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 high‐potency cholesterol‐lowering agent widely prescribed to reduce cardiovascular risk. Precise and rapid quantification of rosuvastatin in human plasma is crucial for pharmacokinetic studies, clinical therapeutic monitoring, and bioequivalence assessments. Advances in sample preparation and mass spectrometric techniques allow laboratories to improve throughput while maintaining sensitivity and robustness.

Study Objectives and Overview

The primary aim of this application note was to demonstrate a streamlined workflow for the determination of rosuvastatin in human plasma using Thermo Scientific SOLA solid phase extraction (SPE) plates paired with an Accucore RP‐MS column and LC‐MS/MS detection. Key goals included:

• Developing a simple, reproducible SPE protocol compatible with 96‐well plates
• Establishing a rapid reversed‐phase LC gradient using Accucore RP‐MS for baseline separation
• Validating method performance parameters: linearity, accuracy, precision, recovery, and matrix effects

Methodology and Instrumentation

Sample Preparation via SPE:

• Condition SOLA plate with methanol and water
• Load plasma sample (100 µL) spiked with d6-rosuvastatin internal standard
• Wash with dilute formic acid and low‐strength methanol
• Elute analytes with 90% methanol, evaporate, and reconstitute in 1:4 methanol‐water (200 µL)

Chromatography and Detection:

• Column: Accucore RP‐MS (2.6 µm, 50 × 2.1 mm) with Defender guard cartridge
• Mobile phase A: water + 0.1% formic acid; B: methanol + 0.1% formic acid
• Gradient: 5% B to 95% B (0.1–1.0 min), hold, then re‐equilibrate; flow rate 0.75 mL/min; runtime 3.6 min
• Detection: TSQ Vantage triple quadrupole MS; HESI positive mode; SRM transitions 482.30→258.15 (rosuvastatin), 488.22→264.17 (d6‐rosuvastatin)

Main Results and Discussion

Linearity and Sensitivity:
The calibration curve over 1–1000 ng/mL displayed excellent linearity (r2 = 0.9984) using 1/x weighting. The lower limit of quantification (LLOQ) was 1 ng/mL with accuracy within ±6% and precision below 6% RSD.

Accuracy, Precision, and Recovery:
QC samples at 3, 400, and 750 ng/mL showed intra‐run accuracy within ±11.6% error and precision ≤5.8% RSD. The mean recovery at the medium QC level was 99.3% with 4.9% RSD, confirming high extraction efficiency and reproducibility.

Matrix Effects and Specificity:
No significant matrix suppression or enhancement was observed (<5% effect). Chromatograms of blanks demonstrated adequate separation of endogenous interferences from the rosuvastatin peak at 1.49 min, ensuring specificity.

Benefits and Practical Applications

The described SPE‐LC‐MS/MS method offers:

• High throughput via 96‐well SPE format and fast 3.6 min analysis time
• Robust sensitivity enabling low‐level detection (1 ng/mL)
• Minimal solvent and sample consumption
• Reproducible performance suited for clinical and bioanalytical laboratories

Future Trends and Applications

Emerging opportunities include:

• Extension to multiplexed statin panels and metabolites
• Further miniaturization with microflow or nanoflow LC-MS systems
• Integration with automated sample handling and data processing workflows
• Adoption of novel SPE chemistries for broader drug classes and biomolecules

Conclusion

A rapid, sensitive, and reliable SPE‐LC‐MS/MS procedure was developed for rosuvastatin quantification in human plasma. The combination of SOLA SPE plates and Accucore RP‐MS columns delivers high recovery, negligible matrix effects, and robust quantitative performance, making it highly suitable for routine bioanalytical applications.

References

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