利用超高效液相色谱和离子色谱法测定注射用磷酸川芎嗪的含量
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摘要
目的:建立超高效液相色谱法(UPLC)和离子色谱法(IC)测定磷酸川芎嗪中川芎嗪和磷酸的含量,为质量评价提供依据。方法:UPLC测定川芎嗪的色谱柱为Waters Acquity BEH C18 (2.1 mm×50 mm,1.7μm);检测波长:300 nm检测川芎嗪,274 nm检测有关物质邻苯二甲酸二甲酯;流动相为0.1%甲酸水溶液(A)-0.1%甲酸乙腈(B),梯度洗脱(0.0～0.8 min,10%B→90%B;0.8～0.81 min,90%B→10%B;0.81～1.00 min,10%B),流速:0.7 m L/min。IC测定磷酸的离子交换色谱柱为Dionex IonPac AS11-HC-4μm (4×250 mm),流动相为30 mmol/L KOH溶液等度洗脱15 min,流速1.0 m L/min,柱温35℃;电导检测器;抑制器电流为50 m A。结果:川芎嗪和磷酸在10～100 g/m L内具有良好的线性关系,相关系数均为1.0,UPLC法测定川芎嗪的回收率为102.0%。IC测定磷酸的回收率为99.8%。7个公司生产的注射剂中川芎嗪的含量均在药典规定的范围90%～110%内。但是其中3个公司生产的注射剂磷酸超出药典规定范围90%～110%。结论:与常规HPLC/UPLC测定磷酸川芎嗪含量方法比较,本文所用方法测定结果更加准确、全面、且重复性好,能够真实反应注射用磷酸川芎嗪的实际含量,对于注射用磷酸川芎嗪的安全性和有效性评估提供了一定的依据。
Objective: To establish a method based on ultra-high performance liquid chromatography(UPLC) and an ion chromatography(IC) for analysis of ligustrazine and phosphoric acid, providing the basis for their qualitative evaluation. Methods:Ligustrazine was separated on Waters Acquity BEH C18(2.1 mm × 50 mm, 1.7 μm). The detection wavelengths were 300 nm for ligustrazine and 274 nm for dimethyl phthalate. Mobile phase A was 0.1% formic acid in water and B was 0.1% formic acid in acetonitrile with gradient elution(0.0～0.8 min,10% B→90% B; 0.8～0.81 min, 90% B→10% B; 0.81～1.0 min, 10% B). The flow rate was 0.7 m L/min. Phosphate anion was separated on Dionex IonPac AS11-HC-4 μm column(4 mm × 250 mm). Equivalent diluted with30 mmol/L KOH for 15 minutes. The flow was 1.0 m L/min. Column temperature was 35 ℃. The conductivity detector suppressor current was 50 m A. Results: Ligustrazine and phosphoric acid had good linearity in the range of 10～100 g/m L and both of the correlation coefficients were 1.0. The recoveries of UPLC method for ligustrazine was 102.0%.The recoveries of IC method for phosphoric acid was 99.8%.The obtained results of the 7 companies' injections of ligustrazine phosphate are all in pharmacopeia range of 90%～110%.But the phosphoric acid contents in 3 companies are out of the range. Conclusion: Compared with the commonly used content of ligustrazine phosphate by HPLC/UPLC, the results of this method are more accurate, comprehensive and reproducible, which can truly reflect the actual content of ligustrazine phosphate in the injection. It provides a better basis for the safety and effectiveness of ligustrazine phosphate for Injection.
Objective: To establish a method based on ultra-high performance liquid chromatography(UPLC) and an ion chromatography(IC) for analysis of ligustrazine and phosphoric acid, providing the basis for their qualitative evaluation. Methods:Ligustrazine was separated on Waters Acquity BEH C18(2.1 mm × 50 mm, 1.7 μm). The detection wavelengths were 300 nm for ligustrazine and 274 nm for dimethyl phthalate. Mobile phase A was 0.1% formic acid in water and B was 0.1% formic acid in acetonitrile with gradient elution(0.0～0.8 min,10% B→90% B; 0.8～0.81 min, 90% B→10% B; 0.81～1.0 min, 10% B). The flow rate was 0.7 m L/min. Phosphate anion was separated on Dionex IonPac AS11-HC-4 μm column(4 mm × 250 mm). Equivalent diluted with30 mmol/L KOH for 15 minutes. The flow was 1.0 m L/min. Column temperature was 35 ℃. The conductivity detector suppressor current was 50 m A. Results: Ligustrazine and phosphoric acid had good linearity in the range of 10～100 g/m L and both of the correlation coefficients were 1.0. The recoveries of UPLC method for ligustrazine was 102.0%.The recoveries of IC method for phosphoric acid was 99.8%.The obtained results of the 7 companies' injections of ligustrazine phosphate are all in pharmacopeia range of 90%～110%.But the phosphoric acid contents in 3 companies are out of the range. Conclusion: Compared with the commonly used content of ligustrazine phosphate by HPLC/UPLC, the results of this method are more accurate, comprehensive and reproducible, which can truly reflect the actual content of ligustrazine phosphate in the injection. It provides a better basis for the safety and effectiveness of ligustrazine phosphate for Injection.
