液相色谱-四极杆/飞行时间质谱法分析29种芬太尼类物质及其碎裂机理北大核心CSCD

芬太尼类物质品种繁多,自我国整类列管后,整类检测是该领域的重点和难点。该文详细研究了29种化合物的二级质谱碎片离子碎裂机理,总结出芬太尼类物质的碎裂规律和特点,为芬太尼物质的整类筛查检测提供参考。建立了分析29种芬太尼类物质的一级和二级质谱库的定性方法,建立了液相色谱-四极杆/飞行时间质谱(LC-QTOF-MS)检测29种芬太尼类物质的定量方法。药品和白色粉末类、蛋白质和乳饮料类样品经乙腈提取,含糖固体或粉末类、饮用水类、果蔬饮料类、保健饮料类、茶饮料类、酒类样品经10%乙腈水溶液提取,提取液经涡旋、离心和过膜后,采用Kinetex C18色谱柱(100 mm×2.1 mm,2.6μm)分离,以乙腈和0.08%甲酸水溶液为流动相进行梯度洗脱,采用四极杆/飞行时间质谱,在正离子模式下,外标法定量检测。结果表明,29种芬太尼类物质在1~20μg/L范围内线性关系良好,相关系数均大于0.995,检出限(LOQ)均为0.01 mg/kg,定量限(LOQ)均为0.05 mg/kg,在降糖药、露露、葡萄糖粉、珍露保健饮料和巧克力样品中3个加标水平平均回收率为85.2%~112.9%,相对标准偏差(RSD)为1.9%~19.8%(n=6)。该方法操作简单,耗时短,灵敏度高,稳定性好,检测品种覆盖范围广,适用于药品类、含糖固体或粉末类、饮料类、饮用水类和酒类等样品中29芬太尼类物质的定性和定量检测。

Analysis of 29 fentanyl analogs and their fragmentation mechanism by liquid chromatography-quadrupole time-of-flight mass spectrometry

Given the wide variety of fentanyl analogs,the test for this entire group tends to be crucial and particularly difficult since all fentanyl-like substances are listed as controlled substances in China.This study meticulously analyzed the fragmentation pathways and mechanisms of 29 fentanyl analogs and summarized the fragmentation pathways and features for the entire group of fentanyl analogs,thus providing a reference for related screening tests.Fentanyl,thiofentanyl,and sufentanil were selected as the representative compounds in this study,and the fragmentation mechanism of their fragment ions was interpreted.The general fragmentation rules for fentanyl analogs were summarized as well.The fragment ions of the three compounds formed by induced cleavage(i)came with high abundance ratios,such as fragment ions of m/z 188,105,194,111,and 238,while the induced cleavage was due to the amide and piperidinyl groups.Moreover,the induction ability of amide group was significantly stronger than that of the piperidinyl group,and induced cleavage was the main fragmentation pathway for most of the fentanyl analogs.Furthermore,the fragment ions with m/z 281 and 287 for fentanyl and thiofentanyl were formed by loss of the propionyl group after single H rearrangement(rH).The fragment ions with m/z 216,146,and 132 for fentanyl and thiofentanyl were formed by double H rearrangement(r2H).Although their abundance ratios were not high,they still had specificity and regularity.Elimination reaction(re)was also a very common fragmentation pathway for these compounds,leading to fragment ions with m/z 134 and 140.Phenylethyl substituents were more prone to the elimination reaction with a higher abundance ratio than thiophenethyl substituents.Compounds such as sufentanil with methoxy substituents at the piperidinyl para-position could produce a large number of fragment ions,which were more susceptible to the rH pathway and loss of methanol neutral molecules,leading to the formation of ions with m/z 355.Similarly,compounds such as remifentanil bearing a methyl formate substituent at the piperidine para-position also produced numerous fragment ions,which were more prone to the rH pathway to lose methyl formate or methanol neutral molecules and furnish fragment ions with m/z 317 or 345.Compounds containing hydroxyl substituents,such asβ-hydroxyfentanyl andβ-hydroxythiofentanyl,produce significant dehydration ions and formed fragment ions with m/z 335(β-hydroxyfentanyl)and m/z 341(β-hydroxythiofentanyl).A method based on liquid chromatography-quadrupole time-of-flight mass spectrometry(LC-QTOF-MS)for the qualitative and quantitative determination of the 29 fentanyl analogs was developed.Drugs and white powder samples were extracted by acetonitrile,as well as protein and milk beverage samples.Sugar-containing solids or powders,drinking water,fruit and vegetable drinks,health drinks,tea drinks,and alcohol samples were extracted by 10%acetonitrile aqueous solution.Following vortexing,centrifugation,and membrane separation,the target compounds were separated on a Kinetex C18 column(100 mm×2.1 mm,2.6μm)with gradient elution at a flow rate of 0.4 mL/min.The mobile phases were composed of acetonitrile and 0.08%formic acid aqueous solution.The target compounds were quantified by LC-QTOF-MS using an external standard method in positive ion mode.The 29 fentanyl analogs showed good linear relationships in the range of 1-20μg/L,and the correlation coefficients were greater than 0.995.The limits of detection(LODs)and limits of quantification(LOQs)were 0.01 mg/kg and 0.05,respectively.The average recoveries were 85.2%-112.9%for hypoglycemic drugs,Lulu drinks,glucose powder,Zhenlu health drink and chocolate,with RSDs of 1.9%-19.8%(n=6).This method is rapid,simple,time-saving,highly sensitivity and stable,and it is applicable to a wide variety of samples.Hence,it is suitable for the identification,confirmation,and quantitative detection of the 29 fentanyl analogs in drugs,solids or powders containing sugar,beverages,drinking water,wine samples,etc.