Shim-pack Velox LC Columns - Maximize LC Separation Performance with Core Shell Technology
Summary
Importance of Topic
Modern liquid chromatography demands high efficiency, rapid analysis, and robust performance to support diverse applications in pharmaceuticals, environmental monitoring, food safety, and clinical research. Core-shell particle technology has emerged as a key advance, offering improved separation power and throughput without requiring ultrahigh pressure systems.
Study Objectives and Overview
This application note describes the design and performance of Shimadzu Shim-pack Velox LC columns featuring core-shell technology. It aims to demonstrate their separation efficiency, method transfer potential, ruggedness, and compatibility across multiple Shimadzu LC platforms. Specific case studies illustrate resolution of complex mixtures, stability under harsh conditions, and compliance with USP method adjustment guidelines.
Used Methodology and Instrumentation
- Shim-pack Velox core-shell columns in chemistries: SP-C18, C18, Biphenyl, PFPP, HILIC with 1.8, 2.7, and 5 µm particle sizes
- Shimadzu LC systems: Prominence HPLC, Prominence-i Plus UHPLC-like, Nexera XR, Nexera-i Plus, Nexera X2 UHPLC
- LC-MS detection: Nexera UHPLC system coupled to LCMS-8050 triple quadrupole mass spectrometer
- Guard column cartridges: Shim-pack Velox EXP Guard and UHPLC Precolumn Filters
Main Results and Discussion
- High-efficiency separations: Core-shell columns delivered increased theoretical plate counts and faster run times compared to fully porous phases.
- Column selection guide: Recommendations for particle size, column dimensions, and system dispersion on different Shimadzu platforms.
- Acid stability: SP-C18 phase maintained consistent retention at pH 1 over extended runtimes, outperforming alternative core-shell and FPP C18 columns.
- Complex mixture resolution: Simultaneous analysis of 22 preservative agents in cosmetic and food products achieved in under 10 minutes.
- USP method transfer: Vitamin B12 assay transferred from 5 µm fully porous to 2.7 µm core-shell columns within allowable L/dp and N adjustments, reducing analysis time and solvent use.
- Isomer separation: Biphenyl and PFPP chemistries successfully resolved structural and cis/trans isomers that are challenging on standard C18 phases.
- HILIC performance: Unbonded core-shell HILIC phase enhanced retention and separation of polar nucleosides, offering orthogonal selectivity to reversed-phase.
- Lot-to-lot reproducibility: Consistent peak shapes and retention times across multiple manufacturing lots demonstrated column reliability.
Benefits and Practical Applications
- Improved resolution and throughput maximize laboratory productivity and reduce analysis costs.
- Wide pH stability and robust ligand protection support demanding LC/MS workflows.
- Versatile chemistries cover a broad spectrum of analyte classes, from small polar molecules to hydrophobic compounds.
- Method transfer capability simplifies scale-up and regulatory compliance in quality control environments.
- Guard column and precolumn filter options further protect columns and extend operational life.
Future Trends and Potential Applications
Ongoing developments in core-shell particle design are expected to drive further improvements in separation speed and efficiency. Integration with high-resolution mass spectrometry, microflow LC, and automated sample preparation will expand applicability in proteomics, metabolomics, and real-time process monitoring. Moreover, emerging stationary phase chemistries tailored for chiral separations and ion-exchange interactions may broaden the utility of core-shell formats.
Conclusion
Shimadzu Shim-pack Velox core-shell columns deliver high-efficiency separations, robust performance, and versatile selectivity across common and challenging LC applications. Their compatibility with existing Shimadzu LC and LC/MS platforms, combined with simplified method transfer and reproducible manufacturing, make them a valuable tool for analytical laboratories seeking to enhance throughput and data quality.
Reference
No specific literature references were cited in the original document.
Content was automatically generated from an orignal PDF document using AI and may contain inaccuracies.
Shim-pack V
elox LC Columns
C190-E219
Maximize LC Separation Performance with Core Shell Technology
Shim-pack Velox LC Columns
© Shimadzu Corporation, 2018
www.shimadzu.com/an/
For Research Use Only. Not for use in diagnostic procedures.
This publication may contain references to products that are not available in your country. Please contact us to check the availability of these
products in your country.
Company names, products/service names and logos used in this publication are trademarks and trade names of Shimadzu Corporation, its
subsidiaries 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, whether or not
they are used with trademark symbol “TM” or “®”.
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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.
First Edition: September 2018, Printed in Japan 3655-08801-20AIK
Free-turning architecture lets you change cartridges by hand without breaking inlet / outlet fluid connections — no tools needed.
Guard column cartridges require Shim-pack Velox EXP Direct Connect Holder (227-32041-01)
Shim-pack Velox EXP Guard Column
Tubing
Nut
Ferrule
Peek End
Ferrule
Peek End
Holder Cap
Holder Body
Guard Column
Cartridge
Ordering Information
Shim-pack Velox 5 µm Columns
Shim-pack Velox EXP Guard Column Cartridge (3 pk)
Shim-pack Velox UHPLC Precolumn Filter (0.2 µ
ｍ)
＊Shim-pack Velox EXP Direct Connect Holder : 227-32041-01
Chemistry
50
100
150
250
SP-C18
227-32006-01
227-32006-02
227-32006-03
227-32006-04
C18
227-32012-01
227-32012-02
227-32012-03
227-32012-04
Biphenyl
227-32018-01
227-32018-02
227-32018-03
227-32018-04
4.6
PFPP
227-32024-01
227-32024-02
227-32024-03
227-32024-04
Type
ID (mm)
2.1
3.0
4.6
UHPLC
SP-C18
227-32028-01
227-32028-02
–
C18
227-32031-01
227-32031-02
–
Biphenyl
227-32034-01
227-32034-02
–
PFPP
227-32037-01
227-32037-02
–
SP-C18
227-32029-01
227-32029-02
227-32029-03
2.7 µ
ｍ
C18
227-32032-01
227-32032-02
227-32032-03
Biphenyl
227-32035-01
227-32035-02
227-32035-03
PFPP
227-32038-01
227-32038-02
227-32038-03
HILIC
227-32040-01
227-32040-02
227-32040-03
Type
ID (mm)
4.6
5 µ
ｍ
SP-C18
227-32030-01
C18
227-32033-01
Biphenyl
227-32036-01
PFPP
227-32039-01
Part No. 1 pack
Part No. 5 pack
Part No. 10 pack
227-32042-01
227-32042-02
227-32042-03
To minimize extra column volume and maximize UHPLC sample throughput with SPE,
SLE, or other sample preparation techniques, pair 1.8 µm Shim-pack Velox UHPLC
columns with an Shim-pack Velox UHPLC Precolumn filter instead of a guard cartridge.
