterization using HPLC, LC/ESI-MSⁿ, NMR and their synthesis[J]. J Pharm Biomed Anal 2016, 120:248
[6] 张得胜,庄波阳,王凌.高效液相色谱法测定瑞舒伐他汀钙片有关物质[J].海峡药学, 2013, 25(10):29 ZHANG DS, ZHANG BY, WANG L. Determination of the related substances in rosuvastatin calcium tablets by high performance liquid chromatography[J]. Strait Pharm J, 2013, 25(10):29
[7] 蔡垠武,利苗艳,周志亮,等.高效液相色谱法测定瑞舒伐他汀钙片中8种有关物质的含量[J].中国药物警戒, 2014, 11(11):657 CAI YW, LI MY, ZHOU ZL, et al. Determination of eight impurities of rosuvastatin calcium tablets by HPLC[J]. Chin J Pharmacovigil, 2014, 11(11):657
[8] TRIVEDI HK, PATEL MC. Development and validation of a stability-indicating RP-UPLC method for determination of rosuvastatin and related substances in pharmaceutical dosage form[J]. Sci Pharm, 2012, 80(2):393
[9] MEHTA TN, PATEL AK, KULKARINI GM, et al. Determination of rosuvastatin in the presence of its degradation products by a stability-indicating LC method[J]. J AOAC Int. 2005, 88(4):1142
[10] REDDY GVR, REDDY BV, HAQUE SW, et al. Development and validation of a stability-indicating uplc method for rosuvastatin and its related impurities in pharmaceutical dosage forms[J]. Quimica Nova, 2011, 34(2):250
[11] YEGOROVA AV, FEDOSENKO GA, SCRYPYNETS YV, et al. Determination of residual amounts of rosuvastatin calcium on the surfaces of pharmaceutical equipment by HPLC and luminescence[J]. J Anal Chem, 2016, 71(1):126
[12] SRI DS, KUMAR TH, RAO KVP, et al. Validated RP-HPLC method for simultaneous determination of rosuvastatin calcium and ezetimibe in pharmaceutical dosage form[J]. Int J Pharm Sci, 2015, 7(4):209
[13] ASSASSI AL, ROY CE, PEROVITCH P, et al. Green analytical method development for statin analysis[J]. J Chromatogr A, 2015, 1380:104
[14] MOSTAFA NM, BADAWEY AM, LAMIE NT, et al. Selective chromatographic methods for the determination of Rosuvastatin calcium in the presence of its acid degradation products[J]. J Liq Chromatogr Relat Technol, 2014, 37(15):2182
[15] SHARMA T, SI SC, SANKAR DG. Development and validation of LC method for the simultaneous estimation of rosuvastatin calcium and olmesartan medoxomil in pharmaceutical dosage form[J]. Int J Chem Tech Res, 2014, 6(2):1115
[16] MUKTHINUTHALAPATI MA, BUKKAPATNAM V, BANDARU SPK. Stability indicating liquid chromatographic method for the simultaneous determination of rosuvastatin and ezetimibe in pharmaceutical formulations[J]. Adv Pharm Bull, 2014, 4(4):405