Waters System Evaluation Matrix 1 User Manual

[ CARE AND USE MANUAL ]

SYSTEM EVALUATION MATRIX #1

CONTENTS

I. INTRODUCTION

II. STORAGE & STABILITY

III. RECONSTITUTION OF THE SYSTEM

EVALUATION MATRIX #1

IV. USE WITH AN ANALYTICAL STANDARD

V. EXAMPLE OF USING THE SYSTEM EVALUATION

MATRIX #1 & DATA AQUISITION

VI. ORDERING INFORMATION

VII. ADDITIONAL RESOURCES

I. INTRODUCTION

This System Evaluation Matrix #1 is intended for use as a

reproducible background signal in the evaluation of LC and MS

system performance.

It is especially suitable for standard small molecule applications

where a complicated matrix is required, but naturally occurring

alternatives, such as urine and plasma, are impractical.

Typically additional application-specific components are added to

the matrix to provide analytical markers, although it may be used

without these.

The matrix provides a background in both positive and negative ion.

II. Storage and Stability.

The lyophilized powder is shipped at ambient temperatures, in

Waters TruView™ Max Recovery Vials (P/N 186005668cv). The

prepared matrix is stable for at least one week. Beyond this time

for performance testing and evaluation, fresh re-preparation of the

matrix is recommended.

System Evaluation Matrix #1 1

[ CARE AND USE MANUAL ]

III. Reconstitution of the System Evaluation Matrix #1

1. The vial contains 33.33 mg of lyophilized matrix. Add 1 mL of

water to the vial to make a concentration of 33.33 mg/mL.

2. Recap and then agitate the vial contents (e.g. via vortexing or

ultrasonic bath), to ensure the lyophilized powder dissolves.

3. For use on SYNAPT® G2-S and detectors of similar sensitivity

dilute 1:20 in water.

IV. Use with an Analytical Standard

The matrix provides an abundance of peaks. By adding an analytical

standard the matrix can be used as a background.

Waters Analgesic Mix (P/N 186006350) can be added to provide

a collection of small molecules suitable for metabolomics method

development. Refer to note 6 in section VII.

• Prepare a 200 ng/µL stock by adding 1 mL acetonitrile to the

Analgesic mix vial.

• Prepare 50 ng/µL working concentration by taking 25 µL of the

stock and diluting with 75 µL of water.

• Add 5 µL of 50 ng/µL Analgesic Mix to 495 µL of the matrix to

make a final concentration of 0.5 ng/µL.

V. Example of Using the System Evaluation Matrix #1 and
Data Aquisition

Figure 1 shows a representative chromatogram of the control matrix

diluted 1:20 on a SYNAPT G2-S.

VI. ORDERING INFORMATION

To order these products, contact your nearest subsidiary, or visit

www. waters.com and click on Order Center.

Description Part number

System Evaluation Matrix #1 186006962

Analgesic Mix 186006350

VII. Additional Resources

1

ACQUIT Y UPLC® I-C lass Documentation CD (715003160)

2

Symmetry® Columns Care and Use Manual (WAT047278).

3

SYNA PT G2-S HDMS online Help (access from the Waters MassLynx® sof tware) .

4

MassLynx online Help (access from t he Waters MassLynx software).

5

ACQUIT Y UPLC I- Class System online Help (access from Waters MassLynx software).

6

TransOmics™ Informatics for Metabolomics and Lipidomics User Guide

For additional, detailed information, refer to the Waters Omics Research Platform Solution for
Metabolomics and Lipidomics System Guide (71500 3815).

You can also consult t he Waters Web site, www.waters.com/omics.

Figure 1. Analysis of Metabolites in System Evaluation Matrix #1.

UPLC Condition s:
Colum n: ACQU ITY UP LC HSS T3, 1.8 μm , 2.1 x 100 mm
Colum n Temp.: 40 ° C
Sampl e Temp.: 6 °C
Flow R ate: 500 µL /min
Injecti on Vol.: 2 µ L, 6 s post injec tion strong wa sh
Run Tim e: 12 mins
Solvent A: w ater/0.1% formic acid
Solvent B: ac etonitrile /0.1% form ic acid
Wash : water/0 .1% formic a cid
Seal Wa sh: 50:50 wa ter/methano l

Source Parameters:

Tune for ma ximum sensi tivity
Capill ary: 3.5 kV
Samplin g cone: 25 V
Sourc e temperatu re: 120 °C
Desolvat ion temperat ure: 500 °C
Desolvation/Nebulizer/C one gas: 1000/6/50 L/hr
Sourc e offset: 80 V

LockS pray Soluti on: 0 .2 ng/µL le ucine enkep halin in 50% met hanol/5 0% water + 0.1% formic ac id (556.2771 Da)

System Evaluation Matrix #1 2

Gradient
Time (min) %A %B Curv e

0.0 9 9 1 Initial

1.0 9 9 1 6

3.0 85 15 6

6.0 5 0 50 6

9.0 5 95 6

10.0 5 95 6

10.1 99 1 6

MS Meth od Parameter s:

Tune for ma ximum sensi tivity
Ionizati on mode: ESI+
TOF acquisition mode: Resolution
Acquis ition method : Continu um MS
TOF ma ss range: 50 -1200 Da
Scan ti me: 0.1 s
Collisio n energy funct ion 2: Trap CE ra mp 20 to 40 V

E