Shim-pack Velox UHPLC Precolumn Filter (0.2 µm)
ID(mm)
Length(mm)
3
2
Maximize LC Separation Performance
with Core Shell Technology
Designed to maximize performance of LC systems, Shimadzu’s Shim-pack
Velox columns with core shell technology enable you to achieve increased
separations and faster analysis times on any LC platform.
Whether developing a high efficiency LC separation method, transferring an
existing method for increased throughput while maintaining resolution, or
are trying to improve the resolution of a complex separation, Shim-pack
Velox columns will satisfy your needs.
Column ruggedness is critical to any LC analysis and Shim-pack Velox
core-shell columns deliver excellent column lifetime for even the most
challenging sample matrices.
Increased resolution with maximum efficiency
improving separation and detection
Faster separation without sacrificing performance
maximizing laboratory productivity and reducing cost of analysis
Increased sample throughput
reducing overall analysis time
Superior ruggedness
reducing cost of analysis
Excellent reproducibility
maintaining analysis and data integrity
Prominence
Prominence-i plus
Nexera XR
Nexera-i plus
Nexera X2
* For maximum lifetime, recommended maximum pressure for 1.8 µm particles is 80MPa.
Shim-pack Velox column will deliver
Combining highly efficient core shell particle technology with a wide range of surface chemistries provides you with the best
opportunity for optimal resolution. With different chemistry characteristics, Shim-pack Velox columns are suitable for use in a wide
variety of applications and challenging separations.
Column Chemistries
We understand that lot to lot consistent performance of columns is required to maximize your laboratory performance. From one lot
to the next, every Shim-pack Velox column you purchase will perform consistently.
Lot to Lot Reproducibility
Column particle sizes and column volumes affect chromatography results significantly if the column configuration does not match
the LC system. As column particle size is reduced, or the column volume (ID and/or the length of the columns) decreases, the
necessity for a lower dispersion system is increased.
Choosing the optimal column configuration for your LC system allows you to achieve improved chromatography. The following
table summarizes the starting recommendations of column configuration for each LC system.
Column Selection Guide for Different LC Platforms
LC System
Particle size
Column I.D.
Column Length
USP Classification
Ligand type
Particle Size (µm)
Pore size (
Å)
SP-C18
L1
Sterically protected C18
1.8, 2.7, 5
90
125 m2/g
130 m2/g
100 m2/g
7 %
7 %
5 %
No
1.0-8.0
100 MPa*
60 MPa
40 MPa
C18
L1
C18
1.8, 2.7, 5
90
125 m2/g
130 m2/g
100 m2/g
9 %
7 %
5 %
YES
2.0-8.0
100 MPa*
60 MPa
40 MPa
Biphenyl
L11
Biphenyl
1.8, 2.7, 5
90
125 m2/g
130 m2/g
100 m2/g
7 %
7 %
5 %
YES
1.5-8.0
100 MPa*
60 MPa
40 MPa
Compounds (50 ng/mL) :
1. Cortisol
2. 11-Deoxycortisol
3. Estradiol
4. Boldenone
5. Testosterone
6. Androstenedione
7. Progesterone
Lot 1
Lot 2
Lot 3
Lot 4
0.0
0.50
1.00
1.50
2.00
Time (min)
2.50
3.00
3.50
4.00
Lot 5
Column: Shim-pack Velox Biphenyl 2.7 µm, 3.0×100 mm
(P/N: 227-32016-03 )
Flow Rate: 0.7 mL/min
Column Temp.: 30 °C
Sample Dilulent: Initial mobile phase
Injection Volume: 5 µL
Mobile Phase A: 0.1 % formic acid in water
Mobile Phase B: 0.1 % formic acid in acetonitrile
Gradient: 40 % B (0 min) 80 % B (3 min)
40 %B (3.01 min – 5 min)
PFPP
L43
Pentafluorophenyl propyl
1.8, 2.7, 5
90
125 m2/g
130 m2/g
100 m2/g
4 %
4 %
3 %
No
2.0-8.0
100 MPa*
60 MPa
40 MPa
HILIC
L3
None
2.7
90
130 m2/g
N/A
No
2.0-8.0
60 MPa
End-Cap
pH range
Suraface Area
Carbon Load
1.8 µm
2.7 µm
5 µm
1.8 µm
2.7 µm
5 µm
1.8 µm
2.7 µm
5 µm
Max Pressure
HPLC
2.7 µm & 5 µm
4.6 mm (3.0 mm)
100-250 mm
UHPLC-like
2.7 µm
3.0 mm (2.1 mm)
50-100 mm
UHPLC
1.8 µm & 2.7 µm
2.1 mm
≤150 mm
Shim-pack Velox LC Columns
Maximize LC Separation Performance with Core Shell Technology
Si
O
Si
O
Si
O
Si
F
F
F
F
F
O
3
2
Maximize LC Separation Performance
with Core Shell Technology
Designed to maximize performance of LC systems, Shimadzu’s Shim-pack
Velox columns with core shell technology enable you to achieve increased
separations and faster analysis times on any LC platform.
