1. Parto Zist Beboud
Seeing is Believing: The Ultrafast LCMS-2020 is the Fastest MS Detector in the World!
Analytical & Measuring Instruments Division, Shimadzu Corporation
Tel:

2. LCMS-2020 Seeing is Believing
Woman: Are you interested in our new LCMS-2020?
Man: Yes. It’s a system equipped with all the functions that are indispensable for ultrafast analysis, isn't it?
Woman: The LCMS-2020 offers an extremely high level of performance. Of course, it can also be used for standard analysis.
The new LCMS-2020 can handle ultrafast analysis.
It incorporates the following capabilities, which are required for ultrafast analysis:
Click: UF scanning
Click: U F switching
Click: U F sensitivity
UF scanning
Ultrahigh scan speed of 15,000 µ/sec
UF switching
High-speed positive/negative polarity switching at 15 msec
UF sensitivity
High sensitivity even in high-speed analysis
Speed is Power
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3. 1. Overview of LC/MS
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4. Electrospray ionization
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5. Atmospheric pressure
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6. Principle of a Mass Spectrometer (MS)
Positive ion
(protonated molecule)
Proton added
Proton removed
Negative ion (deprotonated molecule)
Proton (hydrogen ion)
イオン化したい化合物の分子
Separated in a column
Mixed sample injected
Introduced into MS in order from component with lowest retention
Mobile phase
Column eluate
< ESI >
Ion source
Ion focusing unit
Mass separator
Detector
Atmospheric pressure region
Vacuum region
A high voltage is applied to the column eluate, and it is atomized by nitrogen gas.
The charged droplets gradually become smaller, and ion evaporation occurs.
As the ions pass through the mass separator, they are separated according to mass.
In the detector, the quantity of ions is detected as a current value.
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8. LC-MS: System Configuration and Ionization Method
Note: The LC detector may be removed if it is not required.
Controller
Reservoir tray
Solvent delivery pump (gradient)
Mass spectrometer
Degasser
Mixer
Mobile phase
Column
Mobile phase
LC detector
Nitrogen gas generator
Autosampler cleaning liquid
Autosampler
Column oven
Rotary pump
ESI: Electrospray Ionization
APCI: Atmospheric Pressure Chemical Ionization
This is an extremely soft ionization method, and is suited to the ionization of high-polarity compounds.
This method is suited to the ionization of medium- and low-polarity compounds.
Glass capillary
Charged droplets are formed.
Liquid sample
Nitrogen gas
High voltage (±3 to 5 kV)
+: Positive ions are created.
-: Negative ions are created.
Ion evaporation
Heater
Nitrogen gas
Sample molecule
Corona needle
Solvent molecule
High voltage (3 to 5 kV)
+: Positive ions are created.
-: Negative ions are created.
Liquid sample
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9. Information Obtained from LC and MS
Means of identifying sample
 Retention time
 Retention time + Mass information
Column
Injector
SPD-20A/20AV
SPD-M20A PDA detector
LCMS-2020
Time
UV spectra
The molecular weight of the sample can be ascertained with an MS spectrum. Provides greater selectivity than a UV spectrum.
17.5
mAU
mAU
417
150
421
15.0
125
12.5
100
10.0 75
7.5
5.0
246 50
240
2.5 25
0.0
600
200
300
400
500 nm
200
300
400
500 nm
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10. LC-MS: Analytical Data
Mass Chromatogram
Mass Spectrum
(1)
(2)
int.
(4)
(3)
(1)
TIC
m/z
(2)
m/z 582
int.
(5)
(3)
m/z 193
m/z
time
time
m/z
(4)
(5)
PDA (Photodiode Array)
LC Chromatogram
UV Spectrum nm
(5)
(4) AU
(4)
(2)
(2)
 580 nm nm
time
(3)
 210 nm
(3)
(5)
time
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11. Risks Associated with LC Detectors
Mobile phase preparation errors
Fluctuations in peak retention times
Peak misidentification
If an impurity coincides with the target component:
Changes in area value
Incorrect quantitation A B
If an impurity coincides with the target component: A B
If peak elution is late:
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12. Merits of MS Detectors
The greatest merit in using an MS instrument as an LC detector:
In addition to retention times, mass information for each peak can be obtained simply at the same time.
