Analytix - Sigma-Aldrich · Analytix Equipment and Reagents for UV/VIS Spectroscopy r Reagents and Equipment for UV/Vis Spectroscopy r Cronobacter Detection ... from HACH LANGE The

2011

AnalytixEquipment and Reagents for UV/VIS Spectroscopy

Reagents and Equipment for

UV/Vis Spectroscopy

Cronobacter Detection

Estradiol Analysis

New Solvents for UHPLC

GC Headspace Analysis

Organic CRMs

Pesticide Metabolites

VOLPAC Solution Containers

with New Adapters

2E

dit

ori

al

Celebrating 30 Years of HYDRANAL® Reagents for Karl Fischer Titration

Dear Colleague,

The basis of HYDRANAL is the Karl Fischer titra-

tion method, a well-known technique for

water determination in various substances

such as chemicals, oils, pharmaceuticals and

foodstuf fs. Seizing on an opportunity to

improve the safety and performance of the KF

method, pioneering Riedel-de Haën chemists

Eugen Scholz and Helga Hoffmann replaced

the noxious pyridine with imidazole. Thus began

HYDRANAL, which is now the world-wide leader

in pyridine-free reagents for KF titration and is

setting the industry standard for quality, capac-

ity, speed, safety and reliability.

Over the years, we have built a large knowledge

base. Sigma-Aldrich’s HYDRANAL service labs in

Germany and in the United States provide con-

tinuous customer support in the use of

HYDRANAL reagents and KF techniques, and

their applications for samples as well as for as

analytical requirements.

This technical help includes solving problems

such as sample solubility, side reactions,

selecting the appropriate KF reagent for spe-

cific needs and challenging samples. Support-

ive literature is offered in the form of various

brochures, a manual and a multimedia guide

on CD.

With the broad por t folio of HYDRANAL

reagents to choose from, our top priority has

been compiling, organizing and presenting

our customers with the vast amount of avail-

able information. Our long-time employee,

Helga Hoffmann, has succeeded in this chal-

lenging work and created an image that inspires our cus-

tomers and engages our business par tners. Today,

HYDRANAL is considered to be the undisputed global mar-

ket leader in Karl Fischer reagents, with a well-established

reputation for high quality, consistency and reliability. This is

Helga Hoffmann’s success. Following her retirement earlier

this year, we continue to honor her exemplary work by offer-

ing unparalleled support in serving our customers.

For challenging cases of KF titration, we have expert scien-

tists that would be happy to serve you through our

HYDRANAL Technical Helpline. Call us or email us with

your comments, questions or concerns. We look forward to

sharing our expertise with you! Furthermore, we offer an

extensive array of KF applications with protocols for over

650 applications. For a complete list, please visit our website,

sigma-aldrich.com/hydranal, where you can request

application reports to meet your specific needs.

And don’t forget our Karl Fischer seminars and work-

shops which are regularly organized in countries through-

out the world, giving you the opportunity to personally

discuss special application topics with our experts. New

seminar dates are regularly published on our website

sigma-aldrich.com/events.

As always, our most important HYDRANAL product is our

service. Our goal is your 100% success and satisfaction with

water determinations using HYDRANAL.

Kind regards,

Andrea Felgner

Market Segment Manager HYDRANAL

[email protected]

Andrea Felgner

Market Segment

Manager HYDRANAL

Analytix is published five times per year by Sigma-Aldrich Chemie GmbH,

MarCom Europe, Industriestrasse 25, CH-9471 Buchs SG, Switzerland

Publisher: Sigma-Aldrich Marketing Communications Europe

Publication director: Ingo Haag, PhD

Editor: Daniel Vogler

sigma-aldrich.com/analytix

33

Ta

ble

of

Co

nte

nts

sigma-aldrich.com/analytix

Standards

16 New Primary Reference Standards for the

Analysis of Herbal Medicinal Drugs

17 New Analytical Standards for

Phenethylamine Class Drugs

18 More Reliable Results thanks to Organic

Certified Reference Materials

19 New TraceCERT® ICP Single-Element

Standard Solutions (10 g/L)

20 New Standards for Pesticide Metabolites

Titration

21 Volumetric Titration

VOLPAC® Solution Containers with New Adapter

22 Water Determination in Various Solvents

Karl Fischer titration with HYDRANAL® reagents

Featured Article

4 Reagents and Equipment for UV/VIS

Spectroscopy

Microbiology

6 Recent Developments in our Understanding

of Cronobacter

8 Products and Methods for Cronobacter

Detection

Derivatization

10 Derivatization Agents for LC/MS – An improved

Detection of Estradiol with ESI-MS

Chromatography

12 Exclusively designed for UHPLC:

LC-MS Ultra CHROMASOLV®

13 High Purity Solvents for GC-Headspace

Applications

14 Extensive range of products for Ion

Chromatography

Sigma-Aldrich supplies high-purity water, eluent

concentrates, validation kits and certified reference

materials for highly reliable IC analyses

4

sigma-aldrich.com/spectroscopy

Fe

atu

re A

rtic

le

Reagents and Equipment for UV/VIS Spectroscopy

Novel colorimetric reagents from Sigma-Aldrich

UV/VIS spectroscopy is routinely used in the quantitative

determination of solutions for cations, anions and highly

conjugated organic compounds.

Sigma-Aldrich offers an extensive range of chemicals for

the quantitative photometric analysis of ions and molecules

by UV/VIS spectroscopy. In order to be suitable for this

application, our reagents are guaranteed to have a homo-

geneous appearance, no extraneous color and to be of

reliable quality.

Spectroscopic Cells for Scientific Research –

The variety you need for laboratory success

Sigma-Aldrich offers a full line of spectroscopy accessories,

including highly precise cuvettes for UV, fluorescence and

IR spectroscopy. For a complete product listing, please visit

our website at sigma-aldrich.com/spectroscopy

UV Cuvettes

Sigma-Aldrich offers a full line of optical cells made of

quartz, glass and plastics. All cells are manufactured accord-

ing to high-quality standards and are compatible with all

major spectrometry equipment brands.

In order to guarantee a consistent high quality, Sigma-Aldrich

offers only quartz and optical glass cells produced by Hellma®

Analytics and other selected manufacturers. Every premium-

quality optical cuvette is inspected and gauged to meet

published specifications. The quartz SUPRASIL® cuvettes

provide an excellent spectral performance from below 200

to 2500 nm.

Michael Jeitziner, Market Segment Manager Analytical Reagents & Standards [email protected]

Brand Cat. No. Description Analyte CAS No. Pack Size

FLUKA® 07670 N,N-Diethyl-p-phenylenediamine sulfate salt S2-, Cl2 6283-63-2 25 g, 100 g

FLUKA 08751 4-Amino-3-hydroxy-1-naphthalenesulfonic acid Si 116-63-2 25 g, 100 g

FLUKA 11635 Azomethine-H monosodium salt hydrate B 206752-32-1 5 g, 25 g

FLUKA 11880 Bathophenanthroline Fe 1662-01-7 500 mg, 1 g, 5 g

FLUKA 15100 Bismuthiol I Bi, Cu, Pb, Sb 1072-71-5 10 g

FLUKA 32750 3,3’-Diaminobenzidine tetrahydrochloride hydrate Se, Te 868272-85-9 1 g, 5 g, 25 g

FLUKA 61827 2,6-Bis{[bis(2-pyridylmethyl)amino]methyl}-4-methylphenol PO43- 80528-41-2 250 mg, 1 g

FLUKA 51158 Bis(1-methyl-2-imidazolyl)ketone Fe2+ 62366-40-9 1 g

FLUKA 94979 5a,6-Dihydro-5a,6,6-trimethyl-2,8-dinitro-12H-indolo[2,1-b][1,3]

benzoxazine

CN- 1023640-

20-1

100 mg

FLUKA 87748 3,3’,5,5’-Tetramethylbenzidine Cl2 54827-17-7 1 g, 5 g, 25 g

Product table (Selection)

Please find our complete product list at sigma-aldrich.com/spectroscopy

5

sigma-aldrich.com/spectroscopy

Fe

atu

re A

rtic

le

Novel Spectrophotometers and Turbidity meters

from HACH LANGE

The new instruments from HACH LANGE have been devel-

oped for the analysis of solvents, paints and pharmaceuticals.

For more than 15 years, HACH LANGE has provided expertise

in the spectral measurement of clear liquids with professional

spectrophotometers. The LICO 500 offers simple handling

and fast results with unsurpassed measurement reliability.

Reliable operation through intuitive menu guidance

and archivable user profiles

Includes all important color scales

Correct measurement results thanks to automatic

cuvette identification

High level of measurement reliability through a

comprehensive set of test aids

Only 7 seconds per measurement

Brand Prod. Nr. Description

Fluka® Z800007 LICO 500 Spectral Colorimeter

Fluka Z800236 Set of test filter for LICO 500

Fluka Z800333 HACH DATATRANS Software

Fluka Z800449 2100AN Laboratory Turbidimeter with

tungsten filament light

Fluka Z800996 US version

Fluka Z800562 2100AN IS Laboratory Turbidimeter with

860 nm (infrared) LED

Fluka Z801100 US version

Fluka Z800678 Sample Cell, Lab Turb (pkg of 6 pieces)

Fluka Z800775 HachLink 2000 Software Package

CD-version 2.9

Fluka Z800880 Cable, 2100N/AN – computer

Type Window Material Wavelength Dimension

H x W x D

Volume Material Number

Sigma-Aldrich

Pack Size

100-OS

with PTFE lid

Special optical glass 320 –2500 nm 45 x 12.5 x 12.5 mm 3.5 mL Z600644-1EA 1 cell

101-QS

with PTFE lid

Quartz SUPRASIL® 200 –2500 nm 45 x 12.5 x 12.5 mm 3.5 mL Z600172-1EA 1 cell

111-QS

with PTFE stopper

Quartz SUPRASIL 200 –2500 nm 45 x 12.5 x 12.5 mm 3.5 mL Z600717-1EA 1 cell

104-F-QS

with PTFE lid

Quartz SUPRASIL 200 –2500 nm 45 x 12.5 x 12.5 mm 1.4 mL Z600253-1EA 1 cell

Product table High precision cuvettes (Selection)

Figure 1 Various types of glass and quartz cuvettes (for dimensions, see product table below)

Figure 2 LICO 500 Spectral Photometer

Figure 3 2100 series Laboratory Turbidimeter

100-OS 101-QS 111-QS 104-F-QS

6

www.sigmaaldrich.com/sakazakii

Mic

rob

iolo

gy

methods used in this pre-genomic period for bacterial

characterization.