Whether developing a high efficiency LC separation method, transferring an
existing method for increased throughput while maintaining resolution, or
are trying to improve the resolution of a complex separation, Shim-pack
Velox columns will satisfy your needs.
Column ruggedness is critical to any LC analysis and Shim-pack Velox
core-shell columns deliver excellent column lifetime for even the most
challenging sample matrices.
Increased resolution with maximum efficiency
improving separation and detection
Faster separation without sacrificing performance
maximizing laboratory productivity and reducing cost of analysis
Increased sample throughput
reducing overall analysis time
Superior ruggedness
reducing cost of analysis
Excellent reproducibility
maintaining analysis and data integrity
Prominence
Prominence-i plus
Nexera XR
Nexera-i plus
Nexera X2
* For maximum lifetime, recommended maximum pressure for 1.8 µm particles is 80MPa.
Shim-pack Velox column will deliver
Combining highly efficient core shell particle technology with a wide range of surface chemistries provides you with the best
opportunity for optimal resolution. With different chemistry characteristics, Shim-pack Velox columns are suitable for use in a wide
variety of applications and challenging separations.
Column Chemistries
We understand that lot to lot consistent performance of columns is required to maximize your laboratory performance. From one lot
to the next, every Shim-pack Velox column you purchase will perform consistently.
Lot to Lot Reproducibility
Column particle sizes and column volumes affect chromatography results significantly if the column configuration does not match
the LC system. As column particle size is reduced, or the column volume (ID and/or the length of the columns) decreases, the
necessity for a lower dispersion system is increased.
Choosing the optimal column configuration for your LC system allows you to achieve improved chromatography. The following
table summarizes the starting recommendations of column configuration for each LC system.
Column Selection Guide for Different LC Platforms
LC System
Particle size
Column I.D.
Column Length
USP Classification
Ligand type
Particle Size (µm)
Pore size (
Å)
SP-C18
L1
Sterically protected C18
1.8, 2.7, 5
90
125 m2/g
130 m2/g
100 m2/g
7 %
7 %
5 %
No
1.0-8.0
100 MPa*
60 MPa
40 MPa
C18
L1
C18
1.8, 2.7, 5
90
125 m2/g
130 m2/g
100 m2/g
9 %
7 %
5 %
YES
2.0-8.0
100 MPa*
60 MPa
40 MPa
Biphenyl
L11
Biphenyl
1.8, 2.7, 5
90
125 m2/g
130 m2/g
100 m2/g
7 %
7 %
5 %
YES
1.5-8.0
100 MPa*
60 MPa
40 MPa
Compounds (50 ng/mL) :
1. Cortisol
2. 11-Deoxycortisol
3. Estradiol
4. Boldenone
5. Testosterone
6. Androstenedione
7. Progesterone
Lot 1
Lot 2
Lot 3
Lot 4
0.0
0.50
1.00
1.50
2.00
Time (min)
2.50
3.00
3.50
4.00
Lot 5
Column: Shim-pack Velox Biphenyl 2.7 µm, 3.0×100 mm
(P/N: 227-32016-03 )
Flow Rate: 0.7 mL/min
Column Temp.: 30 °C
Sample Dilulent: Initial mobile phase
Injection Volume: 5 µL
Mobile Phase A: 0.1 % formic acid in water
Mobile Phase B: 0.1 % formic acid in acetonitrile
Gradient: 40 % B (0 min) 80 % B (3 min)
40 %B (3.01 min – 5 min)
PFPP
L43
Pentafluorophenyl propyl
1.8, 2.7, 5
90
125 m2/g
130 m2/g
100 m2/g
4 %
4 %
3 %
No
2.0-8.0
100 MPa*
60 MPa
40 MPa
HILIC
L3
None
2.7
90
130 m2/g
N/A
No
2.0-8.0
60 MPa
End-Cap
pH range
Suraface Area
Carbon Load
1.8 µm
2.7 µm
5 µm
1.8 µm
2.7 µm
5 µm
1.8 µm
2.7 µm
5 µm
Max Pressure
HPLC
2.7 µm & 5 µm
4.6 mm (3.0 mm)
100-250 mm
UHPLC-like
2.7 µm
3.0 mm (2.1 mm)
50-100 mm
UHPLC
1.8 µm & 2.7 µm
2.1 mm
≤150 mm
Shim-pack Velox LC Columns
Maximize LC Separation Performance with Core Shell Technology
Si
O
Si
O
Si
O
Si
F
F
F
F
F
O
5
4
Shim-pack Velox SP-C18
Shim-pack Velox C18
Designed and intended specifically for use under low pH condition, Shim-pack Velox
SP (Sterically Protected)-C18 offers a well balanced retention profile with a long life
time even under harsh, acidic condition needed for LC/MS(/MS) analysis.
Sterically protected to resist strongly acidic (pH 1-3) mobile phase condition
Well balanced retention profile
Suitable for LC/MS(/MS) analysis
Shim-pack Velox C18 is a traditional end-capped C18-bonded phase which offers the
highest hydrophobic retention of any Shim-pack Velox phases and is applicable to a
wide range of applications such as pharmaceutical, food, environmental and clinical
and neutrals at moderately low and mid-range pH.
General purpose column for reversed-phase chromatography
Highest hydrophobic retention among Shim-pack Velox series
Compatible with moderately acidic to neutral mobile phases (pH 2-8)
Sterically protected ligand provides extended low pH stability for the core shell particle. Shim-pack Velox SP-C18 columns maintain a
stable retention profile under strongly acidic mobile phase condition (pH1) .
The assay of cyanocobalamin (a synthetic form of vitamin B12) with 5 µm fully-porous ODS column described in the USP monograph
is transferred to a new method with Shim-pack Velox C18 2.7 µm column, within USP allowable adjustments. Analytical time and
solvent consumption can be saved with transferred methods while meeting the requirements of system suitability.
