Read "A LC-MS/MS Method for Oligonucleotide PK Bioanalysis"
Oligonucleotides are rapidly emerging as promising therapeutic modalities, but their PK bioanalysis poses unique technical challenges. Their highly hydrophilic backbones, negative charges, and tendency for non-specific binding often result in poor retention, weak MS responses, strong matrix interference, and sample loss. Our expert team at Crystal Bio Solutions develops and validates specialized LC-MS/MS workflows that overcome these challenges, enabling reliable, high-throughput PK studies for novel oligonucleotide therapies.
By applying an optimized HFIP/TEA ion-pair buffer system, a BEH C18 column, and a simple PPT sample preparation, our validated method achieved excellent sensitivity, recovery, and stability, delivering robust results with a runtime of just 4.1 minutes per sample.
Quantifying oligonucleotides in preclinical and clinical settings is notoriously difficult. Key challenges include:
Poor retention on ordinary columns due to hydrophilic backbones
Peak tailing and carryover issues
Decreasing MS response with increasing molecular weight
Strong matrix interference and low recovery rates
Sample loss caused by non-specific binding
To address these issues, our experts designed a workflow based on:
HFIP/TEA ion-pair buffer system for improved retention and signal
BEH C18 column (2.1×50 mm) for sharp peak resolution
Gradient elution to optimize separation
Simple protein precipitation (PPT) pretreatment for cleaner samples
This method was fully validated with excellent performance in sensitivity, accuracy, precision, selectivity, recovery, and minimal matrix interference.
The validated method demonstrated:
Standard curve: 2.00 – 2000 ng/mL (R² = 0.9939)
Accuracy (%Bias): -6.5% to 7.0%
Precision (%CV): <8.8%
Recovery: 76.3%
Stability: up to 45 days at -80°C, 144.9h at 4°C
Runtime: ~4.1 minutes per sample
This approach was successfully applied in a Phase 1 clinical study, enabling reliable PK profiling of oligonucleotide drug candidates.
Accurate PK analysis is essential for understanding the absorption, distribution, metabolism, and excretion of oligonucleotide-based drugs. Without validated, high-throughput methods, development can be slowed by unreliable data or regulatory setbacks. Our workflow provides a proven solution to accelerate clinical development while ensuring data quality and compliance.