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<strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong><br />

January 2004 – Issue 18<br />

Inside this Issue<br />

2/3 Detecting "The New PCBs" using GC-<strong>ICP</strong>-<strong>MS</strong> - Challenges<br />

of PBDE Analysis<br />

4 User Profile: ExperTox Inc - Toxicology Laboratory, USA<br />

5 <strong>Agilent</strong> On-line Resources and Improved <strong>ICP</strong>-<strong>MS</strong> User Forum<br />

6 Phosphorus in Phosphorylated Deoxyribonucleotides using<br />

LC-<strong>ICP</strong>-<strong>MS</strong><br />

7 New Team Setup in the Americas<br />

8 Free Series of e-Seminars, Up & Coming Events, Welcome to<br />

New <strong>ICP</strong>-<strong>MS</strong> Users, New Literature


Detecting "New<br />

PCBs" using GC-<strong>ICP</strong>-<br />

<strong>MS</strong> - Challenges of<br />

PBDE Analysis<br />

Steve Wilbur and Emmett Soffey<br />

<strong>Agilent</strong> <strong>Technologies</strong>, Bellevue, WA, USA<br />

Introduction<br />

Brominated flame retardants and<br />

especially polybrominated diphenyl<br />

ethers (PBDEs) are under scrutiny<br />

around the world. PBDEs are widely<br />

used as flame retardants in plastics<br />

and are found in the plastics used<br />

in computers, construction materials,<br />

furniture and textiles. Structurally,<br />

they resemble PCBs, dioxins and<br />

furans, with the chlorines substituted<br />

by bromine. It is this similarity in<br />

structure coupled with recent data<br />

showing significant concentrations<br />

in the environment and human and<br />

animal tissues that has raised<br />

concern. A recent study in Sweden<br />

testing archived human breast milk<br />

showed levels 55 times higher in<br />

1997 than in 1972, with the average<br />

concentrations doubling every 5<br />

years. Further studies indicate that<br />

the US is far more contaminated<br />

than Sweden. For example, sewage<br />

sludge in the US contains 10-100<br />

times more PBDE than European<br />

sludge. While the global demand for<br />

PBDEs totaled 150 million pounds<br />

(68 million kg) in 1997, half of that<br />

was used by North American<br />

industries. Every day, the typical<br />

consumer comes in contact with<br />

dozens, if not hundreds of consumer<br />

goods that contain PBDEs. Since<br />

PBDEs are not covalently bound to<br />

the plastics into which they are<br />

incorporated, they are easily released<br />

into the environment. This can occur<br />

through incineration, leaching of<br />

materials in landfills, dust given off<br />

by degrading textiles and foam<br />

materials or even simple evaporation.<br />

Products containing these compounds<br />

typically contain from 5 to 20% of<br />

the product weight as PBDE.<br />

Because PBDEs are poorly soluble<br />

in water, but highly fat soluble, they<br />

are readily bioaccumulated in fatty<br />

tissues of animals and humans. Recent<br />

research on laboratory animals has<br />

shown that low level exposure to<br />

PBDEs can cause permanent<br />

neurological and developmental<br />

damage. Those most at risk are<br />

pregnant women, developing fetuses<br />

and young children. Already levels of<br />

PBDEs found in some mothers and<br />

fetuses are approaching levels known<br />

to impair learning and development<br />

in mice [1].<br />

Regulation<br />

As early as the mid 1980s to early<br />

1990s bans on the use of some PBDEs<br />

were proposed in Germany, Sweden<br />

and the Netherlands. In 1993, using<br />

its Dioxin Ordinance, Germany<br />

officially restricted the use of PBDEs<br />

because of the possibility of releasing<br />

dioxins when incinerated. In February<br />

2003, the EU announced a ban on<br />

two common PBDEs, Penta and<br />

Octa, in all products from August<br />

2004. The EU is also considering a<br />

ban on the use of Deca PBDE in<br />

electronics products by July 2006.<br />

Despite the much higher levels of<br />

PBDEs in North America, the US<br />

government has not imposed any<br />

regulations on their manufacture or<br />

use. Recently the State of California<br />

introduced legislation which would<br />

ban the use of several types of PBDEs<br />

by 2008. However, the California bill<br />

exempts the deca congener from<br />

the ban which is the most widely<br />

used type in electronic products.<br />

Several other states are considering<br />

similar legislation. Several Japanese<br />

electronic companies will be phasing<br />

PBDEs from their products and<br />

other countries and individual<br />

manufacturers are taking steps to<br />

eliminate their use of PBDEs.<br />

Structure<br />

The general structure of the PBDEs<br />

is given in Figure 1. There are ten<br />

possible sites for bromination; five<br />

on each ring. Similar to PCBs and<br />

dioxins, there are a large number of<br />

structural congeners depending on<br />

the number and location of bromine<br />

