Stability of 5-FU in whole blood and a clinical sampling and delivery procedures for TDM

WEN Yongqing; WANG Wenjuan; BAI Yu; LIU Rufeng; MA Xu

doi:10.6039/j.issn.1001-0408.2025.23.14

您当前的位置：

首页 >

文章列表页 >

Stability of 5-FU in whole blood and a clinical sampling and delivery procedures for TDM

更新时间：2025-12-10

- Stability of 5-FU in whole blood and a clinical sampling and delivery procedures for TDM
- ZHONGGUO YAOFANG Vol. 36, Issue 23, Pages: 2963-2968(2025)
- 作者机构：
  
  北京大学肿瘤医院暨北京市肿瘤防治研究所药剂科/恶性肿瘤发病机制及转化研究教育部重点实验室，北京　100142
- 作者简介：
- 基金信息：
- DOI：10.6039/j.issn.1001-0408.2025.23.14
  CLC： R917
- Received：05 August 2025，
  
  Revised：2025-11-21，
  
  Accepted：22 November 2025，
  
  Published：15 December 2025
- 稿件说明：
移动端阅览
温永青,王文娟,白羽,等.5-FU在全血中的稳定性及其TDM临床采样转运流程研究 [J].中国药房,2025,36(23):2963-2968.

WEN Yongqing,WANG Wenjuan,BAI Yu,et al.Stability of 5-FU in whole blood and a clinical sampling and delivery procedures for TDM[J].ZHONGGUO YAOFANG,2025,36(23):2963-2968.
温永青,王文娟,白羽,等.5-FU在全血中的稳定性及其TDM临床采样转运流程研究 [J].中国药房,2025,36(23):2963-2968. DOI： 10.6039/j.issn.1001-0408.2025.23.14.

WEN Yongqing,WANG Wenjuan,BAI Yu,et al.Stability of 5-FU in whole blood and a clinical sampling and delivery procedures for TDM[J].ZHONGGUO YAOFANG,2025,36(23):2963-2968. DOI： 10.6039/j.issn.1001-0408.2025.23.14.

摘要

目的

考察5-氟尿嘧啶（5-FU）在人全血中的稳定性，并以此为依据建立标准化的5-FU治疗药物监测（TDM）临床采样转运流程。

方法

以EDTA抗凝全血样本为基质制备低（200 ng/mL）、高（5 000 ng/mL）浓度的5-FU全血稳定性考察样品（分别设置无稳定剂组与添加1%体积比的稳定剂组）。将全血稳定性考察样品分别置于室温[（25±2）℃]及冷藏（2～8 ℃）条件下，于0、0.5、1、2、4、7、24 h时间点取样处理，样品经涡旋、离心后取上层血浆，采用甲醇沉淀蛋白后，通过高效液相色谱-串联质谱法测定血浆中5-FU的浓度。基于全血稳定性考察结果制定临床采样转运流程。

结果

未添加稳定剂的空白全血样品中5-FU的浓度显著低于添加了稳定剂的样品（

＜0.05），而不同体积（10、25、50 μL）稳定剂对低、高浓度全血稳定性考察样品中5-FU的浓度测定结果均无显著影响（

＞0.05）。未添加稳定剂时，低、高浓度的5-FU全血样品在室温下分别可稳定保存0.5 h与1 h，冷藏条件下分别可稳定保存2 h与7 h；添加1%体积比的稳定剂后，全血样品在室温与冷藏条件下均可稳定保存24 h。所建5-FU TDM临床采样转运流程为：采样后将全血样本在室温（≤0.5 h）或4 ℃（≤2 h）条件下暂存，并于2～8 ℃冷藏转运；送达实验室后立即添加1%体积比的稳定剂，并于24 h内完成离心，所得血浆于－20 ℃保存。

结论

全血中的5-FU在室温条件下稳定性较差，2～8 ℃冷藏时稳定性略有提升但仍会快速降解；添加1%体积比的稳定剂可显著延长其冷藏保存时间。本研究所建5-FU TDM临床采样转运流程，创新性地将稳定剂添加节点设置于样品接收环节（而非采血后立即添加），在保证样品分析质量的前提下，更贴合临床采血的实际操作条件，显著提升了临床实际操作的可行性。

Abstract

OBJECTIVE

To investigate the stability of 5-fluorouracil （5-FU） in human blood and to establish a standardized clinical sampling and delivery procedure for therapeutic drug monitoring （TDM） of 5-FU.

