Issue 4 • 2014

Analytix

• Proficiency Testing Portal

• NEW Flavor & Fragrance

Standards

• Analytical Standards of

Boswellic Acids

• Environmental Monitoring of

Pharmaceuticals

• Clinical Testing of Hormone

Levels

• Solvents for MS Applications

• Carbonate-selective Sensors

Proficiency Testing – A Critical Tool

22

Proficiency Testing – A Critical Tool

sigma-aldrich.com/analytix

Ed

ito

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l

Dear Colleagues,

Every year billions of measurements are performed at thousands of test-

ing laboratories worldwide. These tests may include stability testing to

ensure the shelf-life of a pharmaceutical product, microbiological test-

ing to ensure the safety of food products or environmental testing to

monitor the quality of drinking water, to mention but a few.

Laboratories have to critically assess their ability to perform these mea-

surements. Inaccurate measurements could lead to potential legal

action and will certainly lead to loss of accredited status, customers and

revenue.

One of the most useful tools available to assess the laboratory’s ability to

perform these measurements is regular participation in a profi ciency

testing program. Profi ciency testing provides an opportunity for labora-

tories to receive an independent appraisal of their data compared

against a reference value and the performance of peer laboratories.

In an eff ort to deliver the best tools to our laboratory customer, we

have developed a new testing portal which can be accessed at

sigma-aldrich.com/proficiencytesting. This portal was designed

with speed and simplicity as the focus, while providing the tools and

features that PT customers need and have come to expect from their

providers. You can read more about the specifics of this new portal

further down in this issue.

To fi nd more information about our profi ciency testing program and to

see the products that Sigma-Aldrich off ers for profi ciency testing please

visit our webpage located at sigma-aldrich.com/profi ciencytesting

Sigma-Aldrich RTC has been operating proficiency testing programs

since 1994 and is an accredited ISO 17043 PT Provider.

Best regards,

Patrick Brumfi eld

Product Manager Profi ciency Testing

Patrick Brumfield

Product Manager Proficiency Testing

pat.brumfield@sial.com

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

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

Publisher: Sigma-Aldrich Marketing Communications Europe

Publication Manager: Michael Jeitziner

Editor: Daniel Vogler

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sigma-aldrich.com/analytix

Feature Article

4 New Data Entry Portal for Proficiency Testing

Standards

5 Flavor & Fragrance Standards

More than 100 NEW Flavor & Fragrance Standards

for the Food and Cosmetic Industry

6 Getting to the Heart of Frankincense

Analytical Standards of Boswellic Acids and Their

Separation

8 Pharmaceutical Environment

Certified Reference Materials for Environmental

Analysis of Pharmaceuticals

10 Short Overview of Sigma-Aldrich’s Isotope

Labeled Pesticide Standards

Internal Standards for Food and Environmental

Analysis

11 Organic TraceCERT® CRMs

The Newest Product Additions to our Organic Neat

Certified Reference Material Portfolio

12 New & Emerging Clinical Testing

Applications: Monitoring Hormone Levels

during Treatment of Pain

Solution CRMs for Corticosteroids, Progestogens

and Other Hormones

Chromatography

14 Choosing the Right Solvent for your

Application

…and Avoiding Signal Suppression in MS

Sensorics

17 A Remarkable Carbonate-Selective

Ionophore for the Determination of

Oceanic Carbon Dioxide Using an Ion-

Selective Electrode

Spectrometry

18 A New Approach for the Detection of Acidic

Pesticides in Water by MS

Increasing the Sensitivity in Pesticide Analysis by

Paired-Ion ESI Detection

Titration

21 Water Determination in Pharmaceutical

Compounds

Karl Fischer Titration with HYDRANAL® Reagents –

Request the new Hydranal Product Line Overview

Brochure

4

New Data Entry Portal for Proficiency Testing

Previous Study Data/Reports

All of your previous study data will be transferred over to the

new system. This will allow for historical trending of results –

a feature that will be available soon on the new portal.

Webinars on the new Portal

A webinar will be off ered to demo the new portal and fi eld

any questions that you might have. If you don’t have the

opportunity to attend the webinar, a copy will be posted

on the homepage to view at your convenience.

Please contact Pat Brumfi eld (pat.brumfi eld@sial.com) to

sign up for our webinar taking place on Monday, October 20,

2PM Greenwich Mean Time.

Pat Brumfield, Product Manager Proficiency Testing pat.brumfield@sial.com

Fe

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sigma-aldrich.com/profi ciencytesting

Sigma-Aldrich® introduces our new proficiency-testing

data-entry portal. The new portal can be accessed at

sigma-aldrich.com/proficiencytesting and will be

available in mid-July 2014. This new portal will replace the

data-entry portal located at www.rt-corp.com

The new portal has been streamlined and will off er superior

speed and ease of use.

Sigma-Aldrich committed resources starting in late 2013 to

design a user interface that was effi cient, simple and some-

thing that would meet the evolving needs of the end user.

The result is our new data-entry portal which is easy to

learn, easy to remember and easy to use.

Some of the features of the new Web portal:

• One-click design that is easy to navigate, even for first

time users

• All open studies prominently displayed on the home

screen

• Easily add new personnel/analysts to your lab

• Add accreditors and enter data all on a single page

• Only the applicable methods are displayed in the

method drop-down box

• Easily copy down method/analyst/analysis date to all

analytes in a sample

• Data is actively saved as you report

• A confirmation screen that shows exactly what you have

entered and will be submitted for evaluation

• The ability to import LIMS files rather than manually

entering your results

5

Flavor & Fragrance Standards More than 100 NEW Flavor & Fragrance Standards for the Food and Cosmetic Industry

Eva Katharina Richter, Product Manager Analytical Standards evakatharina.richter@sial.com

