title: Quantitative determination of five metabolites of aspirin by UHPLC-MS/MS coupled with enzymatic reaction and its application to evaluate the effects of aspirin dosage on the metabolic profile.
abstract: Acetylsalicylic acid (Aspirin, ASA) is a famous drug for cardiovascular diseases in recent years. Effects of ASA dosage on the metabolic profile have not been fully understood. The purpose of our study is to establish a rapid and reliable method to quantify ASA metabolites in biological matrices, especially for glucuronide metabolites whose standards are not commercially available. Then we applied this method to evaluate the effects of ASA dosage on the metabolic and excretion profile of ASA metabolites in rat urine. Salicylic acid (SA), gentisic acid (GA) and salicyluric acid (SUA) were determined directly by UHPLC-MS/MS, while salicyl phenolic glucuronide (SAPG) and salicyluric acid phenolic glucuronide (SUAPG) were quantified indirectly by measuring the released SA and SUA from SAPG and SUAPG after β-glucuronidase digestion. SUA and SUAPG were the major metabolites of ASA in rat urine 24h after ASA administration, which accounted for 50% (SUA) and 26% (SUAPG). When ASA dosage was increased, the contributions dropped to 32% and 18%, respectively. The excretion of other three metabolites (GA, SA and SAPG) however showed remarkable increases by 16%, 6% and 4%, respectively. In addition, SUA and SUAPG were mainly excreted in the time period of 12-24h, while GA was excreted in the earlier time periods (0-4h and 4-8h). SA was mainly excreted in the time period of 0-4h and 12-24h. And the excretion of SAPG was equally distributed in the four time periods. We went further to show that the excretion of five metabolites in rat urine was delayed when ASA dosage was increased. In conclusion, we have developed a rapid and sensitive method to determine the five ASA metabolites (SA, GA, SUA, SAPG and SUAPG) in rat urine. We showed that ASA dosage could significantly influence the metabolic and excretion profile of ASA metabolites in rat urine.
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China.
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Li
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China. Electronic address: njuguo@njucm.edu.cn.
authors.forename: Jian-Ming
authors.initials: JM
authors.lastname: Guo
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China.
authors.forename: Er-Xin
authors.initials: EX
authors.lastname: Shang
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China.
authors.forename: Zhen-Hua
authors.initials: ZH
authors.lastname: Zhu
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Buchang Pharma., Xi'an 710000, China.
authors.forename: Bu-Chang
authors.initials: BC
authors.lastname: Zhao
authors.affiliation: Buchang Pharma., Xi'an 710000, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Shanxi University of Chinese Medicine, Xianyang 712000, China.
authors.forename: Zhi-Shu
authors.initials: ZS
authors.lastname: Tang
authors.affiliation: Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing University of Chinese Medicine, Nanjing 210023, China; Jiangsu Key Laboratory for High Technology Research of TCM Formulae, Nanjing University of Chinese Medicine, Nanjing 210023, China. Electronic address: dja@njucm.edu.cnc.
authors.forename: Jin-Ao
authors.initials: JA
authors.lastname: Duan
mesh_terms: D000818:Animals; D001241:Aspirin / Q000737:chemistry / Q000378:metabolism* / Q000652:urine*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005965:Glucuronates / Q000737:chemistry / Q000652:urine; D005966:Glucuronidase / Q000737:chemistry; D020719:Glucuronides / Q000737:chemistry / Q000652:urine; D006626:Hippurates / Q000737:chemistry / Q000652:urine; D008297:Male; D055442:Metabolome / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012459:Salicylates / Q000737:chemistry / Q000652:urine; D020156:Salicylic Acid / Q000737:chemistry / Q000652:urine; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D005965:Glucuronates; D020719:Glucuronides; D006626:Hippurates; D012459:Salicylates; C000693:salicylurate; C049960:1-salicylate glucuronide; D005966:Glucuronidase; D020156:Salicylic Acid; D001241:Aspirin
keywords: Aspirin; Aspirin dosage; Metabolite; Urinary excretion; β-Glucuronidase