The importance of ultra pure water for analytical research and testing and how

normal handling and storage procedures impact on its purity.

Author: Dr Paul Whitehead PhD, CChem, FRSC, Laboratory Manager, ELGA R&D Facility. Email: paul.whitehead@veoliawater.com www.elgalabwater.com

Introduction Results
Ultra pure water through 50cm of flexible PVC tubing
Methanol
Acetonitrile Acetic acid  Ultra pure Water Ion Chromotography 100
43
(Semiconductor
(TraceSELECT® Ultra) (Trace SELECT®) (PURELAB flex) •A
 nalysis using negative ion chromatography clearly 90
showed that negative ions, particularly nitrite
grade Puranol™)
Pure water is crucial for analytical research and testing applications as Aluminium <1,000 <1,000 <200 <1
Sodium ions, were detected at higher concentrations when splashing had occurred.
80
100 ng/l 70
elements and compounds present in the parts per billion (ppb) range or Antimony <500 <500 <50 <0.2 • When ultra pure water is dispensed impurities in theT
60air can easily reduce sulphonamide
N-butyl-benzene the water’s purity
Arsenic <1,000 <1,000 <500 <2 Ammonium
therefore it is important that air entrainment is minimised. 924
I

lower could affect results by interacting with samples or system components.

50
100 ng/l
C

Cadmium <500 <500 <10 <0.5 40

Calcium <30,000 <20,000 <1,000 <2 To make it more convenient to fill carboys or other large
30 storage containers a length of flexible plastic
Magnesium
One hundred per cent pure water consists solely of water molecules in Chromium <1,000 <1,000 <100 <1 50 ng/l
Calcium
tubing is often attached to fixed tap dispensers on water
20 purifiers. Samples1015
were1149
taken
1394 and analysed
Copper <1,000 <1,000 <200 <2 for plasticizer contamination after passing the ultra pure water through plastic tubing.
10

equilibrium with hydroxyl and hydrogen ions (10 -7M at 25ºC) and has Iron <1,000 <1,000 <500 <2
µS/cm 100 ng/l 0
200 400 600 800 1000 1200 1400 1600 1800 2000
5:09 10:19 15:29 20:39 25:49 31:00 36:10 41:21 46:31 51:41
a characteristic electrical resistivity of 18.2 Mohm.cm. However, the Lead
Magnesium
<500
<5,000
<500
<1,000
<10
<100
<0.2
<1
Potassium
50 ng/l
Scan

Ultra pure water passed through 50cm Ultra pure water

ability of water to dissolve almost every chemical compound to some Manganese <500 <500 <20 <0.5
of flexible PVC tubing
Mercury <200 <1,000 <1,000 <5
extent and to support nearly every form of life results in its purity being
Ultra pure water through 50cm of flexible PVC tubing Ultra pure water
Nickel <1,000 <1,000 <100 <2 43 44
100 100
Potassium <10,000 <10,000 <500 <5
under continual threat from five types of impurities: suspended Sodium <20,000 <20,000 <1,000 <2 Ultra product water
90
80
90
80

particles; inorganic compounds; organic molecules; dissolved gases; Tin <500 <1 <100 <0.5
Time (minutes)
70 70
Titanium <1,000 <1,000 <50 <0.5 60 N-butyl-benzene sulphonamide 60

and microorganisms including their associated biomolecules.

T T
I 924
Vanadium <500 <500 <10 <0.2
I
C 50 C 50
72

Zinc <10,000 <1,000 <200 <2 Figure 4: Ultra-trace cation analysis by pre-concentrating 20ml sample 40 40

