Author(s): Francis Micheal, Balamurali MM, Mohanlal Sayana, Rajendra Prasad M

Email(s): ,

DOI: 10.5958/0974-360X.2020.00381.9   

Address: Francis Micheal1, Balamurali MM1, Mohanlal Sayana2, Rajendra Prasad M3
1Department of Chemistry, School of Advanced Sciences, VIT University, Vellore, Tamilnadu, 632014, India.
2Department of Pharmacokinetic and Drug Metabolism, Strides Arcolab Limited, Bangalore, Karnataka, 560076, India.
3Jeevan Scientific Technology Limited, Hyderabad, Telangana - 500008, India.
*Corresponding Author

Published In:   Volume - 13,      Issue - 5,     Year - 2020

Background: Duloxetine (DUX) is a potent selective serotonin and norepinephrine reuptake inhibitor used to treat depression and anxiety in human. The concentration of DUX in biological matrix is highly variable; hence it is necessary to have a highly sensitive and selective analytical method to measure the anticipated range in the plasma. Thus, there is a necessity to develop a sensitive and practical analytical method for the determination of DUX in human plasma. Objective: The objective of the study is to estimate DUX in human plasma using high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) and its application to clinical pharmacokinetic study by assessing multiple bioequivalence approaches. Method: The analyte was extracted by simple liquid-liquid extraction technique using n-hexane as an extraction solvent. The chromatographic separation was carried out by using Luna® 5 µm C8 (2) 100 Å, LC Column 100 x 4.6 mm with the mobile phase consisting of Milli q water (0.05% formic acid and 0.1% ammonium trifluroacetate solution) and Methanol (23:77%v/v) at 0.5mL/min flow rate. The MRM transitions of 298.00 ? 154.00 and 256.00 ? 148.00 were used for the quantification of DUX and Atomoxetine (internal standard) separately. Results: The method was validated over the concentration range of 0.100-100.000 ng/mL, the coefficient of determination was = 0.9967. The sensitivity of the method was 0.100ng/mL with the accuracy and precision of 108% and 5.12% respectively. All validation parameters are found to be within the acceptable limits as per the USFDA bioanalytical method validation guidelines. Conclusion: We have developed and validated a highly sensitive analytical method for the determination of DUX in human plasma using high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The developed method was successfully applied for analyzing the samples of clinical pharmacokinetic study conducted in healthy human adult subjects and the pharmacokinetic data was analyzed by using average, population and individual bioequivalence approaches. The method was very rugged and is capable of continuously analyzing the clinical pharmacokinetics study samples without any run failures.

Cite this article:
Francis Micheal, Balamurali MM, Mohanlal Sayana, Rajendra Prasad M. Development and Validation of Highly Sensitive HPLC-Ms/Ms Method for the Determination of Duloxetine in Human Plasma and its Application to Clinical Pharmacokinetic Study by Assessing Multiple Bioequivalence Approaches. Research J. Pharm. and Tech 2020; 13(5):2117-2124. doi: 10.5958/0974-360X.2020.00381.9

