Home Mass SpectrometryAnalysis of Remdesivir and other Antiviral Drugs Using LC-MS/MS

Analysis of Remdesivir and other Antiviral Drugs Using LC-MS/MS

Introduction

Remdesivir is a broad-spectrum antiviral drug, typically administered through injections. The drug has been tested in 2020 as a treatment for COVID-19, through the WHO Solidarity Trial. Here we present a fast and robust HPLC-MS/MS method that can be used for Remdesivir and three structural antiviral analogues. Indinavir, Nelfinavir, and Saquinavir are protease inhibitors, typically used in the treatment of the human immunodeficiency virus (HIV).

Several different stationary phases were screened through the development process, with the aim to find and provide the best selectivity for the target analytes. The best overall results were achieved with an RP-Amide column, but also C18, AQ-C18 or Phenyl-Hexyl phases can offer appropriate retention and peak shape for Remdesivir. With a short 30x2.1 mm Ascentis^® Express RP-Amide stationary phase used in gradient mode it is possible to have all compounds well retained and fully resolved under two minutes. Calibration data for Remdesivir (1-250 ng/mL) display adequate linearity (R2 = 0.9995) with a Limit of detection (LOD) and Limit of quantitation (LOQ) determined as 0.7 and 1.3 ng/mL (95% confidence interval), respectively.

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A complex organic molecule structure. It features multiple ring structures, including a benzene ring, and various functional groups such as phosphates, amides, and alcohols. The molecule contains chains of carbon atoms with attached hydrogen atoms, double bonds, and is depicted in a skeletal formula representation where lines represent chemical bonds and vertex ends represent carbon atoms.

Figure 1.Remdesivir, C27H35N6O8P (CAS No. 1809249-37-3)

Organic molecule structure, which is likely of interest due to its relevance in chemistry, particularly in the study of molecular configurations and interactions. The molecule includes multiple ring structures, various functional groups such as hydroxyl (OH), methyl (CH3), and amine (NH2) groups. There are also double bonds present within the rings and between some carbon atoms. The structure is depicted with standard chemical notation, with lines representing bonds between atoms and letters indicating the types of atoms involved (C for carbon, N for nitrogen, O for oxygen).

Figure 2.Nelfinavir, C32H45N3O4S (CAS No. 159989-64-7)

complex organic molecule structure, which is likely of interest due to its relevance in chemistry, particularly in the study of molecular configurations and interactions. The molecule includes multiple ring structures, various functional groups such as hydroxyl (OH), methyl (CH3), and amine (NH2) groups. There are also double bonds present within the rings and between some carbon atoms. The structure is depicted with standard chemical notation, with lines representing bonds between atoms and letters indicating the types of atoms involved (C for carbon, N for nitrogen, O for oxygen).

Figure 4.Saquinavir, C38H50N6O5 (CAS No. 127779-20-8)

Complex organic molecule structure. It consists of multiple ring structures, some of which are hexagonal benzene rings, and others are pentagonal. The molecule includes various atoms connected by single, double, and triple bonds. Atoms are not labeled but can be inferred as carbon and hydrogen in the rings, with functional groups containing oxygen (indicated by red lines) and nitrogen (indicated by blue lines). There are also several hydroxyl (-OH) groups attached to the rings.

Figure 3.Indinavir, C36H47N5O4 (CAS No. 150378-17-9)

Remdesivir Calibration Data (1-250 ng/mL)

Graph with multiple peaks of different heights plotted against two axes. The horizontal axis is labeled from 10.00 to 15.00 in increments of 1, and the vertical axis is labeled from 0 to 7e+6 in increments, with specific values not fully visible. There are four prominent peaks, each a different color: red, blue, green, and purple. The highest peak reaches just below the maximum value on the vertical axis and is colored purple. This graph could be interesting or relevant for analyzing data patterns such as signal intensity over a range of values or for comparing the magnitude of different events or measurements.

Figure 5.Chromatogram showing the separation of Indinavir, Saquinavir, Nelfinavir, and Remdesivir. An example chromatogram showning the baseline separation of Remdesivir and three related antiviral drugs

A graph showing a sharp peak centered around x-value 1.345 with annotations indicating signal-to-noise ratio and peak retention time, labeled as ‘1 ng/mL’ in the upper right corner.

Figure 6.Chromatogram showing the chromatographic response from an injection of 1 ng/mL Remdesivir standard solution. Example LC-MS chromatogram showning the injection of 1 ng/mL standard solution

Figure 7.Remdesivir calibration curve data (1-15 ng/mL). Illustration of the calibration curve linearity at the lower Remdesivir concentration range examined in the current study

Figure 8.Remdesivir calibration curve data (1-250 ng/mL). Illustration of the full calibration range examined in the current study