Cronobacter can grow over a wide temperature range. The

lowest is near refrigeration (~5 °C) and the maximum

growth temperature (44 – 47 °C) is strain-dependent.

The organism’s tolerance to drying has been well noted. For

instance, it can survive for two years desiccated in infant

formula and then rapidly grow on reconstitution. It is the

occurrence of the organism in powdered infant formula

which has been highlighted; however, Cronobacter is

ubiquitous. It has been isolated from a wide range of

sources and asympto matic human carriage has also been

reported. One probable niche for Cronobacter is plant mate-

rial, as it has been isolated from cereals, wheat, corn, soy,

rice, herbs and spices, vegetables and salads. The organism

has been isolated from a range of other foods, including

cheese, meats, milk powder, powdered infant formula and

a large number of food ingredients. The bacterium has

been isolated in the hospital environment and from clinical

samples; cerebrospinal fluid, blood, bone marrow, sputum,

urine, an inflamed appendix, neonatal enteral feeding

tubes and conjunctivae.

Cronobacter spp. have been shown to invade human intes-

tinal cells, replicate in macrophages and invade the blood-

brain barrier. Fatal infant infections have followed cases of

necrotizing enterocolitis (NEC), septicaemia and meningitis.

In Cronobacter meningitis, there is gross destruction of the

brain, leading sadly to either death (40 – 80% of cases) or

severe neurological damage. The pathogenesis of Crono-bacter meningitis is different to Neisseria meningitidis and

neonatal meningitic E. coli, and is similar to that of the

closely related bacterium, Citrobacter koseri.

The use of chromogenic media to differentiate Cronobacter from other Enterobacteriaceae present in samples has

greatly improved the isolation of the bacterium. Unlike

most other members of the Enterobacteriaceae, Crono-bacter constitutively expresses the enzyme α-glucosidase,

and this is exploited as a diagnostic feature in chromogenic

media by incorporating a chromogen such as 5-bromo-4-

chloro-3-indolyl α-D-glucopyranoside (X-α-gluc). Crono-bacter hydrolyze this colorless chromogen to produce

characteristic blue/green colonies for presumptive identifi-

cation on the plate. Nevertheless, presumptive Cronobacter colonies need to be confirmed using phenotyping or geno-

typing methods to avoid the false positive due organisms

such as E. vulneris and E. hermanii.

The newly designated bacterial genus Cronobacter is com-

posed of Gram-negative, facultative anaerobic rods, which

are members of the Enterobacteriaceae family and closely

related to the enterobacter and Citrobacter. It is composed

of C. sakazakii, C. malonaticus, C. turicensis, C. muytjensii, and

C. dublinensis, plus an unnamed sixth species. The organism

came to prominence due to its association with severe neo-

natal infections: necrotizing enterocolitis, septicaemia and

meningitis. The fatality rate following meningitis is 50%,

with the survivors being neurologically damaged for life.

Such infections are rare in infants. However, infections

occur in all age groups, though fortunately with less severe

clinical outcomes.

About 80% of its strains produce a non-diffusible, yellow

pigment on Tryptone Soya Agar at 25 °C, which is why it was

first described as ‘yellow-pigmented Enterobacter cloacae.’ In

the 1980’s, researchers used DNA-DNA hybridization to

show that these strains were a unique taxonomic group

and should be recognized as a separate species Enterobacter sakazakii, named in honor of the Japanese bacteriologist

Riichi Sakazaki. The organism attaches to surfaces, forming

biofilms that are resistant to cleaning and disinfectant agents,

and has also been found as part of the mixed flora biofilm

in enteral feeding tubes from neonatal intensive care units.

However, since the 1980s, bacterial systematics has increas-

ingly used DNA sequencing for its analysis and for deter-

mining relatedness. Analysis of both partial 16S rDNA and

hsp60 gene sequences showed that E. sakazakii isolates

formed at least four distinct genomogroups, which could

be regarded as unique species. However, such a taxonomic

revision required considerable further analysis for substan-

tiation. The Cronobacter genus was defined first in 2007 and

revised in 2008. This slow realization and recognition of

Cronobacter reflects the laborious and time-consuming

Recent Developments in our Understanding of Cronobacter

Prof. Stephen J. Forsythe, Nottingham Trent University

Figure 1 Cronobacter on TSA showing mucoid flat colonies after

three days at 25° C (source: CDC/Dr. J. J. farmer)

7

www.sigmaaldrich.com/sakazakii

Mic

rob

iolo

gy

Members of the Cronobacter genus are diverse and this is

now being investigated at the genomic level. Last year, we

used both whole genome sequence analysis and compara-

tive genomic hybridization-based analysis to describe a

range of virulence traits in Cronobacter [1]. The strain to be

sequenced was C. sakazakii strain BAA-894. This strain had

originally been isolated from powdered formula associated

to a fatal Cronobacter outbreak on a NICU. The genome

comprises a 4.4 Mb chromosome (57% GC content) and two

plasmids: 31 kb (51% GC) and 131 kb (56% GC). The genome

was used to construct a 387,000-probe, oligonucleotide-til-

ing DNA microarray covering the whole genome. Compara-

tive genomic hybridization (CGH) was undertaken on five

other C. sakazakii strains and representatives of the four

other Cronobacter species. Among 4,382 annotated genes,

about 55% of the genes were common to all C. sakazakii strains and 43% were common to all Cronobacter strains,

with 10–17% absence of genes. CGH highlighted 15 clusters

of genes in C. sakazakii BAA-894 that were divergent or

absent in more than half of the tested strains; six of these

are of probable prophage origin. A number of genes

unique to Cronobacter species associated with neonatal

infections (C. sakazakii, C. malonaticus and C. turicensis) were

identified. These included a copper and silver resistance

system known to be linked to invasion of the blood-brain

barrier by neonatal meningitic strains of Escherichia coli. In

addition, genes encoding for multidrug efflux pumps and

adhesins were identified that were unique to C. sakazakii strains from outbreaks in neonatal intensive care units.

OmpA probably has a role in the organism penetrating the

blood-brain barrier, though the mechanism leading to the

destruction of the brain cells is unknown and could, in part,

be a host response. The genes that were shared by the

three strains associated with C. sakazakii outbreaks in NICUs

were compared with the C. sakazakii species type strain

ATCC 29544T, which showed decreased virulence proper-

ties in tissue culture studies. One hundred and forty four

genes present in the three NICU strains were absent in the

type strain. In most of these clusters (genes encoding pro-

teins associated with resistance to different forms of stress)

were identified, including multidrug efflux systems, genes

involved in resistance to oxidative stress and those with a

putative function in resistance to metals.

Typing Cronobacter to understand its diversity has

led to the development of a multilocus sequence typing

( M L S T ) s c h e m e [ 2 ] , w h i c h i s a v a i l a b l e o n l i n e

(www.pubMLST.org/cronobacter). The scheme for C. sakazakii has revealed stable clones, some of which could

be traced over a 50-year period from a wide range of coun-

tries and sources. The MLST scheme is based on seven

housekeeping genes (atpD, fusA, glnS, gltB, gyrB, infB, ppsA;

3036 nt concatenated length). The MLST scheme currently

has 79 defined sequence types covering all Cronobacter species. Earlier this year (2011), it was realized that, although

the clinical isolates were in 10/28 STs defined for C. sakazakii, these were not evenly distributed [3]. Of particular interest

was that half of the strains were ST4. This appears to be a

very stable clone as clinical and non-clinical strains have

been isolated from seven countries for over 50 years.

C. sakazakii ST4 appears to be a highly stable clone with a

high propensity for neonatal meningitis. Therefore, our cur-

rent research is into the genomic analysis of this life-threat-

ening variety of Cronobacter sakazakii and at ASM this year,

we announced the sequencing of eight further Cronobacter genomes, including C. sakazakii ST4. We believe opportuni-

ties of whole genome sequencing and detailed analysis is

necessary in order to further develop reliable and robust

detection schemes and control measures for emergent bac-

terial pathogens such as Cronobacter.

References

[1] Kucerova E., Clifton S.W., Xia X-Q., Long F., Porwollik S.,

Fulton L. et al., 2010: Genome sequence of Cronobacter

sakazakii BAA-894 and comparative genomic hybridization

analysis with other Cronobacter species, PLoS ONE 5, e9556.

[2] Baldwin A., Loughlin M., Caubilla-Barron J, Kucerova E.,

Manning G, Dowson C. et al., 2009: Multilocus sequence

typing of Cronobacter sakazakii and Cronobacter malonaticus

reveals stable clonal structures with clinical significance

which do not correlate with biotypes, BMC Microbiology 9,

page 223.

[3] Joseph S. and Forsythe S., 2011: Association of Cronobacter

sakazakii ST4 with neonatal infections. Emerging Infectious

Disease 2011, in press.

Figure 2 HiCrome™ Cronobacter Agar, Modified (Fluka 14703), with

Cronobacter colonies

8

sigma-aldrich.com/hybriscan

Mic

rob

iolo

gy

Cronobacter is a rod-shaped, motile and facultatively-anaerobic bacterium of the

family Enterobacteriaceae. Originally, Cronobacter was described as yellow-pigmented

Enterobacter cloacae (yellow pigment on a Tryptic Soy Agar (TSA) at 25 °C). However,

today there are also studies that have demonstrated that not all Cronobacter spp. produce yellow-pigmented colonies on Tryptic Soy Agar.

