Raw Material Identification of mRNA Lipid Nanoparticle Components with the Agilent Vaya Raman Spectrometer

Importance of the Topic

Lipid nanoparticles (LNPs) are critical delivery vehicles for mRNA-based therapeutics and vaccines. Verifying the identity and purity of raw materials—lipids, buffers, solvents, and cryoprotectants—ensures consistent product quality, safety, and efficacy. Handheld Raman spectroscopy, particularly spatially offset Raman spectroscopy (SORS), offers rapid, non-destructive material identification through transparent and opaque containers, streamlining quality control in biopharmaceutical manufacturing.

Objectives and Study Overview

This application note demonstrates the capabilities of the Agilent Vaya handheld Raman spectrometer for raw material identity verification of LNP components. The study aims to develop and validate SORS-based methods for distinguishing key lipid and nonlipid excipients used in mRNA LNP formulation, measuring them through their original packaging without sample preparation.

Methodology and Instrumentation Used

Experimental design included four categories of excipients:

• Lipids: cholesterol, DSPC, DMG-PEG 2000
• Buffers: tris, HEPES, citric acid
• Organic solvents: methanol, ethanol, acetonitrile
• Cryoprotectants: sucrose, maltose, trehalose, mannitol, sorbitol

The Agilent Vaya handheld Raman spectrometer with SORS capability was employed. Methods were automatically configured via the built-in development wizard. Raw materials were analyzed in clear or amber glass vials and white HDPE containers. Spectral acquisition parameters were optimized during a performance qualification test, and automated baseline correction was applied.

Main Results and Discussion

Distinct Raman fingerprints were obtained for each excipient category:

• Lipids: Characteristic bands at 1,440 cm⁻¹ (CH2/CH3 deformation), 1,673 cm⁻¹ (C=C stretching), 949 cm⁻¹ (PO stretching), and 1,700 cm⁻¹ (C=O stretching).
• Buffers: Tris exhibited N–H bending at 1,500–1,700 cm⁻¹; HEPES showed SO₃ stretching at 1,046 cm⁻¹; citric acid displayed C=O stretching at 1,700 cm⁻¹.
• Solvents: Methanol (C–O at 1,035 cm⁻¹), ethanol (C–C at 882 cm⁻¹, C–O at 1,050 cm⁻¹), acetonitrile (C–C skeletal at 921 cm⁻¹).
• Cryoprotectants: Unique sugar spectra with torsion, deformation, and stretching bands distinguishing sucrose, maltose, trehalose, mannitol, and sorbitol.

The Vaya system accurately identified all materials through their packaging, eliminating sample handling and reducing contamination risk.

Benefits and Practical Application of the Method

Implementing handheld SORS-based Raman verification:

• Enhances raw material control in quarantine and warehouse areas.
• Eliminates the need to open or transfer samples, preserving sterility.
• Reduces analysis time and labor costs associated with conventional testing.
• Supports compliance with current Good Manufacturing Practice (cGMP) requirements.

Future Trends and Potential Applications

Advancements in handheld Raman and SORS technologies will expand capabilities to:

• Broader excipient libraries and complex formulations.
• Remote or automated in-line quality control in biomanufacturing.
• Integration with artificial intelligence for spectral interpretation and anomaly detection.
• Real-time release testing to accelerate product development cycles.

Conclusion

The Agilent Vaya handheld Raman spectrometer with SORS effectively verifies the identity of mRNA LNP raw materials through original packaging. Its rapid, non-invasive approach supports streamlined quality control workflows and cGMP compliance, offering a robust solution for the growing demands of biopharmaceutical manufacturing.

References

1. Bulbake U et al. Pharmaceutics 2017, 9(2), 12.
2. Challener C. Pharmaceutical Technology 2023, 47(3), 20–22.
3. Prullière F, Presly O. Agilent Technologies white paper 2020, 5994-2091EN.
4. Czamara K et al. J. Raman Spectrosc. 2015, 46(1), 4–20.