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.
Ultra High Performance Liquid Chromatograph
-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
(Max column length: 300 mm)
Compact column oven for LC/MS
front end
(Max column length: 50 mm)
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
Ultra High Performance Liquid Chromatograph
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
Shim-pack XR-ODS II (150 mmL. × 3.0 mmI.D., 2.2 µm)
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
Ultra High Performance Liquid Chromatograph
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
Shim-pack XR-ODS II (75 mmL. × 2.0 mmI.D., 2.2 µm)
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)
Flow rate 0.6 mL/min
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.
Flow rate 0.5 mL/min
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
Flow rate 0.5 mL/min
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.
Ultra High Performance Liquid Chromatograph
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
Flow rate 1.2 mL/min
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
Flow rate 0.4 mL/min
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
Ultra High Performance Liquid Chromatograph
Company names, product/service names and logos used in this publication are trademarks and trade names of Shimadzu Corporation or its affiliates, whether or not they are used with trademark symbol “TM” or “®”.Third-party trademarks and trade names may be used in this publication to refer to either the entities or their products/services. Shimadzu disclaims any proprietary interest in trademarks and trade names other than its own.
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
2