03
Innovation & Technology
(01)
Analytical methods used for batch release qualified/validated in accordance with ICH Q2(R1)
(02)
Appearance, particles, pH, osmolality, water content, melting point, solubility.
Identity, purity and impurity, assay, excipient, Counter ions
Residual solvent, elemental impurities
Forced degradation study, accelerated and long term stability under R&D and GMP conditions
(03)
Molecular weight by LC-MS; sequencing by LC-MS/MS； 2D-HPLC-MS/MS
Tm values between reference standard and test sample agree to whith in a certain degree, the sequence of the sample is confirmed.
Quantify and qualify DNA, RNA in seconds with only 1-2 μL, and obtain full-spectral data
(01)
Molecular mass analysis (LC-MS)
Elemental analysis
Sequence confirmation
FTIR
UV spectrum
Circular Dichroism （CD）
Melting temperature (Tm)
X-ray powder diffraction (XRPD)
Thermogravimetric analysis (TGA)
Differential scanning calorimetry (DSC)
Molar absorption coefficient
(02)
Purity analysis
Impurity identification
Assay
Counter ions
Residual solvent
Elemental impurities
(03)
Single-strand molecular mass
Single strands sequencing by MS/MS
Retention time by IPRP-HPLC
Tm value by UV spec
IR or UV spec
Amino acid analysis
(04)
pH
Osmolality
Extractable volume
Water determination
Subvisible particulate matter
Melting point
Apparance (color, clarity, visible particles)
Dissolution
(05)
Bacterial Endotoxin (Gel-Clot/ Photometric)
Bioburden (Plate count /Membrane filtration)
Sterility (Direct inoculation/Membrane filtration)
| Attribute | Test Item | Method | Instrument |
|---|---|---|---|
| Appearance | Color | Visual Inspection | N/A |
| Clarity | Visual Inspection | N/A | |
| General Properties | Water determination | Karl Fischer Titration | Mettler Toledo C30S |
| Identity | Molecular mass | LC-MS | Agilent 1290 LC and 6135 MS |
| HPLC retention time | HPLC | Agilent 1260/1290, Waters H-Class/I-Class | |
| Sequencing | MS/MS | Waters Acquity premier or 2D-HPLC/Xevo G3 Qtof | |
| Amino acid analysis | HPLC | Agilent 1260/1290, Waters H-Class/I-Class | |
| Infrared spectroscopy | iR | Thermo Nicolet is20 | |
| UV spectroscopy | UV spectroscopy | Thermo E201 | |
| Assay(content) | HPLC | Mass balance | Agilent 1260/1290, Waters H-Class/I-Class |
| Peptide content | Nitrogen determination by Kjeldahl | Kjeldahl | Haineng K1160 |
| Purity | RP-HPLC | RP-HPLC | Haineng K1160 |
| Peptide-Related impurities | RP-HPLC | RP-HPLC | Agilent 1260/1290, Waters H-Class/I-Class |
| Process related impurities | Residual solvents | eadspace GC-FID | Agilent 8890B GC, Agilent 8890B-5977B GCMSD |
| Elemental impurities | ICP-MS | Thermo iCAP RQ | |
| Residual TFA | HPLC | Agilent 1260/1290, Waters H-Class/I-Class | |
| Counter ions | Acetate, Cl-, F-, Na+ | IC | Thermo ICS 6000 |
| Specific tests | Optical rotation | INESA SGW532 | |
| Thiol groups | Ellman's test | N/A | |
| Bacterial endotoxins | Gel-Clot/Photometric | N/A | |
| Safety | Bioburden | Plate/Membrane | N/A |
| Attribute | Test Item | ASO | siRNA | Instrument |
|---|---|---|---|---|
| Appearance | Color Y Y N/A | Y | Y | N/A |
| Clarity | Y | Y | N/A | |
| General Properties | PH | Y | Y | Mettler Toledo S470-K/S400-K |
| Osmolality | optional | optional | YASN Osmo210 | |
| Water determination (lyo) | Y | Mettler Toledo C30S | ||
| Identity | Single-strand molecular mass by IPRP-MS | Y | Y | Agilent 1290 LC and 6135 MS |
| Tm value by UV spec | optional | Y | Thermo E201 with Cell Peltier System | |
| Single strands sequencing by MS/MS | Y | Y | Waters Acquity premier or 2D-HPLC/Xevo G3 Qtof | |
| duplex retention time （Se-HPLC-UV, IPRP-HPLC-UV) | N/A | Y | Agilent 1260/1290, Waters H-Class/I-Class | |
| Counter ions | sodium content by IC | Y | Y | Thermo ICS 6000 |
| Anion content by IC | Y | Y | ||
| Assay(content) | IPRP-HPLC | Y | N/A | Agilent 1260/1290, Waters H-Class/I-Class |
| UV absorption | N/A | Y | Thermo E201. Thermo Nonodrop | |
| Purity | non-denaturing IPRP-HPLC-UV Se-HPLC UV | Y | Y | Agilent 1260/1290, Waters H-Class/I-Class |
| denaturing AX-HPLC UV | N/A | Y | ||
| denaturing IPRP-HPLC UV | N/A | Y | ||
| Process-related impurity | residual solvents by headspace GC-FID | Y | Y | Agilent 8890B GC, Agilent 8890B-5977B GCMSD |
| elemental impurities by ICP-MS | Y | Y | Thermo iCAP RQ | |
| Safety | bacterial endotoxins | Y | Y | N/A |
| bioburden | Y | Y | N/A |