Mass information is a powerful tool for reducing the risks associated with LC analysis, such as the following:
Peak identification (i.e., qualitative) errors
Quantitative errors due to the elution of unpredicted impurities
The peaks (including those that cannot be separated by time) can be separated using mass information. 
This reduces the risk of qualitative and quantitative errors.
m/z 267
m/z 281
m/z 264
m/z 278
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13. 2. Features of the LCMS-2020
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14. From HPLC to UFLC, Then to UFLC-MS
With the introduction of the Prominence Ultrafast LC (UFLC), which offers the ultimate level of speed, superior reproducibility, and comprehensive expandability, ultrafast LC analysis is no utilized in a variety of new fields.
HPLC
UFLC
0.0
3.0
6.0
9.0
12.0
15.0 min.
0.0
0.5
1.0
1.5
2.0
2.5 min.
Shimadzu's Ultrafast LC Lineup
Prominence UFLC: Greatly reduces analysis time without sacrificing separation.
Prominence UFLCXR: Achieves greater separation while maintaining ultrahigh speed.
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15. UFLC-MS
A mass spectrometer responding to the performance of UFLC: LCMS-2020
What makes an MS instrument suitable for UFLC?
The ability to acquire data at high speed without sacrificing data quality is required!
The three things that enable ultrafast analysis:
The ability to perform scan measurement at high speed
UFscanning
The ability to switch between positive and negative ion measurement at high speed
UFswitching
High sensitivity in high-speed measurement
UFsensitivity
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16. Influence of Data Sampling Points
UFLC Data
Influence of Data Sampling Points on Peak Form
20 points 10 points
1.0
min.
If the number of data points decreases, the sensitivity also decreases.
This adversely affects the reproducibility.
4 to 5 points
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17. Scan: Data is acquired in the desired m/z range.
LC-MS: Scan Speed
Scan: Data is acquired in the desired m/z range.
m/z
100
1,100
Scan speed
(scan cycle)
t(s)
Increase the scan speed.
With conventional instruments, the sensitivity decreases.
Decrease the scan speed.
It is difficult to handle high-speed analysis.
With the LCMS-2020, it has become possible to maintain sensitivity when the scan speed is increased.
If the range m/z 100 to 1,600 is scanned,
M = 1,500.
If data is sampled once every second,
Speed = 1,500 µ/sec.
With sharp peaks that span only 1 sec:
Data sampling is required every 0.1 sec (i.e., for 10 data points).
Speed = 15,000 µ/sec
With a sampling speed of 15,000 µ/sec:
Measurement time for 1 m/z = 1  15,000 [sec] = [µ sec].
m/z Transit Time
µ sec
µ sec
µ sec
If the measurement time for 1 m/z is less than the rod transit time, the sensitivity decreases.
A reduction in sensitivity can be prevented by controlling the voltage applied to the quadrupole in accordance with the scan speed and m/z value. - Patent Pending -
m/z
Scan speed
(scan cycle)
1,100
100
t(s)
Quadrupole rod
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18. UFscanning
With measurement at 15,000 u/sec, the sharp peaks of UFLC are captured reliably.
Seeing is Believing.
Samples:
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19. Ultrafast analysis of 16 drugs
UFscanning
With measurement at 15,000 u/sec, the sharp peaks of UFLC are captured reliably.
Ultrafast analysis of 16 drugs 1 2 3 5 4 6 7 8 9 10 12 13 14 15 16 11
Seeing is Believing.
