LCGC0717 - LCGC Millipore - 3

J u ly 2 0 1 7
Water Purification
in the
Chromatography Lab
Sponsored by Presented in partnership with

®
Ionic Hormone Tips: Water Tips: Handling Purification
Contamination Analysis Purification Purified Water System Selection
485F US Highway One South, Suite 210, © 2017 UBM. All rights reserved. No part of this
Iselin, NJ 08830 publication may be reproduced or transmitted in any
(732) 596-0276 form or by any means, electronic or mechanical including
by photocopy, recording, or information storage
PUBLISHING & SALES and retrieval without permission in writing from the
publisher. Authorization to photocopy items for internal/
Michael J. Tessalone educational or personal use, or the internal/educational
Vice President/Group Publisher or personal use of specific clients is granted by UBM for
[email protected] libraries and other users registered with the Copyright
Clearance Center, 222 Rosewood Dr. Danvers, MA
Edward Fantuzzi 01923, 978-750-8400 fax 978-646-8700 or visit http://
Publisher www.copyright.com online. For uses beyond those listed
above, please direct your written request to Permission
Stephanie Shaffer Dept. fax 440-756-5255 or email: Maureen.Cannon@
Sales Manager ubm.com.
Brianne Molnar
Sales Manager UBM Americas provides certain customer contact
data (such as customer’s name, addresses, phone
Oliver Waters numbers, and e-mail addresses) to third parties who
Sales Manager wish to promote relevant products, services, and
other opportunities that may be of interest to you. If
Liz McClean you do not want UBM Americas to make your contact
Sales Executive information available to third parties for marketing
Michael Kushner purposes, simply call toll-free 866-529-2922 between
the hours of 7:30 a.m. and 5 p.m. CST and a customer
Senior Director, Digital Media
service representative will assist you in removing your
name from UBM Americas lists. Outside the U.S., please
SPECIAL PROJECTS phone 218-740-6477.
Kaylynn Chiarello-Ebner LCGC does not verify any claims or other information
Managing Editor, Special Projects appearing in any of the advertisements contained in the
Sabina Advani publication, and cannot take responsibility for any losses
Digital Production Manager or other damages incurred by readers in reliance of such
content.
Vania Oliveira
Project Manager LCGC North America (ISSN 1527-5949 print) (ISSN
1939-1889 digital) is published monthly by UBM Life
Kristen Moore Sciences, 131 West First Street, Duluth, MN 55802-2065.
Webcast Operations Manager LCGC Europe (ISSN 1471-6577) and LCGC Asia Pacific
(ISSN 1754-2715) are published monthly by UBM EMEA,
EDITORIAL Hinderton Point, Lloyd Drive, Cheshire Oaks, Cheshire
CH65 9HQ, UK. Issues are distributed free of charge to
Laura Bush users and specifiers of chromatographic equipment.
Editorial Director
[email protected] To subscribe, call toll-free 888-527-7008. Outside the
U.S. call 218-740-6477.
Megan L’Heureux
Managing Editor, LCGC North America UBM Americas (www.ubmlifesciences.com) is a leading
Stephen A. Brown worldwide media company providing integrated
Group Technical Editor, LCGC North America marketing solutions for the Fashion, Life Sciences and
Powersports industries. UBM Americas serves business
Cindy Delonas professionals and consumers in these industries with its
Associate Editor, LCGC North America portfolio of 91 events, 67 publications and directories,
150 electronic publications and Web sites, as well as
Alasdair Matheson educational and direct marketing products and services.
Editor-in-Chief, LCGC Europe Market leading brands and a commitment to delivering
Kate Mosford innovative, quality products and services enables UBM
Americas to “Connect Our Customers with Theirs.”
Managing Editor, LCGC Europe
UBM Americas has approximately 1000 employees and
Lewis Botcherby currently operates from multiple offices in North America
Assistant Editor, LCGC Europe and Europe.
TOC
Table of contents
Water Purification in the
Chromatography Lab
Ionic Contamination
Why and How to Avoid Ionic Contamina-
tion in Water Used for LC–MS Analyses
Anastasia Khvataeva-Domanov and Stephane
Mabic
4
Hormone Analysis
Hormone Analysis by LC–
MS and Water Impact
A Q&A with Joseph Plurad
12
Tips: Water Purification

Lab Water Hints and Tips for
the Chromatography Lab,
Part 1: Water Purification
16
Tips: Handling Purified Water

Lab Water Hints and Tips, Part 2:
Optimal Handling of Purified Water
20
Purification System Selection

Selecting a Laboratory
Water Purification System
Jean-Francois Pilette
25
Why and How to Avoid
Ionic Contamination in
Water Used for LC–MS
Analyses
Anastasia Khvataeva-Domanov and Stéphane Mabic
Ionic contaminants in the water used in

ultrahigh-pressure liquid chromatography
(UHPLC) analyses with mass spectrometry
(MS) detection lead to adduct formation and
reduced analytical signals because of ion
suppression. In MS, the preferred ion type is the
protonated molecular ion, especially in peptide
analysis, since the partially mobile proton
charge enables more meaningful fragmentation
analysis, as compared to a sodiated peptide
ion. Moreover, the occurrence of protonated
analyte signals demonstrates that solvents and
reagents, as well as the MS instrument used
in analyses, were clean and did not contribute
any contaminating cationic components to
the analytical process. In the experiments
presented here, it was observed that the signal
intensities of the protonated species decreased
as the sodium ion concentration in the water
increased. This was accompanied by an
shutterstock.com/robert_s
increase in the intensity of sodiated adducts..
Water plays an essential role in liquid

chromatography–mass spectrometry (LC–
4 | July 2017 | LCGC Sponsor’s content

Table I: Direct infusion experiments

First set of experiments Second set of experiments
Sample Bradykinin (fragment 1-7) Glu1-fibrinopeptide B
Monoisotopic Monoisotopic
Mass 756.40 Da Mass 1569.65 Da
[M+H] ion (m/z 757) [M+H] ion (m/z 1570)

[M-115+Na] ion (m/z 664) [M+2H] ion (m/z 785)
[M-202] ion (m/z 554) [M+Na+H] ion (m/z 796)
[M+2Na] ion (m/z 807)
[M+3Na-H] ion (m/z 818)
Instrument Applied Biosystems API 2000 Waters Synapt HDMS
Ionization mode ESI+ ESI+
Flow 10 μL/min 10 µL/min
Solvents (a) 96:4 fresh ultrapure water–acetonitrile (a) 50:50 fresh ultrapure water–acetonitrile
(b) 96:4 1 ppm Na+ in ultrapure water/ (b) 50:50 1 ppb Na+ in ultrapure water–acetonitrile
acetonitrile
(c) 50:50 100 ppb Na+ in ultrapure water–acetonitrile
(d) 50:50 1000 ppb Na+ in ultrapure water–acetonitrile

Sample concentration 1 μM 500 pM
MS), where it is used extensively in the data analysis more challenging and
workflow. Contaminants in the water can complicated. Metal ions may come from
affect the quality of data and instrument several possible sources (3) such as
performance; therefore, it is recommended solvent reservoirs, gloves, the analyst,
and prudent to use only the highest purity and the solvents used in preparing the
solvents. Organic contamination of the mobile phase. Therefore, using ultrapure
water used in high performance liquid water free of metal ions will contribute to
chromatography (HPLC) is an important the success of any LC–MS analysis.
issue and has been addressed accordingly
(1), but the ionic purity of the water should Experimental
also be considered, especially when MS Experiments to Evaluate the Effect of
is used as a detection technique (2). Ionic Ionic Contamination in Water on MS
contaminants lead to adduct formation Data
and reduced analytical signals because of Two compounds, bradykinin fragment
ion suppression. 1-7 and Glu1-fibrinopeptide B, were
Electrospray ionization (ESI) remains used to investigate the effect of ionic
the most popular MS technique. In contamination on LC–MS analyses.
positive-ion analyses, it is ideal to have Different samples of peptides were
only protonated peaks of the parent ion prepared and infused directly to a mass
or its fragments in the mass spectrum. spectrometer. The experimental details
The presence of metal adduct peaks, for peptide analyses are described in
such as sodium adducts (M+Na), makes Table I.

(a) (b) 4 105
4 105 3.5 105

[M+H]+
3.5 105 [M-115] +
3 105
3 105 m/z 538 ???
2.5 105
[M-115]+
Intensity (cps)
Intensity (cps)
2.5 105
[M+H]+ 2 105
2 105
1.5 105
1.5 105 [M-115+Na]+
[M-202]+
1 105 m/z 703 ???
1 10 5
[M-202]+
0.5 104 [M+Na]+
0.5 104
0 0
500 550 600 650 700 750 800 850 900 500 550 600 650 700 750 800 850 900
m/z (amu) m/z (amu)
Figure 1: Mass spectra of bradykinin (fragments 1-7). Direct infusion, ESI+, using (a) 96% fresh ultrapure water, 4%
acetonitrile and (b) 96% 1 ppm Na+, 4% acetonitrile.
Experiments to Identify the in Water on MS Data

Levels of Ions in Ultrapure Water The quality of LC–MS data is influenced
To evaluate the level of sodium in ultrapure by many factors, such as instrumentation,
water, water samples from an EMD experimental parameters, sample
Millipore Milli-Q Advantage A10 water preparation, the quality of reagents used,
purification system were analyzed using an and the quality of solvents (5). Water is
Agilent 7700s inductively coupled plasma– used extensively in reversed-phase LC–
mass spectrometry (ICP-MS) instrument MS workflows; therefore, its purity plays
(4). The calibration standards used in a critical role in instrument performance
experiments were a mixture of Agilent and and the quality of data generated.
Spex CertiPrep, and containers were all The purity of water for LC–MS work
perfluoralkoxy (PFA) polymer precleaned is mainly assessed through organic
with ultrapure water. contamination, which can be expressed
by the level of TOC (6), or by certificates
Water Purification Systems of analysis reporting results of suitability
All ultrapure water samples (resistivity of tests.
18.2 MΩ·cm and total oxidizable carbon The second approach to evaluate
[TOC] below 5 ppb) from EMD Millipore water quality for LC–MS analyses is to
water purification systems were analyzed assess its level of ionic purity. Thus, for
immediately after water collection. bottled water, certificates of analysis
usually detail maximum concentrations
Results and Discussion for certain ions because when stored
Effect of Ionic Contamination in a glass bottle, the water will leach