For the classical microbiological tests, a pre-enrichment step is used to recover the

stressed cells, which is followed by a selective enrichment step.

The FDA method recommends Enterobacteriaceae enrichment (EE or Mossel) broth,

which is then streaked onto VRBG Agar; suspect colonies are subcultured onto TSA

where the yellow-pigmented colonies are confirmed by an oxidase test and other bio-

chemical tests. Table 1 lists the diverse biochemical reactions from Cronobacter spe-

cies (not only from FDA).

ISO/TS 22964:2008 methods recommend using buffered peptone water (BPW) as a

pre-enrichment medium and modified lauryl-sulphate broth with vancomycin

(mLST) incubated at 44 °C for the secondary selective enrichment step. The next step

is then a chromogenic agar for isolation and identification (see Table 2).

There are diverse chromogenic agars available, which help to save work and time in

producing results and are more reliable than traditional media. The detection principle

is based on the alpha-glucosidase possessed by Cronobacter spp. (not by most other

Enterobacteriaceae), which cleaves the 5-bromo-4-chloro-3-indolyl-a-D-glucopyrano-

side or similar substrates. In the case of Cronobacter spp., the result is a plate with blue

colonies (see Figure 1); however, biochemical confirmation is still required.

Media Type Cat. No. Name

nonselective pre-enrichment 77187 Peptone Water, phosphate-buffered

selective pre-enrichment 89916 Modified Lauryl Sulfate Tryptose Broth (ISO/TS 22964)

69965 Mossel Broth

isolation and differentiation 92324 HiCrome™ Cronobacter spp. Agar

14703 HiCrome Cronobacter spp. Agar, modified

(ISO/ TS 22964)

22091 Tryptic Soy Agar

79872 Tryptic Soy Agar (ready prepared in Flask)

70189 Violet Red Bile Glucose Agar

Table 2 Media for Cronobacter detection

New Rapid Method

HybriScan®D Cronobacter spp. is a new rapid molecular test system for detection of

bacteria of the genus Cronobacter. It is based on the detection of rRNA by sandwich

hybridization; therefore, no PCR is needed. Further information may be found at

sigma-aldrich.com/hybriscan

Brand Cat. No. Name Assays

Fluka® 12838 HybriScan®D Cronobacter spp. 96 (microplate format)

Biochemical

Test

Cat. No.

of Test

Reaction of

Cronobacter

Gram 77730 -

Oxidase 40560 -

Catalase 88597 +

H2S production 06728 -

Nitrate reduction 51138 +

Citrate utilization 85463 +

Esculin hydrolyzation 06105 +

Arginine hydolyzation D2935 +

Lysine D2935 -

L-ornithine decarboxylation D2935 +

Urease 51463 -

Indole 67309 -

ONPG 49940 +

D-adonitol 55876 -

L-arabinose 80372 +

D-arabitol A3381 -

D-cellobiose 56481 +

Dulcitol 73044 -

D-fructose 53901 +

D-glucose 63367 +

D-galactose 89608 +

Inositol 89614 + (75%)/-

Inulin 90058 +

Lactose 28816 +

Malonate 63290 +/-

D-maltose 77653 +

D-mannitol 94438 +

D-mannose 94445 +

D-melibiose 93196 +

x-methyl-D-glucoside 66940 +

D-raffinose 94226 +

L-rhamnose 93999 +

Salicin 92971 +

Sorbitol 93998 -

D-sucrose 94309 +

D-trehalose 92961 +

Xylose 07411 +

Voges-Proskauer test

(acetoin production)

07689 +

Methyl red test 08714 -

Tryptic Soy Agar at 25 °C 22091 yellow pigment

Table 1 Biochemical reactions of Cronobacter ssp.

Products and Methods for Cronobacter DetectionCronobacter, formerly called Enterobacter sakazakii, is an important consideration for infant formula manufacturers.

Jvo Siegrist, Product Manager Microbiology [email protected]

Figure 1 HiCrome

Cronobacter spp. Agar

(Fluka 92324);

Cronobacter (blue),

E. aerogenes (green)

K. pneumoniae (yellow)

9

sigma-aldrich.com/tlc

Ch

rom

ato

gra

ph

y

Your Day-to-Day Needs for TLC Plates – SolvedSpecial offer of our new Silica Gel matrix on Aluminum TLC plates

Sigma-Aldrich offers you a new quality of TLC plates on aluminum with a standard silica

gel matrix. Easy-to-cut sheets, excellent separation efficiency and an outstanding

wetability ensure optimum use for all your applications. Test them today for your daily

routine work. Visit us at: sigma-aldrich.com/tlc

Material Brand Name Size Fluoresence Indicator Thickness of Layer

49859-50EA Supelco® Silica Gel on TLC-Al foils 4 cm x 8 cm 254 nm 0.20 mm

52038-20EA Supelco Silica Gel on TLC-Al foils 5 cm x 7.5 cm 254 nm 0.20 mm

23478-50EA Supelco Silica Gel on TLC-Al foils 5 cm x 10 cm 254 nm 0.20 mm



55811-20EA Supelco Silica Gel on TLC-Al foils 5 cm x 7.5 cm without 0.20 mm

75196-50EA Supelco Silica Gel on TLC-Al foils 5 cm x 10 cm without 0.20 mm



Silica with a mean pore size of 60 Å, specific pore volume 0.75 mL/g and a particle size of 5 –17 μm, for reliable and

reproducible results. A new binder system offers easy cutting.

Offer valid till December 31, 2011. To take advantage of this special offer, please refer to Promotion Code 971 when

ordering.

Your Day-by-Day Demand of SilicaGet your bulk demand of Silica 60 Å for a special price!

Sigma-Aldrich provides a broad range of silica gel and other separation media from standard qualities to modified

specialities used for dedicated applications. In addition to this extraordinary diversity, we also offer economical prod-

ucts for generic and routine purification in the same spectrum. Silica 60 is one of the most common products and is

used in almost every laboratory. Get your tailor-made quote for Silica 60 Å (premium and standard quality) in bulk

that combines Sigma-Aldrich service and quality with a competitive pricing. Visit us at: sigma-aldrich.com/silicagel

Material Brand Name Purpose Particle Size

60741-1KG; 60741-5KG; 60741-25KG Fluka® Silica Gel 60 Å for column chromatography 0.063 – 0.200 mm

60738-1KG; 60738-5KG; 60738-25KG Fluka Silica Gel 60 Å for column chromatography 0.035 – 0.070 mm

60737-1KG; 60737-5KG; 60737-25KG Fluka Silica Gel 60 Å for column chromatography 0.04 – 0.063 mm

717177-1KG; 717177-5KG; 717177-25KG Aldrich Silica Gel 60 Å for column chromatography, technical grade 0.063 – 0.200 mm

717185-1KG; 717185-5KG; 717185-25KG Aldrich Silica Gel 60 Å for column chromatography, technical grade 0.04 – 0.063 mm

Offer valid till December 31, 2011. To take advantage of this offer, refer to Promotion Code 954 when ordering.

30% off!30% off!

70% off!70% off!

10

1

0.2 0.4 0.6 0.8 1.0 Time [min0.0

0.5

1.0

1.5

2.0

2.5x104

Intens.

171.102

203.095

234.067277.106

301.072354.753

398.151423.152 473.154

+MS2(506.230), 1.7-1.9min #(259-276)

0

50

100

150

200

100 200 300 400 500 600 700 m/z

sigma-aldrich.com/derivatization

De

riv

ati

zati

on

enhance the detection limits significantly. Dansyl chloride is

the most common agent and reacts selectively and quanti-

tatively with E2, testosteron and their derivatives [3– 4]. The

detection is limited to APCI and APPI sources, which have

some disadvantages regarding availability, dopant usage

and the lower sensitivity of the APCI source (Figure 1). The

MS/MS spectra result a large number fragments and a

lower sensitivity on the quantifier.

A more sensitive and versatile derivatization agent for ESI

sources is 4-(Dimethylamino) benzoyl chloride (67954-1G,

DMABC). The reagent can be dissolved in acetone and

applied on the dried residue of the sample extract. An adjust-

ment of the pH is not necessary, only an anhydrous reaction

medium is needed. The high purity of DMABC guarantees

good solubility, and a very selective and quantitative reac-

tion at a moderate temperature between 55– 60 °C (5 min.).

The reagent and possible by-products can be separated

from the analytes using a standard reversed-phase HPLC

column and can be detected down to very low concentra-

tions (s. Figure 3).

Steroid hormones are derivatives of cholesterol and play an

important role in a large variety of organisms, as they can

have direct control over the gene expression. 17β-estradiol

(E2) controls the growth and function of female secondary

sexual characteristics. High blood concentrations inhibit

the formation of further regulatory factors responsible for

ovulation and pregnancy. E2 and its derivatives, e. g. ethinyl

estradiol, are included in commonly used, widespread con-

traceptive pharmaceuticals, which is having a thus far

unconsidered environmental impact: increased concentra-

tions of estradiol and its metabolites in wastewater [1–2].

Clinical and environmental laboratories alike now have a

vital interest in finding the most sensitive method for the

analysis of E2 and other steroid hormones, which are usually

in matrices and so difficult to remove.

E2 is a very unpolar compound and hardly detectable by

ESI. Fortunately, the analyte can be extracted very effi-

ciently with solvents like methylene chloride or acetone. In

addition, this procedure reduces the negative effects of the

matrix, e. g. signal suppression by alkali salts. However, only

the introduction of ionizable moieties by derivatization can

Derivatization Agents for LC/MS – An Improved Detection of Estradiol with ESI-MS

Rudolf Köhling, Senior Scientist, LC/MS Applications [email protected]

Figure 1 Separation and detection of 55 pg E2 as dansyl derivative (EIC, peak 1). Only a short pre-column (Supelco Ascentis Express C18,

2.7 μm, 2.1 x 5mm) is necessary to separate the analyte from excessive reagent and by-products (BPC, magenta). The inset shows the MS/MS

spectrum of [M+H]+=506.235 Da (APCI+).