1: Famotidine
2: Cimetidine
3: Atenolol
4: Lidocaine
5: Atropine
6: Metoprolol
7: Yohimbine
8: Noscapine
9: Bupivacaine
10: Alprenolol
11: Tetracaine
12: Diphenhydramine
13: Erythromycin
14: Dibucaine
15: Isopropylantipyrine
16: Warfarin
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20. LC-MS: Positive/Negative Ion Measurement
Simultaneous Measurement of Positive and Negative Ions
The ease with which positive/negative ions are created depends greatly on the compound characteristics.
With positive/negative polarity switching, both positive and negative ions are measured at the same time.
In simultaneous measurement, the number of sampling points is important.
Peak that readily becomes positive ions
Peak that readily becomes negative ions
Ionization in negative ion mode
Ionization in positive ion mode
20 points
Number of Sampling Points
10 points
4 to 5 points
Chromatograms for both positive and negative ions obtained in a single analysis.
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21. UFswitching
With 15-msec switching during measurement, the sharp peaks of UFLC are captured reliably.
Polarity switching time:
Polarity switching time:
Positive ion measurement
SIM2CH
Negative ion measurement
SIM 2CH
Positive ion measurement
SIM 2CH
Samples:
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22. UFscanning + UFswitching
With measurement at 15,000 u/sec and switching at 15 msec, the sharp peaks of UFLC are captured reliably.
Polarity switching time:
Polarity switching time:
Positive ion measurement
15,000 u/sec
Negative ion measurement
15,000 u/sec
Positive ion measurement
15,000 u/sec
Mass Spectra of Bentazone
Mass Spectra of Dymuron
Mass Spectra of Carpropamid
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23. High-Speed Positive/Negative Switching Analysis of Catechins
Compounds That Give Both Positive and Negative Ions: Catechins
Example of the Analysis of Catechins in Tea (Filtered Sample, 2 µL)
1: (-)-gallocatechin,
2: (-)-epigallocatechin,
3: (+)-catechin,
4: (-)-epicatechin,
5: (-)-epigallocatechin gallate,
6: (-)-gallocatechin gallate,
7: (-)-epicatechin gallate,
8: (-)-catechin gallate
9, 10: methylated catechins 1 2 3 4 5 6
m/z 307 7 8
m/z 291
Positive ions
(-)-epicatechin
m/z 459 1
m/z 443 2 9 10
m/z 473 3 4
m/z 305 5 6
m/z 289 7 8
m/z 457
Negative ions
m/z 441 9 10
m/z 471
Seeing is Believing.
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24. High-Speed Positive/Negative Switching Analysis of Glycyrrhizin
With measurement at 15,000 u/sec and switching at 15 msec, the sharp peaks of UFLC are captured reliably.
LCMS-2020
Measurement time + voltage switching time in each mode:
150 msec
Glycyrrhizin
Seeing is Believing.
Positive
Negative
Measurement time + voltage switching time in each mode:
700 msec
Glycyrrhizin
Positive
Negative
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25. Configuration of the LCMS-2020
Ionization probe
Bottle for standard sample
Dual inlet turbomolecular pump
Mass spectrometric detector
Ion optical system
350 mm
726 mm
553 mm
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26. High Sensitivity Even in High-Speed Analysis
UFsensitivity
High Sensitivity Even in High-Speed Analysis
Qarray
Skimmer
Octopole
Entrance lens
To quadrupole rod
From desolvation line
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27. UFsensitivity UFsensitivity
High sensitivity even in high-speed analysis
Reserpine, 1 pg: S/N > 150 (RMS)
10.0
15.0
20.0
25.0
30.0
35.0
40.0
45.0
8.25
8.50
8.75
9.00
9.25
(x1,000)
(1.00)
Seeing is Believing.
SIM chromatogram of reserpine (1 pg)
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28. Ultrafast Analysis of Erythromycin Impurities
LC-MS: Ion Source CID
Ultrafast Analysis of Erythromycin Impurities
Set quite a high lens voltage, and produce fragment ions.
Normal
718
720
Normal 3 2
158
576
560
CID
158
CID
Normal
716
Normal
750 4 1
158
558
CID
CID
158
592
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29. Features Supporting the Three UFs
Long-Term Stability
A sample was added to blood plasma, acetonitrile was added, centrifugal deproteinization was performed, and the sample was continuously injected in quantities of 1 L.