METHODS

The EDTA-anticoagulated whole blood was used as the matrix to prepare stability assessment samples of 5-FU at both low （200 ng/mL） and high （5 000 ng/mL） concentrations （with groups without stabilizer and with 1% volume ratio of stabilizer）. The stability assessment samples were placed under room temperature [（25±2） ℃] and refrigerated （2-8 ℃） conditions， with sampling at 0， 0.5， 1， 2， 4， 7， and 24 h. After vortexing and centrifugation， the upper plasma layer was collected； proteins were precipitated using methanol， and the concentration of 5-FU in plasma was determined by liquid chromatography-tandem mass spectrometry. Based on the whole blood stability results， clinical sampling and delivery procedures were established.

RESULTS

The concentration of 5-FU in blank whole blood samples without stabilizers was significantly lower than that in samples with stabilizers （

＜0.05）. However， varying volumes （10， 25， 50 μL） of stabilizers had no significant effect on the measured concentrations of 5-FU in stability assessment samples with low and high concentrations （

＞0.05）. Without the addition of a stabilizer， low- and high-concentration 5-FU whole blood samples remained stable at room temperature for 0.5 h and 1 h， respectively， and under refrigeration for 2 h and 7 h， respectively. After the addition of a 1% stabilizer， the whole blood samples remained stable for up to 24 h under both room temperature and refrigerated conditions. Based on these findings， the following procedure was established： after collection， whole blood samples could be temporarily stored at room temperature （≤0.5 h） or at 4　℃ （≤2 h）， and transported at 2-8　℃. Upon delivery to the laboratory， a 1% volume ratio of stabilizer must be added immediately， fo

llowed by centrifugation within 24 h. The resulting plasma should be stored at －20 ℃.

CONCLUSIONS

5-FU in whole blood exhibits poor stability at room temperature. Refrigeration at 2-8　℃ slightly improves stability， but degradation still occurs rapidly. Adding a stabilizer at a 1% volume ratio significantly prolongs the refrigerated storage time. The established sampling and transport procedure for 5-FU TDM innovatively introduces the stabilizer addition step at the laboratory sample reception stage （rather than immediately after blood draw）. This approach ensures analytical quality while offering greater adaptability to real-world clinical sampling conditions， significantly improving practical feasibility.

关键词

Keywords

references

GLEWIS S ， MICHAEL M ， GURNEY H ， et al . Feasibility and population exposure of 5-fluorouracil using therapeutic drug monitoring （PREDICT-5FU）：a multicentre clinical trial [J ] . Br J Clin Pharmacol ， 2025 ， 91 （ 7 ）： 1965 - 1974 .

KALDATE R R ， HAREGEWOIN A ， GRIER C E ， et al . Modeling the 5-fluorouracil area under the curve versus dose relationship to develop a pharmacokinetic dosing algorithm for colorectal cancer patients receiving FOLFOX6 [J ] . Oncologist ， 2012 ， 17 （ 3 ）： 296 - 302 .

BEUMER J H ， CHU E ， ALLEGRA C ， et al . Therapeutic drug monitoring in oncology：international association of therapeutic drug monitoring and clinical toxicology re- commendations for 5-fluorouracil therapy [J ] . Clin Pharmacol Ther ， 2019 ， 105 （ 3 ）： 598 - 613 .

佟志强，吴东媛，张娟，等 . UPLC-MS/MS法同时测定人血浆中5-FU及其代谢产物5-FUH 2 的浓度及应用 [J ] . 中国药房， 2021 ， 32 （ 4 ）： 480 - 484 .