USP requirement of Chromatography <621>
When the column size is changed, the following conditions should be met;
1) L/dp ratio: within -25 % to +50 %
or
Number of theoretical plates (N): within -25 % to +50 % (For SPP)
2) Flow rate: *Based on particle size and internal diameter and ±50%
Low pH Stability
More than 20 compounds used as the preservative agent for industrial products like foods and cosmetics can can be separated by
Shim-pack Velox SP-C18. The simultaneous determination and quantitation of multiple target compounds are possible in a wide
range of commercial product within acceptable analytical times.
Simultaneous Analysis of 22 Preservative Agents
Method Transfer for Cyanocobalamin Analysis within the USP Allowable Adjustment
Flow Rate : 0.6 mL/min
Column Temp. : 60 °C
Mobile Phase A : 0.1 % TFA in water
Mobile Phase B : Acetonitrile
Gradient : 45 % B (0 min – 7.5 min) 5 % B (7.51 min – 50 min)
100 % B (55 min ) 45 % B (55.01 min 60 min)
Preservative agents
Magnified Baseline
LC Conditions
Column: Shim-pack Velox SP-C18, 2.7 µm, 3.0×100 mm (PN: 227-32004-03)
Flow Rate: 1.0 mL/min
Column Temp.: 45
°C
Mobile Phase A: NaH2PO4 aq. (25 mM, pH3.8)
Mobile Phase B: MeOH/ACN = 9/1
Gradient: 8 %B (0 min )
30 % B (0.7 min – 2.7 min) 47 % B (2.71 min – 5min ) 52 %
B (6 min)
80 %B (7 min – 8 min) 8 %B (9.01 min -10 min)
Injection Volume: 1 µL
Conc.: 50 mg/L
1 2-methyl-4-Isothiazolin-3-one
2 2-Bromo-2-Nitro-1,3-propanediol
3 Salicylic Acid
4 Isothiazolinones
5 Benzoic Acid
6 Benzyl Alcohol
7 Sorbic Acid
8 2-Phenoxyethanol
9 Methyl Paraben
10 Ethyl Paraben
11 Methyl Benzoate
12 Isopropyl Paraben
13 Propyl Paraben
14 4-Chloro-3-Methylphenol
15 Ethyl Benzoate
16 Isobutyl 4-Hydroxybenzoate
17 Butyl Paraben
18 Chloroxylenol
19 Phenyl Benzoate
20 Clorofene
21 Triclocarban
22 Triclosan
Original Method
Shim-pack VP-ODS (5 µm, 4.6×150 mm)
Flow rate: 0.5 mL/min
Injection Volume: 100 µL
Column temperature: 25 °C
Detection: UV 361 nm
Column
VP-ODS
(5 µm, 4.6×150 mm)
Velox C18
(2.7 µm, 4.6×100 mm)
L/dp
30,000
37,037
(+23 %)
Flow rate
(mL/min)
0.50
0.50
0.93
5,244
9,497
(+81 %)
4,466
(-15 %)
tR: 0.025 %
Area: 0.175 % (n=6)
tR: 0.035 %
Area: 0.103 % (n=6)
tR: 0.084 %
Area: 0.220 % (n=6)
N
System suitability test result
(Requirement: %RSD < 2.0 %)
Faster Method
Shim-pack Velox C18 (2.7 µm, 4.6×100 mm)
Flow rate: 0.50 mL/min
Injection Volume: 67 µL
Column temperature: 25 °C
Detection: UV 361 nm
Optimized Method
Shim-pack Velox C18 (2.7 µm, 4.6×100 mm)
Flow rate: 0.93 mL/min
Injection Volume: 67 µL
Column temperature: 25 °C
Detection: UV 361 nm
% loss of retention (k) vs. hours at pH 1
5
0
-5
-10
-15
-20
-25
% loss k
Hours
5.163 min
2.368min
1.297 min
0
10
20
30
40
50
60
70
80
90
100
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
min
Shim-pack Velox SP-C18
Brand A FPP C18
Brand B FPP C18
Brand C FPP C18
Brand D Core-shell C18
Brand E Core-shell C18
Brand F Core-shell C18
Shim-pack Velox LC Columns
Maximize LC Separation Performance with Core Shell Technology
Si
O
Si
O
L
：Column length
dp
：Particle size
F
：Flow rate
dc
：Internal diameter of the column
* F2 = F1 ×
dc22 × dp1
dc12 × dp2
254 nm, 280 nm
254 nm, 280 nm
0
40
80
120
mAU
0.0
2.0
4.0
6.0
8.0
min
0
20
40
60
1
2
3 4 6 5 8
7
9
10
1114
12 13
15 18
16 17 19
20
21
22
1
3 4
5 8
7
9
10
1114
1213
15 18
1617 19
20
21
22
1
2
3
4
5 8
7
9
10
1114
1213
15 18
16 17 19
20
21
22
0.0
10.0
20.0
30.0
0.0
2.0
4.0
6.0
8.0
min
1
2
3 4 6
58
7
9
10
11
14
12 13
15
18
16
17
19
20
21
22
0.0
10.0
20.0
30.0
mAU
5
4
Shim-pack Velox SP-C18
Shim-pack Velox C18
Designed and intended specifically for use under low pH condition, Shim-pack Velox
SP (Sterically Protected)-C18 offers a well balanced retention profile with a long life
time even under harsh, acidic condition needed for LC/MS(/MS) analysis.
Sterically protected to resist strongly acidic (pH 1-3) mobile phase condition
Well balanced retention profile
Suitable for LC/MS(/MS) analysis
Shim-pack Velox C18 is a traditional end-capped C18-bonded phase which offers the
highest hydrophobic retention of any Shim-pack Velox phases and is applicable to a
wide range of applications such as pharmaceutical, food, environmental and clinical
and neutrals at moderately low and mid-range pH.
