Owner manual

[ Care and Use ManUal ]
Waters Advanced Purification Columns 8
Replacing an inlet connector O-ring
To replace the inlet connector O-ring:
1. Deactivate the inlet O-ring by turning the seal adjustment knob.
Remove the inlet connector assembly from the column by unscrewing
the plunger.
2. Use a small blunt tool, such as a push pin, to gently pry up the O-ring
so that you may roll it off the connector with your fingers. Carefully
push a new O-ring onto the connector tip. Be sure to select the proper
inlet connector O-ring.
Do not scratch the O-ring groove. If the groove is scratched, the column
will leak.
3. Returntheinletconnectorassemblytothecolumn.Reactivatethe
O-ring.
Replacing a filter O-ring
This small O-ring is located at the edge of the filter skirt. With the filter off,
roll the O-ring off the connector tip and replace it with a new one.
f. Storing the Column
Storage considerations
Store a packed glass column according to the recommendations of the
manufacturer of the packing material. Note the following:
• Storethecolumnwithendplugsinplacetopreventcolumndrying.
• Usebacteriostaticsolutionsinappropriateconcentrations,forexample,
0.2%sodiumazideor30%ethanol.
• Removeallsaltsfromthecolumnbeforeaddinganorganicsolvent.
An organic solvent added to a salt can cause precipitation, resulting in
increased backpressure.
• Extendedstorageinhighsaltbuffersmayresultintheformationof
microcrystals which can lead to high backpressure. (Dissolve microcrys-
tals by slowly introducing a low ionic strength buffer).
• Donotallowthemobilephasetofreeze.Storageat4°Cisacceptable.
IV. sCalIng up separatIons
a. Scaling a Separation
When scaling an analytical separation to a preparative separation, use the
following isolation approach:
1. Define the objective by determining the:
•Complexityofthesamplemixture
•Componenttobeisolated
•Requiredquantity
•Degreeofpurity
•Propertiesofthecomponentsinthemixture
2. Perform crude separations to remove large quantities of extraneous
material or to isolate groups or classes of compounds. Simplify a com-
plex sample through coarse separation techniques, such as liquid-liquid
or solid-liquid extraction, crystallization, or precipitation. Centrifuge or
filter the sample to remove particulates.
3. Developtheseparationmethod.Tooptimizetheseparationforcom-
pounds of interest, make small-scale injections of the sample mixture,
varying the gradient.
4. Perform a loading study. Make progressively larger injections of the
sample mixture on a small scale column, such as the AP-Minicolumn,
to determine the effect of overload on resolution. This study defines the
amount of sample that can be separated in a column of a given volume.
5. Scaleuptheseparation.Translatethesmallscaleparameterstothe
large scale system values. Adjust the:
• Sampleload
• Flowrate
• Gradientvolume
b. Adjusting Sample Load and Flow Rate
When scaling the separation, use analytical and preparative
columns that are:
• Thesamelength
• Packedwiththesameparticlesizematerial