Analytix

Issue 3 • 2011

Water Determination in Milk and Milk Products

• Water Determination in Dairy Products • New UHPLC Solvents • Detection • New Derivatization Reagents • New Solvents for GC-Headspace • Certified Reference Materials • Reference Material for Herbal Medicine 2

Expanded Quality to Meet Your Analytical Needs – Introducing Cerilliant and Resource Technology Corporation (RTC), the two newest additions to Sigma-Aldrich

Dear Colleague, In our eff orts to bring you the highest quality Add the outstanding expertise of Cerilliant and RTC to the and best offering of analytical products, over 40 years of experience at Supelco and 60 years at Fluka Sigma-Aldrich would like to welcome our two and you have an unmatched amount of knowledge and

Editorial newest family members, Cerilliant Corporation products available for your laboratory needs. These compa- of Round Rock, Texas and RTC of Laramie, nies will combine their talents and capabilities with those of Wyoming. The extensive capabilities of these Sigma-Aldrich to create visionary approaches to supply the two companies complement Sigma-Aldrich’s analytical industry with a continually expanding list of new existing Fluka® and Supelco® analytical brands. products that meet the most rigorous quality requirements including cGMP, GLP and ISO requirements. Cerilliant’s quality credentials include accred- itation to ISO Guide 34 and ISO/IEC 17025, certi- Currently the products of Cerilliant and RTC are only avail- fi cation to ISO 9001:2008 and incorporation of able through Cerilliant and RTC themselves or one of Dr. Ingo Haag cGMP and GLP requirements into their everyday their distributors. Please contact them directly or visit Director Marketing activities. They are currently pursuing ISO www.cerilliant.com or www.rt-corp.com to review 13485 and expect to receive their certifi cation products, to ask a question or to place an order. in early 2011. Cerilliant off ers over 2,800 products and more than 30 years of expertise in the Kind regards, design, development, synthesis, characterisation and packaging of analytical standards, calibra- tors, controls and certifi ed reference materials for the pharmaceutical, clinical diagnostics, forensic and clinical toxicology, nutraceutical Dr. Ingo Haag and environmental markets. Director Marketing [email protected] RTC is a premium manufacturer of Certified Reference Materials (CRMs), Quality Control (QC) Samples, and Proficiency Testing (PT) Samples with more than 20 years of experience and a strong expertise in the environmental as well as in the pharmaceutical sector. RTC was one of the original Proficiency Test providers recognised by the US EPA. Its quality systems include ISO/IEC 17025, ISO Guide 34 (CRMs), ISO/IEC 17043 (PT) and ISO 9001:2000.

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 director: Ingo Haag, PhD Editor: Daniel Vogler

sigma-aldrich.com/analytix 3

Feature Article Standards

4 Water Determination in Milk and Milk 17 DAIH- and DNPH- Aldehyde and Ketone Products Standards Karl Fischer Titration with HYDRANAL® Reagents 19 New Analytical Standards for the Analysis of Herbal Medicinal Products

Microbiology of Contents Table 21 New TraceCERT Organic Certified Reference 8 Salmonella’s Tricks and their Detection Materials

Derivatization Titration

11 New Derivatization Reagents 22 Volumetric Titration VOLPAC® Solution Containers with new adapter Chromatography

12 UHPLC – State-of-the-Art in Chromatography Sigma-Aldrich introduces LC-MS Ultra CHROMASOLV®

16 New Headspace Grade Solvents

New Product Corner

15 New SELECTOPHORE® Ionophores with Improved Properties

sigma-aldrich.com/analytix 4

Water Determination in Milk and Milk Products Karl Fischer Titration with HYDRANAL® Reagents

Andrea Felgner, Product Manager Analytical Reagents [email protected] Thomas Wendt, HYDRANAL Technical Service [email protected]

In the production of milk and dairy products, water content plays an important role, technologically as well as economically. Therefore, national and international laws and agreements protect and regulate the defi nition of products and the production processes in dairy technology. Determining the water content in milk and milk pro- ducts can be easily achieved with Karl Fischer (KF) titration.

Whole milk is a colloidal system composed of a continuous aqueous phase in which lactose, whey proteins, and minerals are dissolved. The dispersed phase consists of casein micelles and fat globules. Whole cow’s milk typically contains 90 g/kg of non- fat solids and between 35 to 40 g/kg of fat. Both the fat globules and the casein micelles contribute to the whiteness of milk by light scattering. Cream can be consi- dered a concentrated dispersion of fat globules, while skim milk contains residual Feature Article Feature milk fat of less than 0.1%.

Production of Milk and Milk products An important step in the industrial production of milk products (Figure 1) is the separation of milk into cream and skim milk. Cream can be considered as the fat part of the milk, while skim milk is practically fat-free, containing water, milk proteins, and lactose. Butter is made from cream, whereas buttermilk is the remaining liquid from this process. Raw milk (or cream and skim milk combined as ingredients) is the basic material for many other milk products, such as cheese, milk powder, condensed milk, and fermented milk products.

Drinking Milk – Treatments and classification The European Commission defi nes diff erent types of consumer milks: • Raw milk has not been heated above 40 °C • Whole milk has been heat-treated (pasteurized or sterilized)

Pasteurization is used to destroy all pathogenic microorganisms in milk. The so-called continuous high-temperature short-time (HTST) procedure prescribes a rapid heating of the milk to temperatures from 71-74 °C, and holding the product at this temperature for 15 to 20 seconds.

Sterilization of milk can be achieved by Ultrahigh Temperature heating (UHT). The characteristic temperature range for UHT heating is 135-150 °C with holding times of a few seconds. The goal of UHT processing is to accomplish a sterile product storable at ambient temperature. This heat treatment destroys all microorganisms and the product is then packaged aseptically into sterile, hermetically sealed packages.