General purpose column for reversed-phase chromatography
Highest hydrophobic retention among Shim-pack Velox series
Compatible with moderately acidic to neutral mobile phases (pH 2-8)
Sterically protected ligand provides extended low pH stability for the core shell particle. Shim-pack Velox SP-C18 columns maintain a
stable retention profile under strongly acidic mobile phase condition (pH1) .
The assay of cyanocobalamin (a synthetic form of vitamin B12) with 5 µm fully-porous ODS column described in the USP monograph
is transferred to a new method with Shim-pack Velox C18 2.7 µm column, within USP allowable adjustments. Analytical time and
solvent consumption can be saved with transferred methods while meeting the requirements of system suitability.
USP requirement of Chromatography <621>
When the column size is changed, the following conditions should be met;
1) L/dp ratio: within -25 % to +50 %
or
Number of theoretical plates (N): within -25 % to +50 % (For SPP)
2) Flow rate: *Based on particle size and internal diameter and ±50%
Low pH Stability
More than 20 compounds used as the preservative agent for industrial products like foods and cosmetics can can be separated by
Shim-pack Velox SP-C18. The simultaneous determination and quantitation of multiple target compounds are possible in a wide
range of commercial product within acceptable analytical times.
Simultaneous Analysis of 22 Preservative Agents
Method Transfer for Cyanocobalamin Analysis within the USP Allowable Adjustment
Flow Rate : 0.6 mL/min
Column Temp. : 60 °C
Mobile Phase A : 0.1 % TFA in water
Mobile Phase B : Acetonitrile
Gradient : 45 % B (0 min – 7.5 min) 5 % B (7.51 min – 50 min)
100 % B (55 min ) 45 % B (55.01 min 60 min)
Preservative agents
Magnified Baseline
LC Conditions
Column: Shim-pack Velox SP-C18, 2.7 µm, 3.0×100 mm (PN: 227-32004-03)
Flow Rate: 1.0 mL/min
Column Temp.: 45
°C
Mobile Phase A: NaH2PO4 aq. (25 mM, pH3.8)
Mobile Phase B: MeOH/ACN = 9/1
Gradient: 8 %B (0 min )
30 % B (0.7 min – 2.7 min) 47 % B (2.71 min – 5min ) 52 %
B (6 min)
80 %B (7 min – 8 min) 8 %B (9.01 min -10 min)
Injection Volume: 1 µL
Conc.: 50 mg/L
1 2-methyl-4-Isothiazolin-3-one
2 2-Bromo-2-Nitro-1,3-propanediol
3 Salicylic Acid
4 Isothiazolinones
5 Benzoic Acid
6 Benzyl Alcohol
7 Sorbic Acid
8 2-Phenoxyethanol
9 Methyl Paraben
10 Ethyl Paraben
11 Methyl Benzoate
12 Isopropyl Paraben
13 Propyl Paraben
14 4-Chloro-3-Methylphenol
15 Ethyl Benzoate
16 Isobutyl 4-Hydroxybenzoate
17 Butyl Paraben
18 Chloroxylenol
19 Phenyl Benzoate
20 Clorofene
21 Triclocarban
22 Triclosan
Original Method
Shim-pack VP-ODS (5 µm, 4.6×150 mm)
Flow rate: 0.5 mL/min
Injection Volume: 100 µL
Column temperature: 25 °C
Detection: UV 361 nm
Column
VP-ODS
(5 µm, 4.6×150 mm)
Velox C18
(2.7 µm, 4.6×100 mm)
L/dp
30,000
37,037
(+23 %)
Flow rate
(mL/min)
0.50
0.50
0.93
5,244
9,497
(+81 %)
4,466
(-15 %)
tR: 0.025 %
Area: 0.175 % (n=6)
tR: 0.035 %
Area: 0.103 % (n=6)
tR: 0.084 %
Area: 0.220 % (n=6)
N
System suitability test result
(Requirement: %RSD < 2.0 %)
Faster Method
Shim-pack Velox C18 (2.7 µm, 4.6×100 mm)
Flow rate: 0.50 mL/min
Injection Volume: 67 µL
Column temperature: 25 °C
Detection: UV 361 nm
Optimized Method
Shim-pack Velox C18 (2.7 µm, 4.6×100 mm)
Flow rate: 0.93 mL/min
Injection Volume: 67 µL
Column temperature: 25 °C
Detection: UV 361 nm
% loss of retention (k) vs. hours at pH 1
5
0
-5
-10
-15
-20
-25
% loss k
Hours
5.163 min
2.368min
1.297 min
0
10
20
30
40
50
60
70
80
90
100
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
min
Shim-pack Velox SP-C18
Brand A FPP C18
Brand B FPP C18
Brand C FPP C18
Brand D Core-shell C18
Brand E Core-shell C18
Brand F Core-shell C18
Shim-pack Velox LC Columns
Maximize LC Separation Performance with Core Shell Technology
Si
O
Si
O
L
：Column length
dp
：Particle size
F
：Flow rate
dc
：Internal diameter of the column
* F2 = F1 ×
dc22 × dp1
dc12 × dp2
254 nm, 280 nm
254 nm, 280 nm
0
40
80
120
mAU
0.0
2.0
4.0
6.0
8.0
min
0
20
40
60
1
2
3 4 6 5 8
7
9
10
1114
12 13
15 18
16 17 19
20
21
22
1
3 4
5 8
7
9
10
1114
1213
15 18
1617 19
20
21
22
1
2
3
4
5 8
7
9
10
1114
1213
15 18
16 17 19
20
21
22
0.0
10.0
20.0
30.0
0.0
2.0
4.0
6.0
8.0
min
1
2
3 4 6
58
7
9
10
11
14
12 13
15
18
16
17
19
20
21
22
0.0
10.0
20.0
30.0
mAU
7
6
Shim-pack Velox Biphenyl
Shim-pack Velox PFPP
Shim-pack Velox HILIC
Shim-pack Velox Biphenyl provides enhanced retention of aromatic compounds.