涂碎萍，李获，陈岷，等 . LC-MS测定结直肠癌患者血浆中5-氟尿嘧啶浓度 [J ] . 实用药物与临床， 2018 ， 21 （ 9 ）： 1047 - 1051 .

封敬颖，边静，崔毅轩，等 . 中空纤维离心超滤结合高效液相色谱法测定5-氟尿嘧啶的血药浓度 [J ] . 中国医院药学杂志， 2021 ， 41 （ 13 ）： 1305 - 1308 .

PLEBANI M ， SCOTT S ， SIMUNDIC A M ， et al . New insights in preanalytical quality [J ] . Clin Chem Lab Med ， 2025 ， 63 （ 9 ）： 1682 - 1692 .

NYBO M ， CADAMURO J ， CORNES M P ， et al . Sample transportation：an overview [J ] . Diagnosis （Berl）， 2019 ， 6 （ 1 ）： 39 - 43 .

LIPPI G ， VON MEYER A ， CADAMURO J ， et al . PREDICT：a checklist for preventing preanalytical diagnostic errors in clinical trials [J ] . Clin Chem Lab Med ， 2020 ， 58 （ 4 ）： 518 - 526 .

MURPHY R F ， BALIS F M ， POPLACK D G . Stability of 5-fluorouracil in whole blood and plasma [J ] . Clin Chem ， 1987 ， 33 （ 12 ）： 2299 - 2300 .

LEE J J ， BEUMER J H ， CHU E . Therapeutic drug monitoring of 5-fluorouracil [J ] . Cancer Chemother Pharmacol ， 2016 ， 78 （ 3 ）： 447 - 464 .

LI M ， MINDT S ， LÜCK A ， et al . Drug monitoring detects under- and overdosing in patients receiving 5-fluorouracil-containing chemotherapy-results of a prospective，multicenter German observational study [J ] . ESMO Open ， 2023 ， 8 （ 2 ）： 101201 .

张娟，吴东媛，刘铎，等 . 5-FU治疗药物监测现状及其与结直肠癌化疗毒性相关性的研究进展 [J ] . 中国药房， 2018 ， 29 （ 12 ）： 1724 - 1728 .

李然，焦园园，刘颖，等 . 1例卡培他滨致严重不良反应患者的治疗药物监测 [J ] . 中国药物应用与监测， 2015 ， 12 （ 6 ）： 354 - 356 .

QIN W ， WANG X X ， CHEN W Q ， et al . An in vitro approach to simulate the process of 5-fluorouracil degradation with dihydropyrimidine dehydrogenase：the process in accordance to the first-order kinetic reaction [J ] . Xenobio- tica ， 2021 ， 51 （ 1 ）： 24 - 30 .

RADOVANOVIC M ， SCHNEIDER J J ， MARTIN J H ， et al . Comparison between a single- and a multi-point calibration method using LC-MS/MS for measurement of 5-fluorouracil in human plasma [J ] . J Mass Spectrom Adv Clin Lab ， 2024 ， 33 ： 31 - 37 .

GURJAR M ， PRIYAN K A ， ASIA P ， et al . Optimizing cancer patient care with a robust assay for 5-fluorouracil quantification and in vitro stability in human blood for therapeutic drug monitoring [J ] . Pract Lab Med ， 2024 ， 40 ： e00415 .

范先煜，汪硕闻，范国荣 . LC-MS/MS法同时检测人血浆中氟尿嘧啶、尿嘧啶和二氢尿嘧啶的浓度及其临床应用 [J ] . 药学服务与研究， 2021 ， 21 （ 1 ）： 12 - 18 .

ANDO M ， WATANABE N ， SEIKE R ， et al . Quantification of gimeracil，tegafur，and 5-FU in human plasma via LC-MS/MS with a simplified pretreatment using flow-through extraction [J ] . J Chromatogr B Analyt Technol Biomed Life Sci ， 2025 ， 1251 ： 124424 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

No data

Related Author

No data

Related Institution

No data

⁰