More than 80 aristolochic acids(AAs) and aristololactams(ALs) have been found in plants of the Aristolochiaceae family, but relatively few have been fully studied. The present study aimed at developing and validating ...More than 80 aristolochic acids(AAs) and aristololactams(ALs) have been found in plants of the Aristolochiaceae family, but relatively few have been fully studied. The present study aimed at developing and validating a liquid chromatography tandem mass spectrometry(LC/MS^n) for the analysis of these compounds. We characterized the fragmentation behaviors of 31 AAs, ALs, and their analogues via high performance liquid chromatography coupled with electrospray ionization mass spectrometry. We summarized their fragmentation rules and used these rules to identify the constituents contained in Aristolochia contorta, Ar. debilis, Ar. manshurensis, Ar. fangchi, Ar. cinnabarina, and Ar. mollissima. The AAs and ALs showed very different MS behaviors. In MS1 of AAs, the characteristic pseudomolecular ions were [M + NH_4]^+, [M + H]^+, and [M + H - H_2O]^+. However, only [M + H]^+ was found in the MS1 of ALs, which was simpler than that of AAs. Distinct MSn fragmentation patterns were found for AAs and ALs, showing the same skeleton among the different substituent groups. The distribution of the 31 constituents in the 6 species of Aristolochia genus was reported for the first time. 25 Analogues of AAs and ALs were detected in this genus. A hierarchical schemes and a calculating formula of the molecular formula of these nitrophenanthrene carboxylic acids and their lactams were proposed. In conclusion, this method could be applied to identification of similar unknown constituents in other plants.展开更多
目的采用LC/MSn技术推断拉氧头孢钠原料中杂质的结构。方法采用Shiseido TYPE UG80C18(4.6mm×250mm,5μm)色谱柱,以0.01mol/L醋酸铵溶液-甲醇(99:1,V/V)作为流动相A,以0.01mol/L醋酸铵溶液-甲醇(70:30,V/V)为流动相B,进行梯度洗脱...目的采用LC/MSn技术推断拉氧头孢钠原料中杂质的结构。方法采用Shiseido TYPE UG80C18(4.6mm×250mm,5μm)色谱柱,以0.01mol/L醋酸铵溶液-甲醇(99:1,V/V)作为流动相A,以0.01mol/L醋酸铵溶液-甲醇(70:30,V/V)为流动相B,进行梯度洗脱分离杂质。通过强力试验归属杂质的来源,在正离子模式下获取各杂质的质谱数据,结合头孢菌素的普遍质谱裂解规律和降解反应原理,解析相关杂质的结构。结果推断了12种杂质的化学结构,样品中未检出聚合物杂质。结论LC/MSn法可快速准确推断拉氧头孢钠原料中杂质结构,中国药典的拉氧头孢钠质量标准需要进一步提高。展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.30873466,81173494&81274073)National Science and Technology Major Project(No 2014ZX09304307-001-012)
文摘More than 80 aristolochic acids(AAs) and aristololactams(ALs) have been found in plants of the Aristolochiaceae family, but relatively few have been fully studied. The present study aimed at developing and validating a liquid chromatography tandem mass spectrometry(LC/MS^n) for the analysis of these compounds. We characterized the fragmentation behaviors of 31 AAs, ALs, and their analogues via high performance liquid chromatography coupled with electrospray ionization mass spectrometry. We summarized their fragmentation rules and used these rules to identify the constituents contained in Aristolochia contorta, Ar. debilis, Ar. manshurensis, Ar. fangchi, Ar. cinnabarina, and Ar. mollissima. The AAs and ALs showed very different MS behaviors. In MS1 of AAs, the characteristic pseudomolecular ions were [M + NH_4]^+, [M + H]^+, and [M + H - H_2O]^+. However, only [M + H]^+ was found in the MS1 of ALs, which was simpler than that of AAs. Distinct MSn fragmentation patterns were found for AAs and ALs, showing the same skeleton among the different substituent groups. The distribution of the 31 constituents in the 6 species of Aristolochia genus was reported for the first time. 25 Analogues of AAs and ALs were detected in this genus. A hierarchical schemes and a calculating formula of the molecular formula of these nitrophenanthrene carboxylic acids and their lactams were proposed. In conclusion, this method could be applied to identification of similar unknown constituents in other plants.
文摘目的采用LC/MSn技术推断拉氧头孢钠原料中杂质的结构。方法采用Shiseido TYPE UG80C18(4.6mm×250mm,5μm)色谱柱,以0.01mol/L醋酸铵溶液-甲醇(99:1,V/V)作为流动相A,以0.01mol/L醋酸铵溶液-甲醇(70:30,V/V)为流动相B,进行梯度洗脱分离杂质。通过强力试验归属杂质的来源,在正离子模式下获取各杂质的质谱数据,结合头孢菌素的普遍质谱裂解规律和降解反应原理,解析相关杂质的结构。结果推断了12种杂质的化学结构,样品中未检出聚合物杂质。结论LC/MSn法可快速准确推断拉氧头孢钠原料中杂质结构,中国药典的拉氧头孢钠质量标准需要进一步提高。