It is useful for fast separations in bioanalytical applications due to the increased
retention of early eluting analytes such as dipolar, unsaturated and conjugated
analytes.
Complementary selectivity to alkyl phases
Enhanced separation of aromatic compounds
Ideal for increasing sensitivity and selectivity in LC/MS(/MS) analysis
Amobarbital
5-ethyl-5-(3-methylbutyl)-
1,3-diazinane-2,4,6-trione
(× 1,000,000)
uV
1. cis-Stilbene
2. trans-Stilbene
1
2
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 min
1.0
1.5
2.0
0.0
1.5
1
2
3
4
5
O
O
O
NH
NH
NH2
NH2
NH2
NH
O
O
O
O
O
O
O
HO
HO
HO
HO
OH
OH OH
OH OH
OH OH
OH
N
H
N
N
N
N
N
N
N
N
N
N
1. Toluene
2. Uracil
3. Uridine 4. Adenosine
5. Guanosine
6. Cytidine
6
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
7.5
8.0
8.5
9.0
9.5 min
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
min
2.00
1.75
1.50
1.25
1.00
0.75
0.50
0.25
0.00
200000
150000
100000
50000
400000
300000
200000
100000
Chromatogram of 56 compounds in human urine spiked at the cut off concentration
Pentobarbital
5-ethyl-5-pentan-2-yl-
1,3-diazinane-2,4,6-trione
System: Nexera UHPLC System / LCMS-8050
Column: Shim-pack Velox Biphenyl 2.7 µ
ｍ, 2.1 mm × 100 mm
(PN: 227-32015-03)
Column Temp.: 30 °C
Flow Rate: 0.5 mL/min
Mobile Phase A: 0.15 mM ammonium fluoride in water
Mobile Phase B: Methanol
Gradient: 20 % B (0 min) 48 % B (1.5 min) 53 % B (4 min)
100 %B (6 min - 7.5 min) 20 % B (7.51 min - 9.5 min)
Sample preparation:
Enzymatically hydrolyzed human urine was spiked with target
compounds between 10 - 1000 % of the required cut off concentration.
Samples were subsequently diluted 5x with 0.1 % formic acid.
LC Conditions
Column: Shim-pack Velox HILIC, 2.7 µm, 3.0×100 mm (PN: 227-32026-02)
Column Temp.: 30
℃
Flow Rate : 0.4 mL/min
Mobile Phase : Acetonitrile/20mM AcONH4aq.=9/1
Detection : UV 254 nm
Sample1. Toluene, 2. Uracil, 3. Uridine, 4. Adenosine, 5. Guanosine, 6. Cytidine
Injection Volume : 1 µL
LC Conditions
Column: Shim-pack Velox PFPP, 2.7 µm, 3.0×100 mm (PN: 227-32022-04)
Column Temp.: 40 °C
Flow Rate : 0.4 mL/min
Mobile Phase : Methanol/Water = 9/1
Detection : UV 254 nm
Sample: 1. cis-Stilbene, 2. trans-Stilbene
Injection Volume : 1 µL
Hydrophilic interaction chromatography (HILIC) is an increasingly popular separation
mode that can be used to improve the retention of challenging polar analytes.
Shim-pack Velox HILIC using unbonded core shell particles is specifically designed for
this application.
Orthogonal selectivity to reversed phase chromatography
Increased retention of polar analytes
Increased MS sensitivity
Direct compatibility with sample preparation eluates
Ordering Information
Shim-pack Velox 1.8 µm Columns
Shim-pack Velox 2.7 µm Columns
Shim-pack Velox PFPP (Pentafluorophenylpropyl) provides an alternative selectivity to
C18 columns and is suitable for the analysis of halogenated compounds, positional
isomers and charged bases.
Alternative selectivity to C18 columns
Suitable for positional isomers and halogenated compounds
Offers increased retention for charged bases
Even under the condition where 56 drugs of abuse and metabolites in human urine are quantitated within 10 minutes, two
structural isomers, amobarmital and pentobarbital, which have been historically difficult to separate due to their similarity in
chemical structures, could be relatively well resolved with shim-pack Velox Biphenyl column.
Separating the Structural Isomers
Nucleosides are polar molecules that are not well retained on reversed phase LC columns due to their hydrophilic nature are well
retained and separated with Shim-pack Velox HILIC column.
Retention of Nucleosides
Cis and trans isomers of stilbene that are difficult to resolve with an ODS column due to their similarity in hydrophobicity can be well
separated with Shim-pack Velox PFPP column.
