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

Significance of the Topic

A robust and sensitive analytical method for quantifying rosuvastatin in human plasma is critical in clinical pharmacokinetics, therapeutic drug monitoring and drug–drug interaction studies. Precision and accuracy at low concentration levels ensure patient safety and reliable bioequivalence assessments.

Study Objectives and Overview

This application note presents the development and validation of a rapid solid-phase extraction coupled with liquid chromatography–tandem mass spectrometry (SPE-LC-MS/MS) method. The goal was to isolate rosuvastatin from plasma using Thermo Scientific SOLA plates and to achieve fast chromatographic separation on an Accucore RP-MS column with sensitive detection on a TSQ Vantage triple quadrupole mass spectrometer.

Methodology and Instrumentation

The workflow consisted of:

• Sample preparation: 100 µL plasma loaded onto a SOLA 96-well plate, conditioned with methanol and water, washed with formic acid and low-percent methanol, then eluted in high-percent methanol.
• Extract processing: Evaporation under nitrogen at 40 °C and reconstitution in 1:4 methanol–water.
• Chromatography: Accela 600 pump and Open Autosampler, Accucore RP-MS column (2.6 µm, 50 × 2.1 mm), gradient elution from 5 % to 95 % methanol with 0.1 % formic acid over 3.6 min at 0.75 mL/min, column at 60 °C.
• Mass spectrometry: TSQ Vantage with HESI positive ionization, selected reaction monitoring of m/z 482.30 → 258.15 for rosuvastatin and m/z 488.22 → 264.17 for d6-rosuvastatin IS.

Main Results and Discussion

Calibration was linear from 1 to 1000 ng/mL (r2 = 0.9984) using a 1/x weighted regression. The lower limit of quantification (LLOQ) was 1 ng/mL with acceptable signal-to-noise and precision. Accuracy across quality control levels (3, 400 and 750 ng/mL) ranged within ±11.6 % error, and precision (RSD) was below 5.8 %. Mean extraction recovery was 99.3 % with 4.8 % RSD. Matrix effects were negligible (<5 % deviation) and no significant endogenous interferences were observed.

Benefits and Practical Applications of the Method

• High throughput: 96-well SPE format and short 3.6 min run time.
• Low sample volume: 100 µL plasma per analysis.
• High sensitivity: LLOQ at 1 ng/mL suitable for pharmacokinetic profiling.
• Reliable performance: excellent reproducibility, recovery and matrix tolerance.

Future Trends and Potential Applications

Advances in core-shell column technology and next-generation SPE materials may further reduce analysis time and solvent consumption. Integration with automated sample handling and faster scanning mass spectrometers will support ultra high-throughput bioanalysis. The approach can be extended to other statins and metabolite profiling in drug development programs.

Conclusion

The SPE-LC-MS/MS method using SOLA extraction and Accucore RP-MS chromatography provides a fast, sensitive and precise assay for rosuvastatin in human plasma. Its robust performance makes it suitable for routine clinical and bioanalytical laboratories seeking high throughput and reliable quantification.

References

1. Pelat M, Dessy C, Massion P, Desager J-P, Feron O, Balligand J-L. J. Circ. 2003, 107, 2480.
2. Trivedi RK, Kallem RR, Mullangi R, Srinivas NR. J. Pharm. Biomed. Anal. 2005, 39(3-4), 661–669.