Establishment and clinical application of a method for the determination of tacrolimus concentration in human whole blood

LIU Simin; CHU Yamin; HU Yahui; LONG Guangfeng; CHEN Feng; ZHANG Yuanyuan

doi:10.6039/j.issn.1001-0408.2026.09.12

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Establishment and clinical application of a method for the determination of tacrolimus concentration in human whole blood

更新时间：2026-05-14

- Establishment and clinical application of a method for the determination of tacrolimus concentration in human whole blood
- ZHONGGUO YAOFANG Vol. 37, Issue 9, Pages: 1180-1184(2026)
- 作者机构：
  
  1.南京医科大学附属儿童医院药学部，南京　210008
  2.中国药科大学基础医学与临床药学学院，南京　210009
  3.南京医科大学附属儿童医院检验科，南京　210008
- 作者简介：
- 基金信息：
- DOI：10.6039/j.issn.1001-0408.2026.09.12
  CLC： R969.1
- Received：29 December 2025，
  
  Revised：2026-03-31，
  
  Accepted：31 March 2026，
  
  Published：15 May 2026
- 稿件说明：
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刘思敏,储雅旻,胡雅慧,等.人全血中他克莫司浓度测定方法的建立及临床应用 [J].中国药房,2026,37(09):1180-1184.

LIU Simin,CHU Yamin,HU Yahui,et al.Establishment and clinical application of a method for the determination of tacrolimus concentration in human whole blood[J].ZHONGGUO YAOFANG,2026,37(09):1180-1184.
刘思敏,储雅旻,胡雅慧,等.人全血中他克莫司浓度测定方法的建立及临床应用 [J].中国药房,2026,37(09):1180-1184. DOI： 10.6039/j.issn.1001-0408.2026.09.12.

LIU Simin,CHU Yamin,HU Yahui,et al.Establishment and clinical application of a method for the determination of tacrolimus concentration in human whole blood[J].ZHONGGUO YAOFANG,2026,37(09):1180-1184. DOI： 10.6039/j.issn.1001-0408.2026.09.12.

摘要

目的

建立测定人全血中他克莫司（TAC）浓度的方法，并将其应用于临床。

方法

采用甲醇沉淀蛋白的方法处理全血样品。采用液相色谱-串联质谱（LC-MS/MS）法进行测定，以子囊霉素为内标，使用Kinetex F5 100Å色谱柱，以含0.2 mmol/L甲酸的0.1 mmol/L醋酸铵溶液和甲醇为流动相进行梯度洗脱，流速为0.4 mL/min，进样体积为5 μL；采用电喷雾离子源在正离子模式下进行多反应监测（TAC

821.6→768.6，子囊霉素

809.4→756.1）。以2025年9月1－30日于南京医科大学附属儿童医院接受TAC治疗的83名患儿的86例全血样本为研究对象，采用上述方法测定全血中TAC浓度，并通过Spearman相关性分析、Bland-Altman分析、Passing-Bablok回归分析评估上述方法与传统酶放大免疫分析（EMIT）法的相关性和一致性。

结果

TAC的线性范围为0.5～100 ng/mL，准确度、精密度、提取回收率、基质效应、稳定性试验的考察结果均符合相关要求。临床应用结果显示，LC-MS/MS法和EMIT法测定的患儿全血中TAC浓度的中位值分别为4.4、4.0 ng/mL，且两种方法具有强相关性（相关系数为0.848 1）与良好的一致性（平均相对偏差为6.5%）。

结论

本研究成功建立了测定人全血中TAC浓度的LC-MS/MS法，且该法与EMIT法具有强相关性和良好的一致性，可用于临床治疗药物监测。

Abstract

OBJECTIVE

To develop a method for the determination of tacrolimus （TAC） concentration in human whole blood and to apply it in clinical therapeutic drug monitoring.

METHODS

Whole blood samples were processed by protein precipitation with methanol. The determination was performed using liquid chromatography-tandem mass spectrometry （LC-MS/MS）， with ascomycin serving as the internal standard. Chromatographic separation was carried out on a Kinetex F5 100Å column with a mobile phase consisting of 0.1 mmol/L ammonium acetate containing 0.2 mmol/L formic acid and methanol. Gradient elution was performed at a flow rate of 0.4 mL/min. The injection volume was 5 μL. Detection was conducted using multiple reaction monitoring （

821.6→768.6 for TAC；

809.4→756.1 for ascomycin） with an electrospray ionization source in positive ion mode. The study focused on 86 whole blood samples collected from 83 pedi

atric patients who received TAC therapy at Children’s Hospital of Nanjing Medical University from September 1 to 30， 2025. The aforementioned method was employed to measure the TAC concentration in the whole blood samples. The correlation and agreement between the aforementioned method and the traditional enzyme multiplied immunoassay technique （EMIT） were evaluated through Spearman correlation analysis， Bland-Altman analysis， and Passing-Bablok regression analysis.

RESULTS

The linear range of TAC was 0.5-100 ng/mL； the evaluation results for accuracy， precision， extraction recovery， matrix effect， and stability tests all met the relevant requirements. Clinical application results showed that the median concentration of TAC in pediatric whole blood measured by LC-MS/MS and EMIT methods were 4.4 and 4.0 ng/mL， respectively. Moreover， the two methods exhibited a strong correlation （correlation coefficient of 0.848 1） and good agreement （average relative deviation of 6.5%）.

CONCLUSIONS

A reliable LC-MS/MS method for the determination of TAC concentration in human whole blood is successfully established. This method demonstrates strong correlation and good agreement with the EMIT method， making it suitable for clinical therapeutic drug monitoring.

关键词

Keywords

references

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