Determination of Rosuvastatin in Human Plasma by SPE-LC-MS/MS using SOLA and Accucore RP-MS column

Importance of the Topic

Rosuvastatin is a potent cholesterol‐lowering agent widely prescribed to reduce cardiovascular risk. Accurate quantification of this drug in human plasma is essential for pharmacokinetic studies, therapeutic drug monitoring, and bioequivalence assessments. Developing a robust analytical workflow ensures high confidence in clinical decision‐making and regulatory compliance.

Objectives and Study Overview

The primary aim of this study was to establish and validate a sensitive, rapid, and reliable SPE‐LC‐MS/MS method for measuring rosuvastatin concentrations in human plasma. By combining Thermo Scientific SOLA solid‐phase extraction with Accucore RP‐MS chromatography and triple quadrupole detection, the authors sought to demonstrate linearity over a wide dynamic range, high recovery, minimal matrix effects, and suitability for routine high‐throughput applications.

Methodology and Instrumentation

Sample Preparation and Extraction

• Plasma samples (100 µL) were processed using a Thermo Scientific SOLA 96-well SPE plate under positive pressure.
• The plate was conditioned with methanol and water, followed by loading of plasma, aqueous formic acid wash, low‐organic wash, and elution with 90 % methanol.
• Eluates were evaporated under nitrogen at 40 °C and reconstituted in 25 % methanol aqueous solution (200 µL).

Chromatography and Mass Spectrometry

• LC separation employed an Accela 600 system and Accucore RP-MS column (2.6 µm, 50 × 2.1 mm) with a matching guard cartridge.
• Gradient elution from 5 % to 95 % methanol (each containing 0.1 % formic acid) over 1 min, total run time 3.6 min; flow rate 0.75 mL/min; column at 60 °C.
• Detection used a Thermo Scientific TSQ Vantage triple quadrupole with heated electrospray ionisation (HESI) in positive mode, monitoring SRM transitions 482.3→258.2 for rosuvastatin and 488.2→264.2 for the d6 internal standard.

Main Results and Discussion

Linearity and Sensitivity

• The method exhibited a linear response (1–1000 ng/mL) with r² = 0.9984 using 1/x weighting.
• Lower limit of quantification was demonstrated at 1 ng/mL with acceptable precision and accuracy.

Precision, Accuracy, and Recovery

• Quality control samples at 3, 400, and 750 ng/mL showed intra‐ and inter-day precision (%RSD) below 6 % and accuracy within ±11 %.
• Absolute recovery of rosuvastatin averaged 99.3 % with %RSD of 4.9 % across QC levels.

Matrix Effects and Specificity

• Post-extraction spiking experiments revealed negligible matrix suppression or enhancement (<5.3 %).
• Chromatographic separation resolved an endogenous plasma impurity (retention at 1.92 min) from the analyte peak at 1.49 min, ensuring specificity.

Benefits and Practical Applications

This SPE-LC-MS/MS workflow offers:

• High throughput compatibility for clinical and bioanalytical laboratories.
• Minimal solvent and sample consumption, reducing cost and environmental impact.
• Robust performance with excellent reproducibility, facilitating large pharmacokinetic or therapeutic monitoring studies.

Future Trends and Opportunities

Further developments may include:

• Automation of sample preparation to increase throughput and consistency.
• Extension to other statins or related compounds in multi-analyte bioassays.
• Integration with high-resolution mass spectrometry for metabolite profiling.
• Application to alternative biological matrices such as dried blood spots or cerebrospinal fluid.

Conclusion

A rapid, sensitive, and reliable SPE-LC-MS/MS method was fully validated for rosuvastatin quantification in human plasma. The approach combines efficient SOLA extraction and high-performance Accucore RP-MS chromatography with SRM detection, delivering excellent linearity, recovery, precision, and minimal matrix interference. The protocol is well suited for routine clinical studies and pharmaceutical research.

Reference

1. Pelat M.; Dessy C.; Massion P.; Desager J.-P.; Feron O.; Balligand J.-L. Journal of Circulation 2003, 107, 2480.
2. Trivedi R.K.; Kallem R.R.; Mullangi R.; Srinivas N.R. Journal of Pharmaceutical and Biomedical Analysis 2005, 39(3–4), 661–669.