Fast LC/MS/MS Method for Quantitative Determination of Valsartan in Human Plasma

Importance of the Topic

Valsartan is a key angiotensin II receptor antagonist broadly used to manage hypertension and cardiovascular diseases.
This application note presents a streamlined LC–MS/MS assay to accurately quantify valsartan levels in human plasma, addressing critical needs for speed, sensitivity, and reliability in clinical pharmacokinetic studies and therapeutic drug monitoring.

Goals and Study Overview

The main objective was to develop and validate a rapid, high-throughput UHPLC–MS/MS method for precise measurement of valsartan concentration in plasma samples.
The study compares simple protein precipitation extraction with robust triple-quadrupole mass spectrometry detection, using telmisartan as an internal standard.

Methodology and Instrumentation

Sample Preparation:

• Human plasma (180 µL) spiked with valsartan standards (20–15000 ng/mL) or QC levels (60, 6000, 12000 ng/mL).
  
• Protein precipitation by adding 1000 µL methanol containing internal standard, vortexing, and centrifugation at 13 000 rpm for 10 min.
  
• Supernatant directly injected into UHPLC–MS/MS system.

Chromatographic Conditions:

• Column: Zorbax Eclipse Plus C18 (100 × 2.1 mm, 1.8 µm).
• Mobile phase: 0.2% formic acid in water (A) and acetonitrile (B), isocratic 60:40 v/v.
• Flow rate: 600 µL/min, column temperature 50 °C.

Mass Spectrometry:

• Instrument: Shimadzu Nexera UHPLC coupled to LCMS-8040 triple-quadrupole.
• Ionization: ESI positive mode, interface voltage 4.5 kV, DL block 200 °C.
• MRM transitions: m/z 436→291 (valsartan), m/z 515→276 (telmisartan), optimized CE and bias settings.

Results and Discussion

Calibration and Sensitivity:

• Linear dynamic range: 20–15000 ng/mL, R² = 0.9997.
• Lower limit of quantification (LLOQ): 20 ng/mL with CV <20%.

Accuracy and Precision:

• QC precision (%CV) 1.6–6.5%, accuracy 81.7–101.5% across low, mid, and high levels.

Recovery:

• Extraction recovery of valsartan from plasma: 91.6–114.8%, demonstrating efficient protein precipitation protocol.

Representative MRM chromatograms confirm specificity and reproducibility at LLOQ, LQC, and HQC levels.

Practical Benefits and Applications

This method combines rapid sample processing with high sensitivity and a wide dynamic range, making it suitable for:

• Clinical pharmacokinetic and bioequivalence studies.
• Therapeutic drug monitoring in hospital and research laboratories.
• High-throughput screening in drug development pipelines.

Future Trends and Opportunities

Emerging UHPLC–MS/MS platforms may further reduce run times and solvent consumption.
Integration with automated sample preparation will enhance throughput and reproducibility.
Application of this approach to metabolite profiling and multi-analyte panels could expand its utility in personalized medicine.

Conclusion

A robust, fast, and sensitive UHPLC–MS/MS assay for valsartan quantification in human plasma has been established.
The protocol achieves excellent linearity, precision, accuracy, and recovery, meeting regulatory criteria for bioanalytical methods.
This workflow supports efficient and reliable therapeutic monitoring and pharmacokinetic evaluation of valsartan.

References

No external literature references were provided in the source document.