The document discusses how solvent quality can affect liquid chromatography performance. It explains that different grades of solvents are needed depending on the type of chromatography and detector being used, such as LC-MS, UHPLC-UV, or fluorescence detection. The key factors that determine solvent grade are discussed, including filtration level, metal ion content, UV transmission, fluorescence, and MS background noise. Various solvent grades from companies like Fisher Chemical and Thermo Scientific are presented and their applications and specifications compared. The document provides guidance on choosing the appropriate solvent for different chromatography instrumentation and techniques.

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How solvent quality affects your analysis

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How solvent quality affects your analysis

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Qualities and Sizes

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Liqiud Chromatography
It’s a jungle out there

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How solvent quality affects your analysis
1- Why there is a need for
different grade of solvents ?
2- What can affect the
chromatography performance?
Not all solvents are equal

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Fisher Chemicals – the portfolio

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HPLC Gradient grade
• The economical choice for HPLC
gradient grade for UV
UHPLC-UV Gradient grade
• The good choice for UHPLC-UV
Thermo Scientific
UltiMate™ 3000
UV detector
LC instrumentation Driven Solvent Quality Needs

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HPLC Gradient grade
• The economical choice for HPLC
gradient grade for UV
UHPLC-UV Gradient grade
• The good choice for UHPLC-UV
Thermo Scientific
UltiMate™ 3000
UV detector
Thermo Scientific UltiMate™ 3000
With TSQ or Q Exactive…
Optima LC-MS
• For high sensitivity LC-MS analysis
• Better quality than LC-MS grade
• UV scan from 200-400 nm
• Low metal impurities
LC instrumentation Driven Solvent Quality Needs

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HPLC Gradient grade
• The economical choice for HPLC
gradient grade for UV
UHPLC-UV Gradient grade
• The good choice for UHPLC-UV
Thermo Scientific
UltiMate™ 3000
UV detector
Thermo Scientific UltiMate™ 3000
With TSQ or Q Exactive…
Thermo Scientific Vanquish™
With TSQ or Q Exactive or Orbitrap…& CAD
Optima LC-MS
• For high sensitivity LC-MS analysis
• Better quality than LC-MS grade
• UV scan from 200-400 nm
• Low metal impurities
UHPLC-MS
• For UHPLC-MS applications with highest
sensitivity
• Higher quality than Optima LC-MS
• Lowest metal impurities
• Low baseline noise in MS detector results in higher
signal to noise ratios.
LC instrumentation Driven Solvent Quality Needs

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How solvent quality affects your analysis
Key Questions
 LC or UHPLC
 UV ?
 Fluorescence ?
 CAD ?
 MS ?

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LC-MS
• Low mass noise levels
• Lowest UV absorbing impurity background
• Low metal ions content
• Ultrafiltration level
UHPLC-MS
• Lowest UV absorbing impurity & MS-ionizing impurities
background
• Lowest metal ions content
• Optimal signal to noise ration in MS/MS mode
• Ultrafiltration for demanding UHPLC-MS application
LC-UV
• High UV Transmission
• Low acidity & alkalinity level
• Filtered to 0,2µm
UHPLC-UV
• Outstanding high UV transmission, low acidity & alkalinity level
• Ultrafiltration to 0,1µm
UHPLC-CAD
• Low mass noise levels
• Minimal organic contamination
• Minimal metal content
• Low presence of non volatile dissolved
contaminants
Key criteria required per instruments

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Laboratory Chemicals Division (LCD) - Brands
All examples are with the Fisher Chemical brand

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UV/Vis Detectors
What is important
 High UV transmission
 Ultra filtrated to at least 0,2µm
 Scanned for impurities

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UV/Vis

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UV/Vis
Both of these solvents are LCMS grade

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UV/Vis
Both of these solvents are LCMS grade

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Fluorescence Detectors
What is important
 High UV transmission
 Ultra filtrated to at least 0,2µm
 Low fluorescence between 250nm and
750nm emission & excitation
wavelengths

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Fluorescence

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Fluorescence
Comparison of the fluorescence baseline behavior of different grade water samples
(Pink: purified water, blue: ultra-pure lab water, black: LC/MS grade water

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Fluorescence
Scheme of the influence of different auto fluorescence on the S/N.