substitutions. In the case of PBDEs,<br />

there are 209 possible congeners, with<br />

the individual congeners named 1<br />

through 209. The decabromo congener<br />

is PBDE-209.<br />

Figure 1. General chemical structure of a<br />

polybrominated diphenyl ether<br />

Analytical Challenges<br />

Typically, PBDEs are analyzed and<br />

detected like PCBs or Dioxins using<br />

gas chromatography coupled to a<br />

halogen specific detector such as<br />

electron capture or to a mass<br />

spectrometer. However, unlike PCBs,<br />

PBDEs are much more difficult to<br />

separate and detect using<br />

chromatography. This is due to<br />

several differences between PCBs<br />

and PBDEs. PBDEs are high molecular<br />

weight, high boiling-point compounds<br />

which require high temperatures to<br />

elute from the GC column. However,<br />

unlike PCBs, which are very stable<br />

biphenyl compounds, the diphenyl<br />

ether structure makes PBDEs much<br />

more sensitive to degradation under<br />

high temperature GC conditions. In<br />

addition, the large number of bromines<br />

in the higher congeners places the<br />

molecular weight outside the range<br />

of some mass spectrometers.<br />

Furthermore, since bromine elicits<br />

a much lower response by electron<br />

capture than chlorine, the possibility<br />

of interferences from chlorinated<br />

compounds in some samples can be<br />

troublesome. Much work is ongoing<br />

to determine the optimum GC<br />

column dimensions and phase for<br />

PBDE analysis. At present, the<br />

best inertness for the sensitive 209<br />

congener has been shown to be a<br />

short, thin film 5 meter <strong>Agilent</strong><br />

DB-5 <strong>MS</strong> column [2]. Other columns<br />

tested show significant loss of the<br />

209 congener. However, this column<br />

is incapable of completely resolving<br />

all 209 congeners. Intensive research<br />

is ongoing at <strong>Agilent</strong> to determine<br />

the optimum chromatographic<br />

conditions for this analysis. In the<br />

meantime, only a few of the 209<br />

possible congeners are commonly<br />

used as flame retardants, so<br />

resolution of all 209 is not critical.<br />

The common commercial flame<br />

retardants are called Penta, Octa<br />

and Deca, though, for example, the<br />

Penta product is actually composed<br />

of 45% penta-BDE, 40% tetra and<br />

6% hexa congeners. Worldwide,<br />

the deca product, for which there<br />

is only one congener, is the most<br />

widely used, making up 83% by<br />

weight of the total usage. This<br />

congener is therefore the most<br />

important analytically. It is also<br />

the most difficult to measure.<br />

GC-<strong>ICP</strong>-<strong>MS</strong> Analysis<br />

Because the <strong>ICP</strong>-<strong>MS</strong> measures only<br />

the bromine, molecular weight is<br />

2 <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong> January 2004 - Issue 18 www.agilent.com/chem/icpms


not an issue for detection. Also,<br />

<strong>ICP</strong>-<strong>MS</strong> exhibits extremely high<br />

sensitivity and selectivity for bromine,<br />

eliminating possible interferences<br />

from other halogenated polyaromatic<br />

compounds. In this work [3], a mixture<br />

of 14 PBDE congeners ranging from<br />

#1 to #209 were analyzed by GC-<br />

<strong>ICP</strong>-<strong>MS</strong>. Operating conditions are<br />

given in Table 1. A chromatogram<br />

of the 10 ppb PBDE standard mix<br />

can be seen in Figure 2 with the<br />

identity of the peaks given in Table 2.<br />

GC<strong>Agilent</strong> 6890 with<br />

ALS and ChemStation<br />

software<br />

Column DB-XLB, 30m x 0.25mm<br />

LD x 0.1um (<strong>Agilent</strong><br />

part # 122-1231)<br />

Carrier Helium at 36cm/sec<br />

gas (1.5mL/min) at 100°C,<br />

constant flow mode<br />

Injector 320°C, splitless, 1uL<br />

Detector <strong>Agilent</strong> 7500cs <strong>ICP</strong>-<strong>MS</strong>,<br />

monitoring Br at m/z=81<br />

Table 1 Summary of GC-<strong>ICP</strong>-<strong>MS</strong> configuration<br />

Based on signal to noise measured<br />

in the 10 ppb standard (Figure 3),<br />

approximate detection limits are in<br />

the range of 100 ppt, which is<br />

comparable to those achievable by<br />

GC using a micro-ECD detector.<br />

However the <strong>ICP</strong>-<strong>MS</strong> "detector"<br />

exhibits superior linearity; does not<br />

suffer from non-analyte interferences;<br />

and can definitively identify<br />

brominated compounds. GC-<strong>ICP</strong>-<strong>MS</strong><br />

may well become the technique of<br />

choice for the trace level analysis<br />

of PBDEs in a variety of samples.<br />

Figure 3. Calibration curve (10ppb - 1 ppm) for<br />

PBDE-183 monitoring 79 Br<br />

www.agilent.com/chem/icpms<br />

Figure 2 GC-<strong>ICP</strong>-<strong>MS</strong> chromatogram of 10 ppb PBDE standard mix<br />