A reproducibility of 2.26% was attained over 10 days, indicating superior durability as well as reproducibility.
Peak Area of Nortriptyline
%RSD: 2.26
Internal standard
Analysis time: 6 min
2,500 injections, 10 days
Plasma Sample Injection Number
Ion Source After Continuous Analysis
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30. Features Supporting the Three UFs
Easy Maintenance
The desolvation line can be replaced without stopping the vacuum.
Simple capillary replacement
The ionization probe can be attached and detached with a single operation.
Large source window
Sample bottle for auto-tuning
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31. Features Supporting the Three UFs
A Variety of Ionization Options
ESI probe
Introduction of sample
ESI probe
Molecular weight
10,000
1,000
100
No polarity
Medium polarity
High polarity
ESI
DUIS
APCI
Corona needle
Corona needle
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32. Features Supporting the Three UFs
A Variety of Ionization Options
MS Chromatograms
MS Spectra Obtained with DUIS Measurement
Thiamine
Riboflavin
Calciferol
3 Types of Water-Soluble/Lipid-Soluble Vitamins (Mixed Sample)
1. Thiamine: m/z 265: Cations, water-soluble vitamin created by dissociation
2. Riboflavin: m/z 377: Protonated molecules, water-soluble vitamin
3. Calciferol: m/z 397: Protonated molecules, lipid-soluble vitamin
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33. LCMSsolution ver.5 LCMSsolution Ver. 5
The popular aspects of LCsolution and LCMSsolution Ver. 3 have been retained, and an easier-to-use interface has been added.
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34. New Functions of Data Browser
Comparison of PDA and MS chromatograms
Comparison of chromatograms in different data files
Switching between parallel display and overlap display
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35. Automatic Optimization of MS Parameters
Searches for the optimum lens voltage for each compound
Sets optimum values in method files
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36. 3. Electrospray VS MALDY
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43. 4. LCMS-2020: Influence of Mobile Phase Solvent
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44. Xanthine Derivatives 0.1% Aqueous Formic Acid/MeOH = 80/20 Caffeine
Shim-pack VP-ODS 2  50mm, 5µm
0.2mL/min
Caffeine
Theophylline
Theobromine
Y = ( )X + (612820)
R2 =
Y = ( )X + (880292)
R2 =
Y = ( )X + ( e+006)
R2 =
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45. Xanthine Derivatives Prioritizing maintenance of the separated state
0.1% Aqueous Formic Acid/MeOH = 80/20
XR-ODS 2  75mm, 2.2 µm
0.4 mL/min
Elution Time
9 min  2.2 min
Reduced by 6.8 min
Solvent Volume
1.2 mL  0.88 mL
51% OFF
Theobromine in Green Tea
Area
Theobromine
Concentration
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46. Xanthine Derivatives Specifying the column size
0.1% Aqueous Formic Acid/MeOH = 80/20
XR-ODS 2  50 mm, 2.2 µm
0.4m L/min
Elution Time
9 min  1.6 min
Reduced by 7.4 min
Solvent Volume
1.8 mL  0.64 mL
64% OFF
Theobromine in Green Tea
Area
Theobromine
Concentration
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47. Catechins: ESI(-) 0.5% Aqueous Formic Acid/ACN
XR-ODS 2  50 mm, 2.2 µm
0.5 mL/min
2% (0 min) - 25% (3 min) - 2% ( min) Gradient 1
MAX 19.2 MPa
1: Gallic acid
2: (-)-gallocatechin,
3: (-)-epigallocatechin,
4: (+)-catechin,
5: (-)-epicatechin,
6: (-)-epigallocatechin gallate,
7: (-)-gallocatechin gallate,
8: (-)-epicatechin gallate,
9: (-)-catechin gallate 3 2 5 4 7 8 6 9
0.5% Aqueous Formic Acid/ACN/THF
2% (0 min) - 25% (3 min) - 2% ( min) Gradient 1
MAX 19.2 MPa
* Adding THF improves epimer separation.
