Application Chemist – Shimadzu do Brasil Why reduce solvent consumption ?
1) Price: low cost/analysis
Shortage of acetonitrile increase the price
(October 2008)
2) Eco friendly analytical methods
How to reduce solvent consumption ?
• Methodologies review
• Efficient use of LC systems
• Solvent recycle
• Alternative solvents
• Ultra-fast HPLC systems
Approaches to save solvent
Can you change analytical No Daily conservation
conditions? Recycle of mobile phase Utilization of gradient system Yes
Is use of acetonitrile critical for No
separation? Use of methanol
Yes
Can you modify your instrument
No or purchase a new instrument? Change the column
Yes
Transfer to semi-micro conditions
Transfer to ultra-fast HPLC system Approaches to save solvent
Can you change analytical No Daily conservation
conditions? Recycle of mobile phase Utilization of gradient system Yes
Is use of acetonitrile critical for No
separation? Use of methanol
Yes
Can you modify your instrument
No or purchase a new instrument? Change the column
Yes
Transfer to semi-micro conditions
Transfer to ultra-fast HPLC system Daily Conservation
Use of automation tool
Automatic start-up and shut-down of HPLC system
Use of methanol for column wash
Methanol can be utilized for column wash in most cases
Recycle of Mobile Phase
Principle
Eluate from column is re-used as a mobile phase by returning back to
reservoir bottle automatically
Only “clean” eluate is re-used.
Advantages
Modification of analytical conditions is not necessary
It is easy and effective for simple samples
Application
Isocratic elution only
Recycle of Mobile Phase
Solvent recycle valve kit (option)
For UV-VIS detectors
SPD-20A/20AV
SPD-10Avp/10AVvp Solvent recycle valve kit Operation for SPD-20A/20AV Parameter: threshold and delay time (can be set from LCsolution software or detector front panel) Recycle of Mobile Phase
Possibility of contamination of mobile phase
There is a possibility that contaminants are recycled if the contaminants can’t be detected at measuring wavelength
Countermeasure Risk reduction of contaminants by dual wavelength monitor Use of Gradient System
Utilization of gradient system for isocratic analysis
Reduction of waste of unused mobile phase
Approaches to save solvent
Can you change analytical No Daily conservation
conditions? Recycle of mobile phase Utilization of gradient system Yes
Is use of acetonitrile critical for No
separation? Use of methanol
Yes
Can you modify your instrument
No or purchase a new instrument? Change the column
Yes
Transfer to semi-micro conditions
Transfer to ultra-fast HPLC system Use of Methanol
• Different separation selectivity
• Different elution performance
• Absorption at shorter wavelength
• High viscosity (column pressure)
Approaches to save solvent
Can you change analytical No Daily conservation
conditions? Recycle of mobile phase Utilization of gradient system Yes
Is use of acetonitrile critical for No
separation? Use of methanol
Yes
Can you modify your instrument
No or purchase a new instrument? Change the column
Yes
Transfer to semi-micro conditions
Transfer to ultra-fast HPLC system Change the Column
Short column
tR is reduced
N is reduced Column with small internal diameter
Easy optimization of analysis method
Small adaptation in the HPLC system
Column with reduced particle size
tR is reduced
Pressure is increased Change the Column
Shima-pack VP-ODS (150 x 4.6mm, 4.6 µm)
Peaks:
1)Acetophenone
2)Propiophenone
3)Butilphenone
4)Valenophenone
5)Hexaphenone
6)Heptaphenone
7)Octanophenone
Shim-pack XR-ODS (75 x 3.0 mm, 2.2 µm)
Approaches to save solvent
Can you change analytical No Daily conservation
conditions? Recycle of mobile phase Utilization of gradient system Yes
Is use of acetonitrile critical for No
separation? Use of methanol
Yes
Can you modify your instrument
No or purchase a new instrument? Change the column
Yes
Transfer to semi-micro conditions
Transfer to ultra-fast HPLC system Transfer to Semi-micro conditions
• Solvent saving with maintaining separation
• Column with small internal diameter
• Modification of analytical conditions
(Flow rate, injected volume, etc.)