Peak Congener (2.5 mg/ml)<br />

1 2,2’,4-TriBDE (BDE-17)<br />

2 2,4’,4-TriBDE (BDE-28)<br />

3 2,3’,4’,6-TetraBDE (BDE-71)<br />

4 2,2’,4,4’-TetraBDE (BDE-47)<br />

5 2,3’,4,4’-TetraBDE (BDE-66)<br />

6 2,2’,4,4’6-PentaBDE (BDE-100)<br />

7 2,2’,4,4’5-PentaBDE (BDE-99)<br />

8 2,2’,3,4,4’-PentaBDE (BDE-85)<br />

9 2,2’,4,4’,5,6’-HexaBDE (BDE-154)<br />

10 2,2’,4,4’,5,5’-HexaBDE (BDE-153)<br />

11 2,2’,3,4,4’,5’-HexaBDE (BDE-138)<br />

12 2,2’,3,4,4’,5’,6-HeptaBDE (BDE-183)<br />

13 2,3,3’,4,4’,5,6-HeptaBDE (BDE-190)<br />

14 DecaBDE (BDE-209) (12,5 mg/ml)<br />

Table 2: PBDE peak identification<br />

References:<br />

1. S. Lunder and R. Sharp Tainted Catch, Environmental Working<br />

Group http://www.ewg.org<br />

2. J. Bjorklund, P Tollback and C Ostman, Poster Evaluation of the Gas<br />

Chromatographic Column System for the Determination of<br />

Polybrominated Dephenyl Ethers, Department of Analytical<br />

Chemistry, Stockholm University, Sweden<br />

3. E Kuhn, J. Ellis, S. Wilbur, T. Trainor, A. Gelbin GC Analysis of<br />

Polybrominated Flame Retardants Poster presented at Dioxin 2003<br />

Conference<br />

<strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong> January 2004 - Issue 18<br />

3


User Profile: ExperTox<br />

Inc. Specialists in<br />

Toxicology Testing<br />

Ernie Lykissa,<br />

ExperTox Inc, Texas, USA.<br />

Introduction<br />

ExperTox Inc. was established in<br />

1996 by Environmental Engineer<br />

Loretta M. Anderson and Forensic<br />

Toxicologist Dr. Ernest Lykissa, to<br />

address the increasing demand for<br />

toxicology studies from industry<br />

and State police departments.<br />

They equipped the laboratory with<br />

state-of-the-art instrumentation to<br />

tackle a varied workload including<br />

requests from individuals, industry<br />

or the police for certified analysis<br />

of biological specimens to include<br />

hair, urine blood, tissue and saliva.<br />

The most common request is to test<br />

for any evidence of poisoning or<br />

exposure to certain organics or heavy<br />

metals. ExperTox also conduct a full<br />

range of employment testing, including<br />

drug testing for non-prescribed<br />

narcotics. They will also establish<br />

baseline levels of residual metals<br />

present in contractors and employees<br />

working in heavy industry who may<br />

be exposed to toxic substances in the<br />

course of their work. Other examples<br />

of work undertaken include:<br />

• Analysis of food and nutritional<br />

supplements<br />

• Analysis of date rape drugs,<br />

drugs of abuse and steroids<br />

• Expert witness testimony in<br />

family, civil and criminal courts<br />

A team of chemists, biologists,<br />

medical technicians and toxicologists<br />

use a full range of analytical<br />

equipment, including <strong>ICP</strong>-<strong>MS</strong>, GC-<br />

<strong>MS</strong>, LC-<strong>MS</strong>, EIA (electrothermal<br />

atomization), HPLC and microscopic<br />

analysis, to carry out a full evaluation<br />

and report for each project.<br />

The Role of <strong>ICP</strong>-<strong>MS</strong><br />

ExperTox Inc were one of the first<br />

laboratories in the US to endorse<br />

the <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong>. They purchased<br />