Raw Milk Drinking Milk

Whey

Butter

Cream (Fat) Skim Milk Milk Proteins

Buttermilk

Lactose

Drinking Milk Cheese Milk Powder Condensed Milk Fermented / Cultured Milk Products

Figure 1 Production steps of milk and milk products

sigma-aldrich.com/hydranal 5

Product Water Protein Fat Carbohydrates Organic acids Minerals Whole milk 87.7% 3.3% 3.6% 4.6% 0.2% 0.7% Fat-reduced milk 89.6% 3.4% 1.6% 4.6% 0.2% 0.7% Skim milk 90.9% 3.5% 0.1% 4.8% 0.2% 0.8% Buttermilk 91.2% 3.5% 0.5% 4.0% 0.8% 0.8% Whey (sweet) 93.6% 0.8% 0.2% 4.7% 0.3% 0.6% Condensed milk 74.7% 6.5% 7.6% 9.2% 0.3% 1.5% Coffee cream 81.7% 3.1% 10.5% 4.1% - 0.6%

Table 1 Nutritional composition of milk products

Other Milk Products before taking the sample is recommended, especially for Evaporated / Condensed Milk is produced through the con- quark and yoghurt. For titration of milk powder, a heating of centration and drying of milk. These processing steps lead the reagents to 50 °C may be benefi cial. Since milk powder to products with a smaller volume and a longer shelf life. To at this temperature releases its water content only slowly, further enhance the shelf life of these products, heat treat- the determination can be accelerated at higher tempera- ment for sterilization can be applied. tures. Milk powder with especially high fat content may need the addition of chloroform to the working medium to The partial removal of water from milk or cream leads to improve the solubility of the sample. Coulometric KF titra- production of milk powder or cream powder. The fat and/ tion is not recommended for these samples. Article Feature or protein content of the milk or cream may be adjusted by the addition and/or withdrawal of milk constituents. The Sample sizes and dosage: accurate control and adjustment of a precise value of water Coff ee cream 0.1 g / syringe with needle content is especially signifi cant for milk powders (Table 3). Milk 0.05 g / syringe with needle Milk powder 1.0 g / powder funnel Application Quark 0.1 g / syringe without needle Water determination in coffee cream, quark Yoghurt 0.1 g / syringe without needle (cheese), milk, milk powder, yoghurt Milk consists mainly of water; therefore, its water content Volumetric KF technique determination is straightforward. All of these products Reagent One-component technique release their water easily in the working medium of the KF Titrating agent HYDRANAL®-Composite 5 titration; the determination can be carried out with classical Working medium HYDRANAL-Methanol Rapid or HYDRANAL-Methanol dry KF reagents. However, it is very often necessary to take into Reagent Two-component technique account the inhomogeneous water distribution or separa- Titrating agent HYDRANAL-Titrant 5 tion of components during storage. A homogenization Working medium HYDRANAL-Solvent

EU Regulation USA Regulation Non-standardized whole milk Whole milk Unchanged fat content, not less than 3.5% Minimum fat content of 3.25% Fat-adjusted whole milk Reduced-fat milk Modified fat content, not less than 3.5% Fat content of 2% (actual content indicated on the package) Partially skimmed milk Low-fat milk Adjusted fat content of 1.5%-1.8% Fat content of 1% Skim milk Skim milk Fat content of maximum 0.5% Fat content maximum 0.5% (may be called “fat free”)

Table 2 Classification of Drinking Milk (EU and USA)

Product Milk fat content Water content Milk protein in milk solids-not-fat Cream powder Min. 42% m/m Whole milk powder Min. 26% m/m Max. 42% m/m Max. 5% m/m Min. 34% m/m Partly skimmed milk powder Min. 1.5% m/m Max. 26% m/m Skimmed milk powder Max. 1.5% m/m

Table 3 Composition of milk powders (continued on page 6)

sigma-aldrich.com/hydranal 6 Feature Article Feature

Application Volumetric KF technique Water determination in cheese Reagent One-component technique Cheese does not dissolve in methanol and therefore does Titrating agent HYDRANAL-Composite 5 not release its water content in classical KF solvents. The Working medium HYDRANAL-LipoSolver CM (or addition of formamide to the working medium as a solubil- HYDRANAL-Methanol dry/Rapid and HYDRANAL-Chloroform 1:1) izer is necessary; carrying out the titration at elevated tem- Two-component technique peratures is also recommended. The sample should be Titrating agent HYDRANAL-Titrant 5 finely grated or cut into small pieces; this type of sample Working medium HYDRANAL-Solvent CM (or will give a turbid solution with the working medium after HYDRANAL-Solvent and stirring for 5-10 minutes; the following titration should take HYDRANAL-Chloroform 2:1) about 2-3 minutes (sample size 0.2 g / powder funnel). Coulometric KF titration is not recommended for this Application sample. Water determination in whey fat emulsion This product may coat the electrodes in the titration cell. Volumetric KF technique Fading end-points and erroneously high results are the Reagent One-component technique consequences of this interference. The addition of form- Titrating agent HYDRANAL®-Composite 5 amide to the working medium can prevent this interference, Working medium HYDRANAL-Methanol dry/Rapid and while addition of chloroform is necessary to dissolve the fat HYDRANAL-Formamide dry 1:1 of the sample (sample size 0.1 g / syringe without needle). Two-component technique Coulometric KF titration is not recommended for this Titrating agent HYDRANAL-Titrant 5 sample. Working medium HYDRANAL-Solvent and HYDRANAL-Formamide dry 1:1 Volumetric KF technique Application Reagent One-component technique Water determination in ice cream Titrating agent HYDRANAL-Composite 5 Working medium HYDRANAL-Methanol dry and The sample should be melted and carefully homogenized HYDRANAL-Chloroform and before the analysis. To improve the dissolution of the fat HYDRANAL-Formamide dry 1:1:1 content, the titration should be carried out in HYDRANAL- Two-component technique LipoSolver CM, which contains chloroform (sample size Titrating agent HYDRANAL-Titrant 5 0.05 g / syringe without needle). Coulometric KF titration is Working medium HYDRANAL-Solvent and not recommended for this sample. HYDRANAL-Chloroform and HYDRANAL-Formamide dry 2:1:1