11

1 2 3 4 5 6 Time [min]

DMAB Estradiol Deriv 006.d: BPC 48.9956-2285.9523 +All MS

DMAB Estradiol Deriv 006.d: EIC 420.2551±0.005 +All MS0.00

0.25

0.50

0.75

1.00

1.25

1.50x105

Intens.

0.0

0.5

1.0

1.5

2.0

x104Intens.

reagent, by-products

1

367.6494

376.6544

388.1617

397.1677

406.1734

1. +MS2(420.0000), 3.0-3.3min #(235-252)

0

1000

2000

3000

4000

5000Intens.

260 280 300 320 340 360 380 400 420 m/z

N

O

O

Chemical Formula: C27H33NO3

Exact Mass: 419.2460

m/z: 419.2460 (100.0%), 420.2494 (29.2%), 421.2528 (4.1%)

N

O

Cl

HO

OH

OH

+

4-(Dimethylamino)-benzoylchloride β-Ergosterole

sigma-aldrich.com/derivatization

De

riv

ati

zati

on

References

[1] Birkett J.W. and Lester J.N. (eds.), 2003: Endocrine disrupters in

wastewater and sludge treatment processes, CRC Press.

[2] Metzler M. (ed.), 2001: The handbook of environmental

chemistry: Endocrine Disrupters, Vol. 3, Part 1, Springer Verlag,

Heidelberg.

[3] Nelson R.E., Grebe S.K., O’Kane D.J. and Singh, R.J., 2004:

Clinical Chemistry 50, pp 373 –384.

[4] Zhang F. et al,, 2009: Rapid Communications in ;ass

Spectrometry 23, pp 3637–364.

Figure 2 Derivatization reaction of E2 a nd DMABC. At a high E2 level of 5 ppm, only 0.2 % (rel. area fraction) of DAMBC reacts with the

second hydroxyl moiety (2:1 adduct). At 5 ppb E2 concentration, the 2:1 adduct is below the detection limit.

Figure 3 Injection of 5 pg DMAB-E2 derivative and separation on a UHPLC system using a 2.1 x 50 mm Supelco® Ascentis® Express C18

column (2.7 μm). The mobile phase conditions are water/0.01% formic acid/acetonitrile (30/70) at a flow rate of 0.4 mL/min. The MS/MS

spectrum (inset) shows only four major peaks, which is ideal for quantification and identification using a triple quadrupole mass

spectrometer.

12

sigma-aldrich.com/lc-ms

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 Time [min]

-2.5

0.0

2.5

5.0

7.5

10.0

Intens.[mAU]

-2

0

2

4

6

8

NEW

Ch

rom

ato

gra

ph

y

Positive and negative ion mode testing for all

MS applications

Most narrow and consistent specifications

We ensure UHPLC suitability, which is achieved by considering

parameters of the preparation process far beyond filtration.

The new LC-MS Ultra CHROMASOLV grade solvents are

exclusively tested for high performance with UHPLC gradi-

ent separations plus UV, positive and negative mode MS

detection. Table 1 shows the new products available to

enhance your analytical performance.

Recent innovations in HPLC and mass spectrometry (MS),

including Fused-Core® particles and ultra high perfor-

mance/pressure liquid chromatography (UHPLC) systems,

have pushed the limits of speed (throughput), efficiency

and sensitivity.

Sigma-Aldrich introduces the new LC-MS Ultra CHROMASOLV

product line that provides an outstanding quality and ultra

pure mobile phase, offering advantages such as

Tested specific to UHPLC applications

Extremely small drift in UHPLC gradient analysis

Exclusively designed for UHPLC:LC-MS Ultra CHROMASOLV®

Figure 1 UV drift of solvent baseline resulting from a gradient application of LC-MS Ultra CHROMASOLV Acetonitrile/water (red) compared

with classical HPLC Acetonitrile/water (grey). A gradient drift in UHPLC shows different values, due to an increased sensitivity compared to

classical methods. A regular gradient solvent would result in different chromatograms by running on an HPLC versus UHPLC set up.

Cat. No. Brand Name Description Pack Size

14261 Fluka® Acetonitrile LC-MS Ultra CHROMASOLV, ≥99.9%, gradient-tested for UHPLC, UV & MS 1 L, 2 L

14262 Fluka Methanol LC-MS Ultra CHROMASOLV, ≥99.9%, gradient-tested for UHPLC, UV & MS 1 L, 2 L

14263 Fluka Water LC-MS Ultra CHROMASOLV, gradient-tested for UHPLC 1 L, 2 L

14264 Fluka Trifluoroacetic acid LC-MS Ultra eluent additive, ≥ 99.0%, suitable for UHPLC-MS 1 mL, 2 mL

14265 Fluka Formic acid LC-MS Ultra eluent additive, ≥ 98%, suitable for UHPLC-MS 1 mL, 2 mL

14266 Fluka Ammonium formate LC-MS Ultra eluent additive, suitable for UHPLC-MS 25 g

14267 Fluka Ammonium acetate LC-MS Ultra eluent additive, suitable for UHPLC-MS 25 g

Table 1 New LC-MS Ultra CHROMASOLV solvents and LC-MS Ultra eluent additives.

For further product information, please visit sigma-aldrich.com/lc-ms

13

0 5 10 15 20 25

min.

A = Aldrich/Fluka #44901

B = Other High-Quality DMA

AAABBB

DMA

pA

80

60

40

20

0

sigma-aldrich.com/gc-hs

Ch

rom

ato

gra

ph

y

The following was generated with the assistance of an outside source

using Sigma-Aldrich products. Technical content was generated and

provided by:

Melissa Grella, PhD and Mark Shapiro

PharmaCore, Inc. High Point, NC

Contributed Article

Introduction

GC headspace analysis for residual solvents in pharmaceuticals is an

established practice. ICH guidelines for Class 3 solvents, where permis-

sible levels and thus responses are rather on the larger side, tend to be

the most forgiving when it comes to interfering peaks in the baseline.

However, when analyses are performed on low-responding Class 2 sol-

vents like dimethylformamide, chloroform or dichloromethane, or on

Class-1 solvents, any peaks in the baseline may present analytical issues.

For these reasons, many chromatographers prefer to employ the most

pure diluents for developing and validating their GC headspace residual

solvents methods. One of the most commonly used diluents for GC

headspace residual solvent analysis is N,N-Dimethylacetamide (DMA).

We undertook to compare in our laboratory two different sources of

DMA: Aldrich/Fluka #44901 (GC-HS Grade) and another high-quality

DMA from a leading supplier.

Experimental

The following chromatographic conditions were employed for this

study. Three separate determinations of each DMA material were made.

Results

The chromatograms in Figure 1 clearly demonstrate the superior quality

of the Aldrich/Fluka #44901 DMA. The other high-quality DMA contains

several peaks that could interfere with residual solvent peaks of interest.

These potentially troublesome peaks at RTs ca. 3.5 min., 4.4 min., 5.4 min.,

8.0 min., and 9.8 min. are absent in the Aldrich/Fluka® DMA #44901.

Conclusion

Our laboratory has chosen to use Aldrich/Fluka #44901 DMA for our GC

Headspace residual solvent methods development/validation exercises

and routine release methodology where DMA is the preferred sample

diluent.

Featured Products

Description Pack Size Cat. No.

N,N-Dimethylacetamide 1 L 44901

Related Products

Description Pack Size Cat. No.

Benzyl alcohol 1 L 807083

Cyclohexanone 1 L 68809

1,3-Dimethyl-2-imidizolidinone 100 mL 67484

Dimethyl sulfoxide 1 L 51779

1-Methyl-2-pyrrolidinone 11 L 69337

N,N-Dimethylformamide 1 L 51781

Water 1 L 53463

High Purity Solvents for GC-Headspace ApplicationsComparison of Two N,N-Dimethylacetamide Materials for GC Headspace Analysis

Shyam Verma [email protected]

Column DB-624, 30 m, 0.25 mm I.D., 1.4 μm film thickness

Oven Initial

Temp (°C)

Rate

(°C/min)

Final

Temp (°C)

Hold

Time

Total

Time (min)

35 --- 35 6.00 ---

35 5.00 150 0.00 29.00

35 --- 35 3.00 ---

Inlet Mode: Split

Split Flow: 10.0 mL/min

Split Ratio: 6.5:1

Total Flow: 14.2 mL/min

Inlet/Column Pressure: 16 psi

Temperature: 250 °C

Run Time: 29.00 min.

Carrier Gas: Helium

Headspace

Autosampler

Oven Temperature: 90 °C

Equilibration Time: 25 min. – low shake speed

Loop Temperature: 130 °C

Transfer Line Temperature: 155 °C

GC Cycle Time: 35 min.

Injection Time: 1.0 min.

Injection Volume: 1.0 mL

Pressurization Time: 0.2 min.

Vial Pressure: 14 psi

Loop Fill Time: 0.2 min.

Loop Equilibration Time: 0.05 min.

Detector Type: FID

Temperature: 300 °C

Figure 1 Chromatograms of two DMA materials

14

sigma-aldrich.com/icsigma-aldrich.com/ic

Parameter Specification

Anion traces

Br-, Cl-, F-, I-, NO3-, NO2

-, PO43-, SO4

2- ≤ 1 μg/kg each

Metal and cation traces

Al, Ba, Bi, Cd, Co, Cr, Cu, Fe, Li, Mg, Mn, Mo,

Ni, Pb, Sr, Zn

≤ 5 μg/kg each

Ca, K, Na ≤ 10 μg/kg each

NH4+ ≤ 50 μg/kg each

Organic ion traces

Acetate, formate, glycolate, oxalate ≤ 10 μg/kg each

Conductivity ≤ 2 μS/cm

Table 1 Specifications for IC-grade water (Fluka P/N 00612, package

sizes 2.5 L and 5 L)

Extensive range of products for Ion ChromatographySigma-Aldrich supplies high-purity water, eluent concentrates, validation kits and certified reference materials for highly reliable IC analyses.

