Development and Optimization of an RP-HPLC Method for Atipamezole Quantification to Support Microneedle-Based Drug Delivery
Abstract
There is great potential for the usage of transnasal atipamezole as a treatment for xylazine overdose, particularly given the already well-established success of transnasal microneedle-based drug delivery for rapid absorption of similar agents. With xylazine being continually mixed in with narcotics, the need for a new treatment has become increasingly more prevalent. Thus, the hypothesis of this work is on combining atipamezole with a rapid and effective transnasal delivery method to create a treatment for xylazine toxicity, though the ongoing research in the lab must do so through developing an analytical method to detect atipamezole in environments that simulate physiological conditions encountered during practical application. A reverse-phase high performance liquid chromatography (HPLC) method using a 150x4.6mm, end-capped C18 column was developed with acetonitrile, HPLC water and a buffer as mobile phase and a different citrate-phosphate 5.0 pH buffer solvent for standard preparation. The issues identified during the process were peak splitting, retention time restrictions, and peak symmetry; with the first two having been remedied with the end-capped C18 column and alterations with the mobile phase proportions. Changes in column temperature (25-45 °C), mobile phase buffer (pH 3.2-5 and 20-50mM), flow rate (1-1.3 mL/min), and TEA (0-0.2%) in mobile phases were used to further optimize the method. Resulting from this, the current method involves using a 5.0 pH 50mM sodium-phosphate mobile phase buffer, 45 °C column temperature, no TEA and a flow rate of 1.2mL/min. This method allows for the desired retention time (5-6 min) and acceptable peak symmetry values (~1) for standards below 10 µg/mL. The peak symmetry troubleshooting is ongoing for higher standards. However, the method optimized so far has been found to be reproducible for 0.1-10 ug/mL drug concentrations and can be employed for testing the microneedle’s effectiveness for delivering atipamezole across the porcine nasal mucosa after validation.
Start Time
15-4-2026 1:30 PM
End Time
15-4-2026 4:30 PM
Room Number
Culp Ballroom 316
Poster Number
2
Presentation Type
Poster
Presentation Subtype
Posters - Competitive
Presentation Category
Science, Technology, and Engineering
Student Type
Graduate and Professional Degree Students, Residents, Fellows
Faculty Mentor
Ashana Puri
Development and Optimization of an RP-HPLC Method for Atipamezole Quantification to Support Microneedle-Based Drug Delivery
Culp Ballroom 316
There is great potential for the usage of transnasal atipamezole as a treatment for xylazine overdose, particularly given the already well-established success of transnasal microneedle-based drug delivery for rapid absorption of similar agents. With xylazine being continually mixed in with narcotics, the need for a new treatment has become increasingly more prevalent. Thus, the hypothesis of this work is on combining atipamezole with a rapid and effective transnasal delivery method to create a treatment for xylazine toxicity, though the ongoing research in the lab must do so through developing an analytical method to detect atipamezole in environments that simulate physiological conditions encountered during practical application. A reverse-phase high performance liquid chromatography (HPLC) method using a 150x4.6mm, end-capped C18 column was developed with acetonitrile, HPLC water and a buffer as mobile phase and a different citrate-phosphate 5.0 pH buffer solvent for standard preparation. The issues identified during the process were peak splitting, retention time restrictions, and peak symmetry; with the first two having been remedied with the end-capped C18 column and alterations with the mobile phase proportions. Changes in column temperature (25-45 °C), mobile phase buffer (pH 3.2-5 and 20-50mM), flow rate (1-1.3 mL/min), and TEA (0-0.2%) in mobile phases were used to further optimize the method. Resulting from this, the current method involves using a 5.0 pH 50mM sodium-phosphate mobile phase buffer, 45 °C column temperature, no TEA and a flow rate of 1.2mL/min. This method allows for the desired retention time (5-6 min) and acceptable peak symmetry values (~1) for standards below 10 µg/mL. The peak symmetry troubleshooting is ongoing for higher standards. However, the method optimized so far has been found to be reproducible for 0.1-10 ug/mL drug concentrations and can be employed for testing the microneedle’s effectiveness for delivering atipamezole across the porcine nasal mucosa after validation.