* Peaks 5 and 6 are also separated well. 3 5 2 4 8 7 6 9
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48. Catechins: ESI(-) 0.5% Aqueous Formic Acid/ACN
XR-ODS 2  50 mm, 2.2 µm
0.5 mL/min
2% (0 min) - 25% (3 min) - 2% ( min) Gradient 1
MAX 19.2 MPa 3 2 5 4 7 8 6 9
Same analysis time
Mobile phase B
Acetonitrile
 Methanol
0.5% Aqueous Formic Acid/MeOH
3% (0 min) - 50% (3 min) - 3% ( min) Gradient 1
* Epimer separation is better.
* Sensitivity is also good.
* There is insufficient separation between peaks 3 and 4 although this is not a problem with LCMS. 3 5 2 4 7 8 6 9
MAX 27.5 MPa
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49. Carbamate Pesticides (1) Aldicarb Sulfoxide (2) Aldicarb Sulfone
(3) Oxamyl
(4) Methomyl
(5) Methiocarb Sulfoxide
[M+H]+ : m/z 207
[M+NH4]+ : m/z 240
[M+H]+ : m/z 237
[M+H]+ : m/z 163
[M+H]+ : m/z 242
(6) 3-OH Carbofuran
(7) Methiocarb Sulfone
(8) Aldicarb
(9) Bendiocarb
(10) Carbofuran
[M+H-H2O]+ : m/z 220
[M+H]+ : m/z 258
[M+NH4]+ : m/z 208
[M+H]+ : m/z 224
[M+H]+ : m/z 222
(11) Carbaryl
(12) Thiodicarb
(13) Ethiofencarb
(14) XMC
(15) Pirimicarb
[M+H]+ : m/z 202
[M+H]+ : m/z 355
[M+H]+ : m/z 226
[M+H]+ : m/z 180
[M+H]+ : m/z 239
(16) Isoprocarb
(17) Trimethacarb
(18) Fenobcarb
(19) Methiocarb
(20) Benfuracarb
[M+H]+ : m/z 194
[M+H]+ : m/z 194
[M+H]+ : m/z 208
[M+H]+ : m/z 226
[M+H]+ : m/z 411
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50. Carbamate Pesticides 10-mmol/L Aqueous Ammonium Acetate/ACN
5% (0 min) - 95% (20-25 min) - 5% ( min) Gradient
VP-ODS 2  150 mm, 5 µm, 0.2 mL/min 15 1 3 4 5 2 6 10 7 20 8 9 12 13
* It is difficult to separate peaks 16 and 17 with acetonitrile, so use of methanol is recommended for this analysis.
* Sensitivity is also better with methanol. 19 18 11 14 16 17
10-mmol/L Aqueous Ammonium Acetate/MeOH
5% (0 min) - 95% (20-25 min) - 5% ( min) Gradient 1 15 3 4 2 5 6 7 10 9 18 8 11 13 19 20 12 14 17 16
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51. Carbamate Pesticides 10-mmol/L Aqueous Ammonium Acetate/MeOH
5% (0 min) - 95% (3-4 min) - 5% ( min) Gradient
XR-ODS 2  50 mm, 2.2 µm, 0.4 mL/min 1 6
MAX 25.5 MPa 3 4 5 2 7 15 10 11 9 18 13 19 8 12 17 20 14 16
Analysis Time 40 min  8 min
Reduced by 32 min
Solvent (Methanol) Volume
3.05 mL  1.06 mL
65% OFF
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52. 14 Drugs (1) famotidine (2) cimetidine (3) atenolol (4) ranitidine
[M+H]+ : m/z 338
[M+H]+ : m/z 253
[M+H]+ : m/z 267
[M+H]+ : m/z 315
(5) procaine
(6) lidocaine
(7) yohimbine
(8) alprenolol
(9) doxepin
[M+H]+ : m/z 237
[M+H]+ : m/z 235
[M+H]+ : m/z 355
[M+H]+ : m/z 250
[M+H]+ : m/z 280
(10) imipramine
(11) amitriptyline
(12) furosemide
(13) isopropylantipyrine
(14) warfarin
[M+H]+ : m/z 281
[M+H]+ : m/z 278
[M-H]－ : m/z 329
[M+H]+ : m/z 231
[M+H]+ : m/z 309 [M-H]－ : m/z 307