sigma-aldrich.com/hydranal 7

Application Cat. No. Description Water determination in whey powder Volumetric one-component reagents In our investigations, this sample showed a small side reaction 34805 HYDRANAL-Composite 5 with iodine. These reactions are often pH-dependent; 34806 HYDRANAL-Composite 2 HYDRANAL®-Buff er Base or a working medium containing a 34827 HYDRANAL-Composite 1 portion of salicylic acid helps to suppress this secondary re- 34741 HYDRANAL-Methanol dry action. A titration at elevated temperatures enhances the side 37817 HYDRANAL-Methanol Rapid 34734 HYDRANAL-CompoSolver E reaction and is therefore not recommended. To dissolve the 37855 HYDRANAL-LipoSolver CM high lactose content contained in the sample, the addition Volumetric two-component reagents of formamide is also necessary (sample size 0.5 g / powder 34732 HYDRANAL-Titrant 5 E funnel). Coulometric KF titration is not recommended for 34723 HYDRANAL-Titrant 2 E this sample. 34801 HYDRANAL-Titrant 5 34811 HYDRANAL-Titrant 2 Volumetric KF technique 34730 HYDRANAL-Solvent E Reagent One-component technique 34800 HYDRANAL-Solvent Titrating agent HYDRANAL-Composite 5 34812 HYDRANAL-Solvent CM Working medium HYDRANAL-Buffer Base (or Auxiliary Reagents (Solubilizers) HYDRANAL-Methanol dry/Rapid and 5 g 37863 HYDRANAL-Chloroform HYDRANAL-Salicylic acid) and 34724 HYDRANAL-Formamide dry Feature Article Feature HYDRANAL-Formamide dry 2:1 37865 HYDRANAL-Salicylic acid Two-component technique 34804 HYDRANAL-Buffer Acid Titrating agent HYDRANAL-Titrant 5 37859 HYDRANAL-Buffer Base Working medium HYDRANAL-Solvent and Water standards for titer determination HYDRANAL-Formamide dry 2:1 plus 5 g 34849 HYDRANAL-Water Standard 10.0 HYDRANAL-Salicylic acid 34696 HYDRANAL-Standard sodium tartrate dihydrate

Table 4 Selected Fluka brand HYDRANAL Karl Fischer reagents Application reports (available upon request from [email protected]): References L074 Quark (cheese) [1] Jost, R. Milk and Dairy Products. Ullmann’s Encyclopedia of L077 Whey fat emulsion Industrial Chemistry, 2007. L081 Milk powder (whole milk) [2] Lebensmitteltabelle für die Praxis. Editor: Deutsche L082 Milk powder (skimmed milk) Forschungsanstalt für Lebensmittelchemie, Garching bei München. Wissenschaftliche Verlagsgesellschaft Stuttgart, L085 Milk 2009. L095 Cheese [3] Codex Alimentarius “Codex standard for milk powders and L097 Yoghurt cream powder”. CODEX STAN 207-1999. FAO/WHO. L222 Coff ee cream L493 Whey (powdered) L523 Ice cream

Sigma-Aldrich off ers over 650 application reports. A full list of these reports and all HYDRANAL products can be found on our website sigma-aldrich.com/hydranal To obtain an application report, please contact one of our HYDRANAL laboratories:

Europe and Global Market USA and Canada Mr. Thomas Wendt Mr. Doug Clark Technical Service HYDRANAL HYDRANAL Technical Center D-30926 Seelze, Germany St. Louis MO 63103, USA Tel.: +49 (0) 5137 8238-353 Toll free: +1 800 493-7262 (USA and Canada) Fax: +49 (0) 5137 8238-698 Fax: +1 314 286-6699 E-Mail: [email protected] E-Mail: [email protected]

sigma-aldrich.com/hydranal 8

Salmonella’s Tricks and their Detection

Jvo Siegrist, Product Manager [email protected]

New deadly tools of Salmonella have been discovered, presenting opportunities for novel approaches in controlling Salmonella outbreaks.

Salmonella has the “know how” and the tools to invade into the metabolism of host cells. In a new study, researchers detected a bacterial sorting platform (bacterial type III pro- tein secretion system) that organised the necessary pro- teins. A key role of this cytoplasmic sorting platform is to line up the secretion of proteins in a specific order to be able to take control of the host cell’s function. The platform uses customized chaperones to organise the type III protein secretion. It organises the building up of a needle complex Figure 1 Colorized scanning electron micrograph (SEM) revealing through which the eff ector proteins can later be injected the presence of a single Gram-negative Salmonella typhimurium into the host cells. These new fi ndings may introduce new bacterium that has been isolated from a pure culture (Magnification 20000x; CDC - National Center for Zoonotic, Vector-Borne, and opportunities for novel antimicrobial strategies [1]. Enteric Diseases (ZVED); Janice Haney Carr)

Salmonella contamination is the second leading cause of The media used currently for the diff erentiation and identi-

Microbiology food-borne illness worldwide. Controlling outbreaks of fication of Salmonella are still based on the detection of Salmonella is an important task for food regulators, restau- carbohydrate fermentation indicated by a pH indicator (see rants and the food industry in general. Table 2 for carbohydrate fermentation ability), the detec- tion of proteolytic activity, hydrogen sulphide production The Salmonella family includes over 2,300 serotypes of bacte- and selectivity. Most modern media also combine aspects ria. However, two types, Salmonella enteritidis and Salmonella of this detection system to make the media more reliable. A typhimurium, are responsible for approximately half of all listing of the most common selective and diff erential media infections in humans. Most outbreaks of Salmonella may be appears in Table 3. For a typical microbiological control traced back to dairy, poultry and meat products; however, procedure, please see the ISO method illustrated in Figure 2. Salmonella can grow on nearly any food. Chicken, eggs and their derivative products pose a particularly high risk. In addition, Sigma-Aldrich’s innovative technology has developed a line of chromogenic media products which Kingdom: Bacteria make identifi cation even more reliable and faster as they Phylum: Proteobacteria detect a characteristic enzyme of Salmonella. These reac- Class: Gamma Proteobacteria tions are based on the cleavage of a chromogenic substrate Order: Enterobacteriales which results in a visible color change (see Table 4). Family: Genus: Salmonella