(a) 0 ppb of sodium in the water–acetonitrile mixture TOF MS ES+

100 785.820 1.96e5
786.822
% 776.819 787.835796.826 813.384 816.782 818.784
0
760 765 770 775 780 785 790 795 795 800 805 810 815 820 825 830 835 840 845 850 855 860
m/z
1 ppb of sodium in the water–acetonitrile mixture TOF MS ES+

785.820 1.86e5
100
786.832
%
776.829 796.826 799.605 859.809

0
760 765 770 775 780 785 790 795 795 800 805 810 815 820 825 830 835 840 845 850 855 860
m/z
100 785.830 1.57e5
786.832
%
787.835 796.826 804.812807.815 813.615 827.642 859.798

0
760 765 770 775 780 785 790 795 795 800 805 810 815 820 825 830 835 840 845 850 855 860
m/z
100 785.830 1.37e5
786.832
%
796.826 807.815
771.571 787.835 799.605
827.631 818.805 859.603
0
760 765 770 775 780 785 790 795 795 800 805 810 815 820 825 830 835 840 845 850 855 860
m/z
(b) 0 ppb of sodium in the water–acetonitrile mixture TOF MS ES+
100 785.820 2.20e5
786.315
786.822
%
787.339 787.835
0
785 786 787 788 789 790
m/z
100 785.820 2.20e5
786.326
%
786.832
787.349 787.845
0
785 786 787 m/z 788 789 790
100 785.830 2.20e5
786.326
%
786.832
787.339 787.835
0
785 786 787 m/z 788 789 790
100
785.830 786.326 2.20e5
%
786.832
787.339 787.835
0
785 786 787 788 789 790
m/z
(c) 0 ppb of sodium in the water–acetonitrile mixture TOF MS ES+
100 4.21e4
%
796.826 797.315 786.825 798.314

0 797.825
796 797 798 799 800 801
m/z
100 4.21e4
796.826
%
797.325 799.605
797.835 798.335 800.616
0
796 797 798 799 800 801
m/z
100 796.826 4.21e4
797.325
%
797.825 787.335 799.605 800.606

796.316 798.813
0
796 797 798 799 800 801
m/z
100 796.826
797.325 4.21e4
%
797.825 799.605 800.606

798.335 798.824 799.303
0
796 797 798 799 800 801
m/z
Figure 2: (a) Mass spectra of protonated molecular ion [M+2H] of Glu1-fibrinopeptide B sample, (b) and (c) extracted ion
chromatograms of [M+Na+H] ion as examples of the effect of ionic concentration on MS detection.

200,000
150,000
100,000
50,000
0 785 (M+2H) 796 (M+Na+H) 807 (M+2Na) 818 (M+3Na-H)
0 ppb 1 ppb 100 ppb 1000 ppb
Figure 3: Signal intensities of 500 pmol Glu1-fibrinopeptide B in 50:50 (v/v) acetonitrile–water. The mixture was spiked with
different amounts of sodium ions and injected directly into the mass spectrometer.
out contaminants from the container Table II: Freshly produced ultrapure water sources
even though the water may have been Experiment Water purification system used
of very high purity immediately after Bradykinin direct infu- Elix + Milli-Q Advantage A10 (Q-
sion and ICP-MS Gard and Quantum TEX cartridge,
production. For example, standard glass Millipak final filter)
bottles leach out alkali, contaminating Glu1-fibrinopeptide B Milli-Q Integral (Quantum TEX
direct infusion cartridge)
ultrapure water and leading to a higher
count of adducts. Since the quality of characterize the ionic purity of water.
bottles can vary strongly from one to Thus, at 25 °C, conductivity of 0.055 µS/
another, the nature of ions selected cm, or resistivity of 18.2 MΩ·cm, implies
to report in the certificate of analysis that the water is ultrapure, whereas at 1
depends on the quality of the bottle ppb of Na+ in ultrapure water, resistivity
used to store the water. Table III decreases to 17.6 MΩ·cm, and at 5 ppm,
compares the specifications for some dramatically drops down to 0.093 MΩ·cm;
metal ions in ultrahigh-pressure liquid these values can be calculated based on
chromatography (UHPLC)–MS-grade, the concentration of sodium, its charge, and
LC–MS-grade bottled waters from three mobility (7). Therefore, when fresh ultrapure
vendors, and in fresh ultrapure water. water of 18.2 MΩ·cm resistivity measured
Because ionic impurities present in via an in-line monitor was used to dissolve
water increase its conductivity, the a peptide sample (bradykinin fragment 1-7),
conductivity parameter, or alternatively and infused directly to a mass spectrometer,
its inverse, resistivity, can be used to the resulting spectrum was clean (Figure 1a).

Table III: Specifications for some metal ions in UHPLC–MS-grade or LC–MS-grade water and fresh ultrapure water produced by a
laboratory water purification system specifically for LC–MS practice*
Maximum Concentration (ppb)
UHPLC–MS Grade LC–MS Grade Water LC–MS Grade Water Fresh Ultrapure Water for
Metal Ion
Water Brand X Brand Y Brand Z LC–MS Analyses
Aluminum 20 500 10 1
Calcium 50 100 100 1
Iron 30 100 5 1
Potassium 50 100 10 1
Magnesium 20 100 20 1
Sodium 50 100 200 1
*Specifications of different sources of laboratory water dedicated for MS analyses were collected from official websites of manu-
facturers but their names are not provided to avoid direct comparison.
The parent and fragment peaks represented Specifically, the presence of 1 ppb of
only protonated species. However, when the Na+ decreased the [M+2H] signal intensity
water used was contaminated with sodium by 5%. But the decrease in signal was
ions, the spectrum was more complex 20% with 100 ppb Na+, and 30% when it
with the presence of sodium adduct peaks was 1000 ppb. Altogether the presence
(Figure 1b). of sodium results in more complex
The presence of metal ions can suppress spectra, leading to difficulties in data
the signal of the protonated ion peak of characterization, analyte quantification,
interest. The effect of the sodium ion on the and subsequently more time spent on
signal intensity of Glu1-fibrinopeptide B was data analysis. Therefore, using water in LC–
analyzed by varying the ion concentration in MS analyses that is free of ions is of high
the water–acetonitrile mixture that was used importance. In addition, there are a few
to dissolve the peptide. It was observed key considerations to keep in mind when
that signal intensity of the [M+2H] molecular choosing the source of ultrapure water,
ion decreased with the increase of sodium and also when handling it to minimize
ion concentration in the water. In parallel, contamination.
it was observed that signal intensities of
the sodium adducts increased with the Five Tips to Avoid Ionic
increase in Na+ concentration (Figures 2a, Contamination in LC–MS Practice
2b, and 2c). 1. Choose the Best
Further signal intensities of Glu1- Source of Ultrapure Water
fibrinopeptide B such as the [M+2H], Common choices are freshly produced
[M+Na+H], [M+2Na], and [M+3Na-H] ions ultrapure water from a laboratory water
were recorded for each analyzed sample purification system, and bottled water,
and presented as a function of Na+ such as UHPLC–MS grade or LC–MS
concentration in water (Figure 3). grade. Fresh ultrapure water produced

Table IV: Concentration of sodium ions in fresh ultrapure water It is also recommendable to have
and in the same ultrapure water that was stored in a glass bot-
tle for one day
dedicated glassware for LC–MS practice,
Sample Na+ (ppb) and glassware should be cleaned
Freshly produced ultrapure water 0.020 thoroughly before use.
Ultrapure water in a glass bottle after one day 0.212
3. Keep Your Work Area Clean

using an optimal combination of Moreover, since laboratory air is
purification technologies is usually of very characterized by a lot of contamination and
high purity, with ionic levels below 1 ppb as ultrapure water absorbs contamination
at 18.2 MΩ·cm of resistivity, which can be from the air including volatile molecules (8),
monitored online at the moment of water the working area should be kept clean (9).
collection. In the case of bottled water, Appropriate covers and caps for mobile-
specifications of ionic purity for laboratory phase reservoirs and glassware can help
water are usually provided and can be to avoid contact between ultrapure water
easily consulted to assess the potential risk and the laboratory air, as well. It is also a
of ionic contamination on LC–MS analyses. good idea to wear gloves, and choose
Because ultrapure water is a very ones that are powder-free and have the
aggressive solvent, it tends to absorb fewest metal or ion extractables (for
ions and organic compounds from the example, polyethylene) because simply
container. Thus, maximum concentrations touching glassware with a bare hand can
of ions in final ultrapure product water transfer enough salt to cause a significant
depend significantly on the level of appearance of metal adduct ions.
extractables released by the containers
used to collect freshly produced ultrapure 4. Select Reagents and Organic
water, or to store it, in the case of bottled Solvents of the Highest Purity
water. Table IV shows the concentration Here, to evaluate the purity level necessary
of Na+ in freshly produced ultrapure water for sensitive MS applications, a certificate
and ultrapure water after storage in a of analysis is a good source of information.
glass bottle for one day. Also, it is recommended to obtain details
As shown in Table IV, the Na+ from the supplier beforehand concerning
concentration in freshly produced the level of leachables and extractables
ultrapure water was 0.020 ppb. from solvent and reagent packaging.
After storage in a glass bottle, the
concentration increased to 0.212 ppb. 5. Routine Maintenance
When using a laboratory water purification
2. Choose Glassware of the Highest system, make sure that it undergoes
Quality That Leaches the routine maintenance to ensure the highest
Minimum Amount of Contamination water quality.