Sigma-Aldrich® provides a broad range of flavor and fra-

grance standards for quality control of food and cosmetic

products. Our large portfolio focuses on analytical standards

for aroma compounds from the “EU positive list of autho-

rized fl avoring substances” (EFSA 10/2012) and the “EU-Reg-

ulation on cosmetic products” (1223/2009). Our selection,

which is continuously growing, includes allergenic com-

pounds and flavor enhancers as well as EU banned sub-

stances. Test your products against our Flavor & Fragrance

Standards to ensure they maintain their high quality! Find an

up-to-date product list of all Flavor & Fragrance Standards

arranged according to their occurrence in food and bever-

ages, by substance classifi cation, and in alphabetical order

at sigma-aldrich.com/fl avor

Cat. No. Brand Description Package Size

67369 Fluka® (−)-Ambroxide 100 mg

91034 Fluka (−)-Caryophyllene oxide 100 mg

54328 Fluka (−)-Globulol 10 mg

43201 Fluka (−)-Isoledene 100 mg

93483 Fluka (+)-Cedrol 100 mg

44889 Fluka (+)-Nootkatone 50 mg

18032 Fluka (1R)-(–)-Myrtenal 1 mL

56699 Fluka (3aR)-(+)-Sclareolide 100 mg

04969 Fluka (S)-(−)-Perillaldehyde 1 mL

04174 Fluka 1-Acetonaphthone 1 mL, 5 mL

75544 Fluka 1-Dodecanol 1 mL, 5 mL

18698 Fluka 1-Methylpyrrole 1 mL, 5 mL

76723 Fluka 1-Penten-3-one 1 mL

50714 Fluka 2,3,5-Trimethylpyrazine 1 mL

43042 Fluka 2,3-Benzofuran 1 mL

42038 Fluka 2,3-Butanediol 250 mg

56931 Fluka 2,6-Diisopropylphenol 1 mL

18478 Fluka 2-Acetonaphthone 100 mg

55752 Fluka 2-Ethoxynaphthalene 100 mg

69530 Fluka 2-Ethylfuran 1 mL, 5 mL

14249 Fluka 2-Furyl methyl ketone 1 mL

18245 Fluka 2'-Hydroxyacetophenone 1 mL

94414 Fluka 2-Methoxybenzaldehyde 100 mg

61904 Fluka 2-Methylbutyraldehyde 100 mg

92015 Fluka 2-Naphthalenethiol 100 mg

14672 Fluka 2-Pentyl butyrate 1 mL

74350 Fluka 2-Picoline 500 mg

44638 Fluka 3-(Methylthio)propionaldehyde 500 mg

92588 Fluka 3,4-Dihydroxybenzaldehyde 100 mg

94415 Fluka 3-Carene 1 mL, 5 mL

43212 Fluka 3-Heptanol 1 mL

04649 Fluka 3-Hexanol 1 mL

93856 Fluka 3-Octanol 1 mL

05339 Fluka 3-Picoline 1 mL

43045 Fluka 3-Pyridinemethanol 1 mL

90034 Fluka 4-Hydroxy-3-methoxycinnamic acid 100 mg

43919 Fluka 4-Methylthiazole 1 mL, 5 mL

67397 Fluka 6-Methyl-5-hepten-2-one 1 mL

07056 Fluka Allyl cinnamate 1 mL

12448 Fluka alpha-Humulene 250 mg

30627 Fluka alpha-Terpineol 100 mg

68518 Fluka Amylamine 1 mL

95054 Fluka Azelaic acid 100 mg

80024 Fluka Benzoin 100 mg

89983 Fluka Benzyl isothiocyanate 500 mg

08900 Fluka Benzyl mercaptan 1 mL

78466 Fluka Benzyl propionate 1 mL, 5 mL

75031 Fluka Butyl anthranilate 1 mL

68119 Fluka Butyl propionate 1 mL, 5 mL

90893 Fluka Butylamine 1 mL, 5 mL

94194 Fluka Butyraldehyde 1 mL

52063 Fluka Cadaverine 1 mL

43646 Fluka Citronellyl acetate 1 mL

49673 Fluka Dibenzyl ether 1 mL, 5 mL

91969 Fluka Diethyl carbonate 1 mL, 5 mL

Cat. No. Brand Description Package Size

49919 Fluka Dimethyl anthranilate 1 mL

68986 Fluka Dimethyl disulfide 1 mL

04011 Fluka Dimethyl malonate 1 mL

95563 Fluka Ethyl 2-trans-4−decadienoate 25 mg

63954 Fluka Ethyl acetoacetate 1 mL

72505 Fluka Ethyl benzoate 1 mL, 5 mL

66761 Fluka Ethyl cinnamate 100 mg

08996 Fluka Ethyl lactate 1 mL

73539 Fluka Ethyl linoleate 1 mL

91224 Fluka Ethyl phenylacetate 1 mL, 5 mL

43348 Fluka Farnesol 1 mL

89985 Fluka gamma-Undecalactone 1 mL

40951 Fluka gamma-Valerolactone 1 mL

61696 Fluka Heptaldehyde 1 mL

59964 Fluka Histamine 100 mg

80725 Fluka Hydrocinnamic acid 100 mg

66010 Fluka Hydroxycitronellal 1 mL

95668 Fluka Isoamyl octanoate 1 mL, 5 mL

94888 Fluka Isobutyl butyrate 1 mL, 5 mL

06755 Fluka Isobutylamine 1 mL

07055 Fluka Isoeugenyl acetate 100 mg

68853 Fluka Isoquinoline 1 mL

49599 Fluka Linalyl acetate 100 mg

07406 Fluka m-Anisaldehyde 1 mL

95401 Fluka Menthone, mixture of isomers 1 mL, 5 mL

41263 Fluka Methyl benzoylformate 1 mL

80835 Fluka Methyl cinnamate 250 mg

52466 Fluka Octanal 1 mL, 5 mL

08066 Fluka Oleyl alcohol 100 mg

92819 Fluka o-Tolualdehyde 1 mL

94666 Fluka p-Anisic acid 100 mg

66962 Fluka Pentyl acetate 1 mL

73747 Fluka Phenethyl acetate 1 mL, 5 mL

30311 Fluka Phenyl acetate 1 mL

92528 Fluka Phenyl disulfide 100 mg

91897 Fluka Phloretin 10 mg

61074 Fluka Propiophenone 1 mL, 5 mL

53646 Fluka Propylamine 1 mL

41423 Fluka p-Tolualdehyde 1 mL

49817 Fluka Pyrrole 1 mL, 5 mL

94517 Fluka Quinoline 1 mL

42993 Fluka Theobromine 100 mg

92358 Fluka Thioanisole 1 mL

03416 Fluka Thymoquinone 100 mg

05549 Fluka trans-2,cis-6-Nonadienal 100 mg

90244 Fluka trans-2-Heptenal 100 mg

07592 Fluka trans-2-Nonenal 1 mL

52464 Fluka trans-2-Octenal 1 mL, 5 mL

94575 Fluka trans-2-Pentenal 1 mL

74103 Fluka Triacetin 1 mL, 5 mL

43103 Fluka Triethyl orthoformate 1 mL, 5 mL

73487 Fluka Triethylamine 1 mL, 5 mL

80139 Fluka Undecanal 1 mL

94490 Fluka δ-Octalactone, mixture of isomers 1 mL, 5 mL

Table 1 NEW Flavor & Fragrance Standards

sigma-aldrich.com/fl avor

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6S

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sigma-aldrich.com/medicinalplants

are a group of pentacyclic triterpene acids of which

-boswellic acids are the major biologically active constitu-

ents. Sigma Aldrich® off ers six boswellic acids as analytical

standards and two neat standards of + mixtures for use

in research and quality control.

Figure 2 shows the chromatograms of the ethanolic

extract of a boswellia herbal supplement as well as of a

mixture of the six analytical standards.

Getting to the Heart of Frankincense Analytical Standards of Boswellic Acids and Their Separation

The trees of the genus boswellia have been used in tradi-

tional medicine for thousands of years as an anti-infl amma-

tory agent. Its resin is known as frankincense or olibanum

and is still widely used in dietary supplements and in herbal

medicinal products. The most characteristic components

Matthias Nold, Product Manager Analytical Standards matthias.nold@sial.com

Hugh Cramer, Application Scientist hugh.cramer@sial.com

06813

-boswellic acid

74607

3-O-Acetyl-11-keto- -boswellic acid

49873

3-O-Acetyl- -boswellic acid

78535

11-keto- -boswellic acid

80342

-boswellic acid

56208

3-O-Acetyl- -boswellic acid

Figure 1 Chemical Structures of the Boswellic Acids

Separation of Boswellic Acids

Several conditions, using a variety of solid phases, have been

screened for their suitability to separate six boswellic acids.

The shortest analysis time was achieved using an Ascentis

Express Column (2.7 μm particle size, L × I.D. 10 cm × 3.0 mm)

and acetonitrile/phosphoric acid as eluents.

HOH3C

OHOH

H3C CH3

CH3

H

H3C CH3

CH3

HOH3C

OHOH

H3C CH3

CH3

H

CH3

H

H3CCH3

H3C

O

OH3C

OHOH

H3C CH3

CH3

H

H3C CH3

CH3

HH3C

O

OH3C

OHOH

H3C CH3

CH3

H

CH3

H

H3CCH3

H3C

O

O

HOH3C

OHOH

H3C CH3

CH3

H

CH3

H

H3CCH3

O

HOH3C

OHOH

H3C CH3

CH3

H

CH3

H

H3CCH3

7

sigma-aldrich.com/medicinalplants

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A Boswellia Extract

B Analytical Standards at 50 g/mL of Each Component

Conditions

Column: Ascentis Express Phenyl-Hexyl, 10 cm x 3.0 mm I.D., 2.7 μm (53345-U) Mobile Phase: (A) 0.1% phosphoric acid; (B) acetonitrile Gradient: 50% B for 0.5 min; to 100% B in 4.5 min; held at 100% B for 5 min Flow Rate: 0.6 mL/min Pressure: 2390 psi (165 bar) Column temp.: 35 °C Detector: UV at 210 nm Injection: 5 μL Sample(s): 6 Boswellic acids 50μg/mL in 70:30, water:methanol extract in 50:50, water:ethanol Extraction: 1. Weigh 100 mg Bosewellia Herbal Supplement 2. Add 10 mL ethanol 3. Sonicate for 30 min at 40 °C 4. Dilute: 100 μL extract in ethanol, 400 μL ethanol, 500 μL water

Peak IDs

1) 11-Keto-β-boswellic-acid2) 3-O-Acetyl-11-keto-β-boswellic-acid3) α-boswellic-acid4) β-boswellic-acid5) 3-O-Acetyl-α-boswellic-acid6) 3-O-Acetyl-β-boswellic-acid

Figure 2 Experimental Conditions and Chromatograms of the Separation of Boswellic Acids

Cat. No. Brand Description Package Size

74607 Fluka® 3-O-Acetyl-11-keto-β-boswellic acid 5 mg

56208 Fluka 3-O-Acetyl-α-boswellic acid 5 mg

49873 Fluka 3-O-Acetyl-β-boswellic acid 5 mg

96729 Fluka 3-O-Acetylboswellic acid, mixture of α and β 5 mg

06813 Fluka α-Boswellic acid 10 mg

80342 Fluka β-Boswellic acid 5 mg

63850 Fluka Boswellic acid, mixture of α and β 5 mg

78535 Fluka 11-Keto-β-boswellic acid 5 mg

53345-U Supelco® Ascentis Express Phenyl-Hexyl, 2.7 Micron HPLC Column 10 cm × 3.0 mm

Table 1 Analytical Standards of Boswellic Acids and Column Used

Sigma-Aldrich continuously expands its off ering of analytical

standards for plant constituents, intended for the analysis of

medicinal plants and herbal medicinal products.

On our Web page at sigma-aldrich.com/medicinalplants,

you will fi nd an up-to date list of all phytopharma standards

sorted in alphabetical order, by substance class and by

genus of a large number of the most popular medicinal

plants.

