c196-e079a nexera x2

C196-E079A

Ultra High Performance Liquid Chromatograph

Nexera X2

Maximizing the Potential of

UHPLC/HPLC Analyses

Resolution

Intelligence

Stability

SensitivityIntelligent data solutions, including a new peak calculation method for enhanced separation power, accelerate workflows.

The ultimate platform for high-sensitivity UHPLC and HPLC detection.

Enhanced peak resolution achieved by minimizing external column dispersion.

Superior stability for UHPLC/HPLC analyses.


-PDeAi

Utilizing patented technologies, and driven by customer input, Shimadzu has developed

the Nexera X2 series to deliver superior, real-world performance across a wider

application range, while delivering unparalleled flexibility and reliability in UHPLC/HPLC

analyses. With no limits and no compromises, the Nexera X2 achieves a new milestone in

the evolution of liquid chromatography.

4

Nexera Quaternary system Nexera Method Scouting system

Nexera MP system Nexera TQ system and Nexera SQ system

Nexera SR system Nexera UHPLC/HPLC system

Nexera X2 SeriesA full range of configurations to meet your applications needs…

features superior sensitivity, resolution and stability. * offers new separation technology.

is a flexible quaternary gradient system that is HPLC and UHPLC compatible.

offers comprehensive method development using up to 96 combinations of mobile phases and columns.

offers ultrafast LC/MS and LC/MS/MS analysis.

is an LC/MS front end HPLC for high-throughput analysis of multiple samples.

enables switching from UHPLC to HPLC in a single system using a selection of mixers and columns.

*intelligent Peak Deconvolution Analysis

-PDeAi

The CBM-20A enables system control via a network. The CBM-20A allows for various

system configurations to meet different laboratory workflows.

Nexera X2 Component

Ultra-high-sensitivity and high-resolution detector

The SPD-M30A realizes ultra-high sensitivity and minimized

peak dispersion, supporting the new separation calculation

method, . The temperature-controlled optics provide

stable baseline for genuine UHPLC analysis.

*intelligent Peak Deconvolution Analysis

SIL-30ACMP

SPD-M30A

Upgradable to binary gradient and quaternary gradient, the

LC-30AD also supports solvent blending for automated

mobile phase preparation.

LC-30AD

SIL-30AC SIL-30AC / Rack Changer II

CTO-30ASCTO-30A CTO-20AC

CBM-20A

D e t e c t o r

A u t o s a m p l e r

C o l u m n o v e n

S o l v e n t d e l i v e r y u n i t

Multifunctional autosampler;

pretreatment and loop injection

are available

Features control to a maximum 150°C

(Max column length: 150 mm)

Large space for placing multiple

columns and valves


Compact column oven for LC/MS

front end


Maximum 12 plates can be set with this

combination

Open-access autosampler with

a maximum capacity of 6 plates

5Ultra High Performance Liquid Chromatograph

-PDeAi

Intelligence in UHPLC/HPLC Analysis

Complete Separation of Co-eluted Peaks by ( intel l igent Peak Deconvolution Analysis, Patent pending)

Example 1: Remove tai l ing peak

Example 2: Impurity peak extracted from co-eluted peak

Vertical separationBaseline separation

Removes peak integration ambiguities

Step 2Extraction of single peak signal using

the difference between each spectrum

Step 1Analysis

Step 3Display of single component

representative peak

Component A

Component B

AU

1

2nm

Component A

shows extraction of component B.

shows extraction of component A.

:

:Component B

0.00 0.25 0.50 0.75 1.00 1.25 min

0.5 mg/L spiked

0.488 mg/L measured

(98% accuracy)

0.00 0.25 0.50 0.75 1.00 1.25 min

Target peak

Tailing peak

The target peak is on the tailing peak

This peak looks a single peak.

Remove the tailing peak!!

By removing the main peak, the impurity peak appears on the chromatogram!!0.22 0.24 0.26 0.28 min0.22 0.24 0.26 0.28 min

Peak purity curve

1

2

12

enables the extraction of a single peak from co-eluted peaks by utilizing differences in spectra. The new separation method removes

discussion of integration methods for co-eluted peaks. The also helps detect impurity peaks in a target peak.

-PDeAi-PDeAi

-PDeAi

-PDeAi

-PDeAi

6


Dramatical ly Extended Dynamic Range using ( intel l igent Dynamic Range Extension Calculator, Patent pending)

Simultaneous Quantitative Analysis of Components with Large Differences in Concentrations.