Table V: Water purification technologies used in the production of ultrapure water from tap water
Pretreatment (filter, activated carbon) • Filtration
• Removal of oxidizing agents
Reverse osmosis • Removal of the bulk of contaminants (95–99%): organics, ions, microorganisms, particulates
Electrodeionization • Further removal of ions, and charged organic species
UV (254 nm) • Prevents bacterial contamination
Virgin ion-exchange resins • Removal of trace amounts of ions
Synthetic activated carbon • Removal of trace levels of organics
UV (185, 254 nm) • Photo-oxidation to further remove organics
Point-of-use end filters • Prevents particle and bacterial contamination
• Allows removal of trace levels of organics
Ion Removal in Laboratory Acknowledgment

Water Purification Systems The authors would like to acknowledge
The laboratory water purification Jens Baumgaertner of Merck KGaA in
systems used in this study combine Darmstadt, Germany, and Maricar Tarun-
the technologies shown in Table V to Dube of EMD Millipore Corporation in
efficiently remove different types of Danvers, Massachusetts, for their help
contaminants from tap water feed. The with this article.
removal of ions is carried out by using
reverse osmosis, electrodeionization, and References
(1) M . Tarun, C. Monferran, C. Devaux, and S. Mabic, LCGC The Peak,
ion exchange resins. 7–14 (2009).
To ensure the efficiency of the (2) B.O. Keller, J. Sui, A.B. Young, and R.M. Whittal, Anal. Chim. Acta
6(27), 71–81 (2008).
purification process and make sure (3) M. Oehme, U. Berger, S. Brombacher, F. Kuhn, and S. Kolliker,
that the ultrapure water contains the TrAC, Trend Anal. Chem. 21, 322–331 (2002).
(4) A . Khvataeva-Domanov, G. Woods, and S. Mabic, “Choosing Opti-
lowest possible ionic content, resistivity mal High Purity Water Source in Accordance to ICP-MS Application
Needs and Laboratory Environment,” presented at the Winter Con-
monitoring can be used. Ultrapure ference on Plasma Spectrochemistry, Amelia Island, Florida, 2014.
water of 18.2 MΩ·cm resistivity refers to (5) M.J. Capdeville and H. Budzinsk, TrAC, Trend Anal. Chem. 30,
586–606 (2011).
ultrapure water free of a significant level (6) C . Regnault, I. Kano, D. Darbouret, and S. Mabic, J. Chromatogr. A.
of ions for LC–MS analyses. 1030, 289–295 (2004).
(7) E . Riché, A. Carrié, N. Andin, and S. Mabic, High-Purity Water and
pH Am. Lab. (Boston) 16, 22–23 (2006)
Conclusion (8) N. Nioradze, R. Chen, N. Kurapati, A. Khvataeva-Domanov, S.

Mabic, and S. Amemiya, Anal. Chem. 89, 4836–4843 (2015).
The presence of ions in the water used (9) I. Rodushkin, E. Engstrom, and D.C. Baxter, Anal. Bioanal. Chem.
396, 365–377 (2010).
for LC–MS analyses influences the quality
of data by forming metal adducts and
Anastasia Khvataeva-Domanov and Stéphane Mabic
suppressing analyte signals. To avoid are with Millipore S.A.S. in Saint-Quentin-en-Yvelines, France. Direct
correspondence to: [email protected] ◾
ionic contamination, it is recommended to
use fresh ultrapure water when preparing
mobile phases. It is also important to
apply best laboratory practices.

Hormone Analysis by
LC–MS and Water
Impact
The increased use of hormone-based therapies

in health care throughout the world has
resulted in hormones finding their way into
municipal water supplies. The potential health
risks of unintended consumption of hormones
through drinking water have significantly
increased the interest in identifying these
compounds in our water supplies. The
Milli-Q system incorporates a combination of
purification processes that provides ultrapure,
hormone-free lab water for the LC–MS
techniques used for interference-free analysis of
these contaminants.
LCGC: I know we’ve heard it before, but

why is water quality important in liquid
chromatography and mass spectrometry?
PLURAD: Water is probably the most
used chemical in a laboratory. In liquid
chromatography and mass spectrometry, water
is used throughout the entire workflow. Any
contaminants that remain in the purified water
shutterstock.com/Olga Miltsova
used in an analysis that have a direct impact on

the separation or detection are of concern.
Also, if there are any traces of the molecules
you’re trying to analyze in the water you’re using,
you may have inaccurate results. Water quality is

important to avoid interference with the today? And how did you pick the water
analytes you’re measuring or identifying samples you chose to analyze?
as well as for optimizing instrument PLURAD: In the last 20 years, a lot of
operation. attention has been given to “persistent
organic pollutants” in drinking water
LCGC: Which specific contaminants sources. Much of the original focus
can affect the LC–MS process? in this area was on organic molecules
PLURAD: The most obvious are organics, and species that came from so-called
and in LC–MS, that’s typically what you’re chemical sources such as pesticides and
looking for. Reduction to trace levels is solvents.
key, particularly if the organics are similar With advances in health care, however,
to what you’re analyzing. pharmaceutical sources of these
Water that’s heavy with organics can persistent organic pollutants have
also cause issues with column efficiency become more significant. As many of our
by coating the separation media, resulting listeners may remember, we were once
in poor peak resolution and shifting told to dispose of our expired or unused
peaks. Ions can be a problem. Certain pharmaceuticals by flushing them down
metals can create adducts resulting in the toilet, which in retrospect was not
noisy mass spectra. the best idea because this water, now
Particle-free water is important to carrying these drugs, eventually finds
ensure proper flow through the system. its way back into the municipal drinking
With shrinking columns and tubing, as water supply.
well as improvements and changes to With the escalation of use of hormone-
separation media and higher pressures, based therapies such as topical steroids,
the impact of particles clogging an LC– birth control, and hormone-replacement
MS becomes even more magnified. therapies, there may be long-term effects
Bacterial contamination is a two-headed if these therapeutics exist in our drinking
monster. Bacteria behave as particles, so water, such as effects on human fertility
you run the risk of blocking and clogging and actual embryo development, as well
tubing or columns. But as bacteria die off, as endocrine and other general health
they leech out and reintroduce various issues.
organic and ionic contaminants into the Consequently, there’s extremely high
previously clean water. interest in identifying what’s in the water
and at what levels to determine imminent
LCGC: Regarding the work or long-term health risks. Because our
MilliporeSigma recently completed, lab water systems rely on potable tap
why is there so much interest in water as a feed source and are used
analyzing for hormones in water throughout the world, we felt it was

important to understand what influence higher levels of these persistent

hormones could have on our ability to contaminants you can expect to see
provide ultrapure water. We also wanted some residual contaminants post
to demonstrate that our purification purification.
techniques can provide high-purity water Clearly, a combination of techniques
for the detection of hormones in drinking is required to ensure full removal of
water via LC–MS. these molecules and to have water free
Because this is a global issue, we of hormone residues for your analytical
selected drinking water samples from work. We can consider purification that
various geographies including China, includes activating carbon, reverse
France, and Spain. We’re not stating that osmosis, UV photo oxidation of organics,
these samples reflect the overall water and ion exchange. And if that’s still
qualities in these countries or the safety not enough, we can consider other
of the drinking water sources. These are purification media at the point of use
single points of analysis chosen to show such as additional activated carbon that
that the problem exists to some degree targets specific contaminants.
everywhere.
LCGC: What were the results of your
LCGC: What are the challenges in analyses?
analyzing hormones at trace levels? PLURAD: We found hormones in all of
PLURAD: We found that these the water sources we tested. It stands to
hormones are everywhere and that reason that in highly industrialized and
simple or single-stage purification developed countries you would expect
techniques may not be effective in to see various hormones at various levels.
removing them. Consider deionization, Our R&D team found androsterone and
for example. As a purification technique, estradiol in city water sources in France
it only works on contaminants that have and Spain, and corticosterone was
an electrical charge. Most organics detected in China.
are neutral or very weakly charged. I’d like to reiterate it’s understood that
So deionization is not very effective at these municipal drinking water samples
removing these contaminants. are safe and suitable and approved for
Or, consider reverse osmosis. human consumption. Agencies worldwide
Although this is a workhorse in water recognize the existence of this issue and
purification, a reverse-osmosis system are taking a hard look at the long-term
operating well removes only 95% to effects of having these molecules in the
99% of the contaminants in the water drinking water.
feeding that membrane. This means The combination of purification
that in water systems that have relatively processes embedded in a Milli-Q water

purification system allows us to provide to develop purification packs that are

hormone-free lab water for the LC–MS adapted to remove very specific classes
techniques used to analyze for these of molecules at the actual point of use.
contaminants. Being able to control the purification
process 100% from tap water feeding
LCGC: The sensitivity of analyses the water purification unit gives users the
is constantly improving. How is best chance at managing the impact of
MilliporeSigma responding to this persistent contaminants.
laboratory market demand?
PLURAD: We now have scientists who are
able to analyze and quantify trace levels
of contaminants that are far below the
detection limits provided by traditional
quality measures for ultrapure water; in
fact, by orders of magnitude at this point.
Joseph Plurad,
MilliporeSigma continues to develop North America Field
Marketing Manager,
more efficient means to remove general MilliporeSigma
classes of contaminants and continues