Type 1 ultra pure water is by far the purest substance used in a laboratory
30 30
Bromide <1,000 <2 <20 1015
20 1394 20 1039
Chloride <25,000 <25,000 <20 Fig_3.ai
Results
1149

and is produced from mains drinking water through a series of purification

10 10 1394
Fluoride <10,000 <5 <30 0 0
•S
 ignificant improvements in background for HPLC with UV detection at 210nm were obtained using
steps to remove the different types of impurities. Nitrate <25,000 <25,000 <20 200 400 600 800 1000 1200 1400 1600 1800 2000 200 400 600 800 1000 1200 1400 1600 1800
5:09 10:19 15:29 20:39 25:49 31:00 36:10 41:21 46:31 51:41 5:08 10:19 15:29 20:39 25:49 31:00 36:10 41:20 46:31
Table 1: Comparision of Elemental Impurities in Ultra pure Water and Top Grades of Common Solvents ultra pure water with very low TOC compared to pure water with higher TOC Scan Scan

GC-MS of ultra pure water: effect of plastizer in tubing

•U
 sing ultra pure water minimises background levels, enabling highly sensitive and accurate results Figure 7: GC-MS of ultra pure water: effect of plasticizer in tubing
in analyses using HPLC and ion chromatography even at trace concentrations.
Results:
Ultra pure water
44
Typical laboratory water purification system Results
100

s
d
Fig_7.ai

ect
• All non-gaseous elements were effectively absent from ultra pure water i.e. most had detection

un

ng
ng
90
General laboratory dispensing and storage procedures that affect the purity of

Eff
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 C-MS scans show that ultra pure water passed through flexible PVC tubing can be contaminated

s
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kg
limits of less than 2 ng/l (ppt ) which is orders of magnitude less than all the other solvents tested.

ect
nk

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80

Fo
c

ca
Bla

rsi

Eff
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-Short-Term
ultra pure water

ce
with N-butyl benzene sulphonamide plasticiser illustrating how organic release agents or

mi

dia
70

pe
rfa
gh

gh

w
-Long-Term

Me

Dis

Flo
Ch

Su
Hi

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60
A number of studies were carried out to examine how common laboratory methods used for plasticisers
T
from the tubing can leach into the water.
The impact of ultra pure water contaminants on the reliability and reproducibility
I
Primary Ions C 50
72

Reservoir Polisher dispensing and storing ultra pure water can impact on the water’s purity. • I n addition,
40 a survey of users in a pharmaceutical company, showed that the average total viable
Treatment of ion chromatography Organics
30
Colloids / Particles In normal laboratory use, water is dispensed from a purifier, such as the PURELAB flex, into a vessel bacterial
20
count (TVC) in water from 22 water purifiers without tubing fitted was 0.7 CFU/ml but that
1039
Water may be used in many aspects of an analysis, including preparation of samples, dilutions, this10rose to 26 CFU/ml for seven units1394
with additional tubing on the dispense.
Bacteria where the water may be stirred. The effect of exposing ultra pure water to air over time, with and
Prefilteration Protected Deionisation standards, and blanks, as eluents and for rinsing instruments. Figure 2 summarizes results from an Ultra pure water is often
600 stored in glass
1200 or plastic
1600 wash bottles, therefore to determine the effects of the
0
Gases without stirring, was monitored (Figure 5) as was the effect of collecting water with and without 200 400 800 1000 1400 1800
Activated Carbon Water Storage Activated Carbon investigation into the effects of water impurities on ion chromatography. glass or plastic on the ultra pure
5:08
water samples were taken and monitored for contaminants after 2 days.
10:19 15:29 20:39 25:49 31:00 36:10 41:20 46:31
splashing (Figure 6). Scan
Scavenging (Recirculation) Photo-oxidation GC-MS of ultra pure water: effect of plastizer in tubing

Reverse Osmosis Filtration

Ion-exchange EDI
(a (b Contamination from a wash bottle

y
EDI Recirculation Resistivity of pure water in air

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Fig_7.ai

cib
ity

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ultra pure water stored 2 days in a glass wash bottle (20µl Injection)

itiv

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tec
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18
Eff

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Figure 1: Typical laboratory water purification system.

ect
nk

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Fo
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Higher Blanks
Bla