8

sigma-aldrich.com/pharmametabolites

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Therefore, the occurrence and impact of pharmaceutical

substances in the environment is gaining increasing atten-

tion from researchers and authorities, as well as from com-

panies in the pharmaceutical industry, which have an inter-

est in minimizing their ecological footprint. This increasing

attention was also reflected by the huge quantity of ses-

sions and posters on the topic at this year’s SETAC annual

meeting[1]. Many studies support the presumption that

active pharmaceutical substances (API) do have a detrimen-

tal eff ect on the ecosystem and that their release into the

environment needs to be monitored and controlled.

Authorities such as the EPA (U.S. Environmental Protection

Agency)[2] or the German Umweltbundesamt (UBA)[3] per-

form and/or support studies in this field with the aim of

establishing appropriate guidelines. The Water Framework

Directive[4] of the EU also has an eye on pharmaceuticals, and

in 2012, added diclofenac, which was the fi rst pharmaceutical

to be added to the list of proposed priority substances.

Without a doubt, we can attribute our present-day quality

of life and long life expectancy largely to the achievements

of the medical profession during the last century. Nowa-

days, huge quantities of pharmaceutical substances are

produced and consumed. However, either through excre-

tion or improper disposal via the toilet, a lot of pharmaceu-

ticals end up in sewage treatment plants and eventually in

ground and surface waters. This environmental presence of

highly active chemicals may have unforeseeable eff ects on

the ecosystem.

Towards a Healthier EnvironmentCertified Reference Materials for Environmental Analysis of Pharmaceuticals

Matthias Nold, Product Manager Analytical Standards matthias.nold@sial.com

Cat. No. Brand Description Package Size

PHR1005 Fluka® Acetaminophen 1 g

PHR1127 Fluka Amoxicyllin trihydrate 1 g

PHR1067 Fluka Carbamazepine 1 g

PHR1412 Fluka Chloramphenicol 1 g

PHR1167 Fluka Ciprofloxacin 1 g

PHR1038 Fluka Clarithromycin 500 mg

PHR1058 Fluka Clotrimazole 1 g

PHR1144 Fluka Diclofenac sodium salt 1 g

PHR1145 Fluka Doxycycline hyclate 1 g

PHR1039 Fluka Erythromycin 1 g

PHR1246 Fluka Fenofibrate 500 mg

PHR1049 Fluka Gabapentin 1 g

PHR1004 Fluka Ibuprofen 1 g

PHR1247 Fluka Indomethacin 500 mg

PHR1084 Fluka Metformin hydrochloride 500 mg

PHR1076 Fluka Metoprolol tartrate 1 g

PHR1040 Fluka Naproxen 500 mg

PHR1308 Fluka Propranolol hydrochloride 1 g

PHR1389 Fluka Propyphenazone 1 g

PHR1126 Fluka Sulfamethoxazole 1 g

PHR1041 Fluka Tetracycline hydrochloride 500 mg

PHR1056 Fluka Trimethoprim 1 g

Table 1 Some Pharmaceuticals with Potential Relevance for Environmental Analysis and Proposed CRMs

9

sigma-aldrich.com/pharmastandards

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Inorganic Custom StandardsAn Interactive Online Platform

With a few simple mouse clicks, you can defi ne

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

Several aspects need to be considered for the priorisation

of pharmaceuticals for environmental monitoring: The

ecotoxicological potential, the occurence in the aquatic

environment and the annual consumption of the drug.

To ensure reliable and traceable quantitative results, ISO 17025

accredited labs should use proper certifi ed reference materi-

als (CRMs). For this purpose, Sigma-Aldrich® off ers a compre-

hensive portfolio of pharmaceutical compounds (APIs, related

substances and excipients) of pharmaceutical secondary

standards. Though these products have been developed for

use in pharmaceutical quality control and are traceable to

pharmacopoeia compendial standards, they are also certifi ed

reference materials by the defi nition of ISO, as they are manu-

factured under ISO/IEC17025 and ISO Guide 34 double

accreditation, and a certifi ed value, including uncertainty, is

given in addition to the pharmacopoeia traceable values. The

list on the previous page shows some common pharmaceuti-

cals and proposed certifi ed reference materials. The complete

portfolio comprises over 400 products and can be found at

sigma-aldrich.com/pharmastandards

To complement this offering, Sigma-Aldrich also offers a

comprehensive portfolio of pharmaceutical metabolites

and isotope labeled pharmaceuticals (sigma-aldrich.com/

pharmametabolites) as presented in an article in the pre-

vious issue of Analytix[5].

References

[1] www.basel.setac.eu/?contentid=763

[2] www.epa.gov/ppcp/

[3] Bergmann, A. et al. Zusammenstellung von Monitoringdaten

zu Umweltkonzentrationen von Arzneimitteln UBA-FB 001525;

2011.

[4] http://ec.europa.eu/environment/water/water-dangersub/

lib_pri_substances.htm#prop_2011_docs

[5] Richter, E. Analytix 3/2014, page 8

10

Pesticide StandardsFor Food & Environmental Analysis

• Neat Standards

• Single and Multi-Component Solutions

• Certified Reference Materials (CRMs)

• Matrix CRMs

• Isotopically Labeled Internal Standards

• Pesticide Metabolite Standards

• Proficiency Testing Materials (PT)

D Cl

Cl

D

DCl

Cl

D

D

Cl

ClDNH

S

N

O

CD3

DCD3

O

O N

N

Cl

N

NH

NO2

D D

D

D

Sta

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Short Overview of Sigma-Aldrich’s Isotope Labeled Pesticide StandardsInternal Standards for Food and Environmental Analysis

The Sigma-Aldrich® pesticide standards product line is the most compre-

hensive portfolio available on the market. We proudly off er more than

1300 high-purity pesticide and pesticide metabolite standards and certi-

fi ed reference materials for food and environmental analysis.

Our comprehensive portfolio includes several isotope-labeled pesticide

standards. To determine matrix eff ects and result bias due to a loss of the

analyte during the workup process, isotope-dilution mass spectrometry

is often applied. This technique takes advantage of the fact that isotope-

labeled compounds have nearly the same physical properties as the

non-labeled analogues, and thus the same behavior in the workup and

sample-preparation process. However, they can be distinguished in mass

spectrometry. Therefore, by spiking the sample before workup with an

isotope-labeled analogue, material loss during workup can be deter-

mined and compensated.

Please fi nd below a list of our current products. For more information,

please visit our Web page at sigma-aldrich.com/pesticides or order

our brand NEW brochure at sigma-aldrich.com/lit-request

Eva Katharina Richter, Product Manager Analytical Standards evakatharina.richter@sial.com

Cat. No. Brand Description Package Size

39246 Fluka® Acetamiprid-d3 50 mg

34086 Fluka Alachlor-d13 10 mg

32440 Fluka Amitraz Metabolite BTS 27271

(N/methyl-d3)

10 mg

34053 Fluka Atrazine-d5 10 mg

32965 Fluka Bentazon-d7 10 mg

34164 Fluka Bis(2-ethylhexyl)phthalate-3,4,5,6-d4 25 mg

32413 Fluka Carbendazim-d3 10 mg

34019 Fluka Carbofuran-d3 10 mg

32232 Fluka 6-Chloro-2,4-diamino-1,3,5-triazine-13C3 10 mg

56816 Fluka Clothianidin-d3 50 mg

34021 Fluka 4,4’-DDT-d8 10 mg

34169 Fluka Dibutyl phthalate-3,4,5,6-d4 25 mg

32381 Fluka NEW DEET-(diethyl-d10) 25 mg

48049 Supelco 1,4-Dichlorobenzene-d4 solution;

2000 μg/mL in methanol

1 mL

442247 Supelco 1,4-Dichlorobenzene-d4 1000 mg

442228 Supelco 1,2-Dichloroethane-d4 1000 mg

34233 Fluka Dicamba-d3 10 mg

34186 Fluka Dicyclohexyl phthalate-d4 10 mg, 25 mg

32311 Fluka NEW Diflubenzuron-(chlorophenyl-13C6) 10 mg

32274 Fluka NEW 4-(3,6-Dimethyl-3-heptyl)phenol-

3,5-d2 solution; 100 μg/mL in acetone

1 mL

34201 Fluka Dimethyl-d6 phthalate 25 mg

34018 Fluka Diuron-d6 10 mg

34214 Fluka DNC-d8 10 mg

03811 Fluka α-HCH-d6 5 mg

34170 Fluka Imidacloprid-d4 10 mg

34017 Fluka Isoproturon-d6 10 mg

32300 Fluka NEW Methiocarb-(N-methyl-d3) 10 mg

32668 Fluka Nabam-d4 10 mg

48715-U Supelco Naphthalene-d8 solution;

2000 μg/mL in methylene chloride

1 mL

33452 Fluka Parathion-ethyl-d10 10 mg

34209 Fluka Pirimicarb-d6 10 mg

34054 Fluka Simazine-d10 10 mg

43293 Fluka NEW Tebuconazole-(trimethyl-13C3) 5 mg

38176 Fluka Thiamethoxam-d3 25 mg

33853 Fluka Thionazin-d10 10 mg

34023 Fluka Trichloroanisole-d5 50 mg

Table 1 Neats and Solutions for Isotope-Labeled Pesticides

sigma-aldrich.com/pesticides

Figure 1 Molecular Structure of Deuterated α-HCH Figure 2 Molecular Structure of Deuterated Bentazon Figure 3 Molecular Structure of Deuterated

Imidacloprid

11

Organic TraceCERT® CRMs The Newest Product Additions to our Organic Neat Certified Reference Material Portfolio

Matthias Nold, Product Manager Analytical Standards matthias.nold@sial.com

For accredited testing labs, the availability of reliable and

traceable certifi ed reference materials (CRMs) is crucial since

the use of CRMs for calibration is demanded by ISO/IEC

17025. For the certification of Sigma-Aldrich®’s organic

TraceCERT products, high-performance quantitative NMR

(HP-qNMR®) is applied as a relative primary method to

achieve traceability to NIST SRM. If you would like to learn

more about qNMR and Sigma-Aldrich’s in-house capabili-

ties in this fi eld, please refer to the references cited below.