Jürg Wüthrich, Senior Scientist R&D Europe [email protected]

Ion chromatography (IC) is an analytical technique for

separating and quantifying very low levels of common

anions, like fluoride, chloride, nitrite, nitrate and sulphate,

and cations, like ammonium, alkali and earth alkali ions, in

aqueous samples. Separation occurs via differential inter-

action with an ion-exchange resin. Conductivity is the

most common detection method. The high sensitivity of

IC, which makes it ideal for low-ppm quantification and

trace analysis, requires the use of high-purity eluents and

well-defined standards.

As a leading supplier of high-quality products for all areas

of analytical chemistry, Sigma-Aldrich has expertise in the

production of standards, eluents and water suitable for sen-

sitive ion determinations. Designed to save analysts’ time

and ensure the most sensitive and reproducible IC determi-

nations, our TraceCERT® standards for IC and pre-made

Certified Eluent Concentrates are convenient time and cost-

saving options.

A unique class of IC standards

TraceCERT certified reference materials are produced in a

double-accredited laboratory, fulfilling both ISO/IEC 17025

(General Requirements for the Competence of Testing and

Calibration Laboratories) and ISO Guide 34 (General

Requirements for the Competence of Reference Material

Producers). The certified value of the reference material is

directly traceable to the SI unit kilogram and also measured

against a certified reference material from the U.S. National

Institute of Standards and Technology (NIST). All details

regarding exact content, uncertainty, traceability and expiry

date are described in a comprehensive certificate, which is

available electronically through our website using product

and lot numbers. The complete listing of TraceCERT stan-

dards can be found at sigma-aldrich.com/tracecert

Certified and traceable eluent concentrates

Our certified eluent concentrates are traceable by potentio-

metric titration to NIST Standard Reference Materials and

certified in accordance with ISO Guide 31. A Certificate of

Analysis, an example of which is shown in Figure 1, provides

details regarding exact content, uncertainty, traceability and

expiry date. The certificates for our eluent concentrates are

available in electronic form through our website.

High-purity water for ion chromatography

The purity of the water used in IC-mobile phases is crucial to

permit the ppm to ppb and sometimes even ppt level

determinations of anions and cations. To address this

requirement, we have developed a quality grade of water

specifically for sensitive IC applications. Produced using

state-of-the-art purification technology and ana lyzed with

highly sensitive measurement techniques, our IC-grade

water is suitable for the trace analysis of anions, cations and

organic compounds by IC. Some quality parameters of our

IC-grade water are shown in Table 1. To ensure long-term

quality, it is supplied in special containers that have been

proven in extended storage tests, as demonstrated by the

data presented in Figure 2.

Figure 1 Sample Certificate of Analysis for sodium bicarbonate

solution (Fluka® P/N 36486-1L)

Ch

rom

ato

gra

ph

y

15

sigma-aldrich.com/ic

Traceable and certified standards for

qualification activities

Sigma-Aldrich has developed a test kit (P/N 12674, contains

six certified bromide standards from 5 to 1000 mg/kg) that

is extremely useful for qualifying ion chromatography sys-

tems. The kit is intended for testing detection linearity,

injector precision and injector carry-over, and is also part of

Metrohm’s “IQ/OQ Kit for IC”, which includes all parts for

validation of Professional IC Systems. A special advantage of

the kit is that the certified bromide standards can be used

with conductivity, UV/Vis and electrochemical detectors.

Primary multiion standards (PRIMUS)

Two metrological institutes – the Swiss Federal Institute for

Materials Science and Technology (EMPA, Switzerland) and

the German Federal Institute for Materials Research and

Testing (BAM, Germany) – together certified a set of primary

materials where up to 85 parameters per substance are ana-

lyzed. The purity was then confirmed by high-precision

measurements to additionally demonstrate the accuracy of

the certified content and the correct stoichiometry of the

compound. Using these primary materials, Sigma-Aldrich

produces the PRIMUS standard solutions gravimetrically

under clean-room conditions into precleaned 50 mL HDPE

bottles. PRIMUS standards are primary standards character-

ized by their traceability to SI as well as a uniquely low

degree of expanded uncertainty of <0.2%. These “ready-to-

use,” multi-element standards contain 10 mg/kg of each Br-,

Cl-, F-, NO3-, PO4

3-, SO42- (anion solution, P/N 89886) or Ca2+,

Li+, K+, Mg2+, Na+ (cation solution, P/N 89316).

More technical information and product specifications of

our IC products can be found at sigma-aldrich.com/ic

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75mV

4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 min

1

3

2

4

5

6

7

89

10

11

12

13

r2201527r2221327

Figure 2 Demonstration of long-term product quality of IC-grade water (Fluka P/N 00612). Anion chromatogram from a 4-month leaching

test of 2.5 L HDPE bottle. The black line shows the anions and organics in the concentration of the specification limits.

Cat. No. Brand Description Pack Size*

50439 Fluka® Perchloric Acid concentrate for IC, 0.01 M in water 1 L

36486 Fluka Sodium Bicarbonate concentrate for IC, 0.1 M in water 1 L

56169 Fluka Sodium Carbonate concentrate for IC, 0.1 M in water 1 L

43617 Fluka Sodium Hydroxide concentrate for IC, 0.1 M in water 1 L

50972 Fluka 2,6-Pyridinedicarboxylic Acid concentrate for IC, 0.02 M in water 1 L

68487 Fluka Oxalic Acid concentrate for IC, 0.1 M in water 1 L

16355 Fluka Nitric Acid concentrate for IC, 0.1 M in water 1 L

68279 Fluka Sulfuric Acid concentrate for IC, 0.1 M in water 1 L

50433 Fluka DL-Tartaric Acid concentrate for IC, 0.1 M in water 1 L

61699 Fluka Potassium Hydroxide concentrate for IC, 0.1 M in water 1 L

55517 Fluka Methanesulphonic Acid concentrate for IC, 0.1 M in water 1 L

Product Table Sigma-Aldrich Certified Eluent Concentrates * Packaging: 1L HDPE bottles with 45mm screw thread

The peaks are: (3) acetate, (1) fluoride,

(5) chloride, (7) nitrite, (8) bromide,

(9) nitrate, (10) phosphate, (11) sulphate,

(13) iodide (1 μg/kg each); (4) formate,

(2) glycolate, (12) oxalate (10 μg/kg each);

(6) system peak from carbonate eluent

Ch

rom

ato

gra

ph

y

1616

sigma-aldrich.com/ phytopharma

Sta

nd

ard

s

New Primary Reference Standards for the Analysis of Herbal Medicinal Drugs

Matthias Nold, Product Manager Analytical Standards [email protected]

In the beginning of 2010, Sigma-Aldrich introduced a new

product line of primary reference standards for use in the

quality control of herbal medicinal products. These stan-

dards are produced and qualified by HWI-Analytik in

Rülzheim (Germany) and are distributed exclusively by

Sigma-Aldrich [1]. The absolute content determination of

these standards is performed by quantitative NMR qNMR

[2]. In addition the chromatographic purity is also reported.

We are now complementing our initial portfolio of 23

products with eight new products.

Reference standards used for pharmaceutical analysis in a

GMP environment are standards for qualitative and/or

quantitative determinations within the scope of in-process

controls, batch release analyses and stability studies of

herbal drugs, herbal preparations and finished products.

For all kinds of analytes – constituents with known thera-

peutic activity, active and analytical markers – there is a

need to establish reference standards for quality control

and stability testing of herbal preparations and herbal

medicinal products.

Content assignment for natural products is typically done

by using the 100% minus impurities method by considering

the chromatographic purity as well as the content of resid-

ual solvents, water and inorganic impurities. However, the

use of chromatographic methods for content determination

is only applicable for sufficiently pure (>99.5%) reference

standards. Since the synthesis of natural compounds is very

challenging in most cases, the material used for reference

standards for medicinal herbs is usually isolated from natu-

ral sources. However, a purity exceeding 99.5% is not easily

achieved in the isolation of isolated natural products. In addi-

tion, isolation and purification procedures during production

are performed by preparative chromatography. Consequently,

it is probable that the same techniques are applied for estab-

lishing the composition and for determining the level of

purity within the framework of certification. This redundancy

results in the risk that impurities that could not be separated

during purification may also be overlooked during certifica-

tion. As a result, reference standards isolated from natural

sources can only rarely be certified according to the relevant

guidelines by using the established procedure.

These problems can be avoided by using „relative“ primary

methods of measurement that convey a direct traceability

to the SI units and ensure a higher metrological quality.

These methods are increasingly applied for content assign-

ment of natural products used as pharmaceutical reference

standards. A very powerful relative primary method is

quantitative NMR spectroscopy (see also the article on

page 18), which is applied for the content determination of

the primary standards produced by HWI Analytik.

In the table below, the newest additions to this product

line are shown. A complete listing of the primary refer-

ence standards for medicinal herbs as well as additional

information can be found on our webpage

sigma-aldrich.com/ phytopharma

A list of all analytical standards for ingredients of medicinal

herbs categorized not only alphabetically and by compound

class, but also by plant genus can be found at

sigma-aldrich.com/medicinalplants

Cat. No. Brand Description Pack Size

00130590 Fluka® trans-Anethol 100 mg

00720585 Fluka Apigenin-7-glucoside 10 mg

00020590 Fluka 1,8-Cineol (Eucalyptol) 100 mg

00380590 Fluka (+)-Fenchon 100 mg

00580590 Fluka (-)-Menthol 100 mg

00040590 Fluka -Pinene 100 mg

00670590 Fluka Thymol 100 mg

00820580 Fluka Verbascoside 10 mg

Table 1 New Primary Reference Standards from HWI Analytik exclu-

sively available at Sigma-Aldrich

References:

[1] Förster G, Michel F. and Nold, M, 2010: Analytix 1, page 11.

