User Manual

ManualsBrandsWaters ManualsWater equipmentXSelect HSS XP 2.5 µm Columns

XSelect HSS XP 2.5 µm Columns 1

[ CARE AND USE MANUAL ]

CONTENTS

I. INTRODUCTION

II. GETTING STARTED

a. Column Connection

b. Column Installation

c. Minimizing Band Spread Volume

d. Measuring Band Spread Volume

e. Measuring System Dwell Volume

f. Column Equilibration

g. Solvent Conversion for Normal-Phase Use

h. eCord Installation

i. Initial Column Efficiency Determination

j. VanGuard Pre-Columns

III. COLUMN USE

a. Sample Preparation

b. pH Range

c. Solvents

d. Pressure

e. Temperature

IV. COLUMN CLEANING, REGENERATION AND STORAGE

a. Cleaning and Regeneration

b. Storage after Reversed-Phase Use

c. Storage after HILIC Use

V. eCORD INTELLIGENT CHIP TECHNOLOGY

a. Introduction

b. Installation

c. Manufacturing

d. Column Use

VI. ADDITIONAL INFORMATION

a. Tips for Maximizing XSelect HSS XP 2.5 µm Column Lifetime

b. Troubleshooting Questions

c. Recommended Flow Rates and Anticipated Backpressures for

Reversed-Phase XSelect HSS XP 2.5 µm Columns

XSELECT HSS XP 2.5 µm COLUMNS

I. INTRODUCTION

Thank you for choosing a Waters XSelect

™

High Strength Silica [HSS]

eXtended Performance [XP] 2.5 µm Column. The manufacture of

XSelect HSS XP 2.5 µm Columns begins with ultrapure reagents and

are manufactured in a cGMP, ISO 9001 certified facility to control the

chemical composition and purity of the final product. Well-controlled

manufacturing processes result in industry-leading batch-to-batch

reproducibility. Every column is individually tested. A Performance

Chromatogram and Certificate of Batch Analysis are provided on the

eCord

™

Intelligent Chip.

XSelect HSS XP 2.5 µm Columns are based on the same base

particle technology and bonded phase chemistry as 1.8 µm ACQUITY

UPLC

HSS Columns as well as XSelect HSS 3.5 and 5 µm HPLC

Columns, thus enabling seamless transferability between HPLC,

UHPLC and UPLC

platforms.

XSelect HSS XP 2.5 µm Columns will exhibit maximum

chromatographic performance when used on a member of the

ACQUITY UPLC System family.

Summary of content (11 pages)