Good Separation of Cis / Trans Stilbene
Chemistry
30
50
100
150
SP-C18
2.1
227-32001-01
227-32001-02
227-32001-03
227-32001-04
3.0
–
227-32002-01
227-32002-02
–
C18
2.1
227-32007-01
227-32007-02
227-32007-03
227-32007-04
3.0
–
227-32008-01
227-32008-02
–
Biphenyl
2.1
227-32013-01
227-32013-02
227-32013-03
227-32013-04
3.0
–
227-32014-01
227-32014-02
–
PFPP
2.1
227-32019-01
227-32019-02
227-32019-03
227-32019-04
3.0
–
227-32020-01
227-32020-02
–
Chemistry
30
50
100
150
SP-C18
2.1
227-32003-01
227-32003-02
227-32003-03
227-32003-04
3.0
227-32004-01
227-32004-02
227-32004-03
227-32004-04
4.6
227-32005-01
227-32005-02
227-32005-03
227-32005-04
C18
2.1
227-32009-01
227-32009-02
227-32009-03
227-32009-04
3.0
227-32010-01
227-32010-02
227-32010-03
227-32010-04
Biphenyl
4.6
227-32011-01
227-32011-02
227-32011-03
227-32011-04
2.1
227-32015-01
227-32015-02
227-32015-03
227-32015-04
3.0
227-32016-01
227-32016-02
227-32016-03
227-32016-04
4.6
227-32017-01
227-32017-02
227-32017-03
227-32017-04
Chemistry
30
50
100
150
PFPP
2.1
227-32021-01
227-32021-02
227-32021-03
227-32021-04
3.0
227-32022-01
227-32022-02
227-32022-03
227-32022-04
4.6
227-32023-01
227-32023-02
227-32023-03
227-32023-04
HILIC
2.1
227-32025-01
227-32025-02
227-32025-03
227-32025-04
3.0
–
227-32026-01
227-32026-02
227-32026-03
4.6
–
227-32027-01
227-32027-02
227-32027-03
Shim-pack Velox LC Columns
Maximize LC Separation Performance with Core Shell Technology
(× 100,000)
1.75
1.50
1.25
1.00
0.75
0.50
0.25
0.00
4.3 4.5 4.7 4.9
Amobarbital
Pentobarbital
Si
O
Si
F
F
F
F
F
O
ID(mm)
Length(mm)
ID(mm)
Length(mm)
ID(mm)
Length(mm)
7
6
Shim-pack Velox Biphenyl
Shim-pack Velox PFPP
Shim-pack Velox HILIC
Shim-pack Velox Biphenyl provides enhanced retention of aromatic compounds.
It is useful for fast separations in bioanalytical applications due to the increased
retention of early eluting analytes such as dipolar, unsaturated and conjugated
analytes.
Complementary selectivity to alkyl phases
Enhanced separation of aromatic compounds
Ideal for increasing sensitivity and selectivity in LC/MS(/MS) analysis
Amobarbital
5-ethyl-5-(3-methylbutyl)-
1,3-diazinane-2,4,6-trione
(× 1,000,000)
uV
1. cis-Stilbene
2. trans-Stilbene
1
2
0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 9.0 9.5 min
1.0
1.5
2.0
0.0
1.5
1
2
3
4
5
O
O
O
NH
NH
NH2
NH2
NH2
NH
O
O
O
O
O
O
O
HO
HO
HO
HO
OH
OH OH
OH OH
OH OH
OH
N
H
N
N
N
N
N
N
N
N
N
N
1. Toluene
2. Uracil
3. Uridine 4. Adenosine
5. Guanosine
6. Cytidine
6
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
7.0
7.5
8.0
8.5
9.0
9.5 min
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
6.5
min
2.00
1.75
1.50
1.25
1.00
0.75
0.50
0.25
0.00
200000
150000
100000
50000
400000
300000
200000
100000
Chromatogram of 56 compounds in human urine spiked at the cut off concentration
Pentobarbital
5-ethyl-5-pentan-2-yl-
1,3-diazinane-2,4,6-trione
System: Nexera UHPLC System / LCMS-8050
Column: Shim-pack Velox Biphenyl 2.7 µ
ｍ, 2.1 mm × 100 mm
(PN: 227-32015-03)
Column Temp.: 30 °C
Flow Rate: 0.5 mL/min
Mobile Phase A: 0.15 mM ammonium fluoride in water
Mobile Phase B: Methanol
Gradient: 20 % B (0 min) 48 % B (1.5 min) 53 % B (4 min)
100 %B (6 min - 7.5 min) 20 % B (7.51 min - 9.5 min)
Sample preparation:
Enzymatically hydrolyzed human urine was spiked with target
compounds between 10 - 1000 % of the required cut off concentration.
Samples were subsequently diluted 5x with 0.1 % formic acid.
LC Conditions
Column: Shim-pack Velox HILIC, 2.7 µm, 3.0×100 mm (PN: 227-32026-02)
Column Temp.: 30
℃
Flow Rate : 0.4 mL/min
Mobile Phase : Acetonitrile/20mM AcONH4aq.=9/1
Detection : UV 254 nm
Sample1. Toluene, 2. Uracil, 3. Uridine, 4. Adenosine, 5. Guanosine, 6. Cytidine
Injection Volume : 1 µL
LC Conditions
Column: Shim-pack Velox PFPP, 2.7 µm, 3.0×100 mm (PN: 227-32022-04)
Column Temp.: 40 °C
Flow Rate : 0.4 mL/min
Mobile Phase : Methanol/Water = 9/1
Detection : UV 254 nm
Sample: 1. cis-Stilbene, 2. trans-Stilbene
Injection Volume : 1 µL
Hydrophilic interaction chromatography (HILIC) is an increasingly popular separation
mode that can be used to improve the retention of challenging polar analytes.
Shim-pack Velox HILIC using unbonded core shell particles is specifically designed for
this application.
Orthogonal selectivity to reversed phase chromatography
Increased retention of polar analytes
Increased MS sensitivity
Direct compatibility with sample preparation eluates
Ordering Information
Shim-pack Velox 1.8 µm Columns
Shim-pack Velox 2.7 µm Columns
Shim-pack Velox PFPP (Pentafluorophenylpropyl) provides an alternative selectivity to
C18 columns and is suitable for the analysis of halogenated compounds, positional
isomers and charged bases.
Alternative selectivity to C18 columns
Suitable for positional isomers and halogenated compounds
Offers increased retention for charged bases
Even under the condition where 56 drugs of abuse and metabolites in human urine are quantitated within 10 minutes, two
structural isomers, amobarmital and pentobarbital, which have been historically difficult to separate due to their similarity in
chemical structures, could be relatively well resolved with shim-pack Velox Biphenyl column.
Separating the Structural Isomers
Nucleosides are polar molecules that are not well retained on reversed phase LC columns due to their hydrophilic nature are well
retained and separated with Shim-pack Velox HILIC column.