a 4500 in 1996 and one of the first<br />

7500 systems in 2000. With previous<br />

experience as the head of a drug<br />

testing department for a large<br />

pharmaceutical company, Dr Lykissa<br />

was confident that all the heavy metal<br />

determinations could be handled<br />

directly by <strong>ICP</strong>-<strong>MS</strong> without the need<br />

to invest in additional techniques<br />

like Graphite Furnace Atomic<br />

Absorption (GFAAS). "GFAAS is old<br />

technology, <strong>ICP</strong>-<strong>MS</strong> is a much more<br />

powerful and productive technique<br />

that suits our needs perfectly,"<br />

commented Dr Lykissa.<br />

Analysis of Clinical Samples<br />

Primarily, <strong>ICP</strong>-<strong>MS</strong> is used to<br />

determine heavy metals such as lead,<br />

cadmium, chromium, cobalt, mercury<br />

and platinum. First, all samples of<br />

urine, blood, and other tissues are<br />

hydrolyzed in 5 % HNO3. Gold is<br />

added (10 ug/L) to stabilize any<br />

mercury present in the specimens.<br />

The toxic concentration of most<br />

heavy metals is in the high ppb (ug/L)<br />

range. Anything higher than that is<br />

indicative of an acute exposure and<br />

would require further investigation.<br />

The samples are then analyzed using<br />

the 7500a <strong>ICP</strong>-<strong>MS</strong> fitted with a High<br />

Solids (Babington type) nebulizer<br />

and standard quartz spray chamber.<br />

The instrument is optimized using<br />

a solution containing Li, Y, Ce and<br />

Tl (10ppb) for standard low-oxide/low<br />

interference levels (~0.3% CeO/Ce)<br />

while maintaining high sensitivity<br />

across the mass range. Typical<br />

instrumental conditions are displayed<br />

in Table 1.<br />

Parameter Value<br />

Forward Power 1200 W<br />

Plasma Gas Flow 15.0 L min-1<br />

Auxiliary Gas Flow 1.0 L min-1<br />

Nebulizer Gas Flow 1.2 L min-1<br />

Sampling Depth 0.5 mm<br />

Integration time 3 sec<br />

No. replicates per 3<br />

analysis<br />

Table 1 Instrumental conditions<br />

Results and Discussion<br />

For quantitative results, the<br />

instrument is calibrated using a<br />

mixed concentration multielement<br />

solution containing Ag, As, Cd, Co,<br />

Cr, Ni, Pb, Se, Al, Cu. Hg is calibrated<br />

with its own standard. Analysts<br />

also employ commercial quality<br />

controls for whole blood and urine<br />

that contain the above metals in<br />

normal and elevated concentrations.<br />

Table 1 shows 7500a data obtained<br />

on five different days compared to<br />

reference values obtained from preassayed<br />

blood reference standards.<br />

Element Ref <strong>ICP</strong>-<strong>MS</strong> data<br />

value<br />

ng/mL<br />

over 5 days<br />

As 88 88.2, 83.5, 85.3, 88.9,<br />

88.2<br />

Cd 13 12.4, 12.8, 12.5, 12.8,<br />

12.8<br />

Cr 60 54.6, 58.5, 59,2, 55.6,<br />

56.7<br />

Pb 57 57.2, 57.0, 57.7, 56.8,<br />

57,2<br />

Hg 43 42.4,42.8,44.2,43.5,<br />

43.6<br />

Se 570 562 , 573, 576, 568,<br />

567<br />

Table 1. <strong>ICP</strong>-<strong>MS</strong> control performance data over<br />

5 different days<br />

Case Study Example<br />

ExperTox worked on the case of a<br />

prominent cardiologist who suspected<br />

an estranged girlfriend of poisoning<br />

him. Tests determined the arsenic<br />

level in his urine to be 850 ug/L -<br />

way in excess of normal levels of<br />

20 ug/L and toxic levels of 100<br />

ug/L. Could this be a case of foul<br />

play? Upon close examination and<br />

testing of his house, CCA (copper,<br />

chromium, arsenic salt) treated<br />

wood and insulation material were<br />

discovered in the attic. Recent<br />

flooding in the area had created<br />

damp conditions for mold to thrive.<br />

The mold had absorbed considerable<br />

amounts of the toxic CCA which was<br />

spread throughout the house after<br />

gaining access to the air-conditioning<br />

ducts. The physician abandoned his<br />

house immediately and his urinary<br />

arsenic levels returned to normal,<br />

in a period of 30 days! Case solved.<br />

For more information on Expertox<br />

visit: http://www.expertox.com<br />

7500 Makes TV Debut!<br />

A 7500 <strong>ICP</strong>-<strong>MS</strong> and other <strong>Agilent</strong> equipment,<br />

including a 5973 GC-<strong>MS</strong>, can be seen in the<br />

current series of the US TV show Crime Scene<br />

Investigation (CSI). CSI is a very popular<br />

American primetime series that features a<br />

team of forensic investigators trained to solve<br />

crimes by careful analysis of the crime scene<br />

evidence. Equipping the forensics lab with<br />

state-of-the-art instrumentation adds to the<br />

credibility of the show - if not to its popularity.<br />

4 <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong> January 2004 - Issue 18 www.agilent.com/chem/icpms


<strong>Agilent</strong> On-line<br />

Resources at<br />

Your Fingertips<br />

Karen Morton, <strong>Agilent</strong> UK<br />

Have you seen the latest video<br />

release on the <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> web<br />

site? Follow the link from<br />

www.agilent.com/chem/icpms and<br />

witness the immediate effectiveness<br />

of the Octopole Reactions System<br />

(ORS) via the two video clips. This<br />

is just one reason to go on-line, other<br />

reasons could be to:<br />

•Find and download literature<br />

including application notes,<br />

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issues of the <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong>.<br />

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or instrument question on the<br />

improved <strong>ICP</strong>-<strong>MS</strong> User Forum.<br />

There are now more than 300<br />

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•Overview the principles of <strong>ICP</strong>-<strong>MS</strong><br />

and benefits of the 7500 Series.<br />

•Attend our online e-Seminars.<br />

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•Find support tools.<br />

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at the moment.<br />

The <strong>Agilent</strong> web site is designed to<br />

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resource that users will return to<br />

without hesitation.<br />

Why not register on the <strong>Agilent</strong> web<br />

site using the "log-in" link? You are<br />

invited to complete your profile to<br />

receive the latest news of interest<br />

to you. Be amongst the first to<br />

receive the <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong>.<br />

www.agilent.com/chem/icpms<br />

Reaching these resources is quick and easy.<br />

Web address: www.agilent.com/chem/icpms.<br />

Search for literature in the<br />

"Library"<br />

Find out all you need to know about the<br />

<strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> systems, accessories<br />

