Capillary Electrophoresis SOP
Instrument:
P/ACE MDQ Capillary Electrophoresis System
Instrument Prep:


Turn on the instrument
Let it warm up for 30 minutes
Sample Prep:
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
Prepare chemicals
o Use five 1.5-mL vials
o Add 1.4 mL each of Run Buffer A (x2), Solution A, distilled water, and 1.0 M HCl
 Do not overfill the vials
o Cap these vials using the red caps from the drawer
o Cap 2-3 empty glass vials to be used for waste
o Use a plastic vial for your sample
 Fill with 1mL of Test Mixture B
 Cap using blue cap from the drawer
Load buffer and sample vials according to Dr. Hu’s SOP
o
Direct Control/Load


Wait until buffer trays are sent to the front, lift the cover, and insert vials as
follows:
 BI:A1 – 0.1 M NaOH
 BI:B1 – 1 M HCl
 BI:C1 – distilled water
 BI: D1 – Buffer A
 BO:A1 – Waste
 BO:B1 – Waste
 BO:C1 – Buffer A
 SI:A1 – Test Mix B
 SO:A1 – Waste
Record the position of each vial in your lab notebook
Running the Samples:



Build Method
o Initial Condition: Leave all defaults
o PDA Detector Initial Conditions: Use Channel 1 (check both boxes) at 214 nm, uncheck
Acquisition Enabled (top left quadrant)
o Time Program:
 Rinse 1.0 min (20 psi, forward pressure) from 1.0 M HCl to Waste
 Rinse 1.0 min (20 psi, forward pressure) from distilled water to Waste
 Rinse 2.0 min (20 psi, forward pressure) from 0.1 M NaOH to Waste
 Rinse 1.0 min (20 psi, forward pressure) from distilled water to Waste
 Rinse 2.0 min (20 psi, forward pressure) from Buffer A to Waste
 Inject (0.5 psi pressure, 10 seconds) from sample vial (Test Mixture B) to Waste
 Separate at 0.0 min (Duration: 7 min, Voltage: 25 KV, Ramp: 0.17 min) from Run
Buffer A (BI) to Run Buffer A (BO)
 Autozero at time 1.50 min
 Stop data at 7.0 min
 Rinse 1.0 min at time 8.0 min (20 psi, forward pressure) from distilled water to
Waste
o Be sure to use the SO waste for the injection and be sure to use both of the BO waste
vials throughout the time program
Save and Run Method
o Use Dr. Hu’s SOP for more details on how to do this
Save and print electropherogram with report
o If printer is not working, inform lab instructor or assistant
Turning off the Instrument:

If you are the last group, exit the program, remove your vials and turn off the instrument
o Be sure to leave a buffer vial in the buffer inlet and buffer outlet trays
o
o
o
Choose to inject the tray
Allow the tray to inject, and turn off the instrument
This ensures that the capillary will not dry out
CE Risk Assessment Form
Definitions:
H-Hazard
P-Precautions needed
R-Response to hazard
Hydrochloric Acid
H1.1 Inhalation
P1.1 Work in fume hood
R1.1 If inhaled, move person to fresh air
H1.2 Corrosive-will burn skin
P1.2 Wear gloves
R1.2 Remove contaminated clothing, wash affected area with water for 15 minutes
Sodium Hydroxide
H2.1 Corrosive-will burn skin
P2.1 Wear gloves
R2.1 Remove contaminated clothing, wash affected area with water for 15 minutes