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咖啡因及代谢产物的尿药浓度测定方法建立及临床应用
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| 篇名： | 咖啡因及代谢产物的尿药浓度测定方法建立及临床应用 |
| TITLE： | Method establishment and clinical practice for concentration determination of caffeine and its metabolites in urine |
| 摘要： | 目的 建立同时测定尿液中咖啡因及3种代谢产物茶碱、副黄嘌呤和可可碱浓度的方法并应用于临床。方法以咖啡因-13C3-d3为内标，尿液样本经乙腈沉淀蛋白后，采用高效液相色谱-串联质谱（HPLC-MS/MS）技术测定咖啡因及3种代谢产物的浓度。以WatersACQUITYUPLC®BEHHILIC为色谱柱，60mmol/L乙酸铵溶液（A）-乙腈（B）为流动相进行梯度洗脱，流速为0.5mL/min，柱温为38℃，进样量为2μL。采用电喷雾离子源，以多反应监测模式进行正离子扫描，用于定量分析的离子对分别为m/z195.1→110.0（咖啡因）、m/z181.1→124.0（茶碱）、m/z181.1→124.0（副黄嘌呤）、m/z181.1→138.0（可可碱）、m/z198.1→140.1（内标）。采用上述方法测定19例早产儿呼吸暂停（AOP）患儿尿液中咖啡因及3种代谢产物的浓度。结果咖啡因、茶碱、副黄嘌呤、可可碱检测质量浓度的线性范围分别为0.200～200、0.050～50.0、0.050～50.0、0.100～100μg/mL（r均大于0.990），定量下限分别为0.200、0.050、0.050、0.100μg/mL；日内、日间精密度的RSD均不高于10.37%，基质因子为85.68%～109.90%，提取回收率为93.53%～109.40%（RSD均小于15%），稳定性试验的RSD均小于15%。19例AOP患儿尿液中咖啡因及3种代谢产物的质量浓度分别为（27.346±7.951）、（0.351±0.223）、（0.428±0.395）、（0.472±0.374）μg/mL。结论所建HPLC-MS/MS法操作简单、灵敏度高，可用于AOP患儿尿液中咖啡因及3种代谢产物浓度的测定。 |
| ABSTRACT： | OBJECTIVE To establish a method for concentration determination of caffeine and its three metabolites， theophylline， paraxanthine and theobromine in urine， and apply it in clinical practice. METHODS Using caffeine-13C3-d3 as internal standard （IS）， and the urine samples were protein precipitated with acetonitrile； HPLC-MS/MS method was adopted to determine the concentrations of caffeine and its three metabolites. The determination was performed on Waters ACQUITY UPLC® BEH HILIC column with mobile phase consisting of 60 mmol/L ammonium acetate （A）-acetonitrile （B） （gradient elution） at the flow rate of 0.5 mL/min. The column temperature was set at 38 ℃ ， and the sample size was 2 μL. The electrospray ionization detection was operated in a positive mode by multiple reaction monitoring. The detection ions for quantitative analysis were m/z 195.1→110.0 for caffeine， m/z 181.1→124.0 for theophylline， m/z 181.1→124.0 for paraxanthine， m/z 181.1→138.0 for theobromine， and m/z 198.1→ 140.1 for IS. The above method was used to determine the concentrations of caffeine and its three metabolites in the urine of 19 infants with apnea of prematurity （AOP）. RESULTS The linear ranges of mass concentration of caffeine， theophylline， paraxanthin and theobromine were 0.200-200， 0.050-50.0，0.050 0-50.0， and 0.100-100 μg/mL， respectively. The lower limits of quantification were 0.200， 0.050， 0.050 and 0.100 μg/mL （r＞0.990）， respectively. RSDs of intra-day and intra- day precision were not above 10.37%， and matrix factors were 85.68%-109.90%； extraction recoveries were 93.53%-109.40% （RSD≤15%）， and RSDs of stability tests were all lower than 15%. The concentrations of caffeine and its three metabolites in the urine of 19 cases were （27.346±7.951）， （0.351±0.223）， （0.428±0.395） and （0.472±0.374） μg/mL， respectively. CONCLUSIONS The established HPLC-MS/MS method is simple， sensitive and can be used for the determination of caffeine and its three metabolites in urine samples of AOP. |
| 期刊： | 2023年第34卷第18期 |
| 作者： | 陈相龙；赵杨；黄琼叶；徐铭卿；李悦；陆超；孙鲁宁；王永庆 |
| AUTHORS： | CHEN Xianglong，ZHAO Yang，HUANG Qiongye，XU Mingqing，LI Yue，LU Chao，SUN Luning，WANG Yongqing |
| 关键字： | 咖啡因；代谢产物；早产儿呼吸暂停；尿药浓度；高效液相色谱-串联质谱法 |
| KEYWORDS： | caffeine； metabolites； apnea of prematurity； urinary drug concentration； HPLC-MS/MS |
| 阅读数： | 30 次 |
| 本月下载数： | 4 次 |
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