• Modification of HPLC instruments
Narrow pipe to minimize dispersion, Detector cell and gradient mixer
Approaches to save solvent
Can you change analytical No Daily conservation
conditions? Recycle of mobile phase Utilization of gradient system Yes
Is use of acetonitrile critical for No
separation? Use of methanol
Yes
Can you modify your instrument
No or purchase a new instrument? Change the column
Yes
Transfer to semi-micro conditions
Transfer to ultra-fast HPLC system Ultra Fast system
Ultra Fast Liquid Chromatography Effective reduction of mobile phase consumption (1/5 – 1/10) High throughput analysis Two approaches: Particle size and temperature Particle size and separation
Separation with resolution using small particle size
Flow rate: 0.2 mL/min Flow rate: 0.7 mL/min
Column Plate number Column Plate number
Column A 3,590 Column A 2,320 (down) Column B 5,080 Column B 4,500 (even) Column C 6,810 Column C 8,010 (up) XR-ODS Columns
Substrate Totally porous spherical,
high purity silica gel Particle Size 2.2 um
Stationary Phase C18, C8, Phenyl, SIL
Surface Treatment Endcapping
Carbon Load 18.4%
Maximum Pressure 35MPa*
Shim-pack XR-ODS (2,2 µm)
Temperature effect
Best column efficiency: Increase N and decrease the pressure
Mobile phase: H2O/ACN (3/7, v/v)
Detection: 245 nm.
Peaks: 1: acetophenone
2: propilphenone
3: butilphenone
4: pentilphenone
5: hexanophenone
6: heptanophenone
7: octanophenone UFLC - Prominence
Solvent consumption: 10.5 mL
Peaks 1:actophenone, 2:propiophenone, 3:butyrophenone, 4:valenophenone, 5:hexanophenone, 6:heptanophenone, 7:octanophenone.
Solvent consumption: 2.3 mL
Mobile phase: water/acetonitrile (3/7, v/v)
Temperature: 40℃ UFLC - Prominence Solvent consumption: 12.8 mL
Peaks 1:cefadroxil, 2:cephaprin, 3:cefaclor, 4:cefalexin, 5:cephradine, 6:cefotaxime. 7:cefazolin, 8 :cefuroxime, 9 :cefoperazone, 10 :cefloxitin, 1 1 :cefamandole A, 12 :cephalothin, 1 3: cefamandole B
Temperature : 40℃ Flow rate : 1.0mL/min Method tranfer programm
Method Transfer Program HPLC method: 50 minutes
Parameter Input Sheet This program supports the parameter setup operations required when transferring from a conventional LC to Prominence UFLC. Enter the parameters from the conventional LC into the blank boxes below and click [Optimize] to display the estimated optimal chromatographic conditions and instrument environment for an ultrafast LC using a Shim-pack XR Series column. The suitability of the chromatographic conditions produced by this program is not guaranteed. The estimated parameters may be incorrect due to differences in the column characteristics. Evaluate the suitability of the displayed chromatographic conditions before using them.
A value must be entered in each box with a thick border. When these values are entered, the button at the right changes to [Optimize]. Click this button to display the parameters. Ready
must be filled in.
Chromatographic Conditions
Column : Shim-pack VP-ODS 4.6 mm i .d . 150 mm long 4.6 µ m
Mobile phase A: 20 mmol/L (Sodium) Phosphate <pH 2.5> Init. Conc.; 80 % B: Acetonitrile Init. Conc.; 20 % Gradient program No. Time A Conc. B Conc. 1: min % % 2: min % % 3: min % % 4: min % % 5: min % % 6: min % % 7: min % % 8: min % % 9: min % % Flow rate : 1 mL/min Temperature : 40 °C Detection Wavelength: 254 nm Response: 1000 ms 20 50 100 500 1000 1500
Sample volume : 10 µ L
Instrument Environment Peak Information
Mixer volume : 1.7 mL Minimum elution time : 5 min
0.1 0.5 1.7 2.6
Cell volume : Conventional Analysis time : min
Conventional Semi-micro
2.0 3.0 4.6
1: Maintain separation 30 50 75 100
Transfer Conditions : 2
2: Reduce analysis time Resolution tolerance ; %
3
3: Designate column dimensions and flow rate
mm i .d . mm long mL/min
UFLC method: 9 minutes
Minimized Analysis Cycle time
Prominence UFLC employs a fast autosampler (10sec /10uL injection)
Mobile phase: H2O/ ACN (4/6 to 2/8 in 0.4 min, gradient), Flow rate: 3 mL/min, Temperature: 80C, Detection: 245 nm, Sampe volume: 4L Ultra-fast LC-MS system
LCMS-2020 Ultra-fast LCMS-2020
UF switching technology
High-speed polarity switching
UF scanning technology
Fast scanning speed
UF sensitivity
Superior sensitivity in ultra high speed analysis
UF Scanning Technology
Scanning speed of 15,000 u/sec is achieved without sacrificing
sensitivity or resolution, even during high-speed scans UF Switching Technology
Polarity switching between positive/negative in 15 msec enables
analysis of the fastest LC peaks in both modes UF Sensitivity
Reserpine analysis (1pg) S/N = 350
UF Detection of Catechins Ultra-fast detection of catechins in Positive/Negative modes by LCMS-2020 (x100,000) 2.25 Analysis of catechins in tea
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