16
rsi

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Ba

-Short-Term
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-Long-Term

Resistivity (Mohm-cm)
e

Higher Background
Me

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Ch

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14
Ions ultra pure water stored 2 days in plastic wash bottle (20µl Injection)
Interferences
Objectives Organics
12
Media Fouling
10
The primary objectives of this study were to: Colloids / Particles Surface Coatings
8
•D
 efine the purity of ultra pure water and compare it to other solvents used in analytical research Bacteria Dispersive Effects Static
6 MIX STD (20ppb) 0.2µl Injection
and testing applications. Gases Flow Effects
4
•S
 tudy the impact of ultra pure water impurities on the reliability and reproducibility of ion Stirred
2
chromatography. Figure 2: Effects of water impurities on ion chromatography: (a) on the systemFigure
and 1.(b)
Thethe subsequent
effects potential
of water impurities impact
on the on experimental
ion chromatography technique: (a) 0 5 10 15 20
effects on the system and (b) the subsequent potential impact on experimental 0
• Examine the background levels of ultra pure water for ultra-trace HPLC and ion chromatography. results, (the area of the box provides a qualitative indication of the significance of the
results. Theimpact).
area of the box indicates the significance of the impact (qualitative). 0 5 10 Examples of phthalate ester contamination from wash-bottles (3).
Time (min) Figure 8: LC-MS comparison of phthalate ester contamination from glass versus plastic wash-bottles (ref E)
y

UPW_Fig_1a.ai
ph

•D
 etermine the effects that certain general laboratory dispensing and storage procedures have on
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Fig_6.ai
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the purity of ultra pure water.

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Results
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Results
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UPW_Fig_1.ai
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Figure 5: Effect of exposure to air on resistivity of ultra pure water

De
Ch
Re
Se

• The effects of contamination from ions, organics, colloids, bacteria and gases can all affect sensitivity
Sy