An example of the simultaneous quantitative analysis of acetaminophen, caffeine, and an impurity, which are included in an over-the-counter cold

medicine, is shown below. Calibration curves of high-level acetaminophen and a trace-level impurity can be created from the single analysis data.

is a new analytical method that significantly extends the dynamic range. It enables the analysis of high-concentration samples

without diluting them, and ensures a correct calibration curve.

-DReCi

0

4000mAU wavelength= λa

Due to saturation in the higher concentration region, calibration points shifts.

0

200 λa

0

250*λb

For calibration points in the higher concentration range,area values are calculated by shifting the wavelength automatically using the software. Correct calibration points are plotted after correction.

*

Area values are plotted by conventional calculation.

Automatic correction by

Automatic correction in (*) area

concentration

Area

concentration

Area

concentration

Area

Calibration curve after correction

Conditions

Column

Mobile phase

Shim-pack XR-ODS II (150 mmL. × 3.0 mmI.D., 2.2 µm)

Since three components with large differences in concentrations can be analyzed simultaneously, the effort spent changing the dilution ratio from component to component is eliminated.

Acetaminophen Caffeine Impurity

0.4 AU

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 min

0

250

500

750

1000

1250

1500

1750

2000

mAU

1.25 1.50 1.75 2.00 2.25 2.50 min

0102030405060708090

100mAU

1: 100 mg/L 1: 500 mg/L(Saturation)

2: 42 mg/L

Impurity

0 10 20 30 concentration 0

1.0

2.0

3.0

4.0

5.0

6.0Area (×107)

0 100 200 300 400 concentration 0

0.5

1.0

1.5 R2 = 0.9997416

10 AU

R2 = 0.9999641

0 100 200 300 400 concentration 0

0.5

1.0

1.5Area (×105)Area (×105)

R2 = 0.9998100

0.012 AU

Flow rate 0.9 mL/min (Gradient elution)

A: 10 mM phosphoric acid (sodium) buffer (pH 2.6) / 50 mM Sodium perchlorateB: A / ACN = 30 / 70

Sample 1, Acetaminophen: 10–500 mg/L2, Caffein: 0.84–42 mg/L

Detection PDA 254 nm

-DReCi

Saturated regionLow concentration region

-DReCi

The dynamic range is extended by and the calibration curve from low to high concentration is completed.

-DReCi

Example of calibration curve correction by -DReCi

-DReCi

7

Ultimate Sensitivity

Achieving Maximum Sensitivity

The SPD-M30A photodiode array detector adopts the high-sensitivity, low-dispersion capillary cell, SR-Cell, which achieves a

0.4×10-5 AU noise level. The SPD-M30A offers the ultimate in UHPLC analysis, providing high-sensitivity and high-resolution.

An optional higher-sensitivity cell is also available.

Superior Dynamic Range

The SPD-M30A achieves a superior dynamic range with the standard cell. Although a trace-level peak of 0.005% was added in the

following example, the simultaneous quantitative analysis of the main peak and the trace peak is possible. If a wider dynamic range is

needed, 　　　 makes it possible.

Higher Sensitivity UHPLC Analysis

UHPLC analysis is susceptible to sample diffusion, which occurs inside a cell. However, if using the SR-Cell, sharper peaks can be

obtained. In the following example, the signal intensity of the impurity peaks improved, and the S/N ratio is about double.

If using the SPD-M30A, trace-level impurities can be detected with high-sensitivity.

Example of analysis when adding impurities of 0.005% for a main component

About twice S/N

1.00 1.50 2.00 2.50 min

*

*

*

**

*

*

*

*

* Degradation compound

Conditions

Column

Mobile phase

Flow rate

Sample

Detection

0.9 mL/min

Cefazolin

PDA 245 nm


A: 20 mmol/L phosphoric acid buffer (pH2.5)B: Acetonitrile Gradient elution

: SPD-M30A : Current model (UHPLC system)

-DReCi

µAU

mAU

1750

600

S/N 9.53

0.5 0.7 0.9 min

400

200

0

1250

750

250

0

0.0 0.5 1.0 min

Mai

n pe

ak

Impurity peak

Column Shim-pack XR-ODS lll (50 mmL. × 2.0 mmI.D., 1.6 µm)

Flow rate 0.6 mL/min

Main peak propyl 4-hydroxybenzoate

0.005% Impurity peak ethyl 4-hydroxybenzoate

Conditions

High-sensitivity and high-resolution

SR-Cell

Section of capillary cell

The SR-Cell features high-sensitivity and low-dispersion by adopting a total internal reflection capillary cell.