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You can access the <strong>ICP</strong>-<strong>MS</strong> and ORS Principle<br />

pages from here, or find out about events<br />

where you can meet members of the <strong>Agilent</strong><br />

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Check the Highlights<br />

for the latest<br />

information<br />

Once registered on<br />

the <strong>Agilent</strong> web site,<br />

<strong>ICP</strong>-<strong>MS</strong> users can join<br />

the User Forum - a<br />

great place to get<br />

hints from other users<br />

Access <strong>ICP</strong>-<strong>MS</strong><br />

literature quickly<br />

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Information links<br />

Click here for technical<br />

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place an order or<br />

check the status of an<br />

order here.<br />

<strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong> January 2004 - Issue 18<br />

5


Phosphorus in<br />

Phosphorylated<br />

Deoxyribonucleotides<br />

using LC-<strong>ICP</strong>-<strong>MS</strong><br />

Daniel Profrock, Peter Leonhard<br />

and Andreas Prange, GKSS Research<br />

Centre Geesthacht, Germany.<br />

Introduction<br />

Phosphorus plays an important role in<br />

cell biology for protein phosphorylation,<br />

energy storage and transport at<br />

the cellular level or as one essential<br />

part of the ribose-deoxyribose<br />

phosphorus backbone in the RNA<br />

or DNA chain [1]. The measurement<br />

of phosphorus would provide<br />

information about the phosphorylation<br />

state of a protein, which, in turn, has<br />

a significant effect on different<br />

metabolic pathways. Measuring<br />

phosphorus can also be used for the<br />

detection and quantification of RNA<br />

or DNA due to the fixed stoichiometry<br />

of this element in the above mentioned<br />

macro molecules.<br />

<strong>ICP</strong>-<strong>MS</strong> represents a highly sensitive<br />

technique for the determination of<br />

phosphorus in biological samples.<br />

Moreover, the <strong>Agilent</strong> 7500c Octopole<br />

Reaction System (ORS) <strong>ICP</strong>-<strong>MS</strong><br />

overcomes the limitation of<br />

conventional systems by removing<br />

the interferences caused by<br />

polyatomic ions ( 14 N 16 O 1 H, 15 N 16 O)<br />

on mass 31 P [2].<br />

<strong>ICP</strong>-<strong>MS</strong> Experiments<br />

The 7500c was optimized to minimize<br />

the background of the interfering<br />

ions on phosphorus while maintaining<br />

good overall sensitivity.<br />

Instrumental detection limits for<br />

the simultaneous determination of<br />

phosphorus and other trace elements<br />

in an aqueous solution were<br />

calculated.<br />

Detection limits down to 125 ng/L<br />

were achieved for phosphorus and<br />

from 18 ng/L ( 55Mn) up to 49 ng/L<br />

( 54Fe) for other trace elements<br />

measured simultaneously. The<br />

detection limits for all elements<br />

measured are summarized in Table 1.<br />

Isotope (mass) Detection limit<br />

(ng/L)<br />

P (31) 125<br />

Cr (52) 21<br />

Mn (55) 18<br />

Fe (54) 49<br />

Ni (58) 26<br />

Co (59) 19<br />

Cu (63) 25<br />

Zn (66) 37<br />

Cd (114) 32<br />

Pb (208) 24<br />

Table 1 Detection limits for phosphorus and some<br />

selected simultaneously detected trace elements<br />

in an aqueous, acidified solution, estimated<br />

according to the method outlined in ref. 3.<br />

HPLC-<strong>ICP</strong>-<strong>MS</strong> Experiments<br />

Using UV detection at 254.4nm,<br />

method optimization was undertaken<br />

with a mixture of dUMP, dAMP,<br />

dGMP, dCMP and cTMP. Based on<br />

the results, a 15 mmol L21, pH 5.8<br />

ammonium acetate buffer and 2.5%<br />

methanol (v/v) were used for all<br />

further experiments. The setup<br />

provides baseline separation of the<br />

five investigated compounds in ca. 12<br />

min.<br />

Figure 1a shows a chromatogram of<br />

the element specific determination<br />

of phosphorus in deoxynucleotides<br />

obtained by HPLC-ORS-<strong>ICP</strong>-<strong>MS</strong>.<br />

Single compound samples were used<br />

for peak assignment. The chromatogram<br />

shows one phosphorus- containing<br />

(but not UV active) peak, which<br />

remains unidentified. Detection<br />

limits based on the compound and<br />

on phosphorus were calculated for<br />

each dNMP - see Table 2. An aliquot of<br />

enzymatically digested calf thymus<br />

DNA sample was also separated<br />

under optimized HPLC conditions -<br />

see Figure 1b. The four nucleotides<br />

were baseline separated. Single<br />

compound samples and mixtures of<br />

commercially available deoxynucleotides<br />

were used for peak identification.<br />

Figure 1 Separation and element<br />

specific detection of calf thymus<br />

DNA digested with nuclease P1<br />

analyzed with HPLC-ORS-<strong>ICP</strong>-<strong>MS</strong>.<br />

(a) Mixture of dAMP, dCMP, dGMP<br />

and dTMP (100 mg/L of each compound)<br />

