Establishment and clinical application of a method for the determination of tacrolimus concentration in human whole blood
- VernacularTitle:人全血中他克莫司浓度测定方法的建立及临床应用
- Author:
Simin LIU
1
;
Yamin CHU
1
;
Yahui HU
2
;
Guangfeng LONG
3
;
Feng CHEN
2
;
Yuanyuan ZHANG
2
Author Information
1. Dept. of Pharmacy，Children’s Hospital of Nanjing Medical University，Nanjing 210008，China;School of Basic Medicine and Clinical Pharmacy，China Pharmaceutical University，Nanjing 210009，China
2. Dept. of Pharmacy，Children’s Hospital of Nanjing Medical University，Nanjing 210008，China
3. Dept. of Clinical Laboratory，Children’s Hospital of Nanjing Medical University，Nanjing 210008，China
- Publication Type:Journal Article
- Keywords:
tacrolimus;
LC-MS/MS method;
EMIT method;
whole blood;
therapeutic drug monitoring
- From:
China Pharmacy
2026;37(9):1180-1184
- CountryChina
- Language:Chinese
-
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 （ m / z 821.6→768.6 for TAC； m / z 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.
