Simultaneous Determination of 5-FU and Its Metabolites 5-FUH2 in Human Plasma by UPLC-MS/MS and Appli- cation
- VernacularTitle: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
- Author:
Zhiqiang TONG
1
;
Dongyuan WU
1
;
Juan ZHANG
2
;
Mengfei CHENG
1
;
Shuo ZHANG
1
;
Mei DONG
1
Author Information
1. Dept. of Pharmacy,Harbin Medical University Cancer Hospital,Harbin 150040,China
2. Dept. of Pharmacy,Yantai Yuhuangding Hospital,Shandong Yantai 264000,China
- Publication Type:Journal Article
- Keywords:
UPLC-MS/MS;
5-FU;
5-FUH2;
Plasma
- From:
China Pharmacy
2021;32(4):480-484
- CountryChina
- Language:Chinese
-
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 T 3 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 2>0.990). The quantification limits were 20 ng/mL. RSDs of precision test were all lower than 20%,and relative error 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 simple operation. It can be used for plasma monitoring of 5-FU in patients with advanced colorectal cancer.