Table 1 Scientific classification

Microbiological control in the food industry plays a critical role in preventing Salmonella outbreaks. Tests and media used for the identifi cation of Salmonella take advantage of unique aspects of Salmonella’s physiology or biochemistry relative to other genera within the family Enterobacteriaceae. For example, bacteria from the genus Salmonella are primarily facultative anaerobes, oxidase-negative, catalase-positive and Gram-negative rods. Most strains are motile and fer- ment glucose with production of both acid and gas.

sigma-aldrich.com/salmonella 9

Sample

BPW 08105 Buffered Peptone Water (ISO) 37 °C Non-selective pre-enrichment 18 hrs Annex D

17173 Rappaport Vassiliadis Broth, modified

RVS Broth MKTTn MSRV 43052 Muller-Kauffmann Tetrathionate 41.5 °C 37 °C 41.5 °C Broth, Base (ISO) Selective enrichment 24 hrs 24 hrs 24 hrs 92322 Rappaport Vassiliadis medium (base), (48 hrs if modified, semi-solid negative)

95586 XLD Agar A second media of choice 01993 Salmonella Chromogen Agar Set Salmonella Chromogen Agar, XLD 76721 XLT4 Agar (Base) Selective isolation BGA, XLT4, Hektoen, SS, etc.

70116 No 2 Nutrient Agar Confirmation 37 °C 24 hrs 92499 Triple Sugar Iron Agar (acc. to ISO)

51463 Urea Broth Microbiology Biochemical Test Serological (e.g. TSI) 66304 Lysine Decarboxylase Broth 62915 Lysine Iron Agar

Figure 2 Microbiology of food and animal foodstuffs — Horizontal method for the detection of Salmonella spp.; ISO 6579:2002/A1:2007

Carbohydrate Fermentation Cat. No. of Fluka Acid Gas Carbohydrate Discs Adonitol - - 55876 Arabinose +/- +/- 80372 Cellobiose - - 56481 Dextrose + +/- 63367 Dulcitol +/- +/- 73044 Fructose +/- +/- 53901 Galactose + +/- 89608 Inositol +/- +/- 89614 Lactose--28816 Maltose + +/- 77653 Mannitol + +/- 94438 Mannose +/- +/- 94445 Melibiose++93196 Raffinose--94226 Rhamnose +/- +/- 93999 Salicin--92971 Sorbitol + +/- 93998 Sucrose - - 94309 Figure 3 HiCrome™ MM Trehalose + +/- 92961 Xylose + +/- 07411

Table 2 Typical carbohydrate fermentation ability of Salmonella

(continued on page 10)

sigma-aldrich.com/microbiology 10

Cat. No. Medium Cat. No. Medium Non-selective 08105 Buffered Peptone Water (ISO) Growth and 01477 Nutrient Agar No 2 pre- D3435 Dey-Engley Neutralizing Broth purification (ready to use media in the bottle) enrichment 70123 Nutrient Broth No. 2 media 70116 Nutrient Agar No 2 broth 78104 Nutrient Broth No. 5 N9405 Nutrient Agar pH 6.8

03856 Nutrient Broth No. 4 17209 R-2A Agar 17181 Nutrient Broth pH 6.9 without NaCl 17221 Tryptone Yeast extract Agar 77185 Peptone Water Confirmation A0715 Andrade Peptone Water 94217 Peptone Water, phosphate-buffered media 28943 Andrade Peptone Water, Vegitone (ready-to-use media in the bottle) 36408 Bromcresol Purple Broth 77187 Peptone Water, phosphate-buffered D2935 Decarboxylase Broth Base, Moeller 67331 Peptone Water, phosphate-buffered 60787 Kligler Agar with Ferrioxamine E 66304 Lysine Decarboxylase Broth 40893 Peptone Water, phosphate- 62915 Lysine Iron Agar buffered, Vegitone 39484 Methyl Red Voges Proskauer Broth 07507 Tryptone Water 69150 Methyl Red Voges Proskauer Saline (ready-to-use media in the bottle) Broth 70194 Tryptone water M1053 Motility Test Medium 91366 Universal Pre-Enrichment Broth 72548 Nitrate Broth Selective 69965 Mossel Broth 75315 OF Test Nutrient Agar enrichment 43052 Muller-Kauffmann Tetrathionate 85438 SIM Medium broth Broth, Base (ISO) 44940 Triple Sugar Iron Agar

04584 Rappaport Vassiliadis Broth acc. to 92499 Triple Sugar Iron Agar (acc. to ISO) DIN EN ISO 6579:2002 51463 Urea Broth 17173 Rappaport Vassiliadis Broth, modified Table 3 Salmonella selective and differential media R0773 Rappaport Vassiliadis Medium 92322 Rappaport Vassiliadis medium Brand Cat. No. Name Microbiology (base), modified, semi-solid Fluka® 00563 HiCrome™ MM Agar 84370 Salmonella Enrichment Broth Fluka 90918 HiCrome RajHans Medium, Modified 70153 Selenite Broth (Base) Fluka 78419 HiCrome Salmonella Agar 84922 Selenite Cystine Broth Fluka 05538 HiCrome Salmonella Agar, Improved 86352 TBG Broth Fluka 84369 Salmonella Chromogen Agar 88151 Tetrathionate Broth Fluka 01993 Salmonella Chromogen Agar Set Selective 95388 Table 4 Chromogenic media for Salmonella confirmation B1801 Brilliant Green Agar modified media 70134 Brilliant Green Agar, modified Reference: 16026 Brilliant Green Phenol Red Lactose [1] María Lara-Tejero, Junya Kato, Samuel Wagner, Xiaoyun Liu, Sucrose Agar and Jorge E. Galán. A Sorting Platform Determines the Order 70135 DCLS Agar of Protein Secretion in Bacterial Type III Systems. Science, 90035 DCLS Agar No. 2 3 February 2011. D7809 Deoxycholate Citrate Agar 51490 61792 Leifson Agar 81648 Pril® Mannitol Agar 84368 Salmonella Agar according to Önöz 84369 Salmonella Chromogen Agar 01993 Salmonella Chromogen Agar Set 85640 SS-Agar 95586 XLD Agar 76721 XLT4 Agar (Base)

sigma-aldrich.com/microbiology 11

New Derivatization Reagents

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

The primary use of derivatization is the chemical modification of a compound by derivatizing the functional group (e. g. -OH, -COOH, -NH, and -SH) to promote the use of chromatographic analysis. If left underivatized, these com- pounds can demonstrate poor chromatographic behavior, insuf- fi cient volatility, and poor thermal stability or have inadequate detector response. By chemically modifying these compounds into derivatives, they gain properties amenable for chromatographic separation and accurate analysis.