Retention of Nucleosides
Cis and trans isomers of stilbene that are difficult to resolve with an ODS column due to their similarity in hydrophobicity can be well
separated with Shim-pack Velox PFPP column.
Good Separation of Cis / Trans Stilbene
Chemistry
30
50
100
150
SP-C18
2.1
227-32001-01
227-32001-02
227-32001-03
227-32001-04
3.0
–
227-32002-01
227-32002-02
–
C18
2.1
227-32007-01
227-32007-02
227-32007-03
227-32007-04
3.0
–
227-32008-01
227-32008-02
–
Biphenyl
2.1
227-32013-01
227-32013-02
227-32013-03
227-32013-04
3.0
–
227-32014-01
227-32014-02
–
PFPP
2.1
227-32019-01
227-32019-02
227-32019-03
227-32019-04
3.0
–
227-32020-01
227-32020-02
–
Chemistry
30
50
100
150
SP-C18
2.1
227-32003-01
227-32003-02
227-32003-03
227-32003-04
3.0
227-32004-01
227-32004-02
227-32004-03
227-32004-04
4.6
227-32005-01
227-32005-02
227-32005-03
227-32005-04
C18
2.1
227-32009-01
227-32009-02
227-32009-03
227-32009-04
3.0
227-32010-01
227-32010-02
227-32010-03
227-32010-04
Biphenyl
4.6
227-32011-01
227-32011-02
227-32011-03
227-32011-04
2.1
227-32015-01
227-32015-02
227-32015-03
227-32015-04
3.0
227-32016-01
227-32016-02
227-32016-03
227-32016-04
4.6
227-32017-01
227-32017-02
227-32017-03
227-32017-04
Chemistry
30
50
100
150
PFPP
2.1
227-32021-01
227-32021-02
227-32021-03
227-32021-04
3.0
227-32022-01
227-32022-02
227-32022-03
227-32022-04
4.6
227-32023-01
227-32023-02
227-32023-03
227-32023-04
HILIC
2.1
227-32025-01
227-32025-02
227-32025-03
227-32025-04
3.0
–
227-32026-01
227-32026-02
227-32026-03
4.6
–
227-32027-01
227-32027-02
227-32027-03
Shim-pack Velox LC Columns
Maximize LC Separation Performance with Core Shell Technology
(× 100,000)
1.75
1.50
1.25
1.00
0.75
0.50
0.25
0.00
4.3 4.5 4.7 4.9
Amobarbital
Pentobarbital
Si
O
Si
F
F
F
F
F
O
ID(mm)
Length(mm)
ID(mm)
Length(mm)
ID(mm)
Length(mm)
Shim-pack V
elox LC Columns
C190-E219
Maximize LC Separation Performance with Core Shell Technology
Shim-pack Velox LC Columns
© Shimadzu Corporation, 2018
www.shimadzu.com/an/
For Research Use Only. Not for use in diagnostic procedures.
This publication may contain references to products that are not available in your country. Please contact us to check the availability of these
products in your country.
Company names, products/service names and logos used in this publication are trademarks and trade names of Shimadzu Corporation, its
subsidiaries 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, whether or not
they are used with trademark symbol “TM” or “®”.
Shimadzu disclaims any proprietary interest in trademarks and trade names other than its own.
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.
First Edition: September 2018, Printed in Japan 3655-08801-20AIK
Free-turning architecture lets you change cartridges by hand without breaking inlet / outlet fluid connections — no tools needed.
Guard column cartridges require Shim-pack Velox EXP Direct Connect Holder (227-32041-01)
Shim-pack Velox EXP Guard Column
Tubing
Nut
Ferrule
Peek End
Ferrule
Peek End
Holder Cap
Holder Body
Guard Column
Cartridge
Ordering Information
Shim-pack Velox 5 µm Columns
Shim-pack Velox EXP Guard Column Cartridge (3 pk)
Shim-pack Velox UHPLC Precolumn Filter (0.2 µ
ｍ)
＊Shim-pack Velox EXP Direct Connect Holder : 227-32041-01
Chemistry
50
100
150
250
SP-C18
227-32006-01
227-32006-02
227-32006-03
227-32006-04
C18
227-32012-01
227-32012-02
227-32012-03
227-32012-04
Biphenyl
227-32018-01
227-32018-02
227-32018-03
227-32018-04
4.6
PFPP
227-32024-01
227-32024-02
227-32024-03
227-32024-04
Type
ID (mm)
2.1
3.0
4.6
UHPLC
SP-C18
227-32028-01
227-32028-02
–
C18
227-32031-01
227-32031-02
–
Biphenyl
227-32034-01
227-32034-02
–
PFPP
227-32037-01
227-32037-02
–
SP-C18
227-32029-01
227-32029-02
227-32029-03
2.7 µ
ｍ
C18
227-32032-01
227-32032-02
227-32032-03
Biphenyl
227-32035-01
227-32035-02
227-32035-03
PFPP
227-32038-01
227-32038-02
227-32038-03
HILIC
227-32040-01
227-32040-02
227-32040-03
Type
ID (mm)
4.6
5 µ
ｍ
SP-C18
227-32030-01
C18
227-32033-01
Biphenyl
227-32036-01
PFPP
227-32039-01
Part No. 1 pack
Part No. 5 pack
Part No. 10 pack
227-32042-01
227-32042-02
227-32042-03
To minimize extra column volume and maximize UHPLC sample throughput with SPE,
SLE, or other sample preparation techniques, pair 1.8 µm Shim-pack Velox UHPLC
columns with an Shim-pack Velox UHPLC Precolumn filter instead of a guard cartridge.
Shim-pack Velox UHPLC Precolumn Filter (0.2 µm)
ID(mm)
Length(mm)
Similar PDF
Key words
Key words
Key words
