User Manual

ManualsBrandsWaters ManualsWater equipmentACQUITY UPLC CSH130 C18 1.7 μm and XP 2.5 μm Columns

[ CARE AND USE MANUAL ]

ACQUITY UPLC CSH130 C

Columns 1

Thank you for choosing a Waters ACQUITY UPLC

CSH

™

column.

ACQUITY UPLC CSH130 C

, 1.7 µm and XP 2.5 µm columns feature

Waters Charged Surface Hybrid (CSH) Technology which provides

excellent peak shape, high efficiency and loading capacity for basic

compounds when using acidic, low ionic strength mobile phases.

This same particle technology is used in the XSelect

™

CSH family of

HPLC columns, thus enabling seamless transferability between HPLC

and UPLC

system platforms. The ACQUITY UPLC CSH130 packing

materials were designed specifically for use with the ACQUITY UPLC

systems and are manufactured in a cGMP, ISO 9001 certified

manufacturing facility using ultra pure reagents. Each batch of

ACQUITY UPLC CSH130 C

material is tested chromatographically

with acidic, basic, and neutral analytes as part of qualification for

use in peptide mapping. The results are held to narrow specification

ranges to assure excellent, reproducible performance. ACQUITY UPLC

CSH130 C

batches are also QC tested with a gradient separation

of a tryptic digest of cytochrome c using 0.1% formic acid containing

eluents. Finally, every shipped column is individually tested for packed

bed efficiency and a Performance Chromatogram and Certificate of

Batch Analysis are provided on the eCord

™

intelligent chip. ACQUITY

UPLC CSH130 C

columns were designed and tested specifically for

use on ACQUITY UPLC systems. ACQUITY UPLC CSH130 C

columns

will exhibit maximum chromatographic performance and benefits ONLY

when used on holistically-designed ACQUITY UPLC systems since these

systems and columns were created and designed to operate together.

For these reasons, Waters cannot support the use of ACQUITY UPLC

columns on any system other than an ACQUITY UPLC system.

CONTENTS

I. GETTING STARTED

a. Column Connectors

b. Column Installation

c. Column Equilibration

d. eCord Installation

e. Initial Column Efficiency Determination

f. VanGuard Pre-Columns

II. COLUMN USE

a. Sample Preparation

b. pH Range

c. Solvents

d. Pressure

e. Temperature

III. COLUMN CLEANING, REGENERATING AND STORAGE

a. Cleaning and Regeneration

b. Storage

IV. INTRODUCING ECORD INTELLIGENT CHIP TECHNOLOGY

V. REPRESENTATIVE TEST CHROMATOGRAPH AND CONDITIONS

FOR SEPARATION OF PROTEIN DIGEST

VI. ADDITIONAL INFORMATION

ACQUITY UPLC CSH130 C

1.7 µm AND XP 2.5 µm COLUMNS

Summary of content (9 pages)

PAGE 1
[ CARE AND USE MANUAL ] ACQUITY UPLC CSH130 C18 1.7 µm AND XP 2.5 µm COLUMNS CONTENTS Thank you for choosing a Waters ACQUITY UPLC® CSH™ column. ACQUITY UPLC CSH130 C18, 1.7 µm and XP 2.5 µm columns feature I. GETTING STARTED Waters Charged Surface Hybrid (CSH) Technology which provides excellent peak shape, high efficiency and loading capacity for basic a. Column Connectors b. Column Installation c. Column Equilibration d. eCord Installation e. Initial Column Efficiency Determination f.
PAGE 2
[ CARE AND USE MANUAL ] I. GETTING STARTED 3. When the mobile phase is flowing freely from the column outlet, stop the flow and attach the column outlet to the detector. This Each Peptide Separation Technology ACQUITY UPLC CSH130 C18 column prevents entry of air into the detection system and gives more comes with Certificate of Analysis and a Performance Test Chromatogram rapid baseline equilibration. embedded within the eCord intelligent chip.
PAGE 3
[ CARE AND USE MANUAL ] d. eCord Installation Installation Instructions The eCord button should be attached to the side of the column heater module. 1. The eCord button is magnetized and does not require specific orientation. remove plastic plug. e. Initial Column Efficiency Determination 1. 2. during shipping (collet and ferrule are not yet permanently attached). the “Performance Test Chromatogram”, to analyze the column 3. upon receipt.
PAGE 4
[ CARE AND USE MANUAL ] Table 2. Buffer Recommendations for Using ACQUITY UPLC CSH130 C18 Columns from pH 1 to 11 Additive/Buffer pKa Buffer range Volatility (±1 pH unit) Used for Mass Spec TFA 0.3 - Volatile Yes Ion pair additive, can suppress MS signal, used in the 0.02-0.1% range. Acetic Acid 4.76 - Volatile Yes Maximum buffering obtained when used with ammonium acetate salt. Used in 0.1-1.0% range. Formic Acid 3.
PAGE 5
[ CARE AND USE MANUAL ] Table 3. Column Cleaning Sequence At the time of manufacture, tracking and quality control information will be downloaded to the eCord. Storing this information on the chip will eliminate Polar Samples 1. water 2. methanol 3. isopropanol Proteinaceous Samples the need for a paper Certificate of Analysis.
PAGE 6
[ CARE AND USE MANUAL ] d.Customer Use Information The eCord chip will automatically capture column use data. The top of the screen identifies the column including chemistry type, column dimensions and serial number. The overall column usage information includes the total number of samples, total number of injections, total sample sets, date of first injection, date of last injection, maximum pressure and temperature.
PAGE 7
[ CARE AND USE MANUAL ] V. REPRESENTATIVE TEST CHROMATOGRAPH AND CONDITIONS FOR SEPARATION OF A CYTOCHROME C TRYPTIC DIGEST 1.20 1.10 1.00 0.90 0.80 0.70 0.60 0.50 0.40 0.30 0.20 0.10 0.00 2 3 4 5 6 7 8 9 10 11 12 Min. Peak Identification T1 T13-T14 T14 T4 T9-T10 T10 T8 T15 T19C T19 T12-T13 T12 T5 N-AcGDVEK KYIPGTK YIPGTK IFVQK KTGQAPGFSYTDANK TGQAPGFSYTDANK TGPNLHGLFGR MIFAGIK EDLIAY EDLIAYLK GITWGEETLMEYLENPKK GITWGEETLMEYLENPK CAQCHTVEK (heme attached) Retention Time (min) T19C Result 6.
PAGE 8
[ CARE AND USE MANUAL ] VI. ADDITIONAL INFORMATIONA. • Old aqueous mobile phases. Remember to rinse bottles thoroughly and prepare fresh every 24-48 hours. Tips for maximizing ACQUITY UPLC CSH130 C18 Column Lifetimes • Using phosphate salt buffer in combination with high ACN concentrations (e.g., > 70%) due to precipitation. 1. To maximize ACQUITY UPLC CSH130 C18 column lifetime, pay close 5.
PAGE 9
[ CARE AND USE MANUAL ] Sales Offices Austria and European Export (Central South Eastern Europe, CIS and Middle East) 43 1 877 18 07 Australia 61 2 9933 1777 Belgium 32 2 726 1000 Brazil 55 11 4134 3788 Canada 1 800 252 4752 China 86 21 6156 2666 Czech Republic 420 2 617 11384 Denmark 45 46 59 8080 Finland 358 9 5659 6288 France 33 1 30 48 72 00 Germany 49 6196 400 600 Hong Kong 852 2964 1800 Hungary 36 1 350 5086 Norway 47 6 384 6050 Poland 48 22 101 5900 Puerto Rico 1 787 747 8445 Russia/CIS 7 495 727 4