Waters Quantitative Peptide Retention Standard - Manual (Page 2)

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[ CARE AND USE MANUAL ]
Quantitative Peptide Retention Standard
II. STORAGE AND STABILITY
Lyophilized peptides generally have excellent stabilities, often showing little or no degradation after a few years at -20 °C. Long-term
storage (> 1 year) should be at -80 °C desiccated, medium-term storage (1–12 months) should be at -20 °C desiccated, short-term
storage (< 1 month) may be at 4 °C. Once reconstituted the solution should be used immediately to avoid degradation of peptides which
would compromise the benefit of the standard having been quantitatively formulated.
III. RECONSTITUTION OF THE QUANTITATIVE PEPTIDE RETENTION STANDARD
This standard contains 20 nmoles of each peptide lyophilized. Reconstitute with 133.3 µL of either 0.1% aqueous formic acid or 0.1%
aqueous trifluoroacetic acid to produce a sample concentration of 150 pmol/µL (per peptide). Suggested injection volumes and sample
loads are outlined below.
Table 2: Recommended Mass Load for LC Column Evaluation
Reconstitution Volume Column ID Injection Volume Quantity of Peptide Loaded (each species)
133.3 µL 4.6 mm 19.2 µL 2.9 nmol
133.3 µL 2.1 mm 4.0 µL 600 pmol
133.3 µL 1.0 mm 0.9 µL 140 pmol
IV. EXAMPLE OF USING THE QUANTITATIVE PEPTIDE RETENTION STANDARD
Below is a reference chromatogram with conditions to help provide an example of using the Quantitative Peptide Retention Standard.
Please note retention times are dependent on LC conditions, column and LC system. If the column size is different, the gradient/
injection volume can be scaled accordingly
System: ACQUITY UPLC ® H-Class Bio
Column: ACQUITY UPLC CSH130 C
18
, 130Å, 1.7 µm, 2.1 x 150 mm
Mobile Phase A: Water with 0.1% FA (v/v)
Mobile Phase B: ACN with 0.1% FA (v/v)
Gradient: 0.5% B for 1 min, then to 50% B over 30 min
Flow Rate: 0.3 mL/min
Column Temp. 40 °C
Detector: UV @ 280 nm , 10 Hz
Software: Monoisotopic masses calculated with MassLynx ® 4.1