With our broad portfolio of derivatization reagents, we at Sigma-Aldrich demonstrate our commitment towards seeking new advancements and imple- menting them in improved and innovative products. Check out our latest developments that will

enable you to experience Derivatization improved performance and achieve solutions for challenging chromatography.

Brand Product Description Pack size HPLC Derivatization enhanced sensitivity in electrochemical detection, analysis of steroids FLUKA® 95159 Ferrocenecarboxaldehyde 100 mg FLUKA 56257 Ferroceneboronic acid 100 mg FLUK A 89111 N-Ferrocenyl-maleimide 100 mg FLUKA 76737 3-Ferrocenylpropionic anhydride 100 mg FLUKA 50203 Ferrocenoyl azide 100 mg HPLC Derivatisation for primary amines and diamines FLUKA 49558 2.6-Dimethyl-4-quinolinecarboxylic acid N-hydroxysuccinimide ester 100 mg HPLC-UV Derivatization: synthesis of water-soluble prodrugs of lipophilic alcohols, phenols and amines FLUKA 86546 Dibenzyl chloromethyl phosphate 1 g Derivatization of non steroidal anti-inflammatory drugs

FLUKA 68768 N-Methyl-N-(trimethyl-d9-silyl)trifluoroacetamide 500 μL Silylation for GC-Derivatization without solvent or excess of reagent FLUKA 79271 Trimethylsilyl methallylsulfinate, for GC derivatization 5 mL FLUKA 79264 Triethylsilyl methallylsulfinate, for GC derivatization 5 mL FLUKA 79262 tert-Butyldimethylsilyl methallylsulfinate, for GC derivatization 5 mL

For further information, please visit sigma-aldrich.com/derivatization

sigma-aldrich.com/derivatization 12

UHPLC – State-of-the-Art in Chromatography Sigma-Aldrich introduces LC-MS Ultra CHROMASOLV®

Rudolf Köhling, LC-MS Application Scientist [email protected]

Since the early 1940s, liquid chromatography has pro- obtained from a fast high throughput method using only a gressed through rapid development. Even in the earliest short 2.1 x 5 mm Ascentis® Express C18 column. Although the publications, smaller packing particles and higher column total run-time is only 1 min., all major matrix compounds are pressures were being proposed for greater separation effi - separated from the analyte, which is necessary to optimize ciencies and faster run-times. Years before the introduction the limit of detection and eliminate suppression effects. A

Chromatography of a commercial Ultra High Performance Liquid Chromato- comparable separation on a standard HPLC system is not graphy (UHPLC) system by Waters in 2004, V. Meyer pre- possible or requires extensive modifi cations to the HPLC sys- sented in 1997 a fast chromatography of benzodiazepines tem to minimize peak dispersion and loss of sensitivity. with nonporous 1.5 μm particles, and demonstrated the advantages of increased chromatographic resolution and Figure 2 shows the diff erence between a standard HPLC speed [1]. At the present time, all major HPLC manufactur- column and a Supelco® Ascentis Express column with ers off er UHPLC systems for pressures up to 1200 bar and Fused-Core particles. UHPLC or optimized HPLC systems minimized mixing in tubing connections and fittings. with core-shell particle columns are able to separate com- Another major development focused on the improvement pounds into very narrow fractions having high peak of stationary phases. The combination of small solid mono- responses from UV, MS or other detectors. This advantage disperse particles coated with a porous functionalized shell requires a very high and stable quality for solvents and (Fused-Core®) has led to outstanding column effi ciencies additives. Even a slight impurity may result in additional and has opened up new techniques for standard HPLC and peaks which may create a complicating issue with steep UHPLC users, e.g. very fast separations on short columns. The and fast gradient runs (Figure 3). separation in Figure 1 represents well the clarity of results

sigma-aldrich.com/lc-ms 13

Intens . x104 Intens . 1. +MS, 0.4-0.5min #(60-75), -Peak Bkgrnd 2.5 506.238

300

200

270.128 229.119

298.096

161.082 100 351.149 2.0 409.164 189.104 460.209

134.067 546.399 583.222 661.466 689.499 0 200 2. +MS2(506.230), 1.8-1.8min #(260-274) 171.103 OH

150

100 1.5 O

203.095 SO2 N 50 [M+H]+=171.104 Da

234.067 398.151 427.247 Chemical Formula: C 30H3 NO4S 277.106 301.072 354.753 452.198 0 100 200 300 400 500 600 700 m/z

test run_manual inj_1 ppm.d: BPC 49.000-1201.000 +All MS 1.0 test run_manual inj_1 ppm.d: EIC 171.103±0.01 +MS2(506.230) test run_manual inj_1 ppm.d: EIC 506.235±0.01 +All MS

0.5

1 0.0 0.2 0.4 0.6 0.8 1.0 Time [min]

Figure 1 Estradiol can be derivatized with dansyl chloride (Fluka 39220) and detected by APCI as dansyl derivative [2]. Excessive reagent can be separated from the analyte with a short Ascentis® Express C18 guard column cartridge (Supelco® 53501-5). Using UHPLC allows a complete chromatography even with this short column (2.1 x 5 mm!). The analysis of 10 ppb estradiol resulted in a S/N=60 (MS and MRM).

