串联双柱固相萃取-高效液相色谱-电喷雾多级质谱法测定盐酸二甲双胍制剂中的盐酸二甲双胍及残留物三聚氰胺和双氰胺

Determination of metformin hydrochloride,melamine and dicyandiamide in metformin hydrochloride preparations by tandem dual solid phase extraction cartridges-high performance liquid chromatography-electrospray ionization multi-stage mass spectrometry

建立了串联双柱固相萃取-高效液相色谱-电喷雾多级质谱(HPLC-ESI-MSn)确认及测定盐酸二甲双胍制剂中的盐酸二甲双胍及残留物三聚氰胺和双氰胺的方法。盐酸二甲双胍制剂样品在超声辅助下,采用含0.1%(v/v)乙酸的无水乙醇溶液提取,选择Cleanert PCX-C18串联固相萃取双柱净化富集,以5%(v/v)氨水甲醇溶液为洗脱剂进行洗脱。采用Kromasil-C18色谱柱(100 mm×4.6 mm,3.5μm)分离,梯度洗脱,在电喷雾多级质谱、选择离子模式下检测,外标法定量。通过比较制剂残留物在不同萃取溶剂、固相萃取柱及洗脱液等条件下的回收率,优化了前处理方法。结果表明,在各自的线性范围内,3种目标物的峰面积与质量浓度呈良好的线性关系,相关系数(r^2)≥0.999 2;方法的检出限为1.48~13.61μg/kg,定量限为5.96~45.67μg/kg。3种目标物在低、中、高加标水平下的平均回收率为65.02%~118.33%,相对标准偏差(RSD)≤13.41%(n=5)。该法操作简单,净化效果好,准确度高,重现性好,适用于检测和分析不同盐酸二甲双胍制剂中盐酸二甲双胍及残留物三聚氰胺和双氰胺。

A method for the confirmation and quantification of metformin hydrochloride and its relative substances melamine and dicyandiamide using tandem dual solid phase extraction （SPE） cartridges and high performance liquid chromatography-electrospray ionization multi-stage mass spectrometry （HPLC- ESI-MSn ） was developed. The samples were extracted with anhydrous ethanol containing 0. 1% （v/v ） acetic acid under ultrasound-assisted conditions. The extracts were concentrated and purified using Cleanert PCX and C18 tandem dual solid phaseextraction cartridges, and eluted with 5% （v/v ） ammonia methanol solution. The separation was performed on a Kromasil-C18 column （100 mmx4. 6 mm, 3. 5 ^m） with gradient elution. The detection was performed in selected ion monitoring （SIM） mode using electrospray ioniza-tion multi-stage mass spectrometry. The external standard method was used for quantification. The extraction solvents, types of SPE cartridges and eluents were optimized by comparing the recoveries under different conditions. The results showed that the detector response of each tar-get compound was linear in corresponding mass concentration ranges with the correlation coef-ficients （r2） ^0. 999 2. The limits of detection （LODs） and the limits of quantification （LOQs） of the three analytes were 1. 48-13. 61 ^^^kg and 5. 96-45. 67 ^^^kg, respectively. The recov-eries of the three analytes were 65. 0 2 % -118. 33% spiked at low, medium and high levels. The relative standard deviations （RSDs） were no more than 13. 41%. The method is reliable, easy, and has a better purification effect. The method can be applied to the routine analysis of met-formin hydrochloride and its relative substances melamine and dicyandiamide in different prepa-rations of metformin hydrochloride.