Lab Water Hints and Tips
for the Chromatography
Lab, Part 1: Water
Purification
EMD Millipore recommends evaluating your

unique lab requirements and applications when
choosing a water purification technology
The chromatography lab requires the

highest quality of purified water. Thus, water
purification systems are a vital tool in the life
science laboratory. LCGC recently spoke with
Joseph Plurad, North America Field Marketing
Manager, EMD Millipore, about various
purification technologies that will meet any lab’s
requirements.
LCGC: Why is water important in a

chromatography laboratory?
Joe Plurad: We have to take a big step back
and understand that water is used virtually
everywhere in virtually every laboratory, not just
shutterstock.com/Olga Nikonova
in a chromatography laboratory. Water is used

to prepare samples, and sometimes, it is the
sample. Other times it could be a reagent or a
reactant and often, in the chromatography lab
specifically, it’s used as part of the eluent. Water
is used for many applications in different parts of

the chromatography workflow. chromatography will negatively impact

And think about other places where your work. And a lot of companies, both
water is used in terms of cleaning the in the chromatography side as well as
equipment or cleaning the glassware on the water side, have put a lot of work
and the lab ware, such as bottles that go into understanding what actually is in
on top of an HPLC or beakers used to the water and what it will actually do
prepare the sample. in chromatography. Water quality, in
In the biological world, water is used in sample prep, separation or detection,
the autoclaves. And often will condense can result in a lot of different problems.
or collect on what you are trying to Certainly, contaminants in the water
decontaminate. It is used in humidity can react with the sample, which could
chambers as well. These are all touch change the nature or degrade it, or
points that could impact any experiment, potentially enhance what you’re trying
but there are some specific contaminants to look for. These same contaminants
and specific issues that impact could interact with the solutions that
chromatography. So the impact of water you’re using. It could impact the quality
becomes much more important if it’s not of the eluent, especially as the solutions
properly managed. sit out because water and organics are
Water is a universal solvent, which good nutrient sources for biologics.
means that it can, and will, pick up Particles, and bacteria that behave as
contaminants from the environment, particles, can block columns, and foul
water will dissolve gasses from the air the resin and the media. The impact of
and change the quality. When you think that is poor separations, but if you are
about all of these touch points and all of successful in blocking up a column, that
these possibilities where something can could result not only in column damage
enter the water that is being used in your but instrument damage as well because
experiment, the gravity of its impact on pressure is being placed on the column
chromatography specifically, becomes and these pumps are designed to run at
much more important. very high pressures.
Post separation, if these contaminants
LCGC: What is the impact of poor are allowed to flow through, you will
quality water in chromatography? continue to have issues with detection
Joe Plurad: I don’t think it’s a coincidence techniques and each detection technique
that in the ultrapure water world, will have a specific impact.
chromatography is the number one
application demanding highly purified LCGC: What are the key contaminants
water. So any changes or deviations in water that can affect an experiment?
from water quality that is ideal for And what would happen?

Joe Plurad: In the chromatography achieve the water quality that you need
world, there are generally three classes for your application. From tap water we
of contaminants. The first is particles, or would recommend a pre-treatment to
bacteria as particles in this case. These remove the larger particles.
particles can clog frits, fittings, pre- Once you’ve gone through that
columns, and the columns themselves. prefiltration, we recommend reverse
This will result in column damage; osmosis, which is really the workhorse
damage will occur if you try to over- in water purification. This removes 95%
pressurize the system. of all the contaminants in the incoming
Another contaminant is organics. Trace tap water and across all classes, whether
organics in ultrapure water can foul inorganic, organic, or particulate.
the media of the columns. It will coat The next step is EDI, or
the columns, reducing efficiency. If you electrodeionization, which is similar to
reduce the efficiencies of the columns, our Elix technology. EDI technology uses
they won’t be as good at separating your membranes and ion exchange media and
targets. This results in shifting baselines, electrical current to remove additional
or even tailing and masked peaks. ions from the water purification stream. It
The last contaminant is ions. Within the builds on the work that reverse osmosis
world of ion chromatography, you don’t did and pulls out additional ions. This is
want your target ions in the water that going to provide additional benefit for
you’re using. However, in LC and LCMS, sodium as well as difficult-to-remove ions
alkali metals, and specifically sodium, can like silica.
form adducts that will suppress signals The next step is to pass the water
in the mass spec. Sodium is a particular through ion exchange media again to
concern in water purification and any continue to remove more ions from that
purification has to work extra hard to purified water, and hit the water with UV
reduce sodium levels prior to dispensing photo oxidation to remove organics. This
ultrapure water. will impart a charge to them that can be
removed by downstream ion exchange
LCGC: What technologies in water media. The UV photo oxidation will also
purification can address these deactivate bacteria, and remove them
contaminants? from the equation in the ultrapure water.
Joe Plurad: At EMD Millipore, we want After UV photo oxidation, we will
to educate people that the best way chase it with additional ion exchange
to manage water is to own 100% of media to do its final polishing to get
the process at the point of use. Take to that ultrapure water quality. And
tap water, and bundle or sequence the finally, at the point of use, where you’re
best series of purification techniques to dispensing the water, we recommend a

final stage purification that is specific to you actually collect it for use. Depending
your analytes and your application. At on how often the system is used, we
EMD Millipore, we have specific activated recommend flushing a few liters from
carbon cartridges that are designed to the system if it hasn’t been used in a few
remove volatile organic carbons as well as days. If you are using a water purification
a separate activated carbon filter that is system daily, or several times a day, as
designed for endocrine disruptors. little as 250–500 mL of water can be
Other options include a C-18 reverse- flushed from the system to ensure the
phase silica cartridge, a .2 micron filter freshest, most recently purified water.
at the dispensing point an ultrafilter at The goal is to be prudent about what
the point of use. However, the downside you’re picking in terms of purifying water
to ultrafiltration, is that it generally does and immediately be using it as it’s being
leach out trace organics, which are the made. In doing so, you will most likely get
nature of the membrane. the best results in your chromatography.
The best combination of techniques
specific to what you’re looking for and the EMD Millipore is the U.S. Life Science
specific chromatography technique that subsidiary of Merck KGaA, Darmstadt,
you’re using will determine which are the Germany. Headquartered in Billerica,
best techniques for a specific application. Massachusetts, the global business has
around 10,000 employees, operations in
LCGC: What else can be done to 66 countries, and 2014 revenues of $2.92
optimize results? billion.
Joe Plurad: Step one is again to use the
best technologies to purify specifically for
the analyte in your application. One thing
that we can do is as simple as producing
the right amount of water at the time
you’re about to use it.
Another practical technique includes
trying to avoid plastic containers for Joseph Plurad,
North America Field
sample preparation. Plastics typically Marketing Manager,
MilliporeSigma
use organics as release agents in their
molding process and these release
agents stick to the sides of the containers.
So very fresh glassware, very clean
glassware, is preferred.
Finally, as a point of practice, flush a
little bit of water from your system before

Lab Water Hints and Tips,
Part 2: Optimal Handling
of Purified Water
Following on our previous Dedicated Dialogue

which discussed lab water contaminants and
purification technologies, this Dedicated
Dialogue will consider specific ways to handle
and manage a lab water purification system
and the product water from these systems in
order to ensure consistent and reliable results.
To learn more about the handling of purified
water, LCGC talked with Joseph Plurad, North
America Field Marketing Manager at EMD
Millipore.
LCGC: Can you quickly review some key

contaminants that could be in ultra-pure
water and what they can do to an analysis?
Joseph Plurad: As discussed in our last podcast,
the primary contaminants of focus are organics,
ions and particles. Bacteria are also included
in the particles category because of how they
behave in a chromatography system. It should
shutterstock.com/Yeryomina Anastassiya
be pretty clear the issues that organics can

cause since most chromatographic analyses
are for organic compounds and molecules.
Organic contaminants that will either compete
or mask the organic compounds that you seek
to purify or identify need to be minimized.
Some organics can foul chromatography

media or resins which results in reduced employ multiple purification technologies

column efficiency, unstable baselines, to deliver water that meets technical
and poor peak resolution. Similarly, requirements, regardless of whether
ions can cause competing reactions using tap water or water from de-
along with separation and resolutions ionized, distilled, or reverse osmosis
issues, especially in ion chromatography. systems. In addition to the standard
When chromatography is coupled with ion exchange and UV technologies, for
mass spectrometry (MS) some ions example, companies like EMD Millipore
will create what are known as adducts, have contaminant-specific products to
which suppress the MS signals and remove specific classes of contaminants,
make molecule identification difficult. like trace organics, volatile organics,
In addition, particles, including bacteria persistent organic contaminants like
can clog frits, fittings, pre-columns endocrine disruptors, and/or bacteria and
and the columns themselves. In high particles. There are also filters that can
pressure systems, while it’s not likely remove certain enzymes like nucleases
that particulate contamination will cause and proteases. However it’s important to
the column themselves to rupture or remember that one size does not fit all.
burst—remember that these columns are While a nuclease filter may be perfect
designed to take lots of pressure—there for certain life science applications, it
are real risks of having the fittings and could actually complicate things for basic
the pumps fail as a result. At a more basic organic separation and identification.
level, pre-columns and fritted fittings will Once a system is chosen, operate it
need to be replaced more frequently if regularly and keep it well maintained.
contaminated by particulates, adding cost Purification elements like ion-exchange
and time to your analysis. So clearly, a media and physical filters do need to
well-designed and well-managed water be replaced regularly as they become
system can help reduce the impact of fouled or exhausted. Cartridges and
contaminants in everyday laboratory filters should be rinsed prior to collecting
work. water to ensure that any preservatives
that may remain from manufacturing are
LCGC: What are some good practices flushed out of the system. And of course,
in using a water system to generate proactive cartridge and filter replacement
your chromatography-grade water? ensure that you stay ahead of your
Joseph Plurad: First and foremost, contaminants. Remember that organic
make sure that the water system you’re contaminants can reduce the ion removal
using is optimized for the analysis and efficiency of resin, and once all of the
the experiments you plan on running. ion exchange active sites are consumed
Today’s high purity water systems or blocked, the cartridge provides no