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[ CARE AND USE MANUAL ] XSELECT HSS XP 2.5 µm COLUMNS CONTENTS I. INT RODUC T ION Thank you for choosing a Waters XSelect™ High Strength Silica [HSS] I. INTRODUCTION eXtended Performance [XP] 2.5 µm Column. The manufacture of XSelect HSS XP 2.5 µm Columns begins with ultrapure reagents and II. GETTING STARTED a. Column Connection are manufactured in a cGMP, ISO 9001 certified facility to control the b. Column Installation chemical composition and purity of the final product. Well-controlled c.
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[ CARE AND USE MANUAL ] II. GET T ING STA RT ED c. Minimizing Band Spread Volume Each XSelect HSS XP 2.5 µm Column comes with a Certificate of Band Spreading is a measurement of the system dispersion that Analysis and a Performance Test Chromatogram embedded within impacts the chromatographic performance. Internal tubing diameter the eCord intelligent chip.
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[ CARE AND USE MANUAL ] 6. System Volume At 5 minutes, program a step to 100% B, and collect data for an additional 5 minutes. 7. Measure absorbance difference between 100% A and 100% B. 8. Measure time at 50% of that absorbance difference. 9. Calculate time difference between start of step and 50% point. 10. Multiply time difference by flow rate to calculate system volume. 5 Programmed time = 5 minutes 4.4 %h 50% Absorbance = 0.35852 AU Time = 5.
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[ CARE AND USE MANUAL ] h. eCord Installation Table 3: Column Volumes (mL) eCord Technology represents a significant advancement in column Column Length (mm) Internal Diameter 2.1 mm 3.0 mm 4.6 mm 30 0.10 0.21 0.50 50 0.17 0.35 0.83 75 0.26 0.53 1.25 100 0.35 0.71 1.66 usage tracking management which can be realized if the column is installed on an ACQUITY UPLC System.
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[ CARE AND USE MANUAL ] Note: VanGuard Pre-Columns are shipped with a collet and ferrule that are NOT III. COLUMN USE pre-swaged. This enables the end user to mate the VanGuard Pre-Column to To ensure the continued high performance of XSelect HSS XP 2.5 µm a specific XP 2.5 µm Column and ensures void-free and leak-free connections. Columns, follow these guidelines: Care must be taken when removing the O-ring that holds these two pieces on the pre-column tubing. a. Sample Preparation 1.
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[ CARE AND USE MANUAL ] Table 5: Buffer Recommendations for e. Temperature XSelect HSS XP 2.5 µm Columns XSelect HSS XP 2.5 µm Columns can be used up at intermediate Buffer Additive or pKa Buffer temperatures to enhance selectivity, reduce solvent viscosity and Used for Range Volatility (±1 pH Mass increase mass transfer rates. Comments Spec? unit) Chemistry Ion pair additive, can suppress TFA 0.
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[ CARE AND USE MANUAL ] b. Storage after Reversed-Phase Use For periods longer than four days, store the XP 2.5 µm Column in 100% acetonitrile. For separations utilizing elevated temperature, store immediately after use in 100% acetonitrile. Do not store columns in buffered eluents. If the mobile phase contained a buffer salt, flush the column with 10 column volumes of HPLC grade water (see Table 3 for column volume information) followed by 10 column volumes of acetonitrile.
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[ CARE AND USE MANUAL ] c. Manufacturing d. Column Use The eCord provides the user with the Batch Certificate of Analysis and The eCord Intelligent Chip provides the user with specific column Performance Test Chromatogram. information as well as column use data including: chemistry type, column dimension, serial number and part number.
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[ CARE AND USE MANUAL ] 3. b. Troubleshooting Questions Always remember to: 1. Are you using 100% aqueous mobile phases? 2. What is the age of the mobile phase? 3. Is the mobile phase filtered through a 0.2 µm membrane? Discard and re-prepare aqueous mobile phase every 4. Was the mobile phase prepared fresh or topped off? 24-48 hours (if 100% aqueous mobile phase is required). 5. Is the water source of adequate quality? 6.
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[ CARE AND USE MANUAL ] c. Recommended Flow Rates and Anticipated Backpressures for Reversed-Phase XSelect HSS XP 2.5 µm Columns XP 2.5 µm, 2.1 mm ID Columns (40 °C) Linear Velocity 3 mm/sec 4 mm/sec 5 mm/sec 6 mm/sec Column Dimension Flow Rate [mL/min] Backpressure [psi] Flow Rate [mL/min] Backpressure [psi] Flow Rate [mL/min] Backpressure [psi] Flow Rate [mL/min] Backpressure [psi] 2.1 x 30 mm 0.45 1760 0.6 2350 0.75 2940 0.9 3520 2.1 x 50 mm 0.45 2640 0.6 3520 0.75 4400 0.
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[ CARE AND USE MANUAL ] 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 5094-3788 Canada 1 800 252 4752 x2205 China 86 21 6879 5888 CIS/Russia +497 727 4490/290 9737 Czech Republic 420 2 617 1 1384 Denmark 45 46 59 8080 Finland 09 5659 6288 France 33 1 30 48 72 00 Germany 49 6196 400600 Hong Kong 852 2964 1800 The Netherlands 31 76 508 7200 Norway 47 6 384 60 50 Poland 48 22 6393000 Puerto Rico 1 787