The organic TraceCERT reference materials are character-

ized by:

• Certified content by quantitative NMR (qNMR)

• Manufactured under ISO/IEC 17025 / ISO Guide 34

double accreditation

• Superior level of accuracy, calculated uncertainties, and

lot-specific values

• Traceability to NIST SRM

• Comprehensive documentation delivered with the

product (certification according to ISO Guide 31)

In the last issue of Analytix, we presented 25 new products,

and we are now able to present the latest additions to the

rapidly growing organic TraceCERT product line. You can

fi nd a list of the complete portfolio of over 150 products at

sigma-aldrich.com/organiccrm

Cat. No. Brand Product Package Size

63634 Fluka® Acetophenone 50 mg

14181 Fluka Dimethyl isophthalate 100 mg

97627 Fluka Monoethanolamine 100 mg

80026 Fluka 2,4,6-Tribromophenol 100 mg

67180 Fluka 1,2,4-Trichlorobenzene 100 mg

53766 Fluka 2,4,6-Trimethylphenol 100 mg

30304 Fluka Vanillin 50 mg

50409 Fluka Thymol 1 g

References

[1] Weber, M.; Hellriegel, C.; Rueck, A.; Sauermoser, R.; Wuethrich, J.

Using high-performance quantitative NMR (HP-qNMR®) for

certifying traceable and highly accurate purity values of

organic reference materials with uncertainties <0.1 %,

Accred. Qual. Assur. 18. 2013, 91– 98

[2] TraceCERT® Organic Certified Reference Materials, Analytix,

Vol. 3 2010; Vol. 1 2011; Vol. 3 2011; Vol. 4 2011.

[3] TraceCERT® Organic Certified Reference Materials, Analytix,

Vol. 2 2010.

Sta

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sigma-aldrich.com/organiccrm

NEW FROM SIGMA-ALDRICH®

Certifi ed Reference Materials (CRMs)For the Petrochemical Industry

From ISO/IEC 17025 and ISO Guide 34 Accredited leading producer, Paragon Scientifi c

The Paragon range includes more than 600 products with a wide variety of Petrochemical

Certifi ed Reference Materials.

For more information, visit sigma-aldrich.com/paragon

12S

tan

da

rds

New clinical applications are emerging that monitor hormone levels for

the diagnosis and treatment of pain. Pain management clinics, for

example, are increasingly quantifying neurosteroids, corticosteroids,

and other hormones by LC-MS/MS and other analytical methods for

indication of abnormal hormone production and its relationship to

severe pain.[1,2] According to one study, serum levels of hormones such

as cortisol, pregnenolone, testosterone, thyroid, and dehydroepiandros-

terone (DHEA) represent the most objective biomarkers of severe,

uncontrolled pain.[1]

The study’s author stated that patients taking opioids for treatment of

pain should be screened for hormone abnormalities. Several hormones,

including cortisol, testosterone, and pregnenolone, are suppressed by

opioids with longer-acting opioids the most eff ective at hormone sup-

pression. Replacement hormone therapy may be needed to counteract

the eff ect of opioids on serum hormone levels. For pain patients with

abnormally low hormone levels that are not taking opioids, treatment

with neuroregenerative hormones such as hCG or human chorionic

gonadotropin may provide a benefi t of pain reduction.[1]

Testosterone and cortisol have also been implicated in neonatal[3] and

adult pain-related stressors.[4,5] In these studies, testosterone and cortisol

levels in blood, saliva, or hair were determined using immunoassay and

related analytical techniques. The fi ndings from these studies demon-

strate the importance and infl uence of cortisol and testosterone levels

on pain and pain thresholds during times of high stress.[3,4,5]

To view Cerilliant’s extensive catalog of solution Certified Reference

Materials (CRMs), including hormone categories such as corticosteroids,

DHEA, progestogens, testosterone, and thyroid, visit

sigma-aldrich.com/cerilliant

Register at sigma-aldrich.com/registercerilliant to receive news on

the latest reference standards being developed by Cerilliant.

New and Emerging Clinical Testing Applications: Monitoring Hormone Levels during Treatment of PainSolution CRMs for Corticosteroids, Progestogens and Other Hormones

Derrell Johnson, Manager, New Product Strategy & Tactical Marketing Derrell_Johnson@cerilliant.com

Maximilian Magana, Technical Marketing Specialist Max_Magana@cerilliant.com

sigma-aldrich.com/cerilliant

Cerilliant Hormone Certified Spiking Solutions®

Cat. No. Description Package Size

Corticosteroids

A-096 Aldosterone 100 μg/mL in Acetonitrile

C-106 Cortisol 1.0 mg/mL in Methanol

C-113 Cortisol-D4 100 μg/mL in Methanol

C-117 Corticosterone 1.0 mg/mL in Methanol

C-130 Cortisone 100 μg/mL in Methanol

D-061 11-Deoxycortisol 1.0 mg/mL in Methanol

D-062 21-Deoxycortisol 100 μg/mL in Methanol

D-076 21-Deoxycortisol-D8 100 μg/mL in Methanol

D-078 11-Deoxycortisol-D5 100 μg/mL in Methanol

D-085 Dexamethasone 1.0 mg/mL in Methanol

D-105 11-Deoxycorticosterone 100 μg/mL in Methanol

Progestogens

H-085 17α-Hydroxyprogesterone 1.0 mg/mL in Methanol

H-096 17α-Hydroxyprogesterone-D8 100 μg/mL in Methanol

H-100 17α-Hydroxyprogesterone-2,3,4-13C3 100 μg/mL in Methanol

H-105 17α-Hydroxypregnenolone 100 μg/mL in Methanol

P-069 Progesterone 1.0 mg/mL in Acetonitrile

P-070 Progesterone-D9 100 μg/mL in Acetonitrile

P-104 Pregnenolone Coming Soon

Dehydroepiandrosterone (DHEA)

D-063 DHEA 1.0 mg/mL in Methanol

D-064 DHEA-D5 100 μg/mL in Methanol

D-065 DHEAS 1.0 mg/mL (as free sulfate) in

Methanol

D-066 DHEAS-D5 100 μg/mL (as free sulfate) in

Methanol

Testosterone

D-073 5α-Dihydrotestosterone (DHT) 1.0 mg/mL in Methanol

D-077 5α-Dihydrotestosterone-D3 100 μg/mL in Methanol

H-059 6β-Hydroxytestosterone 100 μg/mL in Methanol

T-034 6β-Hydroxytestosterone-D3 100 μg/mL in Methanol

T-037 Testosterone 1.0 mg/mL in Acetonitrile

T-046 Testosterone-D3 100 μg/mL in Acetonitrile

T-070 Testosterone-2,3,4-13C3 100 μg/mL in Acetonitrile

Thyroid

T-073 L-Thyroxine (T4) 100 μg/mL in 0.1N NH3 in Methanol

T-074 3,3',5-Triiodo-L-thyronine (T3) 100 μg/mL in 0.1N NH3 in Methanol

T-075 3,3',5'-Triiodo-L-thyronine (Reverse T3) 100 μg/mL in 0.1N NH3 in Methanol

T-076 L-Thyroxine-13C6 (T4-13C6) 100 μg/mL in 0.1N NH3 in Methanol

T-077 3,3',5-Triiodo-L-thyronine-13C6 (T3-13C6) 100 μg/mL in 0.1N NH3 in Methanol

T-078 3,3',5'-Triiodo-L-thyronine-13C6

(Reverse T3-13C6)

100 μg/mL in 0.1N NH3 in Methanol

References[1] Hughes, D. Hormone Levels Can Validate Presence of Pain and Affect Treatment.