Tips and Tricks for Handling High Purity Water in the LC-MS Laboratory
Anastasia Khvataeva-Domanov#, Stephan Altmaier§, Martin Lange§, and Stéphane Mabic#
# Merck Millipore, Lab Water, Lab Solutions, 1 rue Jacques Monod, Guyancourt, France, 78280; § Merck Millipore, Advanced Analytics, Lab Solutions, Frankfurter Strasse 250, Darmstadt, Germany, 64293
Introduction
Ultrapure water is highly prone to contamination, e.g. it easily leaches contaminants out of container surfaces and absorbs contamination from the laboratory environment. As ultrapure water is the most frequently used solvent in any LC-MS laboratory, its purity plays a critical role in analyses. There are a number of high purity water handling pitfalls
that result in degradation of its quality. To help analysts critically evaluate the potential risks involved in poor ultrapure water handling, we discuss here (I) the effect of laboratory environment and long high purity water storage, (II) the effect of the container material used to collect ultrapure water, (III) the effect of laboratory ware and equipment
cleaning, and (IV) the effect of poor practices of water purification system usage.
(I) Laboratory Environment and Water Storage (II) Container Material Avoid Plastic Containers (IV) Water Purification System Usage
Avoid long water storage and water exposure to Utilization of plastic devices such as bottles or funnels during Flush before Water Collection
laboratory atmosphere handling and storing of water leads to leaching of ubiquitous
additives (anti-static agents, stabilizers and plasticizers) and Water stagnating in the water purification system may degrade with time. Also contaminants
causes ghost peaks and increased background noise (Figure 3). present in the lab environment can become absorbed by the final polisher membrane and
Different types of contamination are present in laboratory environment (organics, alkali
It is recommended to collect ultrapure water in either surface contaminate water during its collection. Thus, it is recommended to properly flush a system
metals, bacteria, particles), thus water exposure to the lab atmosphere and usage of various
treated brown glass bottles or in borosilicate glass. In standard prior to water collection, e.g. with several liters after the weekend or ~ 250-500 mL when
decanting steps results in water quality degradation. The practice of long-time storage of
glass bottles silica and alkali dissolve and form adducts (see system is used every day.
ultrapure water increases the risk of water contact with the lab environment. This results
in contamination leading to high MS background noise (Figure 1), adduct formation and sodium effect in Figure 2).
signal suppression (Figure 2).
Figure 3. Effect of using polypropylene bottles for ultrapure water storage.
MS profiles of two Milli-Q® water samples stored in surface-treated brown

glass and polypropylene bottles, respectively (top), and TICs of the same
samples (bottom).
The analyses were performed via direct injection of the solvents into the
Bruker Esquire 3000+ ion trap MS operated in ESI+ mode.
Technical Effects
• Ghost peaks and signal suppression in LC-MS Figure 5A. Effect of ultrapure water collection without water purification system flushing when a system
• Decrease in sensitivity and increase of LOD was not used for several days (e.g. after the weekend).
• Increased background noise
• Complexity of MS spectra MS profile of Milli-Q® water collected from an unflushed water purification system directly after the weekend.
Implications of contaminated
Technical Effects high purity water Technical Effects
• Complexity of MS spectra • Complexity of MS spectra
• Decrease in sensitivity • Difficulties in MS data interpretation, e.g. quantification • Decrease in sensitivity
• Noisy MS background • Decrease of chromatography column lifetime • Noisy MS background
• Signal suppression • Increased maintenance of LC-MS instrument • Signal suppression
• Adduct formation • Waste of sample, reagents and solvents • Increase of LOD
Figure 1. Effect of using ultrapure water stored and exposed to the lab atmosphere and its comparison to
• Increase of LOD • Risk of repeated experiments
freshly produced ultrapure water. Figure 5B. Effect of flushing the water purification system prior to ultrapure water collection.
MS profile (top) of LC-MS grade water from competitor stored for four weeks and opened to atmosphere MS profile of freshly produced Milli-Q® water collected from properly flushed water purification system.
several times before analysis and MS profile (bottom) of freshly produced Milli-Q® water analyzed on the
day of water collection.
Analyses: Bruker Esquire 3000+ ion trap MS system, ESI+, 0.2 mL/min via syringe pump at 25°C. Technical Effects
• Decrease in sensitivity and increase of LOD
• Increased background noise
• Complexity of MS spectra
• Ghost peaks in LC-MS
• Adduct formation
(III) Glassware Cleaning

Avoid Dishwashers
Dishwashers are operated utilizing strong bases
and surfactants. The former lead to dissolution
of alkali and silica from glassware, while traces
of the latter remain on the glass surface (see
Technical Effects above).
Figure 4. Effect of dishwashers on glassware cleanness. Figure 5C. Effect of water purification system handling in the laboratory.
Comparison of MS spectra and TICs of Milli- Q® water Comparison of TICs of Milli-Q® water samples collected at different points in time. *Analyses presented in
Figure 2. Effect of Na+ presence on increased adduct formation and signal suppression.
stored in borosilicate bottles cleaned: 1- in dishwasher Figures 5A, 5B, 5C were performed using Bruker Esquire 3000+ ion trap MS system, ESI+, 0.2 mL/min via
1, 2- in dishwasher 2, 3- with a mixture of LC-MS grade syringe pump at 25°C.
A solution of 500 pmol Glu-Fibrinopeptide in 50/50 ACN/water (v/v) was spiked with different amounts
water and ACN. The mixture was analyzed via direct
of sodium (as NaCl) and the mixture was analyzed via direct injection (10 μL/min) into the Synapt® HDMS
injection into the MS, ESI+ mode.
system operated in ESI+ mode.
Conclusion Reference: LW_P 91 EMD Millipore is a division of Merck KGaA, Darmstadt, Germany. EMD
Properly installed and well-maintained Milli-Q® water purification systems meet stringent LC-MS requirements for solvent purity. The best results are obtained when using freshly collected ultrapure water without long storage Millipore, the M mark and Milli-Q are registered trademarks of Merck KGaA, Darmstadt, Germany. All
or exposure to the lab environment. Alternatively, ultrapure water should be collected in clean borosilicate glass not processed in a dishwasher. other trademarks are the property of their owners.
www.emdmillipore.com/labwater © 2014 EMD Millipore Corporation, Billerica, MA, USA. All rights reserved.
purification power or benefit at all, which centimeter resistivity with TOCs less than
can happen relatively quickly. UV lamps five parts per billion are fine for most
should also be replaced when they’re chromatographic techniques.
unable to oxidize organics or kill bacteria,
and depending on the manufacturer, LCGC: What are some best practices
replacement could be required every two for handling the purified water once it’s
years. Finally, make sure you’re regularly been dispensed from the system?
monitoring and reading the water quality Joseph Plurad: One thing to remember
prior to collecting the water for use; both is that water is a universal solvent so it
resistivity, which gives you an idea of the will absorb contaminants from the air
ionic removal, and Total Organic Carbon as well as the container you put it in. To
(TOC), which measures trace organics in minimize impact on experiments, a few
water, can put the system and subsequent things need to be considered. First, only
experiments at risk. Generally, freshly dispense what is needed for a particular
produced water that is 18.2 Mega-ohm/ experiment or “run” to minimize the total

High Purity Water: Hints and Tips

Good Practices in Using a Water Purification System and Handling High Purity Water
Estelle Riché, Stéphane Mabic, Ichiro Kano, Cecilia Regnault, Béatrice Gérion and Julien Bôle
Research and Development, Lab Water, EMD Millipore, St Quentin-Yvelines, France
Introduction Storage of purified water Flushing the water system

The usage of water purification systems is widespread in laboratories. When it comes to utilizing the purification Ultrapure water should not be stored, as it may absorb impurities from the air or the container used for storage. While a water purification system is left idle, the
system and the ultrapure water, however, each laboratory and user develops a number of habits. While some practices However, it is often necessary to store pure water before further purification with a polishing system. The water will quality of the water remaining inside the system
are rational and based on sound experience, others simply arise and set up over time. Working with high purity water be in contact with the reservoir for an extended period of time. Therefore, the reservoir material should be chosen will slowly degrade. Therefore, it is best to discard
actually requires taking a number of precautions and following some simple rules. Some of these “good laboratory carefully as it may leach organic and/or ionic compounds into the water. the first few liters of water produced when water
practices” are described here. High-density polyethylene (HDPE) and polypropylene (PP) were tested. Ultrapure water was stored in carefully rinsed is drawn from an idling system, and collect only
bottles for 24 h, water was analyzed then discarded. The procedure was repeated 3 times. Table 1 and Figure 4 show freshly purified water.
Overview that reservoirs made of HDPE are a better choice than those made of PP. HPLC: Figure 8 demonstrates that some organic
UV254 nm Ion-exchange resins
contaminants may be present in the water initially Figure 8: Effect of flushing on HPLC (A) and LC-MS (B) baselines
Pretreatment pack
+ activated carbon Polishing Unit Day 1 Day 2 Day 3
Ions (ppb)
HDPE 1 PP drawn from a water purification system.
Pretreatment unit (Elix®) (Milli-Q®)
Day 1 Day 2 Day 3 Day 1 Day 2 Day3
Bacteria: Table 2 shows a reduction in bacteria levels Bacteria in water
Fluoride ND ND ND ND ND ND
Chloride 11,7 0,9 0,5 6,0 3,8 2,2
downstream of the membrane when more water Membrane
Tap water Reverse osmosis TOC
Nitrite 2,2 0,5 < 0,1 0,3 ND ND
is drawn. This suggests that the bacteria observed
monitor at the outlet of the filter are of airborne origin.
Phosphate ND ND ND ND ND ND Pharmaceutical-Grade Bacteria in ambiant air
TOC (ppc)
Resistivity TOC To have bacteria-free water, it is recommended

