Author(s): Irina. VM, Praseetha. K, Kathirvel. S, Raja Rajeswari. K


DOI: 10.52711/0974-360X.2024.00329   

Address: Irina. VM1, Praseetha. K1, Kathirvel. S1, Raja Rajeswari. K2*
1National College of Pharmacy, KMCT Group of Institutions, Manassery, Kozhikode - 673602, Kerala, India.
2Department of Pharmaceutical Analysis, Sri Sivani College of Pharmacy, Srikakulam - 532402, Andhra Pradesh, India.
*Corresponding Author

Published In:   Volume - 17,      Issue - 5,     Year - 2024

Plasma slowly loses teriflunomide, an oral immunomodulator used once in a day and majorly it is approved for treating of relapsing-remitting multiple sclerosis. It is not possible to measure the plasma concentrations of teriflunomide at locations without the tools necessary to prepare the processed blood samples. Plasma monitoring can be replaced with DBS technique, dried blood spot sampling, which allows for easier sample storage and travel. A few drops of blood are extracted from the rat tail using a lancet and applied to specially made absorbent filter paper. A UPLC test technique for the measurement of teriflunomide in DBS is created and validated for specificity, accuracy, selectivity, stability and repeatability using blood samples from pharmacokinetic studies. Process efficiency was necessary, method was selective and specific regarding endogenous chemicals, and there was no matrix effect. All concentrations were evaluated for accuracy as well as precision for intra-day and also inter-day analysis. The amount of blood deposited and the punch position within the spot had no bearing on the detection of teriflunomide in the DBS assay, however the hematocrit level had a negligible but tolerable influence on measurement precision. Teriflunomide has a minimum stability of three months at room temperature. With an average ratio of blood to plasma, an association between DBS concentration and plasma concentrations is seen. A straightforward and useful technique for keeping track of teriflunomide concentrations is DBS sampling. The technique has been expanded to the in-vivo determination of teriflunomide in male albino rats and is fully verified in accordance with ICH criteria.

Cite this article:
Irina. VM, Praseetha. K, Kathirvel. S, Raja Rajeswari. K. Ultra-Performance Liquid Chromatography Method for Bioanalysis of Teriflunomide using Dried Blood Spot Methodology. Research Journal of Pharmacy and Technology. 2024; 17(5):2077-6. doi: 10.52711/0974-360X.2024.00329

Irina. VM, Praseetha. K, Kathirvel. S, Raja Rajeswari. K. Ultra-Performance Liquid Chromatography Method for Bioanalysis of Teriflunomide using Dried Blood Spot Methodology. Research Journal of Pharmacy and Technology. 2024; 17(5):2077-6. doi: 10.52711/0974-360X.2024.00329   Available on:

1.    Lakshmy R. Analysis of the use of dried blood spot measurements in disease screening. Journal of Diabetes Science and Technology. 2008; 2: 242–243.
2.    Parker SP. Cubitt WD. The use of the dried blood spot sample in epidemiological studies. Journal of Clinical Pathology. 1999; 52:633–639.
3.    Sharma A, Jaiswal S, Shukla M, Lal J. Dried blood spots: concepts, present status, and future perspectives in bioanalysis. Drug Test Anal. 2014 May;6(5):399-414. doi: 10.1002/dta.1646.
4.    Edelbroek PM. Van der Heijden J. Stolk LM. Dried blood spot methods in therapeutic drug monitoring: methods, assays, and pitfalls. Therapeutic Drug Monitoring. 2009; 31: 327–336.doi: 10.1097/FTD.0b013e31819e91ce.
5.    Schmidt A. Schwind B. Gillich M. Brune K. Hinz B. Simultaneous determination of leflunomide and its active metabolite, A77 1726, in human plasma by high-performance liquid chromatography. Biomedical Chromatography. 2003; 17: 276-281.doi: 10.1002/bmc.244.
6.    Parekh JM, Vaghela RN, Sutariya DK, Sanyal M, Yadav M, Shrivastav PS. Chromatographic separation and sensitive determination of teriflunomide, an active metabolite of leflunomide in human plasma by liquid chromatography-tandem mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Aug 15; 878(24): 2217-2225. doi: 10.1016/j.jchromb.2010.06.028.
7.    Nukendra Prasad N. Venkata Nadh R. Srinivasu N. Quality-by-design-based development and validation of a stability-indicating UPLC method for quantification of teriflunomide in the presence of degradation products and its application to invitro dissolution. Journal of Liquid Chromatography and Related Technologies. 2017; 40(10): 517-527.doi: 10.1080/10826076.2017.1330211
8.    Arun Maruti K. Pranali Prakash P. Kshitija Maruti B. Analytical Method Development and Validation of Teriflunomide Active Pharmaceutical Ingredient by RP-UHPLC. Asian Journal of Pharmaceutical Analysis. 2022; 12(3): 166-172. DOI: 10.52711/2231-5675.2022.00028
9.    Filali-Ansary A. Lunven C. Turpault S. Beyer YJ.Oʼ Brien A. Delfolie al. Dried Blood Spot Methodology in Combination With Liquid Chromatography/Tandem Mass Spectrometry Facilitates the Monitoring of Teriflunomide. Therapeutic Drug Monitoring. 2016; 38(4): 471-482. doi: 10.1097/FTD.0000000000000302.
10.    Eswarudu MM. Chinna Eswaraiah M. Prasanna Kumar K. Sudhakar K. Ultra Performance Liquid Chromatography (UPLC): A Preeminent Technique in Pharmaceutical Analysis. Research Journal of Pharmacy and Technology. 2012; 5(12): 1484-1489.
11.    Food and Drug Administration. Guidance for Industry: Bioanalytical Method Validation Center for Drug Evaluation and Research. Silver Springs, MD: FDA; 2001.
12.    ICH guideline M10 on bioanalytical method validation and study sample analysis. EMA/CHMP/ICH/172948/2019.
13.    Holub M. Tuschl K. Ratschmann R. et al. Influence of hematocrit and localisation of punch in dried blood spots on levels of amino acids and acylcarnitines measured by tandem mass spectrometry. Clinica Chimica Acta. 2006; 373: 27–31.
14.    Adam BW. Alexander JR. Smith SJ. et al. Recoveries of phenylalanine from two sets of dried-blood-spot reference materials: prediction from hematocrit, spot volume, and paper matrix. Clinical Chemistry. 2000; 46: 126–128.
15.    Denniff P. Spooner N. The effect of hematocrit on assay bias when using DBS samples for the quantitative bioanalysis of drugs. Bioanalysis. 2010; 2: 1385–1395.
16.    Evans CA. Bowen CL. Filali-Ansari A. Dried blood spots: challenges. In Microsampling in Pharmaceutical Bioanalysis. London, United Kingdom: Future Science Limited. 2013; 30–46. DOI:10.4155/ebo-13.441
17.    Li W.Tse FL. Dried blood spot sampling in combination with LC-MS/MS for quantitative analysis of small molecules. Biomedical Chromatography. 2010; 24: 49–65.
18.    Hon GM. Hassan MS.Van Rensburg SJ. et al. The Haematological profile of patients with multiple sclerosis. Open Journal of Modern Neurosurgery. 2012; 2: 36–44. DOI: 10.4236/ojmn.2012.23008

Recomonded Articles:

Research Journal of Pharmacy and Technology (RJPT) is an international, peer-reviewed, multidisciplinary journal.... Read more >>>

RNI: CHHENG00387/33/1/2008-TC                     
DOI: 10.5958/0974-360X 

56th percentile
Powered by  Scopus

SCImago Journal & Country Rank

Recent Articles


Not Available