measured on the mass of 31<br />

P for comparison of the retention<br />

times. (b) Enzymatic digest of calf<br />

thymus DNA with nuclease P1<br />

measured on the mass of 31 P.<br />

J. Anal. At. Spectrom., 2003, 18, 708-713 -<br />

Reproduced by permission of The Royal<br />

Society of Chemistry<br />

The four peaks in the chromatogram<br />

could be clearly assigned to dAMP,<br />

dGMP, dCMP and dTMP by comparison<br />

of the retention times. New unknown<br />

peaks were also found during<br />

chromatographic separation of<br />

enzymatically digested DNA samples<br />

which could not be identified by<br />

comparison of the retention times.<br />

Species DL species DL of P DL of P<br />

(ug/L) in dNMP<br />

(ug/L)<br />

absolute (pg)<br />

dAMP 48 5 50<br />

dGMP 56 6 60<br />

dCMP 42 4 40<br />

dTMP 34 3 30<br />

Table 2. Detection limit for phosphorus in<br />

monophosphorylated deoxynucleotides obtained<br />

with HPLC-ORS-<strong>ICP</strong>-<strong>MS</strong><br />

Conclusions<br />

Using the <strong>Agilent</strong> 7500c ORS-<strong>ICP</strong>-<strong>MS</strong><br />

as a sophisticated detector for HPLC<br />

has proved to be a suitable technique<br />

for the separation and element specific<br />

determination of phosphorylated<br />

deoxynucleotides via the phosphorus<br />

located in the sugar backbone of each<br />

nucleotide. Polyatomic ions formed<br />

in the plasma and the interface region<br />

of the <strong>ICP</strong>-<strong>MS</strong> that interfere with the<br />

determination of P at mass 31 were<br />

minimized by the addition of helium<br />

to the collision cell, allowing detection<br />

limits down to 3 ug/L for dTMP.<br />

Furthermore, used as a standalone<br />

<strong>ICP</strong>-<strong>MS</strong>, the <strong>Agilent</strong> 7500c can be used<br />

for the simultaneous determination of<br />

phosphorus and other trace elements<br />

in acidified aqueous solutions down<br />

to the low-ppt level.<br />

References<br />

1 G. Loffler and P. E. Petrides, Biochemie<br />

und Pathobiochemie, Springer, 1998, pp.<br />

23-30.<br />

2 N. Yamada, J. Takahashi and K. Sakata, J.<br />

Anal. At. Spectrom., 2002, 17, 1213-1222.<br />

3 D. R. Bandura, V. I. Baranov and S. C.<br />

Tanner, Anal. Chem., 2002, 74, 1497-1502.<br />

6 <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong> January 2004 - Issue 18 www.agilent.com/chem/icpms


New <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong><br />

Team in North<br />

America<br />

Karen Morton, <strong>Agilent</strong> UK<br />

karen@agilenticpms.com<br />

<strong>Agilent</strong> is investing heavily in<br />

technical support for its <strong>ICP</strong>-<strong>MS</strong><br />

users in North America. Don Potter<br />

has taken over management of the<br />

technical sales and applications<br />

support in the Americas. In addition<br />

to his existing position as <strong>ICP</strong>-<strong>MS</strong><br />

Program Manager in Europe, Don<br />

is now also <strong>ICP</strong>-<strong>MS</strong> Program Manager<br />

for the Americas (Chris Tye performs<br />

a similar role for <strong>Agilent</strong> covering<br />

Japan and Asia/Pacific region).<br />

Don brings 20 years of experience<br />

in <strong>ICP</strong>-<strong>MS</strong> (11 years with <strong>Agilent</strong>)<br />

to the Americas team, and some<br />

new team members to increase the<br />

level of our support.<br />

Technical sales support is now<br />

handled by two product specialists:<br />

Brenda Watson, based in Houston,<br />

TX, has been with <strong>Agilent</strong> for 15<br />

years, formerly working in R&D,<br />

and as a field engineer. Brenda is<br />

responsible for the Western US<br />

states plus TX and LA. We also<br />

welcome back Abe Gutierrez<br />

(Weston, FL), who is responsible<br />

for the Eastern US and Canada.<br />

Abe originally spent 6 years with<br />

<strong>Agilent</strong> as an <strong>ICP</strong>-<strong>MS</strong> applications<br />

chemist and product specialist,<br />

and brings a wealth of experience<br />

to the team. We are also excited to<br />

announce the addition of Lisa<br />

Clark to our team, who will be<br />

working with Abe in a technical<br />

support role. Lisa joins us from<br />

Ashland Chemicals where she was<br />

an inorganic lab manager. While at<br />

Ashland, Lisa set up a state of the<br />

art semiconductor chemicals clean<br />

lab. We will be offering Lisa's<br />

expertise on a consultancy basis to<br />

advise on the set up of a trace<br />

metals analysis lab or laboratory<br />

clean room design.<br />

We have also made significant<br />

additions to our applications support<br />

team. Chris Scanlon (Livonia, MI)<br />

who has 10 years experience in<br />

<strong>ICP</strong>-<strong>MS</strong> as a product specialist with<br />

<strong>Agilent</strong> moves to applications<br />

support, joining Emmett Soffey<br />

(Bellevue, WA), also with 10 years<br />

<strong>ICP</strong>-<strong>MS</strong> experience at <strong>Agilent</strong>.<br />