Bromide ND ND ND ND ND ND Absolute, 0,22 m Filter
cell
UV185/254nm
Nitrade 1,4 0,3 < 0,1 1,3 1,0 0,7
to draw some water (>1 L) through the filter to
Electrodeionization
Sulfate 6,2 0,4 0,1 1,1 1,1 1,0
remove downstream bacteria. Even better results Volume Week 1 Week 2 Week 3 Week 4
Reservoir Calcium 9,0 2,0 1,0 5,0 1,0 1,0 can be obtained by locating the filter outlet in a delivered (L) (CFU/ml) (CFU/ml) (CFU/ml) (CFU/ml)
0,22 µm filter sterile area such as a laminar flow hood. 0,0 73 5 35 86
Magnesium 1,7 0,3 < 0,1 0,4 0,3 0,3
0,5 22 1 2 25
Monitoring the water quality Potassium 2,0 <1 <1 3,0 <1 <1
Rinsing the cartridges High purity water 1,0 0 0 2 3
Flushing the system Sodium 8,0 <1 <1 4,0 <1 <1 3,0 0 0 0 0
Careful storage of purified water HDPE 1 PP
Figure 4: Total organic carbon (TOC) extractables from Table 1: Ionic extractables from polyethylene and polypropylene.
Selection of sampling containers polyethylene and polypropylene. Flushing the water system
Rinsing the cartridges Monitoring the water quality Ultrapure water is an excellent solvent. It has a high affinity for chemicals in its surroundings and can absorb chemicals
Tap water RO cartridge 2
Water purification cartridges often are manufactured with preservatives
RO cartridge 1 RO cartridge 3 readily from storage containers, as well as chemical vapors from the laboratory air. Figure 9 shows that the polyethylene
Monitoring water quality ensures experiment repeatability and quality results. Commonly used monitoring tools are carboys and wash bottles commonly used to store water in laboratories can leach organic molecules into ultrapure
for long-term storage. In particular, this is the case for reverse-osmosis
conductivity and total organic carbon (TOC) meters. water. Glass containers leach less organics, however they may leach inorganic compounds. Polyvinyl chloride (PVC)
(RO) cartridges.
TOC (ppb)
• Conductivity measures the flow of electrons through a fluid, which is proportional to the concentration of and fluoropolymers (PFA) are also commonly used sample containers. Table 3 shows that many compounds may leach
Figure 1 shows the importance of rinsing a newly- installed RO cartridge. from these polymers into ultrapure water, as seen by ICP-MS. Figure 10 shows that chemicals present in laboratory
ions, their charge and mobility.
Initial organic contamination (TOC levels) is high (~500 ppb), then decreases air may also be absorbed by ultrapure water and affect LC-MS results.
• TOC measurement indicates the total amount of organic material present in water.
with time as rinsing occurs (~95 ppb after 300 min).
Polishing cartridges, containing ion-exchange resins and activated carbon, It is therefore important to use freshly produced ultrapure water and to select a sample container compatible with
Many organic molecules do not ionize readily and, as a result, do not influence the conductivity of water. Therefore, the analyses to be done
also must be rinsed initially. Figures 2 and 3 demonstrate the decrease in Time (min)
Figure 1: Initial rinsing of reverse-osmosis purified water may contain some organic compounds and have an excellent resistivity reading. Similarly, a low TOC
resistivity level and TOC level with rinsing time.
cartridges: evolution of TOC levels of permeate value only indicates that there are no organic molecules present in water without indicating the levels of metals or
with time. salts. Combining these two monitoring tools reflects the level of water purity more precisely.
Element (ppt) PFA bottle PVC bottle
B (11) 16,768 68,347
Large amounts of sucrose were added to pure water. Figure 7 shows that it was first retained by the activated carbon
Na (23) 0,782 110,460
of the polishing unit, then broke through the purification system. Resistivity levels remained unchanged, although Mg-2 (24) 0,295 0,800
Resistivity cell (MOhm.cm)
TOC levels rose with time. Al-2 (27) 0,832 0,788

Monitoring
Sucrose K (39) 15,932 28,058 Figure 9: Reverse-phase HPLC chromatogram of ultrapure water stored in polye-
TOC (ppb)
thylene (A) or in glass (B). Absorbance measured at 254 nm.

Ca (40) 0,565 5,834
Polishing
Tap Reverse Storage High purity Fe (56) 9,776 4,428
Unit
Water Osmosis reservoir water
(no UV) Cu (63) 0,883 1,191
Zn-1 (64) 12,092 18,489
Quantum IX # 1 Ag (107) 0,258 0,266
Quantum IX # 1
Quantum IX # 2 Quantum IX # 3 Quantum IX # 2 Quantum IX # 3 Cd (111) 2,417 1,381
Pb-2 (208) 0,055 0,049
Resistivity
Time (min) Time (min) (MOhm.cm)
Table 3: Effect of container material on sample purity. After
Figure 5: Stand-alone A10 TOC meter rinsing each container 5 times with Milli-Q® Element water, Figure 10: Total ESI+ plot (from m/z 150 to 600) of HPLC- grade water kept at room
Figure 2: Initial rinsing of polishing cartridges: changes in Figure 3: Initial rinsing of polishing cartridges: TOC temperature, opened to atmosphere three times a week for 15 minutes during a
resistivity levels of effluent water with time. changes in TOC levels of effluent water with time. fresh Milli-Q® Element water was collected in the containers and
(ppb) 2-month period.
analyzed by ICP-MS.
Figure 6: EMD Millipore Time (min)
flow-through coaxial Figure 7: Evolution of resistivity and TOC levels in ultrapure water
LW_P51 resistivity cell after addition of sucrose in the storage reservoir EMD Millipore is a division of Merck KGaA, Darmstadt, Germany
amount of exposure the water gets to or even dust and plastic fines that result
the air and to the container. Stored high from the manufacturing package or
purity water will degrade with simple storage processes. However, if using glass
absorption of CO2 from the atmosphere, containers, surface-treated amber glass
and if the container itself isn’t well bottles or borosilicate glass containers
managed, organics and particles can are recommended. Regular glass bottles
enter the bottle. Letting water stand for can leach silica and alkali metals which will
days in between runs should be avoided cause problems. So basically, anything
since it introduces the risk of bacteria and that can be done to minimize storage and
other contamination. Also, be mindful exposure to contamination risks should
of the container used. The last podcast become best practice.
discussed avoiding plastic containers.
Plastic containers, depending on the LCGC: Is there anything else you can
material of construction, could include do to minimize the recontamination of
anti-static agents, plasticizers, stabilizers ultra-pure water?

Joseph Plurad: Finding ways to reduce used. The CoA is no longer effective one
exposure is always the best practice. In the lid is removed, and the result is the
addition to picking the right size and same risk of contamination compared to
material of construction, you should water from any water purification system.
really try to use the smallest container
possible should be used to help minimize About EMD Millipore
exposure and contamination and to EMD Millipore is the Life Science division
keep everything clean. Another thing of Merck KGaA of Darmstadt, Germany
to consider is to minimize exposure and offers a broad range of innovative,
and contamination from the tubing performance products, services and
used to introduce ultra-pure water into business relationships that enable
the container, especially if transferring our customers’ success in research,
caps and moving from one container development and production of biotech
to the next. For example, bench top or and pharmaceutical drug therapies.
atmospheric contaminants can easily Through dedicated collaboration on new
be introduced if tubing or caps are scientific and engineering insights, and as
left on the countertop. If experimental one of the top three R&D investors in the
work is sensitive to CO2, bottled Life Science Tools industry, EMD Millipore
vent filters or organic traps should be serves as a strategic partner to customers
considered. Container cleaning should and helps advance the promise of life
also be avoided if possible, especially science. EMD Millipore is known as Merck
in automatic glassware washers. Most Millipore outside of the U.S. and Canada.
commercially available detergents utilize
strong bases that can etch the glass
and dissolve alkali and silica which will
be absorbed into the water. Traces of
surfactants and detergents, which are
organic, could also remain on the surface
of the glassware, only to be leached
off with the ultra-pure water dispensed
into the bottle. The same risks exist for Joseph Plurad
North America Field
bottled water. The certificate of analysis Marketing Manager
EMD Millipore
(CoA) lists the condition of the water
when it was bottled, not necessarily when

Selecting a Laboratory
Water Purification
System
Jean-Francois Pilette
Identifying the most appropriate water

purification system for your laboratory requires
that you determine the types of contaminants
you need to removed, the quality of the
available feed water, the instant and daily
volumes of Type 2 (pure) or Type 1 (ultrapure)
water you need, and the type of water quality
monitoring required. This article discusses
these parameters and describes how they affect
laboratory water use.
Primary Classes of Contaminants

Water contaminants can be divided into
five categories: inorganic salts, dissolved
organic substances, particulates and colloids,
microorganisms, and gases. Certain types of
contaminants are removed to some extent when
natural water is processed into drinking water.
The levels of these contaminants must however
be reduced further and additional contaminants
must be removed when tap water is further
purified to become laboratory-grade water.
shutterstock.com/Africa Studio
Inorganic salts are composed of cations

