Aeris rp hplc column

Peptides Ac-FAEDA, Ac-FAEPA, Ac-FAEAA, Ac-FAQDA, Ac-FAQPA and Ac-FAQAA (1 mg/mL) were incubated at either 37 °C or 60 °C in 50mM MOPS buffer at pH 6.7, 7.0 and 7.4 or 50mM sodium acetate buffer at pH 4.0. Aliquots were collected at various time intervals and were injected onto an Agilent 1100 HPLC using an Aeris RP-HPLC column (2.6 μ, XB-C18, 100 × 2.1 mm, Phenomenex). Breakdown products of peptides Ac-FAQDA and Ac-FAEDA were eluted with the following gradient 2% ACN, 0.1% TFA 0–40 min, 5% ACN, 0.1% TFA 40–60 mins, 10% ACN, 0.1%TFA 60–80 min, 15% ACN, 0.1% TFA 80–100min, 25% ACN, 0.1% TFA 100–120 min, 80 % ACN, 0.1%TFA, 135 min 2% ACN, 0.1%TFA 145min. All other peptides used the following elution gradient: 2% ACN, 0.1% TFA 5 min, 20% ACN, 0.1% TFA 15 mins, 40% ACN, 0.1%TFA 20 min, 25 min 80%ACN, 0.1% TFA, 35 min 80% ACN, 0.1% TFA, 30.1 min 2% ACN, 0.1%TFA, 40 min 2% ACN, 0.1%TFA.
Breakdown of all peptides was monitored at 216nm and 280nm. Peaks were collected and identified by tandem mass spectrometry on an LTQ Orbitrap Fusion Tribrid with a nanoelectrospray ionisation source (Thermo Scientific, San Jose, CA). Data were manually acquired with HCD collision energy set to 15.

Ac-YAQTT and Ac-YAETT (1 mg/mL) were incubated at 60 °C in 50mM MOPS buffer at pH 6.7, 7.4 and 8.0. Ac-FAEAA and Ac-FAEPA were incubated at 60 °C in 50mM MOPS buffer at pH 6.7. Aliquots were injected onto an Agilent 1100 HPLC using an Aeris RP-HPLC column (2.6 μ, XB-C18, 100 × 2.1 mm, Phenomenex). The elution gradient was as follows: 2% ACN, 0.1% TFA 5 min, 20% ACN, 0.1% TFA 15 mins, 40% ACN, 0.1%TFA 20 min, 25 min 80%ACN, 0.1% TFA, 35 min 80% ACN, 0.1% TFA, 30.1 min 2% ACN, 0.1%TFA, 40 min 2% ACN, 0.1%TFA.
Breakdown of Ac-YAQTT, Ac-YAETT, Ac-FAEAA and Ac-FAEPA was monitored at 216nm and 280nm. Peaks were collected and identified by tandem mass spectrometry on an LTQ Fusion orbitrap (Thermo Scientific, San Jose, CA). The above experiments were repeated with peptides incubated in the presence of a 5-molar excess of either phenylethylamine (PE) or the dipeptide GlyAla in 50mM MOPS 6.7 at 60 °C. All experiments were performed in triplicate.