引文
[1]肖衍宇,陈志鹏,平其能,等.磷酸川芎嗪理化性质及生物学性质的考察[J].中国医院药学杂志,2009,29(21):1810-1814
[2]Wang Ya-juan,Zhu Hui-zhi,Tong Jia-bing,et al.Ligustrazine improves blood circulation by suppressing Platelet activation in a rat model of allergic asthma[J].Environmental Toxicology and Pharmacology,2016,45,334-339
[3]许洪亮,李军.注射用磷酸川芍嗓对脑梗死患者梗死灶面积及神经功能的干预作用观察[J].中国药房,2014,25(23):2173-2175
[4]Wei Ying,Liu Jia-qi,Zhang Hong-ying,et al.Ligustrazine attenuates inflammation and the associated chemokines and receptors in ovalbumine-induced mouse asthma model[J].Environmental Toxicology and Pharmacology,2016,46:55-61
[5]Kuang Wen-hua,Zhang Xu,Zhu Wu-fu,et al.Ligustrazine modulates renal cysteine biosynthesis in rats exposed to cadmium[J].Environmental Toxicology and Pharmacology,2017,54:125-132
[6]Zheng Hong,Wang Si-ying,Zhou Ping-kun,et al.Effects of Ligustrazine on DNA damage and apoptosis induced by irradiation[J].Environmental Toxicology and Pharmacology,2013,36(3):1197-1206
[7]Lu Chun-feng,Xu Wen-xuan,Zhang Feng,et al.Ligustrazine prevents alcohol-induced liver injury by attenuating hepatic steatosis and oxidative stress[J].International Immunopharmacology,2015,29(2):613-621
[8]Zeng Ming-fei,Pan Lin-mei,Qi Shun-mei,et al.Systematic review of recent advances in pharmacokinetics of four classical Chinese medicines used for the treatment of cerebrovascular disease[J].Fitoterapia,2013,88:50-75
[9]Wang Peng-long,Zhao Rui,Yan Wen-qiang,et al.Neuroprotection by new ligustrazine-cinnamon acid derivatives on CoCl2-induced apoptosis in differentiated PC12 cells[J].Bioorganic Chemistry,2018,77:360-369
[10]Zhang Hui,Song Yu,Li Zhi-yong,et al.Evaluation of ligustrazine on the prevention of experimentally induced abdominal adhesions in rats[J].International Journal of Surgery,2015,21:115-121
[11]Chen Jian-li,Chen Jun-mao,Wang Xiao-tao,et al.Ligustrazine alleviates acute pancreatitis by accelerating acinar cell apoptosis at early phase via the suppression of p38 and Erk MAPK pathways[J].Biomedicine&Pharmacotherapy,2016,82:1-7
[12]Gao Cheng-jin,Liu Yu-hao,Ma Li-jie,et al.Effects of Ligustrazine on pulmonary damage in rats following scald injury[J].Burns,2012,38(5):743-750
[13]Wang Rong,Ma Chun-hua,Zhou Fan,et al.Siwu decoction attenuates oxonate-induced hyperuricemia and kidney inflammation in mice[J].Chinese Journal of Natural Medicines,2016,14(7):499-507
[14]Cheng Xian-Chao,Liu Xin-yong,Xu Wen-Fang,et al.Design,synthesis,and biological activities of novel Ligustrazine derivatives[J].Bioorganic&Medicinal Chemistry,2007,15(10):3315-3320
[15]Zou Jin-mi,Gao Ping,Hao Xia,et al.Recent progress in the structural modification and pharmacological activities of ligustrazine derivatives[J].European Journal of Medicinal Chemistry,2018,147:150-162
[16]国家药典委员会.中国药典2010年版第二增补本[P].348
[17]国家食品药品监督管理局.国家药品标准新药转正标准第78册[P].148-149
[18]Lin Zheng,Gong Zi-peng,Lu Yuan,et al.A UPLC MSMS method for simultaneous determination of danshensu,protocatechuic aldehyde,rosmarinic acid,andligustrazine in rat plasma,and its application to pharmacokinetic studies of Shenxiong glucose injection in rats[J].Journal of Chromatography.B,2015,997(5):210-217
[19]Chen Hong-ping,Gao Guan-wei,Liu Ping-xiang,et al.Development and validation of an ultra-performance liquid chromatography Q-Exactive Orbitrap mass spectrometry for the determination of fipronil and its metabolites in tea and chrysanthemum[J].Food Chemistry,2018(246):328-334
[20]Dou Li-li,Duan Li,Guo Long,et al.An UHPLC-MS/MS method for simultaneous determination of quercetin 3-O-rutinoside,kaempferol3-O-rutinoside,isorhamnetin 3-O-rutinoside,bilobalide and ligustrazine in rat plasma,and its application to pharmacokinetic study of Xingxiong injection[J].Chinese Journal of Natural Medicines,2017,15(9):710-720
