Source comparison
Verify Thymosin Alpha-1 Purity: Analytical Standards Comparison
Reverse-Phase HPLC Separation purity based on hydrophobicity and retention time ≥98% purity by integrated peak area at 220 nm detection wavelength, no secondary peaks >1% Molecular weight accuracy, sequence identity, or which impurity is present Required but i
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- Reverse-Phase HPLC
- Separation purity based on hydrophobicity and retention time
- ≥98% purity by integrated peak area at 220 nm detection wavelength, no secondary peaks >1%
- Molecular weight accuracy, sequence identity, or which impurity is present
- Required but insufficient alone. HPLC confirms separation, not identity
- ESI-MS or MALDI-TOF MS
- Exact molecular weight and charge state distribution
- Measured mass = 3,108.3 ±0.5 Da with no significant peaks at 3,092 Da (deletion analog) or 3,124 Da (oxidation)
- Amino acid sequence order or positional isomers with identical mass
- Confirms correct molecular weight but not sequence fidelity
- Amino Acid Analysis (AAA)
- Molar ratio of each amino acid after complete hydrolysis
- Detected amino acids match expected sequence ratios: Asx=6, Glx=7, Ser=4, Thr=4, Ala=3, Val=3, Ile=1, Leu=1, Lys=4
- Position of amino acids in the sequence or presence of D-amino acids
- Only method that confirms complete sequence composition independent of molecular weight
- Peptide Sequencing (Edman or MS/MS)
- N-terminal to C-terminal amino acid order verification
- First 10 residues confirmed as N-acetyl-SDAAVDTSSE with no ambiguous peaks
- Low-level impurities below ~5% or C-terminal truncations in minor contaminants
- Gold standard for sequence verification but expensive; rarely done for routine QC