Accessed 11 April 2014 from: www.empr.com/hormone-levels-can-validate-presence-of-pain-and-affect-treatment/article/310528/

[2] Tennant, F. The Use of Hormones in Chronic Pain.Accessed 11 April 2014 from: www.journalofprolotherapy.com/pdfs/issue_08/issue_08_06_hormones_for_pain.pdf

[3] Grunau, R. E.; Cepeda, I. L.; Chau C. M. Y.; Brummelte S.; Weinberg J.; Lavoie P. M.; Ladd M.; Hirschfeld A. F.; Russell E.; Koren G.; Van Uum S.; Brant R.; and Turvey S. E. Neonatal Pain-Related Stress and NFKBIA Genotype Are Associated with Altered Cortisol Levels in Preterm Boys at School Age. Accessed 29 May 2014 from: www.ncbi.nlm.nih.gov/pmc/articles/PMC3774765/#__ffn_sectitle

[4] Choi J. C.; Chung M. I.; and Lee Y. D. Modulation of pain sensation by stress-related testosterone and cortisol. Accessed 29 May 2014 from: http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2044.2012.07267.x/pdf

[5] Karlén J.; Ludvigsson J.; Frostell A.; Theodorsson E.; and Faresjö, T. Cortisol in hair measured in young adults – a biomarker of major life stressors? Accessed 29 May 2014 from: www.biomedcentral.com/1472-6890/11/12#B37

13

sigma-aldrich.com/derivatization

Sta

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New! Brochure for Cosmetic Standards For Beauty with Safety

For precise quality control of raw materials or formulated personal-care and cosmetic products,

we off er analytical standards for several groups of ingredients, including:

• Antibacterial, antifungal substances

• Colorants

• Emulsifiers, thickeners, opacifiers

• Fragrances

• Metals

• PAHs

• Plasticizers

• Preservatives

• Propellants

• UV blockers

Order the brand NEW brochure or visit sigma-aldrich.com/cosmetics for an up-to-date product list.

New TLC Staining Solution for Boronic Acids Curcumin solution 28982 eff ectively stains various types of boron containing species,

such as:

• Free boronic acids

• Boronic acid pinacol esters

• Potassium trifluoroborate salts

For rapid monitoring of your Suzuki-Miyaura cross-coupling, or any other organic reaction

involving boronic acids and related derivatives, try curcumin solution 28982.

For more information on TLC derivatization, visit sigma-aldrich.com/derivatization

Reference:

[1] Lawrence, K.; Flower, S. E.; Kociok-Kohn, G.; Frost, C. G.; James, T. D. A simple and effective

colorimetric technique for the detection of boronic acids and their derivatives. Anal. Methods

2012, 4, 2215–2217.

1 691615 (+)-Vinylboronic acid pinanediol ester;

2 674710 3-Methoxy-1-propyn-1-ylboronic acid

pinacol ester;

3 346225 (2-Methylpropyl)-boronic acid;

4 683590 Potassium trans-3-methoxy-1-

propenyltri-fluoroborate

14

sigma-aldrich.com/lc-ms

Introduction

In routine LC-MS analysis, reliability of methods is crucial in

order to avoid false analytical data or increase in time per

sample due to necessary corrections or repetition of deter-

minations. Besides instrument sensitivity, which generally

improves with every generation, solvent quality plays an

increasingly important role. Lesser-grade solvents may con-

tain impurities, and are not tested for LC-MS applications,

resulting in interferences or signal suppression. Impurities

that most commonly cause problems are high alkali-ion

concentrations, plasticizers and detergents.

Experimental

A mixture containing 6 alkylresorcinols (phenolic lipids

used as food intake biomarkers) at 0.1 μg/mL was separated

on a Titan C18 UHPLC column, using a previously published

method (Ross 2012). [1]

The following conditions were employed, with determina-

tions using fi rst LC-MS CHROMASOLV® methanol, then stan-

dard HPLC grade methanol, before switching back to LC-MS

methanol. The standard mixtures were created in neat

methanol.

Column Supelco Titan™ C18 1.9 μm 2.1x100 mm

Mobile phase A = mQ water, B = MeOH (LC-MS

CHROMASOLV or standard HPLC grade)

Gradient min %B

0 89

6 99

7 99

8 89

10 89

Flow 0.65 mL/min

Injection volume 1 μL

MS APCI+ operated in MRM mode

Instrument Shimadzu LCMS-8030plus including Nexera

LC-30AD pumps, Nexera SIL-30AC autosampler,

Nexera CTO-20AC column oven, Prominence

DGU-20A 5r degasser and Shimadzu 8030plus

triple quadrupole

Choosing the Right Solvent for Your Application…and Avoiding Signal Suppression in MS

Matthias Drexler, Product Manager, Analytical Reagents and Spectroscopy matthias.drexler@sial.com

Experimental data courtesy of Otto Savolainen and Alastair Ross at Chalmers University of Technology, Gothenburg, Sweden

Results

Figure 1 clearly shows all key analytes are separated, ionized

and detected with the selected method when utilizing a

gradient of mQ water and LC-MS CHROMASOLV methanol.

The instrument was then run with two blanks containing

standard HPLC grade methanol, after which the same ana-

lyte mixture was analyzed again. Immediately strong signal

suppression can be seen in the resulting chromatogram

(see Figure 2). The eff ect is especially apparent for the ana-

lytes with longer retention time. After 10 injections working

with HPLC grade methanol, the suppression effects are

even higher (Figure 3) leading to complete suppression of

some of the analyte signals after a few dozen injections.

The instrument becomes “blind” to the analytes at these

concentrations.

After switching back to a gradient containing LC-MS grade

methanol, two blanks were run before injection of the analyte

mix and running the method again. In Figure 4, only minimal

(if at all) remaining signal suppression can be seen and all

6 analytes can again be clearly detected and quantifi ed.

Conclusion

These results show the importance of choosing the right

solvent quality to support the analytical task at hand. High

purity and sensitive UV gradient methods can give only a

rough picture of a solvent and its suitability for MS detec-

tion. Relying on sub-par solvents, especially during method

development, may lower the apparent cost per sample, but

have a serious negative impact on limit of quantification

(LOQ), while also negatively affecting the lifetime of the

instrument.

Our LC-MS CHROMASOLV solvents are application tested,

off er low cation impurities, and high reliability to help sci-

entists to avoid the hidden costs resulting from signal sup-

pression. More information can be found at

sigma-aldrich.com/lc-ms

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sigma-aldrich.com/uhplc

Cat. No. Featured and Related Products Package Size

Solvents

34967 Acetonitrile, LC-MS CHROMASOLV 250 mL, 1 L, 2.5 L, 4x4 L, 6x1 L, 4x2.5 L, 20 L

34966 Methanol, LC-MS CHROMASOLV 1 L, 2.5 L, 4x4 L, 6x1 L, 4x2.5 L

39253 Water, LC-MS CHROMASOLV 1 L, 4x4 L, 20 L

14261 Acetonitrile, LC-MS Ultra CHROMASOLV, for UHPLC-MS 1 L, 2 L

14262 Methanol, LC-MS Ultra CHROMASOLV, for UHPLC-MS 1 L, 2 L

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

Columns and Accessories

577124-U Titan C18 UHPLC Column, 1.9 micron, 10 cm x 2.1 mm 1 ea

577127-U Titan C18 UHPLC Guard Cartridge, 1.9 micrometer, 1.9 μm particle size, L × I.D. 5 mm × 2.1 mm 3 ea

577133-U Titan C18 UHPLC Guard Cartridge Holder, 1.9 micron, for use with Titan Guard Cartridges 1 ea

Analytical Standards of Alkylresorcinols

56453 5-Tridecylresorcinol 10 mg

91822 5-Pentadecylresorcinol 10 mg

97001 5-Heptadecylresorcinol 10 mg

49519 5-(Nonadecyl-1,1,2,2-d4)resorcinol 10 mg

57981 5-Nonadecylresorcinol 10 mg

53503 5-Eicosylresorcinol 10 mg

50851 5-Heneicosylresorcinol 10 mg

03422 5-Tricosylresorcinol 10 mg

0,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 4,5 5,0 5,5 6,0 6,5 7,0 7,5 8,0 8,5 9,0 9,5 min

0,0

0,5

1,0

1,5

2,0

2,5

3,0

3,5

4,0

(x1,000)

0,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 4,5 5,0 5,5 6,0 6,5 7,0 7,5 8,0 8,5 9,0 9,5 min

0,00

0,25

0,50

0,75

1,00

1,25

1,50

1,75

2,00

2,25

2,50

2,75

3,00

3,25

3,50

3,75

4,00

(x1,000)

0,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 4,5 5,0 5,5 6,0 6,5 7,0 7,5 8,0 8,5 9,0 9,5 min

0,00

0,25

0,50

0,75

1,00

1,25

1,50

1,75

2,00

2,25

2,50

2,75

3,00

3,25

3,50

(x1,000)

0,5 1,0 1,5 2,0 2,5 3,0 3,5 4,0 4,5 5,0 5,5 6,0 6,5 7,0 7,5 8,0 8,5 9,0 9,5 min

0,00

0,25

0,50

0,75

1,00

1,25

1,50

1,75

2,00

2,25

2,50

2,75

3,00

3,25

3,50

3,75

4,00

(x1,000)

Figure 1 Chromatogram Using LC-MS CHROMASOLV Methanol in the Gradient Figure 2 Chromatogram after 2 Blanks Run and First Injection using Standard

HPLC Grade Methanol

Figure 4 Chromatogram after Switching back to a Gradient with LC-MS

CHROMASOLV Methanol and Running Two Blanks

Figure 3 Chromatogram after 10 Injections using a Gradient Containing Standard

HPLC Grade Methanol

For very sensitive UHPLC-MS instruments, we recommend using LC-MS

Ultra solvents, which are tested on UHPLC-(qTOF)MS both in positive

and negative mode, are fi lled in leach-resistant clear borosilicate bottles

for very low cation concentrations, and exhibit an extremely strong lot-

to-lot consistency to get the most out of today’s sensitive UHPLC-MS

instruments.