Emmett, like Chris, also has many<br />

www.agilent.com/chem/icpms<br />

The <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> Team at the Winter Plasma Conference, January 2004, Ft. Lauderdale, USA<br />

years experience with GC and LC<br />

and supports our fast expanding<br />

GC-<strong>ICP</strong>-<strong>MS</strong> and LC-<strong>ICP</strong>-<strong>MS</strong> userbase.<br />

Due to the increased demand for<br />

on-site <strong>ICP</strong>-<strong>MS</strong> consulting and<br />

classroom training, we have also<br />

formed an applications consultant<br />

team. We currently have three fully<br />

trained consultants whom we<br />

contract with to provide on-site<br />

training. Dr. Johan Schijf of the<br />

University of S. Florida has been<br />

delivering our training classes at our<br />

Wilmington, DE facility for the past<br />

two years, and we welcome Dr Tom<br />

Rettberg, formerly VG/Thermo<br />

<strong>ICP</strong>-<strong>MS</strong> manager, into the team as<br />

applications consultant. Tom has over<br />

15 years experience in <strong>ICP</strong>-<strong>MS</strong> and<br />

is very well known in the industry.<br />

We also welcome back former <strong>Agilent</strong><br />

application chemist Mike Radle as<br />

a consultant for semiconductor<br />

applications.<br />

In addition to this very strong team,<br />

we also have Steve Wilbur (Bellevue,<br />

WA), worldwide environmental<br />

<strong>ICP</strong>-<strong>MS</strong> specialist based in the US.<br />

Steve is one of the leading authorities<br />

on GC-<strong>ICP</strong>-<strong>MS</strong>, having co-developed<br />

the <strong>Agilent</strong> GC interface and written<br />

extensively on the subject.<br />

For hardware support, we now have<br />

22 factory trained <strong>ICP</strong>-<strong>MS</strong> hardware<br />

engineers in N. America and we<br />

will add more as needs arise. This<br />

team is supported and trained by<br />

<strong>ICP</strong>-<strong>MS</strong> product support engineer<br />

Ron Sanderson (Dayton, OH) to<br />

ensure our <strong>ICP</strong>-<strong>MS</strong> support is<br />

maintained at the highest standards<br />

available in the industry.<br />

<strong>Agilent</strong> is absolutely committed to<br />

supporting its N. American <strong>ICP</strong>-<strong>MS</strong><br />

customers and with this new team<br />

now in place we are confident we<br />

are delivering the highest quality<br />

support for our <strong>ICP</strong>-<strong>MS</strong> customers.<br />

We'll bring you news of additions<br />

to our <strong>ICP</strong>-<strong>MS</strong> support teams in<br />

Latin America, Europe and<br />

Asia/Pacific in future issues of the<br />

<strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong>.<br />

Some contact e-mail addresses are<br />

given below:<br />

Don Potter<br />

don_potter@agilent.com<br />

Chris Scanlon:<br />

chris_scanlon@agilent.com<br />

Abe Gutierrez:<br />

abe@agilenticpms.com<br />

Emmett Soffey:<br />

emmett_soffey@agilent.com<br />

Brenda Watson:<br />

brenda_watson@agilent.com<br />

Tom Rettberg:<br />

trett@agilenticpms.com<br />

Dr. Johan Schijf:<br />

johan@agilenticpms.com<br />

Lisa Clark:<br />

lisa@agilenticpms.com<br />

Mike Radle:<br />

mike@agilenticpms.com<br />

<strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong> January 2004 - Issue 18<br />