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53. 14 Drugs 0.1% Aqueous Formic Acid/ACN Analysis Time 6.5 min  4 min
5% (0 min) - 80% (3-4 min) - 5% ( min) Gradient
XR-ODS 2  50 mm, 2.2 µm
0.4 mL/min 2 3 1 4 5 6 7 8 13 9
Analysis Time
6.5 min  4 min
Reduced by 1.5 min
Solvent Volume
1.06 mL  0.36 mL
66% OFF 10 11 14 12
5% (0 min) - 80% (1-1.5 min) - 5% ( min) Gradient
* Analysis can be performed in the same way even if the gradient conditions are changed; as a result, the analysis time can be reduced. 2 3 1 4 5 6 7 13 8 9 10 11 14 12
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54. 14 Drugs 0.1% Aqueous Formic Acid/ACN 0.1% Aqueous Formic Acid/MeOH
5% (0 min) - 80% (3-4 min) - 5% ( min) Gradient
XR-ODS 2  50 mm, 2.2 µm
0.4 mL/min 2 3 1 4 5 6 7 8 13 9 10 11 14 12
Same analysis time
Mobile phase B
Acetonitrile
 Methanol
0.1% Aqueous Formic Acid/MeOH
5% (0 min) - 85% (3-4 min) - 5% ( min) Gradient 3 2 1 6 7
* It is difficult to separate peaks 1 to 5. 4 8 13 5 9 10 11 14 12
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55. 14 Drugs 0.1% Aqueous Formic Acid/ACN
5% (0 min) - 80% (3-4 min) - 5% ( min) Gradient
XR-ODS 2  50 mm, 2.2 µm
0.4 mL/min 2 3 1 4 5 6 7 8 13 9
Same analysis time
Mobile phase B
Acetonitrile
 Methanol
Mobile phase A
0.1% aqueous formic acid
 10-mmol/L aqueous ammonium acetate 10 11 14 12
10-mmol/L Aqueous Ammonium Acetate/MeOH
5% (0 min) - 85% (3-4 min) - 5% ( min) Gradient 2 3 4 5 7 6 1
* Peaks 1 to 5 are separated well.
* Sensitivity is at least as good.
* The elution order changes but, with LC/MS, identification is simple. 8 13 9 10 11 14 12
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56. 14 Drugs 10-mmol/L Aqueous Ammonium Acetate/ACN
5% (0 min) - 65% (1-1.5 min) - 5% ( min) Gradient
XR-ODS 2  50 mm, 2.2 µm
0.4 mL/min 3 2 4 5 1 7 6 8 9 13 10 11 14 12
10-mmol/L Aqueous Ammonium Acetate/MeOH
5% (0 min) - 85% (1-1.5 min) - 5% ( min) Gradient 2 3 4 5 1
* The elution order changes but, with LC/MS, identification is simple. 7 6 8 9 13 10 11 14 12
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57. A mass spectrometer responding to the performance of UFLC: LCMS-2020
Summary
A mass spectrometer responding to the performance of UFLC: LCMS-2020
With the LCMS-2020, the peaks output from the Prominence UFLC at high speed can be captured.
High sensitivity can be attained even in high-speed measurement.
It is easy to change from acetonitrile to methanol.
- Sensitivity
- Pressure
- Changes in separation patterns
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58. Related Technical Reports
Technical Reports on Ways of Saving Solvent
No. 24
Reduced Solvent Consumption and Operational Costs with Ultra High Speed Analysis
No. 25
Reduced Solvent Consumption and Operational Costs with Existing LC Systems
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