To learn more about LC-MS Ultra, visit sigma-aldrich.com/uhplc

References:

[1] Ross, AB. Analysis of alkylresorcinols in cereal grains and products using

ultra high-pressure liquid chromatography with fluorescence, ultraviolet,

and CoulArray electrochemical detection. Journal of Agricultural and Food

Chemistry 60, 2012, 8954 – 8962.

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16C

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sigma-aldrich.com/analytix

GC-Headspace SolventsSpecifically developed and optimized for sensitive static

GC-Headspace analysis of volatile organics.● High purity – microfiltered (0.2 μm)● Improved analyte recoveries● Longer shelf life – ed under inert gas● Specifications matching USP, Ph.Eur. & ICH guidelines

For additional information, visit us at sigma-aldrich.com/gc-hs.

Description Size Cat. No.

1,3-Dimethyl-2-imidizolidinone 1 L 67484

N,N-Dimethylacetamide 1 L 44901

Dimethyl sulfoxide 1 L 51779

N,N-Dimethylformamide 1 L 51781

Water 1 L 53463

1-Methyl-2-pyrrolidinone, for GC-HS 1 L 69337

Benzyl alcohol 1 L 80708

LC-MS Ultra Solvents and Additives Designed for UHPLC Quality exceeding your expectations

Purity for low detection limits

• Suitability tested by UHPLC-MS/TOF

• Lot-to-lot reproducibility

• Microfiltered (0.1 μm), filled in clear borosilicate glass containers

For more information or to request a LC-MS Ultra CHROMASOLV Solvents

and Additives Brochure (OUX), please visit sigma-aldrich.com/uhplc

17

Se

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A Remarkable Carbonate-Selective Ionophore for the Determination of Oceanic Carbon Dioxide Using an Ion-Selective Electrode

Potentiometric detectors based on ion-selective electrodes

are used in a wide range of applications for detecting vari-

ous ions important in the clinical, environmental and indus-

trial areas because they offer advantages such as high

selectivity, sensitivity, good precision, simplicity, portability,

non-destructive analysis, and low cost[1].

To map out the global carbon cycle, it is very important to

know detailed information about the spatial and temporal

variations in inorganic carbons in oceans, especially in rela-

tion to the air-sea redistribution of the rapidly increasing

anthropogenic carbon dioxide in the atmosphere and its

infl uence on the greenhouse eff ect[2].

A frequently measured parameter to quantify the oceanic

carbon system is the total CO2 in seawater. It is estimated

from the total alkalinity determined by potentiometric

titration or from the concentration of dissolved CO2 mea-

sured directly with coulometric methods, infrared spectros-

copy, and gas chromatography, which, in seawater, is not

trivial and often requires rather elaborate instrumentation.

Under normal seawater conditions, CO2(aq) is a minor species

(<1%), and the concentration of CO32- is about 10 –11% of the

total inorganic carbons. Hence, considering the relative

abundance of CO32- ion in seawater, carbonate-selective

electrodes should be used in seawater analysis.

Nam et al. synthesized a new carbonate-selective iono-

phore (93206) that behaves like a molecular tweezer; two

trifl uoroacetophenone groups were attached to a rigid cho-

lic acid derivative (Figure 1). The observed carbonate selec-

tivity of log K PotCO3,Cl

= -6.8 of the electrode over chloride and

other anions (incl. lipophilic ones) appeared to be high

enough to apply to seawater analysis.

Daniel Weibel, Product Manager Analytical Reagents daniel.weibel@sial.com

Using 93206 as ionophore in the electrode, the total CO2

level was measured in Yellow Sea water, providing a fast,

stable, and reproducible potentiometric response. In this

manner, the level of total CO2 was determined to be

1.94 ± 0.03 (n = 15) mmol/kg seawater, which was found to

be in good agreement to values determined by a CO2 gas

sensor and a standard potentiometric titration method.

Recommended Membrane Composition (% by wt.)

Cat. No. Description wt%

93206 Carbonate ionophore VII 5.1%

91661 Methyltridodecylammonium chloride 1.2%

02138 Bis(1-ethylhexyl)adipate 56.8%

81392 Poly(vinyl chloride) 36.9%

Dissolved in ethyl acetate and THF (3:5 v/v)

Recommended Cell Assembly

Reference || sample solution || liquid membrane |

0.1 M NaH2PO4, 0.1 M Na2HPO4, 0.01 M NaC | AgCl, Ag

Electrode Characteristics and Function

Selectivity Coef ficients log K PotNa,M

as obtained by the

matched potential method at pH 8.0 (0.1 M Tris-H2SO4):

log K PotCO3,Cl : -6.8 log K Pot

CO3,NO3 : -4.5 log K PotCO3,Salicylate : -1.3

log K PotCO3,Br

: -6.7 log K PotCO3,NO2

: -4.8

log K PotCO3,ClO : -2.0 log K

PotCO3,SCN : -2.8

Slope: -26.0 mV/dec

Detection level: 5.8*10-7 mol/L CO32-

For more information on our sensoric applications, visit

sigma-aldrich.com/selectophore

OCH3

O

N(C8H17)2

H3C

CH3

O

O

O

O

H

H

H H

H

F3COC

CH3

O

O

N(C8H17)2CH3

CH3

O

O

COCF3

COCF3

F3COC

References:

[1] Selectophore® Ion Sensor Materials; Sigma-Aldrich Co.: St. Louis, MO, 2011.

[2] Choi, Y. S.; Lvova, L.; Shin, J. H.; Oh, S. H.; Lee, C. S.; Kim, B. H.; Cha, G. S.; Nam, H. Determination of Oceanic Carbon Dioxide Using a

Carbonate-Selective Electrode. Anal. Chem. 2002, 74, 2435 –2440.

sigma-aldrich.com/selectophore

Figure 1 Structural Representations of the Carbonate-Selective Ionophore 93206, Indicating the Tweezer-Like Shape of the Molecule on the Right.

18

sigma-aldrich.com/ipc

Spec

trom

etry

In previous publications, di- and tri-cationic ion-pair reagents were used to detect inorganic compounds, such as chlorides and sulfates.[1-2]. A recent publication by C. Xu et al [3] demonstrates the versatility of such ion-pair reagents, which are added post-column to the mobile phase and increase the ESI-MS sensitivity extraordinarily by forming positively charged adducts of reagent and anionic analytes (Figure 1).

Figure 1 The Instrumental Setup of HPLC-PIESI-MS.

Since the aim of water analysis is the detection of the small-est amounts of pesticides in water or other biological matri-ces to understand the effect on health and the environ-ment, this new method supports analytical laboratories in this challenge.

LC-MS MethodThe different pesticides are separated on a Supelco Ascentis Express C18 (15 cm x 2.1 mm i.d., 2.7 µm particle size) column using a gradient from 95% 5 mmol/L formic acid/water and 5% 5 mmol/L formic acid/methanol to 5% aqueous buffer within 20 min at 0.4 mL/min. The ion-pair reagent is mixed to the LC flow post-column via a mixing tee with a flow rate of 0.1 mL/min. Injection volume of water samples is 25 µL (fixed loop). This method demonstrates a very good sensi-tivity with a standard LC-MS system. Table 1 shows the list of analytes analyzed with this technique.

A New Approach for the Detection of Acidic Pesticides in Water by MSIncreasing the Sensitivity in Pesticide Analysis by Paired-Ion ESI Detection

Jens Boertz, Product Manager Analytical Reagents jens.boertz@sial.comRudolf Köhling, Sr. Scientist rudolf.koehling@sial.com

Chengdong Xu, Graduate Student1, Zachary S. Breitbach, Research Scientist1, Daniel W. Armstrong, R.A. Welch Professor, 1Department of

Chemistry & Biochemistry, University of Texas at Arlington sec4dwa@uta.edu

In Table 1 also the results for 19 acidic pesticides using three different ion-pairing reagents are presented. The lim-its of detection (LOD) of 19 pesticides obtained were in the range from 0.6 to 19 pg. The fourth ion-pairing reagent applied (1,9-Nonanediyl-bis(3-methylimidazolium) difluo-ride) did not show any improvement in terms of LOD. In total, this approach shows an increased sensitivity of one to three orders of magnitude for the pesticides analyzed.