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CAD Detectors
What is important
 Ultra filtrated to at least 0,1µm

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CAD
This is just a tiny
bit of what is tested

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Corona Sources of Noise: Methanol
min0.25 0.5 0.75 1 1.25 1.5 1.75
mV
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ADC1 A, ADC1 CHANNEL A (CO010407120479.D)
ADC1 A, ADC1 CHANNEL A (CO010407120476.D)
ADC1 A, ADC1 CHANNEL A (CO010407120475.D)
3 Injections of 20% Methanol : 80% Water.
All injections represent different methanol
suppliers

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Corona Sources of Noise: Methanol
Manufacturer Grade Bottle Volume (L) Background Range (pA)
Riedel-de Haën LC/MS Chromasolv 1 0.042 – 0.060
JT Baker Microelectronics 2 0.066 – 0.130
EMD Omnisolv for GC 4 0.125 – 0.255
EMD Omnisolv- Gradient 1 0.044 – 0.047
Burdick and Jackson HPLC Grade 4 0.261 – 0.264
Brands of Methanol* and their effects on background current.
(20% methanol / 80%water solution @ 1.0 ml/min, no column)
*Lot to lot variations may exist, these are only guidelines
 Recommend Fisher Optima brand LC/MS grade methanol.

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Acetonitrile – HPLC grade
Acetonitrile – Trace Analysis Grade
Corona Sources of Noise: Acetonitrile

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MS Detectors
What is important
 Ultra filtrated to at least 0,1µm
 Scanned for impurities

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MS
• Depending on application
• UHPLC-MS Optima™
• Optima LC-MS
• LC-MS

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LC-MS applications?
Without MS Grade With MS Grade

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MS requirement
Application Grade
LC-UV Gradient Test
Diode array detector
Signal to Noise Metal tested Packaging
LC-MS
Optima LC-MS
UHPLC-UV Suitability
test
Scan from 200 to 400nm
No test
17 Amber Glass bottle
UHPLC-MS
UHPLC-MS
UHPLC-MS suitability
test
Scan from 200 to 400nm
Signal to noise ratio of
production peak (MS/MS)
250 ppt propazine
(m/z 188) > 10 S/N
17
Borosilicate glass bottle
Reducing significantly
leaching of metal cations
( Na+ & K+)
• Lowest UV absorbing impurity & MS-ionizing impurities background
• Lowest metal ions content
• Optimal signal to noise ration in MS/MS mode

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How solvents quality affect your LC-MS performance
 Black is the LCMS ( Optima LC-MS Fisher Chemical )
 Blue line is UHPLCMS ( UHPLC-MS Thermo Scientific )
UHPLC-MS quality is much better vs LCMS , background noise is lower meaning much
better quality for the Corona detector sensitivity

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Summary
HPLC or UHPLC
• UV absorbance characteristics
• particles content and size
LC-MS or UHPLC-MS
• UV & MS-ionizing impurities background
• Metal ion content
• Particle content and size of the particles

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System cleaning
What can affect the
chromatography
performance?
Phtalates
Polypropylene glycol -PPG
Polyethylene glycol - PEG
Siloxanes

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System cleaning
Clean up contaminated LC & LC-MS instruments
Reduce background noise
Facilitate instrument installation, preventative maintenance
Help to avoid troubleshoot instruments
• Effective removal of common background contamination such as:
• Polyethylene glycol usually observed in ESI+ as a series of ions separated by 44 Da
• Polypropylene glycol usually seen in ESI+ as a series of ions separated by 58 Da
• Phthalates observed as [M+H]+ in ESI+ at m/z = 391, 419 and 447 which are attributed to
• diisooctylphthalate, dinonylphthalate, diisodecyl phthalate, respectively
• Siloxanes observed at m/z = 445, 519, 593, 667 and 741

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Thermo Scientific ChromaCare™ LC-MS Flush Solution
• Assay by GC-TCD is 25%  2%
• Color ≤ 10 APHA
• Residue ≤ 1mg/L
• Optical Absorbance at 210nm ≤ 1.0 AU
• Optical Absorbance at 220nm ≤ 0.5 AU
• Optical Absorbance at 254nm ≤ 0.05 AU
• Calcium (Ca) ≤ 20ppb
• Potassium (K) ≤ 10ppb
• Sodium (Na) ≤ 50ppb
New code T11110 ( 1L)
• Formulation:
Acetonitrile 25%
Methanol 25%
Isopropanol 25%
Water 25%

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Common Background Contamination Ions in Mass Spectrometry
Cleaning out:
Polyethylene glycol, polypropylene
glycol, phthalates, organic solvent
clusters, solvent modifiers, fatty
acids, metal ions, tritons, tweens
and siloxanes.
Metal ions form adducts with
varying numbers of substrates to
give characteristic ESI+ ions.

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Thank You For Your Attention!
Questions?