8


High-sensitivity Cell (optional)

An optional high-sensitivity SR-Cell features a light path of 85 mm, longer than the standard cell, allowing it to detect the most

minute peaks.

Example of Pharmaceutical Impurity

The high-sensitivity cell can detect impurities in pharmaceuticals which are difficult to detect using the standard cell. Valsartan and

decomposition product were analyzed using both cells by the Nexera SR system. As shown, analysis of a trace-level impurity is possible using the

high-sensitivity cell.

0.0 2.5 5.0 min

1

2

3

4

5

6

7

: High-sensitivity cell : Standard cell

Conditions

Column

Mobile phase

Flow rate

Sample

Detection


50 % Acetonitrile Solution

1.0 mL/min

Alkylphenone

PDA 245 nm

S/N comparison of standard cell and high-sensitivity cell

Peak 1 2 3 4 5 6 7

High-sensitivity cell 62684 49113 39355 28628 18415 10061 4936

Standard cell 13121 9357 7287 5066 3178 1721 830

Relative difference 4.8 5.2 5.4 5.7 5.8 5.8 5.9

0.0 0.5 1.0 1.5 2.0 2.5 min0

500

1000

1500

2000

mV

Valsartan

* Degradation compound

*

1.0 1.5 2.0 2.5 min

0

1

2

mV

** *

Peak intensity: 6.9 times higher

: High-sensitivity cell : Standard cell

Conditions

Column

Mobile phase

Flow rate

Kinetex 2.6u XB-C18 100A (100 mmL. × 3.0 mmI.D., 2.6 µm)

Water / Acetonitrile / Acetic acid = 500 / 500 / 1

1.5 mL/min

9

10

Maximized Resolution

Superior peak shape for maximum peak resolution

In ultrafast analysis, external column dispersion affects peak resolution. The SR-Cell in the SPD-M30A has minimized the cell volume to

realize excellent peak shape with higher peak resolution, supporting genuine UHPLC analysis.

Superior spectrum resolution and l inearity

The SPD-M30A provides unrivaled spectrum resolution by adopting an optimized optic system. In addition, the new signal treatment

technology realizes excellent spectrum linearity across a wide area from low concentration to high concentration. The SPD-M30A supports

analyses, such as purity analysis, that require a wide dynamic range.

Nexera SR system for high-sensitivity and high-resolution

Nexera SR system is configured with the new SPD-M30A photodiode array detector.

The new peak calculation technology, , proposes a new approach for separation

of co-eluted peaks and extraction of impurity peaks from main peaks. The superior

sensitivity and resolution expand the range of UHPLC analyses.

A benzene spectrum is often used for evaluation of spectrum resolution.

The SPD-M30A shows world’s highest* spectrum resolution, B/A=2.90.

The SPD-M30A shows excellent spectrum linearity in the analysis of

caffeine samples with different concentrations.

Column Shim-pack XR-ODS III(50 mmL. × 2.0 mmI.D., 1.6 µm)


Peak No. Nexera X2 Other vendor

1

2

3

1.309

1.297

1.273

1.546

1.486

1.432

0.20 0.25 0.30 0.35 0.40 0.45 0.50 min

peak

1

peak

2

peak

3

Nexera X2

Other vendor's UHPLC

230 240 250 260 270 280 nm

0

B

A

B/A=2.90

Caffeine 100 mg/L

Caffeine 1000 mg/L

250 300 350 nm(Normalize)

mAU

Similarity 0.9998

-PDeAi

Nexera SR system* As of November 2012, according to Shimadzu survey.

Tailing factor (Tf5%)

11

Unrivaled Stability

Baseline stabil ization by temperature-controlled TC-Optics

The SPD-M30A adopts the new temperature-controlled TC-Optics with the SR-Cell that has an optimized heat exchange inlet pipe.

These features achieve faster stabilization with low external dispersion for UHPLC analysis.

The faster stabilization shortens the “wait time” for analysis, thereby increasing total analytical throughput.

Stable performance against room temperature shift

The SPD-M30A incorporates TC-Optics to achieve a more stable baseline against room temperature shifts.


Mobile phase Methanol

Detector PDA 254 nm

Temperature RT

Other vendor's PDA detector

SPD-M30A

0 10 20 30 40 50 min

-40

-30

-20

-10

10

0

mAU

SPD-M30A

Baseline after instrument startup


Mobile phase Methanol

Detector UV 254 nm

0 100 200 300 400 500 600 min-20

-15

-10

-5

0

5

10

15

20

25

mAU

254 nm

Room temperature

Other vendor's PDA detector

SPD-M30A25°C

30°C

The SPD-M30A shows a stable baseline faster after

startup while the detector from other vendor

shows baseline drift during 1 hour.