1.    Lantz, RJ.; Gillespie, TA.; Rash, TJ.; Kuo, F.; Skinner, M.; Kuan, HY.; Knadler MP. Metabolism, excretion, and pharmacokinetics of duloxetine in healthy human subjects. Drug Metab Dispos., 2003; 31 (9): 1142-50.
2.    Cymbalta (duloxetine hydrochloride) Delayed-Release Capsules for Oral Use, The prescribing information, 2016; Eli Lilly and Company Indianapolis, IN 46285, USA
3.    Cymbalta 60 mg hard gastro-resistant capsules, SMPC, 2017; Eli Lilly and Company Limited Lilly House, Priestley Road, Basingstoke, Hampshire, RG24 9NL.
4.    Li, H.; Li, T.; Li, Y.; Shen, Y. Pharmacokinetics and Safety of Duloxetine Enteric-coated Tablets in Chinese Healthy Volunteers, A Randomized, Open-label, Single- and Multiple-dose Study. Clin Psychopharmacol Neurosci., 2013; Apr;11 (1): 28-33.
5.    Zhao, RK.; Cheng, G.; Tang, J.; Song, J.; Peng, WX. Pharmacokinetics of duloxetine hydrochloride enteric-coated tablets in healthy Chinese volunteers: a randomized, open-label, single- and multiple-dose study. Clin Ther., 2009; 31 (5): 1022-36.
6.    Tiwari, G.; Tiwari R. Bioanalytical method validation: an updated review. Pharmaceutical Methods., 2010; 1 (1): 25–38.
7.    Waldschmitt, C.; Vogel, F.; Maurer, C.; Hiemke, C. Measurement of duloxetine in blood using high-performance liquid chromatography with spectrophotometric detection and column switching. Ther Drug Monit., 2007; 29 (6):767-72.
8.    Tatar Ulu S. Determination and validation of duloxetine hydrochloride in capsules by HPLC with pre-column derivatization and fluorescence detection. J Chromatogr Sci., 2012, Jul; 50(6): 494-8.
9.    Kaza, M., Gilant, E., Filist, M., Szlaska, I., Pawiński, T., Rudzki, PJ. Determination of duloxetine in human plasma with proven lack of influence of the major metabolite 4-hydroxyduloxetine. Clin Biochem., 2014; Sep; 47 (13-14):1313-5.
10.    Johnson, JT.; Oldham, SW.; Lantz, RJ.; DeLong, AF. High performance liquid chromatographic method for the determination of duloxetine and desmethyl duloxetine in human plasma. Journal of Liquid Chromatography and Related Technologies., 2006; 19:10, 1631-1641, DOI: 10.1080/10826079608005497.
11.    Mercolini, L.; Mandrioli, R.; Cazzolla, R.; Amore, M.; Raggi, MA. HPLC analysis of the novel antidepressant duloxetine in human plasma after an original solid-phase extraction procedure. J Chromatogr B Analyt Technol Biomed Life Sci., 2007; 856 (1-2):81-7.
12.    Thejaswini J. C.; Gurupadayya, BM.; Ranjith Kumar, K. Quantitative determination of duloxetine HCL in human plasma by GC-FID method. International Journal of Pharmacy and Pharmaceutical Sciences, 2013; 5 (2).
13.    Senthamil Selvan, P.; Gowda, KV.; Mandal, U.; Sam Solomon, WD.; Pal TK. Determination of duloxetine in human plasma by liquid chromatography with atmospheric pressure ionization-tandem mass spectrometry and its application to pharmacokinetic study. J Chromatogr B Analyt Technol Biomed Life Sci., 2007; 858 (1-2):269-75.
14.    Udhaya, R.; Naidu, PY.; Reddy, AR.; Vidya Sagar, TV. Validation of an UPLC-MS/MS method for simultaneous Quantification of two Selective Serotonin and nor Epinephrine Reuptake Inhibitors and one Selective Serotonin Reuptake Inhibitor in human plasma. Der Pharma Chemica., 2012; 4 (3): 1164-1173
15.    Reddy, DC.; Bapuji, AT.; Rao, VS., Himabindu,V., Raju, DR., Syedba, S.,   Ravikiran, HLV. Development and Validation of a LC/MS/MS Method for the Determination of Duloxetine in Human Plasma and its Application to Pharmacokinetic Study. E-Journal of Chemistry., 2012;   9 (2), 899-911. ISSN: 0973-4945; CODEN ECJHAO.
16.    Ma, N.; Zhang, BK.; Li, HD.; Chen, BM.; Xu, P.; Wang, F.; Zhu, RH.; Feng, S.; Xiang, DX.; Zhu, YG. Determination on of duloxetine in human plasma via LC/MS and  subsequent  application to a pharmacokinetic study in healthy Chinese volunteers. Clin Chim Acta., 2007; 380 (1-2):100-5.
17.    Tianmei, S.; Knadler MP.; Lim,MT.; Yeo, KP.; Teng, L.; Liang, S.; Pan, AX.; Lobo, ED. Pharmacokinetics and tolerability of duloxetine following oral administration to healthy Chinese subjects. Clin Pharmacokinet., 2007; 46 (9):767-75.
18.    Veeragoni1, A.; Sindgi, V.; Satla, S.Validated LC- MS Bioanalytical Method for the Estimation of Duloxetine Hydrochloride in Human Plasma. Der Pharmacia Lettre., 2016; 8 (8): 355-360.
19.    Miller, V. Ask the Experts: Automation: Part II. Bioanalysis., 2013; 5 (17), 2085–2094.
20.    Guidance for industry: bioanalytical method validation. Draft Guidance, U. S. Department of Health and Human Services, Food and Drug Administration, Centre for Drug Evaluation and Research (CDER), Centre for Biologics Evaluation and Research (CBER): May 2018 ( ) (Accessed Aug 09, 2019).

Recomonded Articles:

Author(s): A. S. K. Sankar, B. Datchayani, N. Balakumaran, Mohammed Rilwan, R. Subaranjani

DOI: 10.5958/0974-360X.2017.00047.6         Access: Open Access Read More

Author(s): Sonte Sushmitha, Sedimbi Revathi Priyanka, L. Mohan Krishna, M. Srinavasa Murthy

DOI: Not Available         Access: Open Access Read More

Author(s): Rekha Rajendran, R Hemachander, T Ezhilarasan, C Keerthana, DL Saroja, KV Saichand, Mohamed Gasim Abdullah

DOI: Not Available         Access: Open Access Read More

Author(s): Vandana Gautam, Dhriti Kapoor, Saroj Arora, Renu Bhardwaj*

DOI: 10.5958/0974-360X.2016.00166.9         Access: Open Access Read More

Author(s): Manju Rawat, SJ Daharwal, Deependra Singh

DOI:         Access: Open Access Read More

Author(s): Mukta D. Naykode, Durgacharan A. Bhagwat, Swapnil D. Jadhav, Harinath N. More

DOI: 10.5958/0974-360X.2017.00133.0         Access: Open Access Read More

Author(s): Mayanka Singh, Manoj Charde, Rajesh Shukla, Rita M. Charde

DOI: Not Available         Access: Open Access Read More

Author(s): Y. Prem Kumar, K. Vinod Kumar, V. Sai Kishore

DOI: Not Available         Access: Open Access Read More

Author(s): Anket Sharma, Vinod Kumar, Parminder Kaur, Ashwani Kumar Thukral, Renu Bhardwaj

DOI: 10.5958/0974-360X.2015.00299.1         Access: Open Access Read More

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