[2] Veith, M., 2010: Analytix 1, page 14.

17

sigma-aldrich.com/drugstandards

Sta

nd

ard

s

New Analytical Standards for Phenethylamine Class Drugs


For the forensic and clinical analysis of illegal drug substances,

Sigma-Aldrich offers a wide portfolio of analytical standards.

As part of this product range, we present new standards for

three ill icit drugs of the Cathinone (amphetamine;

phenethylamine) class and their deuterated analogues.

Butylone (also bk-MBDB), 4-Methoxymethcathinone (also

bk-PMMA or Methedrone) and 4-Methylmethcathinone

(also 4-MMC or Mephedrone) are synthetic designer drugs

that produce stimulating effects. Recently, these substances

have been increasingly consumed as drugs of abuse.


06525 Fluka® Butylone hydrochloride 10 mg

36988 Fluka Butylone-d3 hydrochloride 10 mg

92481 Fluka 4-Methylmethcathinone hydrochloride 10 mg

73903 Fluka 4-Methylmethcathinone-d3 hydrochloride 10 mg

73961 Fluka 4-Methoxymethcathinone hydrochloride 10 mg

76881 Fluka 4-Methoxymethcathinone-d3 hydrochloride 10 mg

For a complete product listing of Fluka drug standards, please visit our webpage at sigma-aldrich.com/drugstandards.

Please also visit www.cerilliant.com, a new member of the Sigma-Aldrich family, for additional drug standards.

O

O

O

H3C

HN

H3CO

O

O

H3C

HN

D3C

O

CH3

HN

CD3

H3Cl

O

CH3

HN

CH3

H3COl

O

CH3

HN

CD3

H3COl

O

CH3

HN

CH3

H3Cl

06525

92481

73961

36988

73903

76881

18S

tan

da

rds

sigma-aldrich.com/organiccrm

More Reliable Results Thanks to Organic Certified Reference Materials

Matthias Nold, Product Manager, Analytical Standards [email protected]

Alexander Rück, Senior Scientist, R&D Europe [email protected]

Christine Hellriegel, Senior Scientist, R&D Europe [email protected]

There are many factors that determine the quality of an

analytical measurement: Reliable analytical methods,

sophisticated instruments and modern laboratory equip-

ment are as important as the proficient skills and experi-

ence of the scientists that perform the measurements.

However, even if all these requirements are fulfilled, the

reliability of the analytical measurements still ultimately

depend on the reliability of the reference material that is

used for calibration. Thus, trusting your analytical

results means trusting your reference material.

Traceability of your reference material to an international

standard (such as a reference material from NIST) is an effec-

tive means of achieving highly reliable and comparable

results. However, for organic molecules, it is often not easy

to find a traceable reference material. Most commercially

available reference materials are not NIST-traceable, since

for most analytical techniques, traceability can only be

achieved if an international standard of the same com-

pound is available.

By using a relative primary method whose results are inde-

pendent of the chemical structure, traceability can be

established between two completely different compounds.

A highly potent, relative primary method is quantitative 1H-NMR (qNMR).

In the last year, the innovative new product line of organic

TraceCERT® standards has been introduced [1]. These CRMs

are certified in a double accredited laboratory, fulfilling

both ISO/IEC 17025 and ISO Guide 34 using high perfor-

mance quantitative NMR (HP qNMR®) for content deter-

mination with direct traceability to NIST and SI.

The organic TraceCERT reference materials are character-

ized by:

Certified Content by quantitative NMR (qNMR)

Superior level of accuracy, calculated uncertainties and

lot-specific values

Traceability to NIST

Comprehensive documentation delivered with the

product (certification according to ISO Guide 31)

The primary advantage of 1H quantitative NMR as a relative

primary method is that the integrals of the proton signals

are completely independent of the chemical structure.

Therefore, with a small set of NIST traceable internal stan-

dards, we are able to certify basically any organic compound

by measuring a gravimetrically produced solution of the

analyte and the internal standard. The exactly known mass-

to-mass ratio then allows for the calculation of the analyte

content with a very high precision. Typical expanded

uncertainties range from 0.5% down to 0.1%.

New organic TraceCERT products

Sigma-Aldrich is continually expanding the product offer-

ing of organic TraceCERT CRMs. Our current portfolio

comprises over 60 products, including amino acids, natural

products, PAHs, fatty acids/FAME, pesticides and pharma-

ceuticals, as noted in previously published articles [1].

The table below lists the most recent product additions. On

our website (sigma-aldrich.com/organiccrm), you can

find a complete up-to-date list of all products or download

example certificates and technical articles.

Cat. No. Brand Produkt Pack Size Product Group

90606 Fluka® Methyl Heptadecanoate 100 mg FA/FAME

94386 Fluka Oleic acid 100 mg FA/FAME

91827 Fluka Biotin 50 mg Natural Products

11474 Fluka Fluoranthene 100 mg PAH

05995 Fluka Deltamethrin 50 mg Pesticide

75451 Fluka Benz[a]anthracene 50 mg PAH

30958 Fluka Benzo[b]fluoranthene 50 mg PAH

03323 Fluka Benzo[k]fluoranthene 10 mg PAH

51968 Fluka Benzo[a]pyrene 50 mg PAH

68486 Fluka Diazinon 50 mg Pesticide

90935 Fluka Atrazine 50 mg Pesticide

Table NEW organic neat CRMs TraceCERT

References:

[1] TraceCERT® Organic Certified Reference Materials, Analytix 2/2010, 3/2010 and 1/2011.

19

sigma-aldrich.com/tracecert

Sta

nd

ard

s

Single-Element Standard Solutions (10 g/L)

Matthias Nold, Product Manager, Analytical Standards [email protected]

Jürg Wüthrich, Senior Scientist, R&D Europe [email protected]

The Fluka brand inorganic TraceCERT® products are highest-quality

certified reference materials (CRMs) that are developed and produced

according to ISO/IEC 17025 and ISO Guide 34 [1,2]. For the production,

very well characterized highest-purity starting materials are used.

TraceCERT CRM are traceable to at least two independent references

(i.e., NIST, BAM or SI unit kg) and are delivered together with comprehen-

sive documentation, including a proper uncertainty calculation, expiry

date and storing/handling instructions.

The portfolio of 1g/L single-element standard solutions for ICP, as well as

for AAS, includes standards for more than 50 different elements so far

and is continually expanding. Furthermore, last year we introduced 12 of

the most commonly analyzed elements as concentrated solutions at

10 g/L for ICP [3]. Now we present an additional 12 concentrates that

complement this product line. Similar to the 1 g/L ICP solutions, these

products are supplied in 100 mL HDPE bottles sealed in an aluminum

bag. Due to this sophisticated packaging, the standards have low uncer-

tainties and a shelf life of four years. The printed certificates are delivered

with the product and list up to 70 trace impurities.

The new ICP concentrates are listed in the table below. For the complete

TraceCERT product portfolio and for further information on the inorganic

TraceCERT line, please visit sigma-aldrich.com/inorganiccrm or order

the inorganic TraceCERT brochure.

Element (10 g/L) Composition Cat. No. Pack Size

Antimony 91482 100 mL

Cesium 79261 100 mL

Chromium 93104 100 mL

Cobalt 01488 100 mL

Germanium 73439 100 mL

Lead 39082 100 mL

Palladium 50719 100 mL

Tin 42991 100 mL

Titanium 44973 100 mL

Tungsten 50938 100 mL

Yttrium 02312 100 mL

Zirconium 51971 100 mL

References:

[1] TraceCERT Traceable Certified Reference Materials: Analytix 5/2006 and

1 to 4/2007.

[2] Double accreditation brings a new class of CRMs, 2008: Analytix 2.

[3] New 10g/L TraceCERT Single Element Standards for ICP, 2010: Analytix 1.

Inorganic Custom StandardsAn Interactive Online Platform

With a few simple mouse clicks, you can define

your own multi-component standards for ICP and IC

in TraceCERT quality, using our Inorganic Custom

Standards Online Platform:

sigma-aldrich.com/csp

For all TraceCERT Custom Standards we guarantee:

Certification under double accreditation following

ISO/IEC 17025 and ISO Guide 34

Highest level of accuracy and reliability

Minimized uncertainties and lot-specific values

Traceability to at least two independent references

(i.e., NIST, BAM or SI unit kg)

Printed certificate according to ISO Guide 31

Light and gas-tight aluminum foil bag packaging

allowing up to four years’ shelf life

20S

tan

da

rds

sigma-aldrich.com/standards

New Standards for Pesticide Metabolites

When analyzing pesticide residues in environmental or food

samples, not only the native pesticide actives, but also their

metabolites should be tested. At Sigma-Aldrich, we are con-

tinually complementing our comprehensive product portfo-

lio of PESTANAL® standards with the addition of new

metabolite standards. Recently, the metabolites of Dimetha-

chlor, Spirotetramat and Metolachlor have been added.


