Waters ACQUITY UPLC BEH Glycan, 1.7 µm Columns, Glycan Performance Test Standard - Manual

[ CARE AND USE MANUAL ]
ACQUITY UPLC BEH Glycan, 1.7 �m Columns 1
CONTENTS
I. INTRODUCTION
II. GETTING STARTED
a. eCord ™ Installation
b. Column Connectors
c. Column Installation
d. Column Equilibration
e. Initial Column Efficiency Determination
f. Useful Functional Tests for Benchmarking a New Column
III. COLUMN USE
a. Sample Preparation
b. Operating pH Limit
c. Solvents
d. Pressure
e. Temperature
IV. TROUBLESHOOTING
V. COLUMN CLEANING, REGENERATION, AND STORAGE
a. Cleaning and Regeneration
b. Storage
VI. INTRODUCING eCORD INTELLIGENT CHIP TECHNOLOGY
a. Introduction
b. Installation
c. Manufacturing Information
d. Column Use Information
VII. ORDERING INFORMATION
ACQUITY UPLC BEH GLYCAN, 1.7 �m Columns
I. INTRODUCTION
Thank you for choosing Waters ACQUITY UPLC ® BEH Glycan column
designed for HILIC-mode separations of 2-aminobenzamide (2-AB)
labeled glycans.* This column chemistry and the UPLC
® instrument
conditions recommended in this manual are capable of separating
both neutral and charged species. Retention of 2-AB-labeled
oligosaccharides is based on the hydrophilicity of the molecule, a
parameter that is broadly related to hydrodynamic volume or
molecular size. The high resolving power of this column is due in part
to the small particle size (1.7 µm) of the fully porous packing material.
Chemical and mechanical stability of the column are the consequence
of Waters ethylene bridged hybrid (BEH) particle composition.
The column may be calibrated using a labeled dextran hydrolysate
ladder, such that elution may be expressed in terms of glucose units
(GU). Under the suggested chromatographic conditions, the retention
of 2-AB labeled oligosaccharides on the ACQUITY UPLC BEH Glycan
column may be predicted based on the hydrophilic contributions of
the individual constituent monosaccharides.
The ACQUITY UPLC BEH Glycan column is based on the 1.7 µm
particles that are characteristic of UltraPerformance LC
® . The small
particle size packing reduces dispersion and band broadening so
that improved resolution, sensitivity, and speed are obtained in
glycan separations. It is commonly expected that very high system
backpressures will be observed with such small particles. The high
pressure capability of the ACQUITY UPLC, ACQUITY UPLC H-Class,
and ACQUITY UPLC H-Class Bio systems is required for UPLC glycan
analysis. With HILIC mode, back pressure increases with the increasing
water content during the gradient. If an aqueous wash is chosen
before re-equilibration, the flow rate must be lowered to prevent
excessive back pressure.