A novel method based on paired-ion electrospray ioniza-tion (PIESI) mass spectrometry has been developed for determination of acidic pesticides at ultratrace levels in surface and ground waters. The proposed approach pro-vides greatly enhanced sensitivity for acidic pesticides and overcomes the drawbacks of the less sensitive negative ion mode ESI-MS. The limits of detection (LODs) of 19 acidic pesticides were evaluated with four types of dicationic ion-pairing reagents (IPR) in both single-ion monitoring (SIM) and selected-reaction monitoring (SRM) mode. The LOD of 19 pesticides obtained with the use of the optimal dica-tionic ion-pairing reagent ranged from 0.6 pg to 19 pg, indicating the superior sensitivity provided by this method. The transition pathways for different pesticide-IPR com-plexes during the collision-induced dissociation (CID) were identified. To evaluate and eliminate any matrix effects and further decrease the detection limits, off-line solid-phase extraction (SPE) was performed for DI water and a river water matrix spiked with 2000 ng/L and 20 ng/L pesticide standards respectively, which showed an average percent recovery of 93%. The chromatographic separation of the acidic pesticides was conducted by high-performance liquid chromatography (HPLC) using a C18 column (15 cm x 2.1 mm i.d., 2.7 µm particle size) in the reversed- phase mode using linear gradient elution. The optimized HPLC–PIESI-MS/MS method was utilized for determination of acidic pesticides at ng/L level in stream/pond water samples. This experimental approach is one to three orders of magnitude more sensitive for these analytes than other reported methods performed in the negative-ion mode.

[Dicat I]2+

MS pump water/

methanol

ESI-MS

Sample Solution

Supe

lco

Asc

entis

® Ex

pres

s C18

col

umn

LC pump 40 µM paired ion reagent

in water Mixing tee

[Analyte + Dicat I]+

Analyte™

19

sigma-aldrich.com/ipc

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Sample 1,5-Pentanediyl-bis(1-butylpyrrolidinium)

difluoride

1,3-Propanediyl-bis(tripropylphosphonium)

difluoride

1,5-Pentanediyl-bis(3-benzylimidazolium)

difluoride

SIM SRM SIM SRM SIM SRM

LOD (pg) m/za LOD (pg) m/zb LOD (pg) m/za LOD (pg) m/zb LOD (pg) m/za LOD (pg) m/zb

2,4-D 45 543.3 7.0 416.2 10 581.3 8.0 187.2 16 605.2 8.0 385.3

MCPA 3.6 523.4 1.1 396.3 15 561.3 15 187.2 10 585.3 10 385.3

2,4,5-T 25 577.3 3.7 450.2 16 615.3 24 187.2 25 639.2 3.7 385.3

2,4-DB 37 571.4 6.7 444.3 700 609.3 700 187.2 2000 633.3 2000 385.3

MCPB 30 551.4 9.0 424.3 1100 589.4 720 187.2 1300 613.3 1200 385.3

2,4,5-TB 32 605.3 6.0 478.3 1800 643.3 6000 187.2 5300 667.2 2500 385.3

Cloprop 9.0 523.4 2.7 396.3 10 561.3 10 187.2 7.0 585.3 3.5 385.3

Dichlorprop 30 557.3 4.5 430.3 20 595.3 12 187.2 10 619.2 4.0 385.3

Fenoprop 20 591.3 4.0 464.3 20 629.3 20 187.2 30 653.2 10 385.3

Mecoprop 30 537.4 12 410.3 3.8 575.4 2.8 187.2 15 599.3 4.7 385.3

Dicamba 15 543.3 3.0 416.3 6.5 581.3 6.5 187.2 6.0 605.2 2.6 385.3

2,3,6-TBA 110 547.3 6.3 420.2 60 585.2 60 187.2 17 609.2 3.8 385.3

Clopyralid 20 514.3 1.5 387.2 4.5 552.3 4.5 187.2 5.0 576.2 5.0 385.3

Quinclorac 6.0 564.3 0.60 437.2 7.5 602.3 7.5 187.2 10 626.2 3.0 385.3

Quinmerac 15 544.4 2.8 417.3 7.0 582.3 5.2 187.2 3.5 606.3 3.5 385.3

Flupropanate 5.0 469.4 5.0 342.2 3.5 507.3 1.1 187.2 3.1 531.3 0.088 385.3

MCA 6.0 417.3 3.0 290.2 10 455.3 35 187.2 17 479.2 15 385.3

TCA 160 485.2 19 359.3 1.5 523.2 2.0 187.2 15 547.1 7.5 385.3

Dalapon 10 465.3 0.75 359.3 5.0 503.3 5.0 187.2 15 527.2 2.9 385.3

Table 1 Limits of Detection of Pesticide Standard Solutions Obtained by PIESI-MS in the SIM Mode and SRM Mode with the Use of Dicationic Ion-Pairing Reagent.

a Indicates the mass-to-charge ratio of the complex monitored in the SIM mode.

b Indicates the mass-to-charge-ratio of the SRM fragment monitored in the SRM mode.

Cat. No. Name (Abbreviation) Exact Mass Package Size

31518 2,4-dichlorophenoxyacetic acid (2,4-D) 219.97 250 mg

45555 4-chloro-o-tolyloxyacetic acid (MCPA) 200.02 250 mg

45667 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) 253.93 250 mg

45420 4-(2,4-dichlorophenoxy)butyric acid (2,4-DB) 248.00 250 mg

36145 4-(4-chloro-o-tolyloxy)butyric acid (MCPB) 228.06 100 mg

S412325 4-(2,4,5-trichlorophenoxy)butyric acid (2,4,5-TB) 281.96 25 mg

233013 2-(3-chlorophenoxy)propionic acid (cloprop) 200.02 50 g

45436 2-(2,4-dichlorophenoxy)propionic acid (dichlorprop) 233.99 250 mg

45691 2-(2,4,5-trichlorophenoxy)propionic acid (fenoprop) 267.95 250 mg

36147 2-(4-chloro-o-tolyloxy)propionic acid (mecoprop) 214.04 100 mg

45430 3,6-dichloro-2-methoxy-benzoic acid (dicamba) 219.97 250 mg

R169676 2,3,6-trichlorobenzoic acid (2,3,6-TBA) 223.92 25 mg

36758 3,6-dichloro-2-pyridine-carboxylic acid (Clopyralid) 190.95 250 mg

36521 3,7-dichloro-8-quinoline-carboxylic acid (Quinclorac) 240.97 250 mg

36522 7-chloro-3-methyl-8-quinoline-carboxylic acid (Quinmerac) 221.02 250 mg

402923 2-chloroacetic acid (MCA) 93.98 100 g

T6399 2,2,2-trichloroacetic acid (TCA) 161.90 5 g

35562 2,2-Dichloropropionic acid (dalapon) 141.96 250 mg

56618 1,5-Pentanediyl-bis(1-butylpyrrolidinium) difluoride solution 100 mL

75128 1,9-Nonanediyl-bis(3-methylimidazolium) difluoride solution 100 mL

42341 1,3-Propanediyl-bis(tripropylphosphonium) difluoride solution 100 mL

76507 1,5-Pentanediyl-bis(3-benzylimidazolium) difluoride solution 100 mL

53802 Supelco Ascentis Express ™ C18 (15 cm x 2.1 mm i.d.,

2.7 μm particle size)

1 EA

94318 Formic acid 50 mL, 250 mL

34485 Methanol 2.5 L, 4x2.5 L

Table 2 Chemicals Used to Carry Out HPLC-PIESI-MS Experiments .

References

[1] Köhling, R.; Reichlin, N.; Wille, G. Analytix No. 9.

[2] Köhling, R.; Reichlin, N.

Analytix No. 3.

[3] Chengdong X.; Armstrong, D. W.

Analytica Chimica Acta, 792. 2013 1–9.

20

sigma-aldrich.com/analytix

Do You Use IC High-Purity Certifi ed Reference Standards?In that case, check out our IC TraceCERT®!

• Produced under double accreditation, fulfilling ISO/IEC 17025 and ISO Guide 34

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• Certificates can be accessed on the Web by using the product and lot numbers

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Water Determination in Pharmaceutical CompoundsKarl Fischer Titration with HYDRANAL® Reagents

Water-content determination is mandatory for many mate-

rials used in the manufacturing of medicines. Karl Fischer

(KF) titration is the long-standing standard method for this

analysis prescribed by the leading pharmacopoeias, such as

the European (Ph.Eur.), the United States (USP) and the Japa-

nese (JP).