45447 Fluka® Dimethachlor 250 mg

32632 Fluka Dimethachlor 3.4 Metabolite CGA 373464 100 mg

32635 Fluka Dimethachlor 3.6 Metabolite SYN 530561 100 mg

32497 Fluka Dimethachlor 3.3 Metabolite SYN 528702 Sodium 50 mg

32713 Fluka Spirotetramat 100 mg

32484 Fluka Spirotetramat Metabolite BYI08330-cis-enol 10 mg

32485 Fluka Spirotetramat Metabolite BYI08330-cis-keto-hydroxy 10 mg

32486 Fluka Spirotetramat Metabolite BYI08330-mono-hydroxy 10 mg

32487 Fluka Spirotetramat Metabolite BYI08330 enol-glucoside 10 mg

36163 Fluka Metolachlor 100 mg

32637 Fluka S-Metolachlor 12.4 Metabolite CGA 357705 100 mg



32646 Fluka S-Metolachlor 12.7 Metabolite NOA 413174 100 mg



Table Analytical Standards (PESTANAL®) for the Metabolites of Dimethachlor, Spirotetramat and Metolachlor


Dimethachlor and its Metabolites

45447 32632 32497 32635

Dimethachlor 3.4 Metabolite CGA 373464 3.3 Metabolite SYN 528702 3.6 Metabolite SYN 530561

Spirotetramat and its Metabolites

32713 32484 32485 32486 32487

Spirotetramat Metabolite Metabolite Metabolite Metabolite

BYI08330-cis-enol BYI08330-cis-keto-hydroxy BYI08330-mono-hydroxy BYI08330 enol-glucoside

Metolachlor and its Metabolites

36163 32637 32641 32645 32646 32648 32650

Metolachlor Metabolite Metabolite Metabolite Metabolite Metabolite Metabolite

CGA 357705 CGA 368209 CGA 37736 NOA 413174 CGA 50268 CGA 50721

N

O

Cl OCH3

CH3H3CN

O

S

OO

O

ONa

CH3

CH3 OHN

O

SHO

OH

ONa

OH3C CH3 N

H3C OH

OCH3

OH

O

O

NH OCH3

O

H3C

CH3

O

O

CH3OCH3

CH3

NH

HO

O

OCH3

CH3

CH3

NH

O

O

OCH3

OH

CH3

CH3

NH

HO

O

OCH3

OH

CH3

CH3

NH

O

O

OCH3

O

OH

OH

OH

HO

N

O

CH3

Cl

CH3

H3CO CH3

CH3

CH3

N

O

CH3

O

HO

HO

O

CH3

CH3 HN

O

S

O

ONa

O

HN

O

OH

CH3

H3C

CH3

CH3

N

O

CH3

O

NaO

NaO3S

CH3

CH3

NH

H3CO

OHHN

O

OH

O

CH3

H3C

21

sigma-aldrich.com/volpac

Tit

rati

on

Volumetric TitrationVOLPAC® Solution Containers with new Adapter

Our 5L and 10L VOLPAC solution containers are perfect

when larger volumes of ready-to-use titration solutions are

needed. VOLPAC containers consist of a flexible polyethylene

bag with an outlet tap and a cubic cardboard supporting

frame. Because of VOLPAC’s clever design, air cannot enter

the container during the dispensing operation. As a result,

VOLPAC containers can be drained completely without

contamination for less waste and more reliable analyses.

Advantages of VOLPAC containers:

High quality of content and packing

Easy handling

No contamination during dispensing

Reduced storage footprint and packaging material

New! All VOLPAC containers are now supplied with two

different-sized adapters, allowing direct connection of

the titrating tube to the VOLPAC container.

Cat. No. Description

33665-5L-VP

33665-10L-VP

Buffer solution pH 4.0 (20 °C) with fungicide (citric acid/sodium hydroxide/

sodium chloride): red colored

pH 4.0

33666-5L-VP

33666-10L-VP

Buffer solution pH 7.0 (20 °C) with fungicide (potassium dihydrogen phosphate/

disodium hydrogen phosphate): green colored

pH 7.0

33667-5L-VP

33667-10L-VP

Buffer solution pH 9.0 (20 °C) (borax/hydrochloric acid): blue colored pH 9.0

33668-10L-VP Buffer solution pH 10.0 (20 °C) (borax/sodium hydroxide): violet colored pH 10.0

35102-5L-VP

35102-10L-VP

IDRANAL® III solution (EDTA-Na2) 0.2 mol/L

34277-5L-VP-R

34277-10L-VP-R

Sodium carbonate solution 0.5 mol/L

35233-5L-VP Sodium thiosulfate solution 0.2 mol/L

35328-5L-VOLPAC

35328-10L-VP

Hydrochloric acid solution Reag. Ph. Eur. 1 mol/L

Table 1 Selected VOLPAC containers (a complete list may be found at sigma-aldrich.com/volpac). VOLPAC products can be easily

recognized by the tag ‘-VOLPAC’ or ‘-VP’ in their product number.

22

sigma-aldrich.com/hydranal

Tit

rati

on

Strong basic amines must be neutralized by adding ben-

zoic or salicylic acid to the KF working medium. Details are

described in application report L288.

A titrating agent with a water equivalent (titer) of 2 is gener-

ally preferred for volumetric titration due to the primarily low

water content of solvents, such as HYDRANAL-Composite 2

or HYDRANAL-Titrant 2 (E). For very low water contents, titer 1

(HYDRANAL-Composite 1) may be preferred, provided that

the working conditions and parameters are ideal and stable.

Coulometric KFT for solvents without side reactions

However, all of the aforementioned techniques do not

apply to the sensitive coulometric titration method. For

example, the double bond of vinyl groups often has a neg-

ative effect in the case of coulometric KFT, whereas such

samples are analyzed easily with volumetric titration. The

coulometric KF titration method is well-suited for solvents

that do not cause any side reaction.

Volumetric standard procedure

30 mL HYDRANAL-Methanol Rapid, HYDRANAL-Methanol

dry, or HYDRANAL-CompoSolver E are placed in the titration

vessel and titrated to dryness with HYDRANAL-Composite 2

or HYDRANAL-Composite 1.

Depending on the expected water content of the sample,

2–10 g are weighed in exactly by difference, and the water

content is titrated with HYDRANAL-Composite 2 or

HYDRANAL-Composite 1.

The two-component reagents HYDRANAL-Titrant (E) and

HYDRANAL-Solvent (E) can be used in the same way.

Coulometric standard procedure

5 mL HYDRANAL-Coulomat CG are placed in the cathodic

compartment of a coulometric cell with diaphragm; the

anodic compartment is filled to the same level with approxi-

mately 100 mL of HYDRANAL-Coulomat AG. A coulometric

cell without diaphragm only requires 100 mL of HYDRANAL-

Coulomat AG.

The machine is switched on and titrates automatically to

dryness. If the drift is low (<10 μg H2O/min) and stable, the

sample can be injected. Instead of HYDRANAL-Coulomat

AG, HYDRANAL-Coulomat AD (cell without diaphragm) or

HYDRANAL Coulomat E (cell with and without diaphragm)

may also be used.

Determining the water content in different solvents is usually

problem-free if the solvent does not cause any side reactions

with the Karl Fischer (KF) reagent. Due to the chemical prop-

erties or impurities from the production process contained in

the solvents, side reactions can occur and interfere with the

KF reaction; for example, reactions with iodine or methanol

are possible. These side reactions can often be recognized

by looking at the titration curve; sluggish end points or no

end point at all can reveal such interferences. But even if the

titration indicates an end point after sufficient time, the

determined water contents is most often too high.

A side reaction with iodine can be avoided in only a few

cases. Lowering the pH value of the working medium in the

titration vessel can help suppress this interference. In this

case, 5–7 g of salicylic acid are added to the titration vessel

and diluted in the respective working medium (HYDRANAL-

Solvent or Methanol dry).

Side reactions with methanol can in many cases be avoided

by using a special medium for determining the water con-

tent in aldehydes and ketones. These compounds require

special methanol-free reagents due to pronounced side

reactions, such as the formation of acetals and ketals (which

produce additional water), or the bisulfite addition (which

consumes water from the sample and thereby produces

results that are too low). HYDRANAL-Composite 5K and

HYDRANAL-Medium K or HYDRANAL-KetoSolver are suit-

able reagents for such samples.

Subtle complications occur with the use of DMSO as a sol-

vent. The titration process proceeds routinely, but the deter-

mined result is too low, since DMSO affects the stoichiometry

of the KF reaction. A detailed application report on DMSO is

available (L 141).

Another group of solvents requiring special attention are

halogenated solvents such as 1,2-dichloroethane. If they con-

tain free chlorine, the water content cannot be determined

easily for the following reason: once the KF working medium

in the titration vessel is titrated to dryness, it contains iodide.

Since chlorine has a strong oxidizing effect, it oxidizes the

iodide to iodine after the sample addition, and the KF reac-

tion takes place. Depending on the quantity of chlorine, the

resulting water content will either be too low or the working

medium will turn brown due to an excess of iodine, and so

the titration is no longer possible. Solutions for this problem

are described in detail in application reports L001 and L003.

Water Determination in various SolventsKarl Fischer titration with HYDRANAL® reagents

Andrea Felgner, Market Segment Manager HYDRANAL [email protected]

Thomas Wendt, HYDRANAL Technical Service [email protected]

23

sigma-aldrich.com/hydranal

Tit

rati

on

It would appear more suitable to use a reagent with lower

titer such as HYDRANAL-Composite 2 because of the low

water content of the sample. However, the side reaction is

not sufficiently suppressed with Composite 2. The coulo-

metric titration appears to be unsuitable due to its sensitiv-

ity. Recommended sample size is approx. 5 g.

Reagent Volumetric 1-comp. Technique

Titrating agent HYDRANAL-Composite 5

Working medium HYDRANAL-Medium K or

HYDRANAL-Working Medium K

Application – Water determination in

n-vinyl-2-pyrrolidone (L371)

The determination of the water content by KFT proceeds

sluggishly in methanol-based working media due to the

occurrence of a side reaction. Replacing methanol in the titra-

tion vessel with methanol-free HYDRANAL-Working Medium

K results in a quick and interference-free determination of the

water content. Recommended sample size is approx. 5 g.