Intens. 7 x10 40 min run-time with conventional C18 column, 1.0 4.6x100 mm, 3.5 μm, 5 μL injection volume.

0.8 1 4 3 5 0.6

0.4

0.2

0.0 Chromatography 0 5 10 15 20 25 Time [min] Intens. 6 x10 7.7 min run-time with Supelco Ascentis Express C18 6 2.1x50 mm, 2.7 μm, 0.6 μL injection volume(!)

4 3 5 1 4

2

0 0 1 2 3 4 5 6 Time [min]

Figure 2 BPC and UV traces of an in-process control sample. The use of Fused-Core® particles leads to much shorter run-times and less sample and solvent consumption while maintaining peak resolution.

(continued on page 14)

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Intens. [mAU]

10.0

7.5

5.0

2.5

0.0

-2.5

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

Figure 3 Standard gradient grade acetonitrile quality (blue) compared to our new LC-MS Ultra CHROMASOLV® Fluka® 14261 (red) under equal conditions (gradient from 0 to 100% acetonitrile within 2 min). In both cases, LC-MS Ultra CHROMASOLV Water, Fluka 14263, is used as aqueous phase.

In combination with very sensitive detection systems, e.g. UV detectors with fi ber optics, focusing quadropols, or time-of- fl ight or ion trap mass spectrometers, UHPLC sets new limits for the purity of solvents and the additives used as mobile phase buff ers. Generally, this is a much more challenging task than the removal of particles. Chromatography

Brand Product Name Description Pack size Fluka 14261 Acetonitrile LC-MS Ultra CHROMASOLV, ≥99.9%, tested for UHPLC-MS 1 L, 2 L Fluka 14262 Methanol LC-MS Ultra CHROMASOLV, ≥99.9%, tested for UHPLC-MS 1 L, 2 L Fluka 14263 Water LC-MS Ultra CHROMASOLV, tested for UHPLC-MS 1 L, 2 L Fluka 14264 Trifluoroacetic acid LC-MS Ultra eluent additive, ≥ 99.0% suitable for UHPLC-MS 1 mL, 2 mL Fluka 14265 Formic acid LC-MS Ultra eluent additive, ≥ 98% suitable for UHPLC-MS 1 mL, 2 mL Fluka 14266 Ammonium formate LC-MS Ultra eluent additive, suitable for UHPLC-MS 25 g Fluka 14267 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.

Reference: [1] Meyer, V. Praxis der Hochleistungs-Flüssigchromatographie, 1. Aufl., Wiley-VCH, Weinheim, 2004. [2] Nelson, R. E., et al. Clinical Chemistry 2004, 50 (2), 373–384.

sigma-aldrich.com/lc-ms 15

New SELECTOPHORE® Ionophores with Improved Properties

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

Selectivity The performance of chemical sensor devices depends on three “S” factors: Selectivity, Figure 1 Magic Sensitivity and Stability. The ionophore plays the most critical role in optimizing Triangle: Key these factors (see Figure 1). Sigma-Aldrich has successfully developed and intro- Performance duced many new ionophores with improved characteristics over previously existing Parameters of ones. The Fluka® branded ionophores are application tested using the electrode Chemical Sensors body IS-561, Fluka 45137. Two examples of ionophores with improved properties are Sensitivity Stability listed below.

Magnesium Ionophore VI, Selectophore function tested, Fluka 63112 ETH 5506, 1,3,5-Tris[10-(1-adamantyl)-7,9-dioxo-6,10-diazaundecyl]benzene

This magnesium-selective ionophore is highly lipophilic O O 2+ HN N (log PTLC=8.5) and obtains sufficient selectivities for Mg

against the main interfering cations; therefore, it can be H H used for magnesium measurement even in serum samples. N N N N This ionophore has a linear response between 1•10-6 M and O O O O 6•10-2 M magnesium ion. Figure 2 Magnesium ionophore VII

Nitrate ionophore VI, Selectophore function tested, Fluka 07295 (9-Hexadecyl-1,7,11,17-tetraoxa-2,6,12,16-tetraazacycloeicosane) HN NH Nitrate ionophore VI is the best nitrate-selective ionophore O O C H (C H ) C H available. The PVC membrane stability is improved by this 2 2 14 3 O O ionophore due to the high lipophilicity of the hexadecyl HN NH group (logP =8.3 (calc.)). The membrane obtains a linear response between 5•10-6 M and 0.1 M nitrate ion. Figure 3 Nitrate ionophore VI

Recommended Membrane Composition: 10.0 wt% Nitrate ionophore VI (07295) 0.5 wt% Tetraoctylammonium chloride (87985) 46.0 wt% Dibutyl phthalate (80100) 43.5 wt% Poly(vinyl chloride) high molecular weight (81392)

Lead ionophore VIII, Selectophore function tested, Iron ionophore VI, Selectophore function tested, Fluka 61796 Fluka 42942 This ionophore obtains excellent selectivity properties and The new Iron(III) ionophore VI obtains an extended linear

is therefore used within the diagnosis of clinical disorders as range and superior selectivity properties. It can be there- Product Corner New well as in the monitoring of environmental samples. fore used for clinical analysis as well as for environmental analysis.