[21]Izabela Matysiak,Maria Balcerzak,Rajmund Michalski.Ion chromatography with conductometric detection for quantitation of formic acid in Polish bee honey[J].Journal of Food Composition and Analysis,2018,73(7):55-59
[22]Qiu Hui-min,Geng Jin-ju,Han Chao,et al.Determination of Phosphite,Phosphate,Glyphosate and Aminomethylphosphonic Acid by Two-Dimensional Ion Chromatography System Coupled with Capillary Ion Chromatography[J].Chinese Journal of Analytical Chemistry,2013,41(12):1910-1914
[23]Han C,Geng J,Xie X.Determination of phosphite in a eutrophic freshwater lake by suppressed conductivity ion chromatography[J].Environmental Science and Technology,2012,46(19):10667-10674
[24]Segawa H,Yamanaka S,Ito M,et al.Internalization of renal type IIc Na-Pi cotransporter in response to a highphosphate diet[J].American journal of physiology-heart and circulatory physiology,2005,288(3):587-596
[25]Miyamoto K,Ito M,Tatsumi S,et al.New aspect of renal phosphate reabsorption:the type IIc sodium-dependent phosphate transporter[J].Journal of the American Society of Nephrology,2007,27(5):503-515
[26]Narchi H.Hyperostosis with hyperphosphatemia:evidence of familial occurrence and association with tumoral calcinosis[J].Journal of Pediatrics,1997,99(5):745-748
[27]Mc Clatchie S,Bremner AD.Tumoral calcinosisean unrecognized disease[J].British Medical Journal,1969,1(5637):153-155
[28]Hacihanefioglu U.Tumoral calcinosis.A clinical and pathological study of eleven unreported cases in Turkey[J].journal of bone and joint surgery,1978,60(8):1131-1135
[29]Adams WM,Laitt RD,Davies M,et al.Familial tumoral calcinosis:association with cerebral and peripheral aneurysm formation[J].Journal of Neuroradiology,1999,41(5):351-355
[30]Goldenstein PT,Neves PD,Balbo BE,et al.Dialysis as a treatment option for a patient with normal kidney function and familial tumoral calcinosis due to a compound heterozygous FGF23 mutation[J].American Journal of Kidney Diseases,2018,72(3):457-461
[2]Wang Ya-juan,Zhu Hui-zhi,Tong Jia-bing,et al.Ligustrazine improves blood circulation by suppressing Platelet activation in a rat model of allergic asthma[J].Environmental Toxicology and Pharmacology,2016,45,334-339
[3]许洪亮,李军.注射用磷酸川芍嗓对脑梗死患者梗死灶面积及神经功能的干预作用观察[J].中国药房,2014,25(23):2173-2175
[4]Wei Ying,Liu Jia-qi,Zhang Hong-ying,et al.Ligustrazine attenuates inflammation and the associated chemokines and receptors in ovalbumine-induced mouse asthma model[J].Environmental Toxicology and Pharmacology,2016,46:55-61
[5]Kuang Wen-hua,Zhang Xu,Zhu Wu-fu,et al.Ligustrazine modulates renal cysteine biosynthesis in rats exposed to cadmium[J].Environmental Toxicology and Pharmacology,2017,54:125-132
[6]Zheng Hong,Wang Si-ying,Zhou Ping-kun,et al.Effects of Ligustrazine on DNA damage and apoptosis induced by irradiation[J].Environmental Toxicology and Pharmacology,2013,36(3):1197-1206
[7]Lu Chun-feng,Xu Wen-xuan,Zhang Feng,et al.Ligustrazine prevents alcohol-induced liver injury by attenuating hepatic steatosis and oxidative stress[J].International Immunopharmacology,2015,29(2):613-621
[8]Zeng Ming-fei,Pan Lin-mei,Qi Shun-mei,et al.Systematic review of recent advances in pharmacokinetics of four classical Chinese medicines used for the treatment of cerebrovascular disease[J].Fitoterapia,2013,88:50-75
[9]Wang Peng-long,Zhao Rui,Yan Wen-qiang,et al.Neuroprotection by new ligustrazine-cinnamon acid derivatives on CoCl2-induced apoptosis in differentiated PC12 cells[J].Bioorganic Chemistry,2018,77:360-369
[10]Zhang Hui,Song Yu,Li Zhi-yong,et al.Evaluation of ligustrazine on the prevention of experimentally induced abdominal adhesions in rats[J].International Journal of Surgery,2015,21:115-121
[11]Chen Jian-li,Chen Jun-mao,Wang Xiao-tao,et al.Ligustrazine alleviates acute pancreatitis by accelerating acinar cell apoptosis at early phase via the suppression of p38 and Erk MAPK pathways[J].Biomedicine&Pharmacotherapy,2016,82:1-7