Working medium HYDRANAL-Working Medium K

Cat. No. Description

Volumetric one-component reagents

34805 HYDRANAL-Composite 5



34816 HYDRANAL-Composite 5K

34741 HYDRANAL-Methanol dry

37817 HYDRANAL-Methanol Rapid

34734 HYDRANAL-CompoSolver E

34698 HYDRANAL-Medium K

34817 HYDRANAL-Working Medium K

34738 HYDRANAL-KetoSolver

Volumetric two-component reagents

34732 HYDRANAL-Titrant 5 E

34723 HYDRANAL-Titrant 2 E

34801 HYDRANAL-Titrant 5

34811 HYDRANAL-Titrant 2

34730 HYDRANAL-Solvent E

34800 HYDRANAL-Solvent

Coulometric reagents

34836 HYDRANAL-Coulomat AG

34810 HYDRANAL-Coulomat AD

34826 HYDRANAL-Coulomat E

34840 HYDRANAL-Coulomat CG

Water standards

34849 HYDRANAL-Water Standard 10.0



34694 HYDRANAL-Water Standard Oil

Table Selected Fluka brand HYDRANAL Karl Fischer reagents

Sigma-Aldrich offers over 650 application reports. A full list

can be found on our website. To obtain application reports in

PDF form, please contact one of our HYDRANAL laboratories

(sigma-aldrich.com/hydranal).

Application – Water determination in vinyl

acetate (L271)

Vinyl acetate contains a double bond, causing a side reaction

during the KF titration (presumably iodination). However, this

side reaction takes place very slowly, and under certain

conditions, the water determination is still possible. Thus a

volumetric determination method with a limited sample

amount and a stop time of 10 sec. may still be carried out.

The coulometric determination is not recommended since

an end-point is not obtained.


Titrating agent HYDRANAL®-Composite 5

Working medium HYDRANAL-Methanol dry or HYDRANAL-

Methanol Rapid

Volumetric 2-comp. Technique

Titrating agent HYDRANAL-Titrant 5

Working medium HYDRANAL-Solvent and HYDRANAL-

Methanol dry (2:1) (mixture with lowered

pH value)

Application – Water determination in vinyl chloride

(L344)

Using HYDRANAL reagents, the determination of water in

vinyl chloride can be accomplished by KFT without prob-

lems. Both coulometric and volumetric determination can

be carried out for this product. Because of the low water

content of the sample, a titrant with a titer of 1 or 2, for

example HYDRANAL-Composite 2, is recommended for

volumetric determination.

When using the coulometric technique, the water capacity

of 100 mL anolyte is limited to approx. 20 g of this sample,

due to conductivity interferences. An extraction time of

2– 3 minutes and a sample size of 20 g is recommended for

the volumetric titration; 1 minute extraction time and sam-

ple size of 5 g is recommended for the coulometric

titration.



Working medium HYDRANAL-Methanol dry or

HYDRANAL-Methanol Rapid

Coulometric Technique

Anolyte HYDRANAL-Coulomat AG or

HYDRANAL-Coulomat AD

Catholyte HYDRANAL-Coulomat CG

Application – Water determination in vinyl methyl

acetamide (L367)

This material is inclined to exhibit a strong side reaction that

appears to originate not only from the double bond of the

vinyl group, but also from the methanol in the titration vessel.

If methanol is replaced by HYDRANAL-Working Medium K or

HYDRANAL-Medium K, the side reaction can be sufficiently

suppressed to allow the water determination to proceed.

HYDRANAL-Composite 5 is used as the titration reagent.

ArgentinaFree Tel: 0810 888 7446Tel: (+54) 11 4556 1472Fax: (+54) 11 4552 1698

AustraliaFree Tel: 1800 800 097 Free Fax: 1800 800 096Tel: (+61) 2 9841 0555Fax: (+61) 2 9841 0500

AustriaTel: (+43) 1 605 81 10Fax: (+43) 1 605 81 20

BelgiumFree Tel: 0800 14747Free Fax: 0800 14745Tel: (+32) 3 899 13 01Fax: (+32) 3 899 13 11

BrazilFree Tel: 0800 701 7425Tel: (+55) 11 3732 3100Fax: (+55) 11 5522 9895

CanadaFree Tel: 1800 565 1400Free Fax: 1800 265 3858Tel: (+1) 905 829 9500Fax: (+1) 905 829 9292

ChileTel: (+56) 2 495 7395Fax: (+56) 2 495 7396

ChinaFree Tel: 800 819 3336Tel: (+86) 21 6141 5566Fax: (+86) 21 6141 5567

Czech RepublicTel: (+420) 246 003 200Fax: (+420) 246 003 291

DenmarkTel: (+45) 43 56 59 00Fax: (+45) 43 56 59 05

FinlandTel: (+358) 9 350 9250Fax: (+358) 9 350 92555

FranceFree Tel: 0800 211 408Free Fax: 0800 031 052Tel: (+33) 474 82 28 88Fax: (+33) 474 95 68 08

GermanyFree Tel: 0800 51 55 000Free Fax: 0800 64 90 000Tel: (+49) 89 6513 0Fax: (+49) 89 6513 1160

HungaryIngyenes telefonszám: 06 80 355 355Ingyenes fax szám: 06 80 344 344Tel: (+36) 1 235 9063Fax: (+36) 1 269 6470

IndiaTelephoneBangalore: (+91) 80 6621 9400New Delhi: (+91) 11 4358 8000Mumbai: (+91) 22 2570 2364Hyderabad: (+91) 40 4015 5488Kolkata: (+91) 33 4013 8003FaxBangalore: (+91) 80 6621 9650New Delhi: (+91) 11 4358 8001Mumbai: (+91) 22 4087 2364Hyderabad: (+91) 40 4015 5488Kolkata: (+91) 33 4013 8000

IrelandFree Tel: 1800 200 888Free Fax: 1800 600 222Tel: (+353) 402 20370Fax: (+ 353) 402 20375

IsraelFree Tel: 1 800 70 2222Tel: (+972) 8 948 4100Fax: (+972) 8 948 4200

ItalyTel: (+39) 02 3341 7310Fax: (+39) 02 3801 0737

JapanTel: (+81) 3 5796 7300Fax: (+81) 3 5796 7315

KoreaFree Tel: (+82) 80 023 7111Free Fax: (+82) 80 023 8111Tel: (+82) 31 329 9000Fax: (+82) 31 329 9090

MalaysiaTel: (+60) 3 5635 3321Fax: (+60) 3 5635 4116

MexicoFree Tel: 01 800 007 5300Free Fax: 01 800 712 9920Tel: (+52) 722 276 1600Fax: (+52) 722 276 1601

The NetherlandsFree Tel: 0800 022 9088Free Fax: 0800 022 9089Tel: (+31) 78 620 5411Fax: (+31) 78 620 5421

New ZealandFree Tel: 0800 936 666Free Fax: 0800 937 777Tel: (+61) 2 9841 0555Fax: (+61) 2 9841 0500

NorwayTel: (+47) 23 17 60 00Fax: (+47) 23 17 60 10

PolandTel: (+48) 61 829 01 00Fax: (+48) 61 829 01 20

PortugalFree Tel: 800 202 180Free Fax: 800 202 178Tel: (+351) 21 924 2555Fax: (+351) 21 924 2610

RussiaTel: (+7) 495 621 5828Fax: (+7) 495 621 5923

SingaporeTel: (+65) 6779 1200Fax: (+65) 6779 1822

SlovakiaTel: (+421) 255 571 562Fax: (+421) 255 571 564

South AfricaFree Tel: 0800 1100 75Free Fax: 0800 1100 79Tel: (+27) 11 979 1188Fax: (+27) 11 979 1119

SpainFree Tel: 900 101 376Free Fax: 900 102 028Tel: (+34) 91 661 99 77Fax: (+34) 91 661 96 42

SwedenTel: (+46) 8 742 4200Fax: (+46) 8 742 4243

SwitzerlandFree Tel: 0800 80 00 80Free Fax: 0800 80 00 81Tel: (+41) 81 755 2828Fax: (+41) 81 755 2815

United KingdomFree Tel: 0800 717 181Free Fax: 0800 378 785Tel: (+44) 1747 833 000Fax: (+44) 1747 833 313

United StatesToll-Free: 800 325 3010Toll-Free Fax: 800 325 5052Tel: (+1) 314 771 5765Fax: (+1) 314 771 5757

VietnamTel: (+84) 3516 2810Fax: (+84) 6258 4238

Internetsigma-aldrich.com

Order/Customer Service (800) 325-3010 Fax (800) 325-5052

Technical Service (800) 325-5832 sigma-aldrich.com/techservice

Development/Custom Manufacturing Inquiries (800) 244-1173

Safety-related Information sigma-aldrich.com/safetycenter

Sigma-Aldrich Worldwide Offi ces

©2011 Sigma-Aldrich Co. LLC. All rights reserved. SIGMA, SAFC, SIGMA-ALDRICH, ALDRICH, and SUPELCO are trademarks of Sigma-Aldrich Co. LLC, registered in the US and other countries. Sigma brand

products are sold through Sigma-Aldrich, Inc. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see product

information on the Sigma-Aldrich website at sigma-aldrich.com and/or on the reverse side of the invoice or packing slip.

Fluka, TraceSELECT, TraceSELECT ULTRA, PERDROGEN, CHROMASOLV, FIXANAL, HYDRANAL, Riedel-de Haën, IDRANAL, SPECTRANAL, and VOLPAC are trademarks of Honeywell Specialty Chemicals Seelze GmbH. Sigma-Aldrich Corp. is a subsidiary of Merck KGaA, Darmstadt, Germany.

World Headquarters3050 Spruce St.

St. Louis, MO 63103(314) 771-5765

sigma-aldrich.com

Enabling Science to Improve the Quality of Life

Date: 09/2011

Sams Code: NUR

Documents

Analytix - Sigma-Aldrich · Analytix Equipment and Reagents for UV/VIS Spectroscopy r Reagents and Equipment for UV/Vis Spectroscopy r Cronobacter Detection ... from HACH LANGE The