N NH HN O N O N H N O H CH3 N N O Figure 5 Iron(III) ionophore VI CH3

N N H References: H N N 63112 Magnesium ionophore VI O’Donnell, J., et al., Anal. Chim. Acta 281, 129-134, 1993, Figure 4 Lead ionophore VIII 07295 Nitrate ionophore VI unpublished. 61796 Lead ionophore VIII W.-J. Zang et al., Analytical Order a free copy of our guide on ”Selectophore – Ion Sensor Materials” at our Letters 40, 2007, 1023–35. website: sigma-aldrich.com/selectophore or reply using the enclosed BRC. 42942 Iron ionohore VI R. Maleki et al., Turk. J. Chem. 33, 2009, 1–10.

sigma-aldrich.com/selectophore 16

New Headspace Grade Solvents

Michael Kiselewsky, Product Manager Analytical Reagents [email protected]

When developing a GC-HS method, parameters such as and USP monographs. In addition, Sigma-Aldrich has now sample solvent, extraction temperature, extraction time, expanded its GC-HS portfolio with two new products, sample volume and headspace volume are optimized. GC-HS Cyclohexane and 1-Methyl-2-pyrrolidinone. Sigma-Aldrich has developed solvents specifi cally for GC-HS applications. The purity and handling specifications for All solvents are microfi ltered at 0.2 μm and packed under these solvents meet the requirements of the European inert gas for extended shelf life. Pharmacopoeia (Ph.Eur.) and United States Pharmacopeia (USP), as well as ICH guidelines. The new GC-HS product line Literature includes water and three of the most commonly used [1] United States Pharmacopeia, 31st Edition (2008), <467> Residual Solvents. organic solvents: dimethyl sulfoxide (DMSO), N,N-dimethyl- [2] Ph.Eur. 6.0 (2008) Method 2.4.24, Identification and control formamide, and N,N-dimethylacetamide. N,N-dimethylfor- of residual solvents. mamide and dimethyl sulfoxide are specifi ed in Ph.Eur. and [3] ICH Guideline Q3C, Impurities: Guideline for Residual USP for water-insoluble substances. Water is the preferred Solvents, The Fourth International Conference on solvent for water-soluble solutions, as described in Ph.Eur. Harmonization, July 17, 1997.

Cat. No. Brand Product Name Abbreviation BP Pack size NEW 68809 Fluka® Cyclohexanone, for GC-HS - 155 °C 1 L 44901 Fluka N,N-Dimethylacetamide, for GC-HS DMA 166 °C 1 L 51781 Fluka N,N-Dimethylformamide, for GC-HS DMF 153 °C 1 L 67484 Fluka 1,3-Dimethyl-2-imidazolidinone, for GC-HS DMI 225 °C 100 mL, 1 L 51770 Fluka Dimethyl sulfoxide, for GC-HS DMSO 189 °C 1 L NEW 69337 Fluka 1-Methyl-2-pyrrolidinone, for GC-HS NMP 202 °C 1 L 53463 Fluka Water, for GC-HS - 100 °C 1 L

Product table Solvents for GC Headspace Analysis

For more information, please visit our website: sigma-aldrich.com/gc-hs Chromatography

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sigma-aldrich.com/gc-hs 17

DAIH- and DNPH- Aldehyde and Ketone Standards

Ingrid Hayenga, Senior Chemist, R&D Europe [email protected]

Formaldehyde and acetaldehyde are key components of The oxidative coupling of aldehydes with two molecules of photochemically generated air pollution. Oxidation of vola- N-methyl-benzothiazolon-(2)-hydrazone (MBTH) leads to tile organic compounds (VOC) by reaction of OH radicals is an intensively colored product. Although it is possible to considered to be their predominant source, but they are determine different aldehydes and ketones with this also emitted directly into the urban atmosphere during the method, its selectivity depends heavily on the analyte incomplete combustion of fossil fuels. It has been assumed structure and the experimental conditions. that these two important intermediate species account for up to 90% of the total aldehydes in air [1]. Therefore, to achieve speciation information, a chromato- graphic separation is required. For atmospheric detection For short chain aldehydes and ketones, a number of of short chain aldehydes and ketones, the use of impreg- derivatization reagents have been developed. Their detection nated silica-gel cartridges has provided good sensitivity, is performed by photometry or fl uorescence, e.g. reaction the impregnating reagents being 2-diphenylacethyl- with chromotropic acid or pararosaniline. These reactions 12.3-indandione-1-hydrazone (DAIH) or 2.4-dinitrophenyl- are valid if a sum parameter is to be obtained; however, hydrazone (DNPH). These reagents are also applicable as their major drawback is the inability to determine diff erent precolumn derivatizing reagents for liquid phase analysis. [2] carbonyls simultaneously within one sample.

The following table provides a summary of the most important derivatizing agents as well as their advantages and disadvantages [2].

Reagents Analytical Advantage Disadvantage Air sampling method techniquesa 2.4-Dinitrophenylhydrazine HPLC-UV, International standard procedure; large Interferences with airborne I, Cb, D, PSD

(DNPH) HPLC-MS, variety of applications available; reagents oxidants (Nox, O3); limited (GC-FID, CE) and derivatives stable sensitivity Hantzsch reagent UV, fluorescence, Well suitable for on-line analysis; Slow and non-quantitative I, flow-through HPLC-UV, fluorescence detection reaction; limited to aliphatic analyzers HPLC-fluorescence carbonyls Chromotropic acid + UV/Vis Only readily available instrumentation Suitable only for I pararosaniline required formaldehyde 4-nitrophenyl-hydrazine HPLC-UV Slightly red-shifted absorption maxima Replaced by DNPH because C, I (pNPH) of the derivatives compared to the DNP- of its faster derivatization hydrazones reaction Halogenated aromatic GC-ECD, GC-FID, Improved chromatographic resolution Slow reactions with C, I hydrazines and GC-MS compared to HPLC carbonyls; E and Z isomers hydroxylamines separated N-methyl-benzothiazolon- UV/Vis, HPLC-UV Only readily available instrumentation Selective for formaldehyde I (2)-hydrazone (MBTH) required but cross-selectivity to 4-Amino-3-hydrazino- other aldehydes 5-mercapto-1,2,4-triazol (Purpald)

Dansylhydrazine (DNSH) HPLC- Low limits of detection Instability of DNSH and C, I Standards fluorescence its derivatives towards hydrolysis N-Methyl-4-hydrazino-7- HPLC-UV, Slightly faster reaction with carbonyl C, I, PSD nitrobenzofuran (MNBDH) HPLC-MS compounds, higher selectivity and lower detection limits compared to DNPH as well as other N-alkylated hydrazine reagents

a: C = cartridge, I = impinger, D = denuder, PSD = passive sampling devices b: reagent coated cartridges are commercially available

