Page 106 - 《中国药房》2021年第4期

UPLC-MS/MS 法同时测定人血浆中 5-FU 及其代谢产物 5-FUH2
的浓度及应用 Δ

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佟志强 ，吴东媛 ，张 娟 ，程梦飞 ，张 硕 ，董 梅 （1.哈尔滨医科大学附属肿瘤医院药学部，哈尔滨
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150040；2.烟台毓璜顶医院药学部，山东 烟台 264000）
中图分类号 R917；R735.3 文献标志码 A 文章编号 1001-0408（2021）04-0480-05
DOI 10.6039/j.issn.1001-0408.2021.04.15
摘 要 目的：建立同时测定人血浆中5-氟尿嘧啶（5-FU）及其代谢产物5-氟-5，6-二氢尿嘧啶（5-FUH2 ）浓度的方法，并应用于临
床。方法：血浆样品经乙酸乙酯二次萃取处理后，以5-溴尿嘧啶（5-Bru）为内标，采用超高效液相色谱-串联质谱技术（UPLC-MS/
MS）测定。色谱柱为Acquity UPLC HSS T3，流动相为甲醇-水（30 ∶ 70，V/V），流速为0.3 mL/min，柱温为20 ℃，进样量为5 μL。采
用电喷雾离子源，以多反应监测进行负离子扫描，毛细管电压为 1.5 kV，锥孔电压为 20 V，脱溶剂温度为 450 ℃，脱溶剂气流为
850 L/h，锥孔气流速为50 L/h；用于定量分析的离子对分别为m/z 129.00→41.90（5-FU）、m/z 130.87→82.92（5-FUH2 ）、m/z 189.00→
42.10（5-Bru）。选择2020年8－10月哈尔滨医科大学附属肿瘤医院接受5-FU持续静脉滴注46 h为基础化疗方案治疗的晚期结直
肠癌患者10例，测定其持续静脉滴注18～30 h时的5-FU稳态血药浓度及5-FUH2血药浓度，计算5-FU药-时曲线下面积（AUC）和
5-FUH2/5-FU浓度比值。结果：5-FU、5-FUH2检测质量浓度的线性范围均为20～1 000 ng/mL（R 均大于0.990），定量下限均为20
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ng/mL；精密度试验的RSD均小于20％，相对误差在±10％范围内；提取回收率、基质效应均不影响待测物的测定。10例晚期结直
肠癌患者中，5-FU 稳态血药浓度为 180.04～622.83 ng/mL，AUC 为 8.28～28.65 mg·h/L；5-FUH2 血药浓度为 336.48～948.43
ng/mL；5-FUH2/5-FU浓度比值为0.93～4.21；10例患者的5-FU AUC存在约3～4倍的个体差异。结论：所建方法精密度、准确性均
较好，灵敏度较高且操作简便，可用于晚期结直肠癌患者体内5-FU血药浓度的监测。
关键词 超高效液相色谱-串联质谱法；5-氟尿嘧啶；5-氟-5，6-二氢尿嘧啶；血药浓度

Simultaneous Determination of 5-FU and Its Metabolites 5-FUH2 in Human Plasma by UPLC-MS/MS and
Application
TONG Zhiqiang ，WU Dongyuan ，ZHANG Juan ，CHENG Mengfei ，ZHANG Shuo ，DONG Mei（1. Dept. of
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Pharmacy，Harbin Medical University Cancer Hospital，Harbin 150040，China；2. Dept. of Pharmacy，Yantai
Yuhuangding Hospital，Shandong Yantai 264000，China）
ABSTRACT OBJECTIVE：To establish the method for simultaneous determination of 5-fluorouracil（5-FU）and its metabolites
5-fluoro-5，6-dihydrouracil （5-FUH2 ） in human plasma，and apply it in the clinic. METHODS：After plasma samples were
processed twice by ethyl acetate，UPLC-MS/MS method was adopted using 5-bromouracil （5-Bru） as internal standard. The
determination was performed on Acquity UPLC HSS T3 column with mobile phase consisted of methanol-water（30 ∶ 70，V/V）at
the flow rate of 0.3 mL/min. The column temperature was 20 ℃，and sample size was 5 μL. An electrospray ion source was used
to carry out negative ion scanning with multiple reaction monitoring. The capillary voltage was 1.5 kV；the taper hole voltage was
20 V；the desolvent temperature was 450 ℃；the desolvent air flow was 850 L/h；the cone hole gas velocity was 50 L/h. The ion
transitions for quantitative analysis were m/z 129.00→41.90（5-FU），m/z 130.87→82.92（5-FUH2 ），m/z 189.00→42.10（5-Bru），
respectively. From Aug. to Oct. 2020，10 patients with advanced colorectal cancer were treated with continuous intravenous drip of
5-FU for 46 hours were collected from Harbin Medicinal University Cancer Hospital. Steady-state plasma concentration of 5-FU and
plasma concentration of 5-FUH2 were determined at 18-30 h of continuous intravenous drip. The area under the curve（AUC）for
5-FU and concentration ratio of 5-FUH2/5-FU were calculated. RESULTS：The linear range of 5-FU and 5-FUH2 were 20 to 1 000
μg/mL（R ＞0.990）. The quantification limits were 20 ng/mL. RSDs of precision test were all lower than 20％，and relative error
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ranged ±10％. The extraction recovery and matrix effects didn’t affect the determination of substance to be measured. Among 10
patients with advanced colorectal cancer，the steady-state concentration of 5-FU were 180.04-622.83 ng/mL，and AUC of 5-FU
ranged from 8.28 to 28.65 mg·h/L. The concentration of 5-FUH2 ranged 336.48-948.43 ng/mL，and concentration ratio of 5-FUH2/
5-FU ranged 0.93-4.21. AUC of 5-FU in 10 patients had about 3-4 fold individual differences. CONCLUSIONS：The established
method has good precision and accuracy，high sensitivity，and
Δ 基金项目：黑龙江省卫生计生委科研课题（No.2016-095）
simple operation. It can be used for plasma monitoring of
＊硕士研究生。研究方向：抗肿瘤药物治疗药物监测。电话：
5-FU in patients with advanced colorectal cancer.
0451-86298698。E-mail：1078408734@qq.com
KEYWORDS UPLC-MS/MS； 5-FU； 5-FUH2； Plasma
# 通信作者：主任药师，硕士生导师，博士。研究方向：医院药
concentration
学。电话：0451-86298880。E-mail：13804567370@163.com
·480 · China Pharmacy 2021 Vol. 32 No. 4 中国药房 2021年第32卷第4期