(positively charged ions) and anions (negatively
charged ions). The cations present in natural
water are typically sodium, calcium, iron,
manganese, lead, and aluminum. The most

common anions in natural water are Ionic contamination can also be an

chloride, nitrate, sulfate, phosphate, and issue in HPLC analyses involving mass
carbonate. The presence of unknown spectrometry detectors, especially when
or unwanted ions, or an excessive using electrospray positive (ESI +) mode.
concentration of ions, in laboratory-grade Instead of providing the conventional
water can adversely affect laboratory [M+H]+ response, sodium and potassium
processes. For example, if lead ions are ion contamination can result in [M+Na]+
being analyzed and the water used to peaks (called sodiated peaks) and [M+K]+
wash glassware or prepare standard peaks. These unintended peaks produce
solutions contains lead, the results will be incorrect mass assignments. Achieving
inaccurate. Ion concentrations therefore multiple peaks for a single molecule
have to be minimized in laboratory-grade makes data analysis more complicated
water. and difficult.
Silica, a negatively charged mineral, Dissolved organic substances, which
may also be classified as an anion. As a contain carbon atoms, are another
contaminant, silica is a special case. It is source of water contamination. Organic
weakly charged and exists in different substances can have a natural origin. For
forms. Monomeric silica comprises a example, when leaves fall on the ground
single silica atom, dimeric silica is made in autumn, rain water starts dissolving
up of two atoms, and polymeric silica is them. In time, components of the leaves
composed of many silica atoms, as well as and wood are released and dissolve in
silica sheets and silica crystal forms. Silica the rain water. The long cellulose chains
therefore encompasses a multitude of and smaller molecules such as lignin
contaminants with common behaviors but monomers, gallic acid, and catechin (a
also small differences. These differences component of the tannins in the wood)
make silica difficult to remove from water. vary widely in nature and in molecular
Removing ionic contaminants from weight.
laboratory water avoids positive bias, Natural water may also be contaminated
interferences, and variations in the by dissolved organic substances such
ionic strength of solutions during as the following generated by human
analysis. Ionic contaminants can also activities:
affect chromatographic results. In high- •S olvents called volatile organic
performance liquid chromatography compounds (VOC), such as
(HPLC) and ion chromatography (IC), ionic perchlorethene, used in the dry cleaning
contaminants can result in additional of clothing
peaks, increased background levels •M olecules such as benzene found in
that can affect sensitivity, and artificially gasoline
increased values. •P esticides such as herbicides,

fungicides, and insecticides used by the additional inorganic and organic

farming industry and in gardening contaminants.
• Polyaromatic hydrocarbons (PAHs), Colloids (such as colloidal silica) are
which are molecules released, for stable suspensions of organic and
example, from coal tar used as a inorganic particulates (0.1 to 0.001 µm in
pavement sealcoat size). The particulates in colloids usually
• Polychlorobiphenyls (PCBs), a persistent have the same electric charge, and
industrial contaminant whose use has the repulsion prevents the liquid from
been forbidden since 1976 settling (unless a coagulating agent of
• Plasticizer substances added to the opposite charge is added). Hard
polymers to make them softer, for particulates can damage or impair the
example, bisphenol-A (BPA), widely operation of laboratory instruments
used with polyvinyl chloride (PVC), a such as pumps and injectors, and soft
polymer used to make most of the particulates can plug chromatography
plastic pipes used to carry tap or columns and frits. Particulates can also act
deionized water as a shelter for microorganisms and may
operate as a catalyst to accelerate certain
As with ionic contaminants, organic reactions.
contaminants must be removed Microorganisms such as bacteria and
in order to avoid positive bias and viruses are also common natural water
interferences during organics analyses. contaminants. Bacteria vary in size and
Organic contaminants can also affect shape. Escherischia coli, a microorganism
chromatography by coating stationary commonly found in the human intestine
phases. This results in slower mass are 2 μm, while Pseudomonas diminuta,
transfer (shifting retention times), reduced is the smallest bacteria. When grown
resolution, baseline issues, and shorter in a very poor culture medium with
column lifetimes. stirring, Pseudomonas diminuta grows
Particulates and colloids contaminants as a sphere 0.27 μm in diameter.
can be divided into two classes: hard Because it is the smallest bacterium
particulates (such as sand and clay) and known, this microorganism is typically
soft particulates (such as vegetal debris). used to challenge membrane filters for
Hard and soft particulates can offer bacteriological retention.
protection against ultraviolet rays and Bacteria can be a nuisance even when
chemical agents, while acting as a surface dead because they release byproducts
for bacteria to adhere to. Particulates and that can cause problems with scientific
colloids are typically between 0.1 and 100 experiments. Gram negative bacteria
µm. Hard and soft particulates can slowly can release pyrogens from their walls,
dissolve in natural water, generating and other bacteria can release nucleases

Figure 1: DNA Separation Column: Injection 1
Retention HPLC Grade Bottled Water – TOC > 50 ppb
Figure 2: DNA Separation Column:– Injection 1950
such as RNases and DNases. Bacteria contaminants that can alter biological
and byproducts need to be removed experiments such as cell cultures. Bacteria
from laboratory water sources because byproducts such as pyrogens and
they are sources of ions and organic nucleases can affect cell operation and

Milli-Q® Water – TOC = 4 ppb
Figure 3: DNA Separation Column:– Injection 6400
molecular biology mechanisms. Bacteria Other gases can dissolve in rain water
also effect chromatography in much the and contribute to its acidification,
same way as organic contaminants. including sulfide dioxide (SO2, from
Dissolved gases are also natural combustion plants, industrial processes,
water contaminants. The primary transportation, and natural origins),
gases dissolved in water are typically nitrates (originating primarily from road
the main constituents of the air that is transportation and combustion plants),
in equilibrium with the water source: and ammonia (NH3, typically originating
oxygen, nitrogen, and carbon dioxide from agriculture activities). Removing
(which is in equilibrium with carbonic dissolved gases from water may be
acid). The solubility of these gases is required for certain experiments to
a function of temperature. Another avoid the formation of bubbles. Bubble
common dissolved gas contaminant is formation can affect the measure of
radon, a water-soluble gas produced optical density or result in outgassing,
by the decay of radium isotopes. It which can cause pump and detector
is naturally found in ground water problems in chromatography.
and results from granite formations,
phosphate deposits, and uranium Water Monitoring Techniques
deposits. In addition to being a Conductivity is the most accepted means
contaminant, radon may also cause of determining the purity of the water.
human health problems, including cancer. By definition, conductivity is a material’s

18.2 MOhm x cm water.

DNA Sep Number of
The conductivity of a material is directly
Column Water Source TOC (ppb) Injections
related to the temperature; conductivity
1 Bottle Brand B 777 408
increases—and resistivity decreases—as
2 Bottle Brand E 100 555
temperature rises. The formula used to
3 Bottle Brand C 87 2103 calculate conductivity is directly related
4 Bottle Brand A 32 1235 to the temperature of the material so
5 Bottle Brand D 16 2167 a normalized or compensated value
6 Milli-Q® Gradient 4 6394 is often reported in order to have a
7 Milli-Q® Gradient 4 10685 standard point of reference. Typically this
Bottle = HPLC Grade Bottled Water – Various Brands temperature is 25°C. Therefore, while 18.2
Figure 3: DNA Separation Column:– Injection 6400 MOhm x cm water is the accepted value
for Type 1 (ultrapure) water, it is only valid
if it is measured at or compensated to
ability to conduct electricity. However, 25°C. Since many laboratory experiments
since Type 1 (ultrapure) water itself does can be affected by ions, it is important to
not conduct electricity, conductivity, is measure resistivity to assess the presence
by proxy a measurement of all of the ions of ions.
that remain after the purification process. Because there can be multiple sources of
As the conductivity value becomes very organic contamination and because it can
small and approaches the requirements be difficult to completely removeremove
of Type 2 (approaching pure) water, the trace organics due to theirthe complex
purified water is said to be characterized nature, it is important to ensure that
by resistivity (the inverse of conductivity). the concentration of organics in water
Resistivity is a technique that attempts remains low and constant over time. This
to assert the absence of ions in Type 1 can only be achieved if a trusted measure
(ultrapure) water. Resistivity detects ions of total organic carbon (TOC) is regularly
present in water with a high degree of performed on the water prior to use.
sensitivity: A concentration as low as Figure 1 through Figure 3 demonstrate
1 ppb (1ug/L) of a salt causescause a the effects TOC can have on a
significant drop in the value of resistivity. chromatographic separation. Using
Assuming that water fully and equally HPLC-grade bottled water with a TOC
dissociates intoto H+ and OH– in greater than 50 ppb, resolution can be
equilibrium (resulting in a pH of 7), the seen to degrade significantly (Figure
theoretical conductivity of a solution of 1 and Figure 2) over the course of
H+ and OH– is 0.055 uS/cm at 25°C, or approximately 2000 injections. However
18.2 MOhm x cm at 25°C. For this reason, using Milli-Q® water, which has a TOC of
Type 1 (ultrapure) is often referred to as 4 ppb, the separation remains virtually

UV Tank
Progard ®
Reverse EDI Lamp for
for RO Osmosis (Elix )
®
254 Type 2
protection cartridge Module nm Water
Figure 4: Water Purification Techniques: Pure Water
unchanged after more than 6000 filter to retain particulates.

injections. Table 1, which compares the The conductivity of the treated water
effects of TOC levels on column lifetime is then measured before it enters the
for bottled water and Milli-Q® water, reverse osmosis cartridge. The reverse
further illustrates this point. osmosis cartridge removes more than
97% of the ions in tap water, as well
Water Purification Techniques as 99% of bacteria, particulates, and
The production of Type 2 (pure) water organics larger than 200 Dalton. A
from tap water is achieved using a series scanning electron microscope picture of a
of purification technologies, as illustrated reverse osmosis membrane cross-section
in Figure 4. Tap water entering a water is shown in Figure 5. The membrane
purification system such as the Milli-Q® is separated into two layers: a one-
Integral system first passes through a micron-thick active layer that removes
Progard pretreatment cartridge, whose contaminants on top, and a thicker (100
goal is to protect the reverse osmosis µm) porous support underneath that
cartridge. The pretreatment cartridge provides mechanical support to the
contains activated carbon to remove thinner active layer. The reverse osmosis
chlorine, polyphosphate to prevent membrane is able to reject contaminants
calcium carbonate precipitation, and a using two combined mechanisms related