Higher Blanks Fig_4.ai • A

 fter two days storage in glass or plastic water bottles, analysis of ultra pure water by LC-MS
and reproducibility to some degree, thereby compromising and potentially negating results
Materials and Methods Higher Background showed, that among other impurities, di-n-octyl phthalate was present in the water from the
• Contaminating ions tend to have a significant but short-term effect, producing high blanks, high Results:
Type I ultra pure water used in the various tests was obtained from a PURELAB Ultra Analytic (ELGA Interferences plastic wash-bottle at ppb levels but at much lower levels in the water from the glass bottle.
background and chemical interferences that directly impact results. •W
 ithin seconds the ultra pure water starts to absorb CO2 from the air forming carbonic acid. Therefore for the analysis of organic compounds glass bottles are preferable for storing water to
LabWater) fed from a PURELAB Option R. Type II pure water was produced from PURELAB Option R. Media Fouling
• Organics, colloids and bacteria affect background/blanks but also tend to have a longer-term impact
Surface Coatings
This procedure occurs more rapidly with stirring and reduces the resistivity of the water from plastic wash-bottles (ref E).
The inorganic purity of Type I ultra-pure water was measured by Balazs Analytical Services. ICP-MS 18.2 Mohm.cm to a minimum of about 1.3 Mohm.cm. (The high conductance of the hydrogen ions
through media
Dispersive Effects fouling and surface coating that can affect parts of the instrumentation, such as the •T
 o maximise its purity, ultra pure water should be used soon after it is dispensed. In a survey of
was used to measure elemental concentrations, silica was determined colourimetrically and ions by enables this large change to occur at a CO2 concentration of only 0.5 mg/l).
chromatography
Flow Effects column, the detector or inner surfaces of the system itself. wash-bottle use, Kuroki (Ref D) found that over 80% of users did not refill their wash-bottles every
ion chromatography after pre-concentration. Data on the organic solvents were also obtained by ICP-MS.
•W
 hile carbon dioxide does not degrade the water for most applications, its effect on resistivity day with ultra pure water.
The assessments of the effects of impurities on ion chromatography are based on extensive contacts can mask the contamination of the water by other ions.
with equipment suppliers and users and in-house investigations. TheFigure
impact water
1. The effects purity
of water impurities on
on the backgrounds
ion chromatography technique:for
(a) HPLC and ion chromatography • S
 imilar contamination problems can occur if commercially bottled purified water for high sensitivity
effects on the system and (b) the subsequent potential impact on experimental
The importance of water purity for obtaining highly sensitive results using HPLC and ion chromatography
results. The area of the box indicates the significance of the impact (qualitative). analysis is stored and reused after opening.
Ultra-trace IC data was obtained in-house, concentrating 20 ml samples of Type I ultra pure water.
The HPLC traces were provided courtesy of T. Enami and N. Nagae of Nomura Chemicals. Detection at techniques was investigated. For HPLC, the background level of pure water, Type II, TOC <50ppb
210nm was used after a gradient from 0 to 100% acetonitrile over 30 minutes at 1 ml/min using a (Figure 3a) was compared with ultra pure water, Type I, TOC 1ppb (Figure 3b) and for ultra-trace cation Contamination from room atmosphere
Develosil RPAQUEOUS -5 4.6x150mm column. analysis using ion chromatography a 20ml sample was pre-concentrated. (Figure 4)
UPW_Fig_1.ai Conclusion
The detection of plasticisers in ultra pure water was carried out by GC-MS of methanol solutions of uS • The extremely high purity of ultra pure water enables accurate results to be obtained from high
residues after low temperature evaporation. sensitivity analyses such as ion chromatography and HPLC.
HPLC using Pure Water HPLC using Ultra Pure Water 0.8
CI – • Analytical accuracy is dependent on the use of water from a well designed water purification
HPLC using Pure Water HPLC using Ultra Pure Water NO2– system which maintains and monitors water purity.
0.6
The purity of ultra pure water compared to other solvents typically used in analytical • Easy to use ultra pure water dispensing and good laboratory practice during collection and use are
research and testing applications 0.02 0.02 0.4 b
essential to maintain the purity of ultra pure water and prevent results being compromised.
AUB AUB
Ultra pure water needs to be free from all 5 types of impurities if it is to be used across the whole
range of analytical and experimental applications. Measuring the levels of impurities in ultra 0.01 0.01
0.2
pure water is limited mainly by the sensitivity of the measurement techniques and the testing
environment. From current ultra-trace data ultra pure water is at least 99.99999975 % pure as the a References
PURELAB Ultra Analytic 0
Pure Water (TypeII) A. Suzuki, Kawaguchi O., Enami T. and Kuroki Y. (2006) Abstract of Proceedings of 15th Environmental Chemistry Forum; 182-183 (3)
maximum total levels of non-gaseous impurities present are less than 1.5 µg/l (ppb) for organic TOC <50ppb TOC 1ppb
B. C
 linical and Laboratory Standards Institute. Preparation and Testing of Reagent Water in the Clinical Laboratory;
compounds and less than 1.0 µg/l for other elements and ions. TIME (min) TIME (min) 0 2.5 5 7.5 10 12.5 15 17.5 Approved Guideline-Fourth Edition. CLSI document C3-A4 ( 2006)
C. Kuroki Y. (2006) Chromatography; 27(3), 125-9
The level of elemental impurities in ultra pure water were compared with data on the purest grades
D. Kuroki Y. (2003) Industrial Water; 541, 24-30 (2)
of three commercially available solvents; methanol, acetonitrile and acetic acid, used widely in
Improvements in background for HPLC with UV detection at 210mm using ultra pure water with veryImprovements
low TOC in background for HPLC with UV detection at 210mm using ultra pure water with very low TOC
Figure 6: Negative ion chromatography example of contamination from the atmosphere due to differences in water collection method:
Fig_5.ai E. H
 orikiri S., Fujita N., Kuroki Y. and Enami T. (2006) Abstracts of Proceedings of 54th Mass Spectrometry
analytical research and testing. Figure 3: HPLC: UV detection at 210nm using (a) pure water and (b) ultra pure water (a) water collected with splashing; (b) water collected by flowing it along the vessel i.e no splashing Analysis General Forum; 458-459

Fig_2a.ai Fig_2b.ai