International Journal of Chemical and Biochemical Sciences (ISSN 2226-9614)[/vc_column_text][/vc_column][/vc_row]
VOLUME 25(19) (2024)
High throughput LC-MS/MS method for the quantitation of baclofen in human plasma by solid phase extraction using 96 well plate format
- Bhanu Prakash*1, A. Krishnamanjari Pawar2, B. Geetha Madhuri3,
1College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, Andhra Pradesh, India
2College of Pharmaceutical Sciences, Andhra University, Visakhapatnam, Andhra Pradesh, India
3QPS Bioserve India Pvt. Ltd, IDA Balanagar, Hyderabad, India
Abstract
Oral baclofen is indicated for the treatment of spasticity resulting from multiple sclerosis and is very useful for the relief of flexor spasms and concomitant pain, muscular rigidity, and clonus. It can also be used to treat patients with spinal cord injuries and other spinal cord diseases. In present work, a sensitive and high throughput LC-MS/MS method was developed and validated to estimate Baclofen in Human plasma. The developed LC-MS/MS method was sensitive enough to detect Baclofen in human plasma with a high degree of accuracy and precision, using Baclofen-d4 as an internal standard. Using BDS Hypersil C8 5µm, 100 X 4.6mm column and a mobile phase consisting of 2mM Ammonium formate with 0.02% v/v formic acid: Acetonitrile (85:15 v/v) was used to achieve best chromatographic separation. A flow rate of 1 mL/min was used to optimize the run time of 3.2 min. Plasma sample extraction was performed by Solid Phase extraction using 96 well plates. The developed bioanalytical method was validated successfully. This high throughput bioanalytical method developed and validated can be applied to routine sample analysis. this method can accommodate 80 study samples in single run excluding the CC and QC samples. Since the method uses 96 well plate formate it gives flexibility in assigning the CC, QC, and study samples in fixed well before the initiation of the process which in turn avoids the risk of sample mix up during processing. The method validation results were found to be within the acceptance range. It can be concluded that the developed high throughput LC-MS/MS method is sensitive enough to measure Baclofen up to an LOOQ 5.023 ng/mL in Human plasma and cater to high throughput analysis using 96 well plates.
Keywords: Baclofen, Baclofen D4, Solid Phase Extraction LC-MS/Ms method, 96 well plates.
Full length article – PDF *Corresponding Author, e-mail: tummuru1@gmail.com Doi # https://doi.org/10.62877/69-IJCBS-24-25-19-69