Organics Particulates, Rejection is based on

Colloids & Size (~ MW)
Ions Bacteria Electric Charge
Active layer (1 m)
Porous
Support
Mechanical
Backing
SEM Picture of a RO
membrane cut through
Figure 5: RO Membrane: Structure & Properties
to the size and electric charge of the 15 L/hour to the drain and recycles 27 L/
contaminants. The reverse osmosis hour upstream of the reverse osmosis
membrane therefore effectively rejects cartridge. As a result, the system only
ions, organics (even if they are not needs an additional 20 L/hour of tap
electrically charged), particulates, colloids water to provide the 47 L/hour required
and bacteria. to feed the reverse osmosis cartridge.
The rejection performance of the reverse At the outlet of the reverse osmosis
osmosis membranes depends on the cartridge, a conductivity meter measures
nature of their material; most are made the quality of the water produced and
of cellulose acetate or polyamide. the reverse osmosis cartridge rejection
Rejection performance also depends on efficiency.
the manufacturer’s process and quality. The permeate water then enters the
For instance, in the RiOs™ 5 water Elix® module, which removes most of
purification system, the reverse osmosis the remaining ions in order to produce
cartridge is fed by a flow rate of 47 L/hour Type 2 (pure) water with a resistivity
to produce 5 L/hour permeate water. greater than 5 MOhm/cm, and a TOC
Consequently, the cartridge rejects 42 L/ greater than 30 ppb. A resistivity
hour. However, of the 42 L/hour rejected meter at the outlet of the EDI module
by the cartridge, the system sends only measures the quality of the water

UV 185 nm AC / IonEx Polisher
MILLIPORE
Ultra
Pure
Water
Pure
Water Recirculation
Figure 6: Purification Techniques to Ultrapure Water
produced by the Elix®. Elix®’s capabilities The water then moves through a
are based on electrodeionization Quantum™ cartridge, which contains
technology developed and patented activated carbon and an ion-exchange
by EMD Millipore that uses permanent resin to remove the remaining traces of
regeneration by a weak electric current. inorganic and organic ions to produce
Elix® modules can last for several years Type 1 (ultrapure) water. The synthetic
and the water does not need to be activated carbon, produced from the
softened upstream. The resulting Type 2 pyrolysis of styrene beads, has a small
(pure) water is stored in a reservoir and (<150 Å) pore size, has a surface area of
can be either used directly or further 1100m2/g, and is relatively homogeneous.
processed to produce Type 1 (ultrapure) Consequently, adsorption kinetics are
water. rapid for low-molecular-weight organics,
The production of Type 1 (ultrapure) water and very few mineral ions are released.
from Type 2 (pure) water is achieved using Activated carbon cartridges are used
the system outlined in Figure 6. The Type in these polishing cartridges mainly to
2 (pure) water first flows through a 185 nm remove trace organics. Small organic
UV lamp module to oxidize organics into molecules diffuse inside the pores and
electrically charged organic ions and to link to binding sites by weak van der
destroy bacteria. Waals forces. The quality of the Type 1

Figure 7: Polisher Packs
(ultrapure) water produced is monitored and calibration of the resistivity and

by an inline resistivity meter and an online TOC monitors are key to ensuring
TOC monitor. Typically, the sum of all ions water quality. Daily usage should also
is lower than 1 ppb and the sum of all be considered, as well as ergonomy,
organics is lower than 5 ppb. bench space, volumetric dispensing,
The produced Type 1 (ultrapure) water and flow rate (so that the water level can
can then be dispensed though a Q-POD® be accurately adjusted). A laboratory
with a POD-Pak polisher (Figure 7 at water system should also have intuitive
the outlet in order to further adjust the interface.
water quality to meet the user’s needs. Quality assurance (ISO9001 certification
The POD-Pak polisher can be adapted as and cGMP compliance) is also important.
needed for each application. Certificates for calibration, conformity,
and consumables quality must be
Selecting, Operating, and Maintaining available, and established norms of
a Water System ASTM (resistivity) and USP (resistivity and
Several parameters are important to TOC suitability tests) must be followed.
consider when choosing a laboratory CE certification, cUL certification, and a
water system. Quantity is a practical consumables validation guide should also
consideration, but perhaps more be included.
important is the quality of the water Qualification, installation, calibration,
needed—Type 2 (pure) water or Type maintenance, repair, and technical
1 (ultrapure) water. If Type 1 (ultrapure) services should be provided. Operating
water is required, the type, sensitivity, costs over the system’s lifespan—not only

the original capital expenditure—should and tackling some of the world’s most
be considered. Finally, impact on the pressing scientific challenges.
environment should be considered. As Merck KGaA, Darmstadt, Germany’s
During routine operation, storage of Type Life Science business, EMD Millipore is
1 (ultrapure) water should be avoided, as proud to be part this legacy of discovery
it can be quickly and easily contaminated. and innovation. With the company’s
For best results, avoid storing water other businesses in the United States,
in carboys. Produce water on demand we’re celebrating our collective 125-year
to prevent container extractables U.S. history and the boundless curiosity
and bacteria buildup. Unnecessary behind our past—and future—success.
connections—such as tubing at the outlet We invite you to join us in this
after the consumable filter—should be celebration and learn how you can be
avoided. Manufacturer-recommended part of our journey of discovery. Because
maintenance schedules should be Smarter, Together describes how we
followed for consumable replacements to collaborate not only internally, but
reproducibly obtain high water quality. also with our customers, vendors and
Since purified water is the most common partners across the global life science
reagent found in most laboratory industry.
facilities, a properly configured and
maintained laboratory water system is
critical to the success of almost every
laboratory experiment. The appropriate
grade of water is essential for everything
from washing glassware to buffer
preparation and cell culture analyses to
complex analytical techniques such as
HPLC or LC-MS. Appropriate purified Jean-Francois Pilette
water ensures the success of projects Group Product

Manager (Retired)
and maintain productivity. EMD Millipore
In 1890, Merck KGaA, Darmstadt,

Germany established its U.S. presence
in New York City. Today, the company
employs approximately 5,000 people
in the United States across three
businesses—Healthcare, Performance
Materials and Life Science—that
together are asking very big questions

LCGC0717 - LCGC Millipore - 3

Uploaded by

Copyright:

Available Formats

LCGC0717 - LCGC Millipore - 3

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

LCGC0717 - LCGC Millipore - 3

Uploaded by

Copyright:

Available Formats

J u ly 2 0 1 7

Sponsored by Presented in partnership with

Tips: Water Purification

Tips: Handling Purified Water

Purification System Selection

Ionic contaminants in the water used in

increase in the intensity of sodiated adducts..

Water plays an essential role in liquid

4 | July 2017 | LCGC Sponsor’s content

Table I: Direct infusion experiments

[M+H] ion (m/z 757) [M+H] ion (m/z 1570)

(d) 50:50 1000 ppb Na+ in ultrapure water–acetonitrile

5 | July 2017 | LCGC Sponsor’s content

(a) (b) 4 105

4 105 3.5 105

Experiments to Identify the in Water on MS Data

6 | July 2017 | LCGC Sponsor’s content

(a) 0 ppb of sodium in the water–acetonitrile mixture TOF MS ES+

1 ppb of sodium in the water–acetonitrile mixture TOF MS ES+

776.829 796.826 799.605 859.809

787.835 796.826 804.812807.815 813.615 827.642 859.798

796.826 797.315 786.825 798.314

797.825 787.335 799.605 800.606

797.825 799.605 800.606

7 | July 2017 | LCGC Sponsor’s content

0 785 (M+2H) 796 (M+Na+H) 807 (M+2Na) 818 (M+3Na-H)

0 ppb 1 ppb 100 ppb 1000 ppb

8 | July 2017 | LCGC Sponsor’s content

9 | July 2017 | LCGC Sponsor’s content

3. Keep Your Work Area Clean

10 | July 2017 | LCGC Sponsor’s content

Ion Removal in Laboratory Acknowledgment

Conclusion (8) N. Nioradze, R. Chen, N. Kurapati, A. Khvataeva-Domanov, S.

11 | July 2017 | LCGC Sponsor’s content

The increased use of hormone-based therapies

LCGC: I know we’ve heard it before, but

used in an analysis that have a direct impact on

12 | July 2017 | LCGC Sponsor’s content

13 | July 2017 | LCGC Sponsor’s content

important to understand what influence higher levels of these persistent

14 | July 2017 | LCGC Sponsor’s content

purification system allows us to provide to develop purification packs that are

classes of contaminants and continues

15 | July 2017 | LCGC Sponsor’s content

EMD Millipore recommends evaluating your

The chromatography lab requires the

LCGC: Why is water important in a

in a chromatography laboratory. Water is used

16 | July 2017 | LCGC Sponsor’s content

the chromatography workflow. chromatography will negatively impact

17 | July 2017 | LCGC Sponsor’s content

18 | July 2017 | LCGC Sponsor’s content

19 | July 2017 | LCGC Sponsor’s content

Following on our previous Dedicated Dialogue

LCGC: Can you quickly review some key

be pretty clear the issues that organics can

20 | July 2017 | LCGC Sponsor’s content

media or resins which results in reduced employ multiple purification technologies

21 | July 2017 | LCGC Sponsor’s content

(d) 50:50 1000 ppb Na+ in ultrapure water–acetonitrile