Transfer of the HPLC Method for Related Substances Analysis of Metoclopramide HCl from an Agilent 1260 Infinity LC System to an ACQUITY Arc System

This application note demonstrates transfer of an HPLC method for related substances of metoclopramide HCl from an Agilent 1260 Infinity LC System to an ACQUITY Arc System by comparing chromatographic separation and system suitability results.

Benefits

The ACQUITY Arc System successfully replicated chromatographic separation obtained on the Agilent 1260 Infinity LC System, generating reliable, robust, and reproducible results.

Analysis of an active pharmaceutical ingredient (API) and its related substances must be conducted by every manufacturer to ensure safety and efficacy of the end pharmaceutical drug product. Often, these assays are performed across the organization or sent out to contract partners that utilize instruments from different vendors. Therefore, it is essential for the analytical laboratories to successfully  transfer methods between different instruments to ensure product consistency and compliance with regulations. Effective method transfer generates equivalent results for the same analysis independent of the instrument, laboratory, or resources. This is important to eliminate revalidation of the method, which is time consuming and costly.

In this study, we demonstrated the transfer of the HPLC method for related substances analysis of metoclopramide HCl from an Agilent 1260 Infinity LC System to an ACQUITY Arc System.

The success of the method transfer to the new instrumentation was measured by examining the chromatographic separation for comparable results and verifying that the system suitability results met the specifications defined in the USP General Chapter <621> Chromatography.¹

The related compounds of metoclopramide HCl used in this study are listed in Table 1. Separate stock solutions were prepared in methanol at 1.0 mg/mL. A metoclopramide stock solution was diluted with water to 0.5 mg/mL and spiked with related substances at 1.0%. For system suitability, each stock solution was transferred to one vial and diluted with water to 0.05 mg/mL concentration of each analyte. The mixture was then diluted with standard diluent (50:50 methanol/water) to 15 μg/mL concentration for system suitability determination.

The HPLC method was first run on an Agilent 1260 Infinity LC System and then transferred to an ACQUITY Arc System. The ACQUITY Arc System offers a novel Multi-flow path Technology, which enables efficient method transfer of legacy methods developed on previous generations of LC instrumentation. Instrument conditions for transfer of the HPLC method are listed in Figure 1.

The chromatographic data of a sample containing metaclopramide API with 1.0% of related substance acquired on both the Agilent 1260 Infinity LC and ACQUITY Arc systems is displayed in Figure 2. The chromatographic separation produced on an ACQUITY Arc System was comparable with the results obtained on the Agilent 1260 Infinity LC System.

The mass spectral data acquired using an ACQUITY QDa Detector coupled to an ACQUITY Arc System was used to confirm identity of metoclopramide and related substances by mass detection. The mass analysis window from Empower 3 Software (Figure 3) shows UV and mass spectral data from a single result. This UV detection enhanced with MS spectral data was used to quickly confirm identity of the sample components.

Performance of the HPLC method on the ACQUITY Arc System was verified by evaluating system suitability of five replicate injections of the system suitability solution according to the requirements defined in the USP General Chapter, <621>, Chromatography¹ and compared to the results obtained on the Agilent 1260 Infinity LC System. Results of method suitability acquired on both systems passed the USP specifications and are summarized in Table 2. The repeatability of retention times and peak areas of the method run on the ACQUITY Arc System were substantially lower than the USP specifications of less than 2.0% RSD and comparable to the Agilent 1260 Infinity LC System. The USP resolution values between all the peaks were comparable on both systems, indicating no loss in resolution for method transfer. The USP peak tailing factors were also comparable.

Enabled by a unique Multi-flow path technology, the ACQUITY Arc System, easily accepts and replicates methods from a variety of platforms without compromising method integrity. The ACQUITY Arc System provides powerful LC performance and secures the laboratory’s investment by ensuring integration with new technologies such as the ACQUITY QDa Detector and Empower Chromatography Data System (CDS) Software.

The HPLC method for metoclopramide and its related substances was successfully transferred from an Agilent 1260 Infinity LC System to an ACQUITY Arc System. The ACQUITY Arc System successfully replicated the quality of chromatographic separation obtained on the Agilent 1260 Infinity LC System and met the USP specifications for system suitability. Meeting the system suitability requirements is essential for laboratories to remain in compliance with the current Good Laboratory Practices regulations. Moreover, the ACQUITY QDa Detector, in conjunction with UV detection, enabled quick confirmation of peak identity by mass detection.

Overall, the ACQUITY Arc System delivers reliable, robust, and reproducible results, enhancing throughput and laboratory efficiency for routine testing in the QC laboratory.

1. USP General Chapter <621> Chromatography, USP 37-NF 32, The United States Pharmacopeia Convention, Official December 1, 2014.