12

Measured pH of mobile phase prepared by solvent blending

Combination of UHPLC / HPLC Switching system and solvent blending

Blending 1 3.010

Blending 2 7.192

3.004

Solvent blending Manual preparation

7.191Blending 1 A / B = 6 / 4

Blending 2 A / B = 2 / 8

Pump Y solvent Methanol

A: 20 mmol/L phosphoric acid aqueous solutionB: 20 mmol/L disodium hydrogen-phosphate

Pump X solvent

Ultimate Flexibility

The modular nature of Nexera X2 allows a

solvent blending function that can mix

solvents according to desired ratios. This

function enables dilution of solvent, buffer

preparation and modifier addition for

mobile phase preparation.

Buffer pH can be optimized using the

solvent blending function as shown to the

right chromatograms. The function is a

useful tool for optimizing mobile phase

conditions in method development.

Automation of mobile phase preparation by solvent blending function

Solvent deliveryunit 1

Solvent deliveryunit2

A

B

C

D

A

B

C

D

Y

X

85%

10%

50%

20%

5%

25%

5%

0%

Solvent BlendingAutomated mobile phase preparation

Binary GradientGradient elution by prepared solvents

0.0 1.0 2.0 3.0 4.0 min

Blending 1

1, 2, 3

pH 3.0A / B = 6 / 4

0.0 1.0 2.0 3.0 4.0 min

Blending 2

1

23

pH 7.2A / B = 2 / 8

1. Benzoic ac id2. Sorbic ac id3. methyl 4-hydroxybenzoate

Nexera UHPLC/HPLC system

The solvent blending function can be added to a Nexera UHPLC/HPLC switching system by

upgrading the configuration. The system supports all processes from method development to

method transfer from HPLC to UHPLC.


Both the SIL-30AC and SIL-30ACMP support the use of multiple

rinse solvents for rinsing the needle’s outer surface and inner

surface to thoroughly minimize carryover.

The carryover level was extremely low even after 30,000 injections.

13

Nexera X2 autosamplers support ultrafast injections to increase analytical throughput. In particular, the SIL-30ACMP achieves a 7-second

injection time and a 14-second cycle time, making it ideally suited for ultrafast analysis.

Ultrafast 7-second injection and 14-second cycle t ime

The Nexera X2 autosamplers, SIL-30AC and SIL-30ACMP, provide near-zero carryover. The improved design maintains a low carryover

level, even during long-term usage, to ensure reliable analysis.

Minimizing carryover with multiple r inse solvents

Enhanced UHPLC Reliability

0

25000

50000

75000

100000

125000

1:235.40>86.10(+)2:256.10>167.10(+)3:281.10>86.10(+)

0.0 0.25 0.5 0.75 1.0 min

14 sec

0.0 0.5 1.0 1.5 2.0 2.5 min

-25

0

25

50

75

100

125

150

175

200

mAU

2.25 2.50 2.75 min

0

250

500

750uV

0.00018%

Shim-pack XR-ODS II (30 mmL. × 1.5 mmI.D., 2.2 µm) Column


Event #

1

2

3

Q1 m/z

235.4

256.1

281.1

Q3 m/z

86.1

167.1

86.1

Compound

Lidocaine

Diphenhydramine

Imipramine

Shim-pack XR-ODS III (150 mmL. × 2.0 mmI.D., 2.2 µm)Column

Mobile phase Water / Methanol = 7 / 3


Sample Caffeine 4000 mg/L

Contact surfaces of needle and needle port

where carryover is likely to occur

Port section is rinsed by any of the rinse solutions(1 to 3 solutions)

Discharge to drain port

Needle port(schematic diagram)

Drain

Multi-rinse mechanism canaccommodate up to 4 solutions

Sample loop

Needle

Rinse ports

For rinsing needle surface andinner surface (3 types)

For rinsing needlesurface (1 type)

R0 R1 R2 R3


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For Research Use Only. Not for use in diagnostic procedures. The contents of this publication are provided to you “as is” without warranty of any kind, and are subject to change without notice. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication.

© Shimadzu Corporation, 2013www.shimadzu.com/an/Printed in Japan 3655-01302-30ANS

Nexera X

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c196-e079a nexera x2

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