7


<strong>ICP</strong>-<strong>MS</strong> e-Seminar<br />

Series 2004<br />

Join our free 60 minute e-seminars<br />

designed to help you get the most<br />

from your <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> from the<br />

comfort of your office.<br />

Applying Reaction Cell <strong>ICP</strong>-<strong>MS</strong> to<br />

Routine Multielement Analysis<br />

Ed McCurdy (Manchester, UK)<br />

February 18 - 2pm UK, 3pm<br />

Europe, 9am EST, 6am PST<br />

February 25 - 2 pm EST, 11am PST<br />

Environmental Analysis:<br />

A Comparison of Cell and Non-cell<br />

<strong>ICP</strong>-<strong>MS</strong><br />

Emmett Soffey (Bellevue, WA, USA)<br />

March 16 - 2pm UK, 3pm Europe,<br />

9am EST, 6am PST<br />

March 23 - 2 pm EST, 11am PST<br />

Developments in Speciation<br />

Measurement with <strong>ICP</strong>-<strong>MS</strong> as a<br />

Detector<br />

Steve Wilbur (Bellevue, WA, USA)<br />

March 18 - 2pm UK, 3pm Europe,<br />

9am EST, 6am PST<br />

March 25 - 2 pm EST, 11am PST<br />

Semiconductor Applications of<br />

Octopole Reaction Cell <strong>ICP</strong>-<strong>MS</strong><br />

Abe Gutierrez (Miami, FL, USA)<br />

March 24 - 2pm UK, 3pm Europe,<br />

9am EST, 6am PST<br />

March 31 - 2 pm EST, 11am PST<br />

Reaction Cell <strong>ICP</strong>-<strong>MS</strong> for<br />

Environmental Metals Analysis<br />

Steve Wilbur (Bellevue, WA, USA)<br />

April 8 - 4pm UK, 5pm Europe,<br />

11am EST, 8am PST<br />

To register visit<br />

www.agilent.com/chem/icpms<br />

and click on the e-Seminar<br />

Highlight for more information.<br />

Toll-free or local rate phone numbers<br />

are now available in the following<br />

countries: Austria, Belgium, Canada,<br />

Denmark, France, Germany, Ireland,<br />

Italy, Netherlands, Norway, Spain,<br />

Sweden, Switzerland, UK and USA.<br />

Events<br />

Pittcon<br />

March 7-12 2004, Chicago, USA<br />

http://www.pittcon.org/<br />

Forum Labo<br />

March 23-26 2004, Paris, France<br />

http://www.forumlabo.com<br />

This information is subject to change<br />

without notice<br />

© <strong>Agilent</strong> <strong>Technologies</strong>, Inc. 2004<br />

Printed in the U.S.A. January 27, 2004<br />

5989-0588EN<br />

New Members of the <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> Club<br />

A very warm welcome to the following companies and institutions<br />

that have recently added an <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> to their analytical facilities.<br />

• North Creek Analytical 5, USA • Texas DOH, USA • Westinghouse<br />

Savannah River 2, USA • UCLA, USA • USDA Alameda, USA • Ministerio<br />

de Salud, Panama • USDA Riverside, USA • Batelle 2, USA • GLA Labs,<br />

USA • BSI Inspectorate, USA • Univ of Delaware, USA • Chiron, USA •<br />

Georgia EPA, USA • Instituto Mexicano del Petróleo, Mexico • Jamaica<br />

Bureau of Standards, Jamaica • CEPIS, Peru • Policia de Investigaciones<br />

de Chile, Chile • ETH Zurich, Switzerland •ARPA Perugia, Italy • Agri<br />

Paradigma, Italy • STUA Lunebourg, Germany • Paks, Hungary • Uni<br />

Graz 4, Austria • Labor Dr Wessling 3, France •Labor Dr Wessling 2,<br />

Germany • Uni Essen 2, Germany • Uni Seville, Spain • Special Metals,<br />

UK • AES Howden, UK • LGC 2, UK • ATU, Germany • Inst of Food<br />

Research, UK • ASM Terni, Italy • Amdel Industrial & Environmental,<br />

Australasia • Chinese University HK, Hong Kong • KT & G Central<br />

Research Institute, Korea • Nongshim Anyang Factory, Korea • Ministry<br />

of Agriculture, PR China • Qingdao Ocean University, PR China •<br />

Shanghai Jintong University, PR China • UMC (Hsinchu) 2, Taiwan, •<br />

UMC (Hsinchu) 3, Taiwan<br />

Easier to Use <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> User Forum<br />

The <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> User Forum is now much easier to use and more<br />

intuitive thanks to its new and more powerful platform. For those not<br />

familiar with the User Forum, it's an on-line tool designed strictly for<br />

<strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> users wishing to share information relating to their <strong>ICP</strong>-<strong>MS</strong>.<br />

Questions can be applications, hardware, software, model/accessory,<br />

support specific or anything you like! New features include:<br />

• Identify new postings and discussions of interest to you.<br />

• Elect to "subscribe" to receive e-mail notification of new postings.<br />

• Search previous postings for key words or phrases using the basic or<br />

advanced search facility.<br />

• Rate your interest in the discussion, and even choose to ignore a topic<br />

that isn't relevant to you.<br />

Why not take a look?<br />

To access the <strong>ICP</strong>-<strong>MS</strong> Users Forum, simply go to www.agilent.com/chem/icpms<br />

and click on the <strong>ICP</strong>-<strong>MS</strong> User Forum link. First time visitors will need to<br />

register on the <strong>Agilent</strong> web site. New subscribers will also be prompted to<br />

enter their instrument serial number on their first visit only. Existing<br />

members will simply need to log in, as usual. Then simply click through<br />

to join the Forum.<br />

Please feel free to share your opinions on the <strong>ICP</strong>-<strong>MS</strong> User Forum with<br />

us at any time by sending an e-mail to karen@agilenticpms.com.<br />

Recent <strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> Publications<br />

To view and download these latest publications, go to<br />

www.agilent.com/chem/icpms and look under "Key Information"<br />

Technical Overview: ChemStation Software Suite for <strong>Agilent</strong> <strong>Technologies</strong> 7500 Series<br />

<strong>ICP</strong>-<strong>MS</strong>, 5989-0205EN<br />

Application Note: Fast and Accurate Determination of Arsenobetaine (AsB) in Fish<br />

Tissues using HPLC-<strong>ICP</strong>-<strong>MS</strong>, 5988-9893EN<br />

Application Note: Determination of Mercury in Microwave Digests of Foodstuffs by<br />

<strong>ICP</strong>-<strong>MS</strong>, 5989-0027EN<br />

Application Note: Speciation of Volatile Selenium Species in Plants using GC-<strong>ICP</strong>-<strong>MS</strong>,<br />

5988-9461EN<br />

<strong>Agilent</strong> <strong>ICP</strong>-<strong>MS</strong> <strong>Journal</strong> Editor<br />

Karen Morton for <strong>Agilent</strong> <strong>Technologies</strong><br />

e-mail: editor@agilent.com

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