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References
This is the reference part of the
presentation

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Liqiud Chromatography
• UHPLC-MS Optima™
• Optima LC-MS
• LC-MS
• Advanced HPLC Gradient grade
• UHPLC Gradient grade
• HPLC Gradient grade
• HPLC Fluorescence
• HPLC Electrochemical

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LC instrumentation Driven Solvent Quality Needs
Chromatography Application Instrument and Detector Type
Fisher Chemical & Thermo
Scientific Solvent Grade
UHPLC-MS UHPLC coupled with Mass detector or CAD UHPLC-MS Optima
HPLC-MS LC coupled with Mass detector Optima LC-MS HPLC
UHPLC-UV UHPLC coupled with UV detector UHPLC Gradient Grade
HPLC gradient analysis LC gradient coupled with UV detector HPLC Gradient Grade
HPLC Fluorescence UHPLC/LC gradient coupled with FL detector HPLC Florescence

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Filtration requirement in chromatography
Application Filtered to
Fisher Chemical
Thermo Scientific
UHPLC-MS
0,05µm (acetonitrile & methanol )
0,03µm water
UHPLC-MS grade Optima
HPLC-MS
0.1 µm (acetonitrile & methanol )
0.03 µm (Water only) Optima LC-MS grade
UHPLC-UV To 0,1 µm UHPLC Gradient Grade
HPLC gradient analysis To 0,2µm HPLC Gradient Grade

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Laboratory Chemicals | Portfolio Overview
Organic and
inorganic chemistry
Life science Screening libraries
Reagents used to
synthesize a
compound or material.
Focused on high purity
inorganics, metals and
materials, building
blocks, and moisture
sensitive reagents.
Eliminates the need
for testing with
essential reagents
prequalified for
molecular biology,
protein chemistry, and
cell biology
World-leading
screening libraries
and compound
collections of diverse
drug-like molecules
designed to
accelerate the drug
discovery process
For Research Use Only. Not for use in diagnostic procedures
Analytical
chemistry
A full portfolio of dry
reagents, salts, acids,
solutions, and high-
purity solvents ready
to analyze and
purify a compound or
material.
Specialized
chemical services
Custom chemical,
testing, or packaging
solutions in research
to scaling up/
development for
production

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UHPLC-MS Optima™
Grade
UHPLC-MS Optima™
Application
UHPLC-MS
Definition
Ultra high-purity solvents specifically qualified for UHPLC-MS instrumentation. Specification based on higher
ionization efficiency to detect organic contaminants in full scan MS with the absence of an additive. Signal to
noise specification greater than ten when measured with 250 ppt Propazine using MS/MS. Filtered at 0.1μm,
packaged in borosilicate glass and tightened metal specifications minimizes metal ion adduct formation
Designed to ensure:
• low metal ion adduct formation
• reduce column clogging
• improve peak profiles

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Optima LC-MS
Grade
Optima LC-MS
Application
LC-MS
Definition
Optima LC-MS grade products meet stringent purity requirements of LC/MS and UHPLC by addressing
the need for minimal organic contamination with 0.1μm filtration to make particle free. Evaluated for 17
metal impurities at ppb concentrations for minimal metal mass adduct formation. High ionization
efficiency to detect organic contaminants at 50 ppb max (positive) and 300 ppb max (negative) in full
scan MS. Screened for UV-absorbing contaminants at every wavelength in the 200 to 400 nm range to
afford smooth baselines and to reduce interferences

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Grade
LC-MS
Application
LC-MS
Definition
Ideal mobile phase for routine LC-MS applications. Guaranteed for low level of trace metals and nonvolatile
residue. Low level of absorbance, performance under gradient conditions. Filtered at 0.2μm
LC-MS

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Advanced HPLC Gradient grade
Grade
Advanced HPLC Gradient grade
Application
HPLC Gradient grade
Definition
Advanced HPLC gradient grade specifically manufactured to guarantee a very low level of gradient baseline
drift. Includes lot analysis and absorbance curve on the label. Filtered at 0.2μm.

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UHPLC Gradient grade
Grade
UHPLC Gradient grade
Application
UHPLC-UV
Definition
Advanced HPLC gradient grade specifically manufactured to guarantee a very low level of gradient baseline
drift. Includes lot analysis and absorbance curve on the label. Filtered at 0.2μm.

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HPLC Gradient grade
Grade
HPLC Gradient grade
Application
HPLC Gradient grade
Definition
HPLC solvents suitable for gradient analysis. Guaranteed for low absorbance/high UV transmission and low
concentration of non-volatile impurities. In some instances may be suitable for fluorescence detection.
Includes lot analysis and absorbance curve on the label. Filtered at 0.2μm.

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HPLC Fluorescence
Grade
HPLC Fluorescence
Application
HPLC with Fluorescence and UV detectors
Definition
HPLC solvents suitable for Fluorescence and UV detectors. Guaranteed for low fluorescence between 250nm
and 750nm emission & excitation wavelengths.

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HPLC Electrochemical
Grade
HPLC Electrochemical
Application
HPLC with Electrochemical and UV detectors
Definition
HPLC solvents suitable for Electrochemical and UV detectors. Guaranteed for low electrochemical activity
and low UV absorbance/high transmission. Includes lot analysis and absorbance curve on label.

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Solvent overview

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More details on Thermo Scientific™ ChromaCare Flush solution