Volumetric Karl Fischer Technique

The volumetric KF technique can be used for samples with

high water content (approx. 0.1–100%). The sample is added

to the titration vessel containing a suitable working medium,

such as Hydranal Methanol dry or Hydranal Solvent. With

the corresponding iodine-containing titrating agent,

Hydranal Composite or Hydranal Titrant, the water content

of the sample is determined by titration. End-point deter-

mination is carried out by applying a constant current and

measuring the voltage via double platinum electrode; or

vice versa, applying a constant voltage and measuring the

current. The water content is calculated from sample

weight, consumption of titrating agent, and water equiva-

lent (titer) of the titrating agent. Pharmaceutical com-

pounds such as ethosuximide, used for epilepsy treatment,

(application L510), or the cytostatic drug cyclophospha-

mide (application L463), can be analyzed according to stan-

dard procedures without interference. These titrations can

also be carried out in less toxic ethanol-based reagents, the

Hydranal E-types.

Coulometric Karl Fischer Technique

Samples with lower water content (up to 1%) can be ana-

lyzed using the coulometric KF method. The required

iodine is generated electrochemically in the titration vessel

by anodic oxidation of iodide, which is contained in the

Hydranal Coulomat reagents. The water content is calcu-

lated from the amount of the produced (consumed) elec-

tric current (which equals iodine consumption) over time.

Karl Fischer Oven Technique

Insoluble samples, samples that undergo side reactions

with the KF reagents, or samples that release their water

only at high temperatures, may be analyzed using the KF

oven. The water of the sample is driven out at a variable

temperature and transferred by a suitable carrier gas into

the KF titration vessel. The water content is determined

Andrea Felgner, Market Segment Manager andrea.felgner@sial.com

according to standard procedures. 5-Aminolevulinic acid-

HCl (lyophilisate) shows a strong side reaction in methanol;

an end point cannot be reached with direct KF titration

methods. For this compound, we recommend determina-

tion with the KF oven in combination with the coulometric

titration technique, due to the low water content of this

substance. Suitable application parameters are (application

L506):

• Evaporation temperature 80 °C (decomposition of the

sample starts at 120 °C)

• Determination time 600 seconds

• Anolyte Hydranal Coulomat AG or Hydranal Coulomat

AG-Oven

• Catholyte Hydranal Coulomat CG

Side Reactions and pH-Influencing Samples

Substances containing nitrogen compounds may cause

interference with the pH value of the working medium.

Side reactions occur during the KF titration, possibly lead-

ing to coated electrodes, fading end points or no end

points at all, and erroneous results. These side reactions can

be suppressed through the addition of suitable buff er sub-

stances to the working medium in the titration vessel.

Benserazide hydrochloride, an active ingredient used in the

treatment of Parkinson’s disease, is an example of a nitro-

gen-containing substance. For correct determination of its

water content, Hydranal Buff er Base or a mixture of Hydranal

Methanol dry and salicylic acid should be used as a working

medium, and then be titrated with Hydranal Composite 2

(application L416). Profl avine hemisulfate, a topical antisep-

tic, also increases the pH value of the working medium and

no end point is achieved. Hydranal Buff er Base or a mixture

of Hydranal Methanol dry and benzoic acid can be used to

lower the pH value and prevent the side reaction; titrate

with Hydranal Composite 5 (application L354). Water-con-

tent determination in penicillin can also be disturbed by pH

infl uences; a side reaction occurs: penicillin derivatives such

as penicilloic acid and other hydrolysis products are oxi-

dized by iodine. By conducting the titration in weakly acidic

conditions, this side reaction can be suppressed. The KF

one-component technique with Hydranal Methanol dry or

Hydranal Methanol Rapid and Hydranal Composite 2 gives

a pH value of approximately 5 in the titration vessel. This is

Tit

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(continued on page 22)

22

sigma-aldrich.com/hydranal

suffi cient for titrating the water content in penicillin with-

out any side reactions. If an end point is not easily reached,

addition of salicylic acid to the working medium (before

pre-titration) is recommended. Titration of penicillin with

the KF two-component technique is also possible. A mix-

ture of Hydranal Solvent and salicylic acid is recommended

as a working medium and can be titrated with Hydranal

Titrant (application L166).

Enhancement of Sample Solubility

For samples that are only poorly soluble in the KF working

medium, Hydranal-Buffer Base or a mixture of Hydranal

Methanol dry and salicylic acid can be recommended as

working media to enhance solubility and therefore yield

correct determination results (titrating agent Hydranal

Composite). Examples are the beta-lactam antibiotics

amoxycillin-3-hydrate (application L352) or ampicillin

(application L422).

However, other substances such as riboflavin phosphate

sodium (biochemical cofactor, also used as a food dye)

prove insoluble in the alcohol-based KF media. As a suit-

able solubilizer for this compound, formamide is recom-

mended. It can be added to the working medium (volu-

metric KF technique) in a ratio of 1:1 (application L509).

Application Reports

All application reports can be obtained by contacting our

Hydranal technical service team (hydranal@sial.com). Visit

our Web site sigma-aldrich.com/hydranal for complete

product listings and more information on KF titration.

On our Web site sigma-aldrich.com/hydranal you will

fi nd more information about the Ph. Eur. requirements for

KF titration. Our Hydranal technical service team has carried

out suitability tests for a range of substances, which are

available upon request.

Take advantage of both our expertise gained from over

thirty years of experience and our extensive applications

database on KF titration. For any questions, help, or feed-

back, please visit our Web site or contact us (details below).

Figure 1 Application Report for Polysorbate 80 (L608)

Tit

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on

Cat. No. Description

L166 Penicillin-G sodium and penicillin-G-potassium

L230 Dobutamine hydrochloride

L242 Erythromycin

L249 Adenosine-5’-triphosphoric acid disodium salt

L297 Glycerine monostearate

L307 Benzyl penicillin procaine

L326 Magnesium stearate

L352 Amoxycillin-3-hydrate

L354 Proflavine hemisulfate

L416 Benserazide hydrochloride

L422 Ampicillin

L448 Gentamicin sulfate

L463 Cyclophosphamide

L489 Disodium hydrogen phosphate-12-hydrate

L506 Aminolevulinic acid-HCl (Lyophylisate)

L509 Riboflavin phosphate sodium

L510 Ethosuximide

L604 Povidon

L607 Sorbitol solution (70%)

L608 Polysorbate 80

L660 Ibuprofen (film tablets)

L661 Zineb

L664 Niclosamide hydrate

L670 Atrazine desisopropyl

L675 Atrazine desisopropyl-2-hydroxy

L698 Asulam

Table 1 Selected Application Reports for the Pharmaceutical

Industry

23

sigma-aldrich.com/hydranal

Cat. No. Description

Reagents for Volumetric KF Titration

34805 Hydranal Composite 5, reagent for one-component KF titration

34806 Hydranal Composite 2, reagent for one-component KF titration

37817 Hydranal Methanol Rapid, working medium for fast one-component KF titration

34741 Hydranal Methanol dry, working medium for one-component KF titration

34801 Hydranal Titrant 5, reagent for two-component KF titration

34811 Hydranal Titrant 2, reagent for two-component KF titration

34800 Hydranal Solvent, working medium for two-component KF titration

34724 Hydranal Formamide dry, solvent for KF titration

34804 Hydranal Buffer Acid, buffer substance for KF titration

37859 Hydranal Buffer Base, buffer substance for KF titration

37865 Hydranal Salicylic Acid, buffer substance for KF titration

32035 Hydranal Benzoic Acid, buffer substance for KF titration

Reagents for Coulometric KF Titration

34836 Hydranal Coulomat AG, anolyte reagent

34739 Hydranal Coulomat AG-Oven, anolyte reagent for KF oven

34840 Hydranal Coulomat CG, catholyte reagent for cells with diaphragm

Water Standards for KF Titration

34849 Hydranal Water Standard 10.0, 1 g contains 10.0 mg = 1.0% water, tested against NIST SRM 2890 (exact value on CoA)

34828 Hydranal Water Standard 1.0, 1 g contains 1.0 mg = 0.1% water, tested against NIST SRM 2890 (exact value on CoA

34847 Hydranal Water Standard 0.1, 1 g contains 0.1 mg = 0.01% water, tested against NIST SRM 2890 (exact value on CoA)

34696 Hydranal Standard sodium tartrate dihydrate, solid standard for volumetric KF titration, water content ~15.6% (exact value on CoA)

Table 2 Selected Hydranal Reagents for KF titration

Request the new Hydranal Product Line Overview brochure

• Volumetric Titration with One-Component Reagents

• Volumetric Titration with Two-Component Reagents

• Coulometric Titration with Hydranal Coulomat

• Hydranal Water Standards

• Hydranal E-Type Reagents

• Reagents for Ketones and Aldehydes

• Technical Support

Order this brochure by returning the attached business reply card or visit

sigma-alrich.com/hydranal for an electronic version.

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