Region

Applications

Standards and IS, Clarity OTX Central Nervous System (CNS) Tissue

Sample Preparation Details

Part Number :8E-S103-EGA
Description :Clarity® OTX
Condition 1 :Methanol
Load :Pre-treated sample
Wash 1 :50 mM Ammonium Acetate, pH 5.5/50% ACN
Elution 1 :100mM Ammonium Bicarbonate, 1mM TCEP, pH 9.5/40 % THF
Evaporate: :Evaporate to dryness
Condition Methanol
Equilibrate 50 mM Ammonium Acetate, pH 5.5
Load Pre-treated sample
Wash 50 mM Ammonium Acetate, pH 5.5/50% ACN
Elute 100mM Ammonium Bicarbonate, 1mM TCEP, pH 9.5/40 % THF
Notes :Aliquot 75 µL calibration standards, QC samples, study samples onto 2 mL 96-well round bottom plate Add 30 µL IS (1 µg/mL) to all wells except blank controls (add 30 µL water to blanks) Condition 96-well plate with 1 mL MeOH Equilibrate with 1 mL Equilibration Buffer (50 mM Ammonium Acetate, pH 5.5) Add 600 µL of Lysis Loading Buffer to all samples Vortex mix samples for ~5 min; incubate at RT for additional 5 minutes Transfer solution to TOMTEC or equivalent Wash OTX plate 3X with 1 mL Equilibration Buffer Wash plate 3X Wash Buffer (50 mM Ammonium Acetate, pH 5.5: ACN, 50:50) Add 0.5 mL Elution Buffer (100 mM Ammonium Bicarbonate, pH 9.5:ACN:THF (50:40:10, v/v/v) and elute into collection plate Dry samples to ~400 µL by Nitrogen

Application Detail (App ID: 26262)

Part No:00D-4790-AN
Dimensions:100 x 2.1 mm ID
Elution Type:Gradient
Elution A:1.0 % HFIP, 0.1% DIPEA in Water with 10 µm EDTA
Elution B:0.075% HFIP, 0.0375% DIPEA in Water:ACN (35:65) with 10 µm EDTA
Gradient Profile:
Step No. Time (min) Pct A Pct B
1 0 95 5
10 6.2 80 20
11 6.3 20 80
12 7.5 5 95
13 7.8 5 95
14 7.9 95 5
2 0.1 95 5
3 2.5 80 20
4 4 40 60
5 4.1 5 95
6 4.4 5 95
7 5 80 20
8 5.8 5 95
9 6.1 5 95

Flow Rate:0.5 mL/min
Col. Temp: 80 °C
Detection: LC/MS/MS @ (0°C)

Analyte Details

Analyte Detail not available