Analytical Methods
Method Specifications
All analytical methods are adapted from United States Pharmacopeia (USP), European Pharmacopoeia (EP), and International Council for Harmonisation (ICH) guidelines. Full method validation packages available to audit clients on request.
Purity
Reverse-Phase HPLC
- Reference Method
- USP <621> Chromatography
- Instrument
- Agilent 1260 Infinity II with Diode Array Detector
- Column
- C18 reverse-phase, 4.6 × 250 mm, 5 μm particle
- Mobile Phase
- Water / acetonitrile / 0.1% TFA, gradient elution
- Detection
- UV at 214 nm and 280 nm
- Limit of Detection
- 0.05% (relative to main peak)
- Limit of Quantitation
- 0.1%
- Precision (RSD)
- ≤ 2.0% at 100% target
- Linearity Range
- 0.1% – 120% of target concentration
- Typical Run Time
- 15 – 45 minutes
Identity
LC-MS Identification
- Reference Method
- USP <736> Mass Spectrometry
- Instrument
- Waters Xevo TQ-S with Acquity UPLC
- Ionization
- Electrospray (ESI), positive and negative modes
- Mass Accuracy
- ± 0.01 Da (monoisotopic mass match)
- Mass Range
- 50 – 4000 m/z
- Sample Preparation
- Dissolution in water or mobile phase, 0.5 – 1.0 mg/mL
- Low-Level Detection
- ng/mL range (compound-dependent)
Safety
Bacterial Endotoxins Test (LAL)
- Reference Method
- USP <85> Bacterial Endotoxins Test
- Method Type
- Kinetic Chromogenic LAL
- Instrument
- Charles River Endosafe Kinetic-QCL Reader
- Limit of Detection
- 0.005 EU/mL
- Reporting Range
- 0.005 – 50 EU/mL
- Typical Precision (RSD)
- ≤ 10% at standard curve mid-range
- Sample Preparation
- Per client-supplied dilution protocol or default per-mg dilution
Stability
Karl Fischer Water Content
- Reference Method
- USP <921> Method 1c (coulometric) / 1a (volumetric)
- Instrument
- Metrohm 915 KF Ti-Touch
- Detection Range
- 10 ppm – 100% water
- Sample Size
- 20 – 200 mg (compound-dependent)
- Typical Precision
- ± 0.05% at 1.0% water
Safety
Residual Solvents (GC-MS)
- Reference Method
- USP <467> Residual Solvents
- Instrument
- Shimadzu GCMS-QP2020 NX with HS-20 Headspace
- Coverage
- ICH Q3C Class 1, 2, and 3 solvents
- Limit of Detection
- Sub-ppm for most Class 1 / 2 solvents
- Sample Preparation
- Headspace vial, DMSO or DMF dissolution