[12]Gao Cheng-jin,Liu Yu-hao,Ma Li-jie,et al.Effects of Ligustrazine on pulmonary damage in rats following scald injury[J].Burns,2012,38(5):743-750
[13]Wang Rong,Ma Chun-hua,Zhou Fan,et al.Siwu decoction attenuates oxonate-induced hyperuricemia and kidney inflammation in mice[J].Chinese Journal of Natural Medicines,2016,14(7):499-507
[14]Cheng Xian-Chao,Liu Xin-yong,Xu Wen-Fang,et al.Design,synthesis,and biological activities of novel Ligustrazine derivatives[J].Bioorganic&Medicinal Chemistry,2007,15(10):3315-3320
[15]Zou Jin-mi,Gao Ping,Hao Xia,et al.Recent progress in the structural modification and pharmacological activities of ligustrazine derivatives[J].European Journal of Medicinal Chemistry,2018,147:150-162
[16]国家药典委员会.中国药典2010年版第二增补本[P].348
[17]国家食品药品监督管理局.国家药品标准新药转正标准第78册[P].148-149
[18]Lin Zheng,Gong Zi-peng,Lu Yuan,et al.A UPLC MSMS method for simultaneous determination of danshensu,protocatechuic aldehyde,rosmarinic acid,andligustrazine in rat plasma,and its application to pharmacokinetic studies of Shenxiong glucose injection in rats[J].Journal of Chromatography.B,2015,997(5):210-217
[19]Chen Hong-ping,Gao Guan-wei,Liu Ping-xiang,et al.Development and validation of an ultra-performance liquid chromatography Q-Exactive Orbitrap mass spectrometry for the determination of fipronil and its metabolites in tea and chrysanthemum[J].Food Chemistry,2018(246):328-334
[20]Dou Li-li,Duan Li,Guo Long,et al.An UHPLC-MS/MS method for simultaneous determination of quercetin 3-O-rutinoside,kaempferol3-O-rutinoside,isorhamnetin 3-O-rutinoside,bilobalide and ligustrazine in rat plasma,and its application to pharmacokinetic study of Xingxiong injection[J].Chinese Journal of Natural Medicines,2017,15(9):710-720
[21]Izabela Matysiak,Maria Balcerzak,Rajmund Michalski.Ion chromatography with conductometric detection for quantitation of formic acid in Polish bee honey[J].Journal of Food Composition and Analysis,2018,73(7):55-59
[22]Qiu Hui-min,Geng Jin-ju,Han Chao,et al.Determination of Phosphite,Phosphate,Glyphosate and Aminomethylphosphonic Acid by Two-Dimensional Ion Chromatography System Coupled with Capillary Ion Chromatography[J].Chinese Journal of Analytical Chemistry,2013,41(12):1910-1914
[23]Han C,Geng J,Xie X.Determination of phosphite in a eutrophic freshwater lake by suppressed conductivity ion chromatography[J].Environmental Science and Technology,2012,46(19):10667-10674
[24]Segawa H,Yamanaka S,Ito M,et al.Internalization of renal type IIc Na-Pi cotransporter in response to a highphosphate diet[J].American journal of physiology-heart and circulatory physiology,2005,288(3):587-596
[25]Miyamoto K,Ito M,Tatsumi S,et al.New aspect of renal phosphate reabsorption:the type IIc sodium-dependent phosphate transporter[J].Journal of the American Society of Nephrology,2007,27(5):503-515
[26]Narchi H.Hyperostosis with hyperphosphatemia:evidence of familial occurrence and association with tumoral calcinosis[J].Journal of Pediatrics,1997,99(5):745-748
[27]Mc Clatchie S,Bremner AD.Tumoral calcinosisean unrecognized disease[J].British Medical Journal,1969,1(5637):153-155
[28]Hacihanefioglu U.Tumoral calcinosis.A clinical and pathological study of eleven unreported cases in Turkey[J].journal of bone and joint surgery,1978,60(8):1131-1135
[29]Adams WM,Laitt RD,Davies M,et al.Familial tumoral calcinosis:association with cerebral and peripheral aneurysm formation[J].Journal of Neuroradiology,1999,41(5):351-355
[30]Goldenstein PT,Neves PD,Balbo BE,et al.Dialysis as a treatment option for a patient with normal kidney function and familial tumoral calcinosis due to a compound heterozygous FGF23 mutation[J].American Journal of Kidney Diseases,2018,72(3):457-461