The most relevant DNPH- and DAIH-aldehyde and ketone standards are available at Sigma-Aldrich either as solutions or as pure substances, as analytical standards or as BCR certifi ed reference materials. (continued on page 18)

sigma-aldrich.com/standards 18

NO2 H NO2 R 1 O N H NH2 N N R 2 R 1 R 2 O2N O2N Figure 1 Derivatization of carbonyl compounds with 2.4-dinitrophenylhydrazone (DNPH) O O O O O

R 1 R 2 N N H 2N N

R 1 R 2 Figure 2 Derivatization of carbonyl compounds with 2-Diphenylacetyl-1.3-indandione-1-hydrazone

Cat. No. Product Pack size Cat. No. Product Description Pack 14423 Acetaldehyde DAIH derivative 50 mg size 02819 Acetone, DAIH derivative 50 mg 33852 Decanal 2.4-DNPH > 98.5% 10 mg, 13173 Acroleine, DAIH derivative 50 mg 100 mg 55556 Crotonaldehyde, DAIH derivative 50 mg 442597 Formaldehyde-2.4-DNPH ampule of 91547 Cyclohexanone, DAIH derivative 50 mg 100 mg 06947 Formaldehyde, DAIH derivative 50 mg 33848 Heptanal 2.4-DNPH > 98.5% 100 mg 51299 Propionaldehyde, DAIH derivative 50 mg 442614 Hexaldehyde-2.4-DNPH ampule of 100 mg 442321-U 2.5-Dimethylbenzaldehyde ampule of 100 mg -2.4-DNPH 442639 Methacrolein-2.4-DNPH ampule of 100 mg 442339 2-Butanone-2.4-DNPH ampule of 100 mg 33851 Nonanal 2.4-DNPH > 98.5% 100 mg 442434 Acetaldehyde-2.4-DNPH ampule of 100 mg 33849 Octanal 2.4-DNPH > 98.5% 100 mg 442436 Acetone-2.4-DNPH ampule of 50 mg 442722 o-Tolualdehyde-2.4-DNPH ampule of 442441 Acrolein-2.4-DNPH ampule of 25 mg 100 mg 442469 Benzaldehyde-2.4-DNPH ampule of 100 mg 442768 Propionaldehyde-2.4- ampule of 442504 Butyraldehyde-2.4-DNPH ampule of 100 mg DNPH 100 mg 442529 Crotonaldehyde-2.4-DNPH ampule of 100 mg 442735 p-Tolualdehyde-2.4-DNPH ampule of 442533 Cyclohexanone-2.4-DNPH ampule of 100 mg 100 mg Table 1 DAIH- and DNPH – standards, pure substance 34396 Succinaldehyde > 98.5% 50 mg 442834 Valeraldehyde-2.4-DNPH ampule of 100 mg

Cat. No. Product Description Pack Size 47344 2-Butanone-2.4-DNPH solution 100 μg/ml in acetonitrile ampule of 1 ml 47340-U Acetaldehyde-2.4-DNPH solution 1000 μg/ml in acetonitrile ampule of 1 ml 4M7340-U Acetaldehyde-2.4-DNPH solution 1000 μg/ml in acetonitrile pkg of 5 ampules (1 ml each) 47341 Acetone-2.4-DNPH solution 1000 μg/ml in acetonitrile ampule of 1 ml 4M7341 Acetone-2.4-DNPH solutions 1000 μg/ml in acetonitrile pkg of 5 ampules (1 ml each) 47342 Acrolein-2.4-DNPH solution 1000 μg/ml in acetonitrile ampule of 1 ml 4M7258-U Aldehyde/ketone-DNPH TO11/IP-6A Mix 15 μg/ml each component in acetonitrile pkg of 3 ampules (1 ml each) 47343 Benzaldehyde-2.4-DNPH solutions 100 μg/ml ampule of 1 ml Standards 47345-U Butyraldehyde-2.4-DNPH solution 1000 μg/ml ampule of 1 ml 47649-U CARB carbonyl-DNPH Mix 1 In acetonitrile (varied) ampule of 1 ml 47650-U CARB Method 1004 DNPH Mix 1 3 μg/ml in acetonitrile ampule of 1 ml 47651-U CARB Method 1004 DNPH Mix 2 2 μg/ml in acetonitrile ampule of 1 ml 47672-U Carbonyl DNPH Mix 1 20 μg/ml in acetonitrile ampule of 1 ml 47671-U Carbonyl DNPH Mix 2 2 μg/ml in acetonitrile ampule of 1 ml 47175-U Crotonaldehyde-2.4-DNPH solution 100 μg/ml in acetonitrile ampule of 1 ml 47674-U Cyclohexanone-2.4-DNPH solution 500 μg/ml in acetonitrile ampule of 1 ml 48149-U EPA TO-11A Six Component Carbonyl-DNPH Mix 15 μg/ml each component in acetonitrile pkg of 1.5 ml 47177 Formaldehyde-2.4-DNPH solution 100 μg/ml in acetonitrile ampule of 1 ml 47564-U Glutaraldehyde-2.4-DNPH solution 100 μg/ml in acetonitrile ampule of 1 ml (continued on page 19)

sigma-aldrich.com/standards 19

Cat. No. Product Description Pack Size 47178-U Hexaldehyde-2.4-DNPH solution 1000 μg/ml in acetonitrile ampule of 1 ml 47886 Isobutyraldehyde-2.4-DNPH solution 100 μg/ml in acetonitrile ampule of 1 ml 47179 Isovaleraldehyde-2.4-DNPH solution 1000 μg/ml in acetonitrile ampule of 1 ml 47180-U Methacrolein-2.4-DNPH solutions 100 μg/ml in acetonitrile ampule of 1 ml 47183 m-Toluoaldehyde-2.4-DNPH solution 100 μg/ml in acetonitrile ampule of 1ml 47032-U o-Phthaldialdehyde-(DNPH)2 solution 10 μg/ml in acetonitrile:DMSO (7:3) pkg of 3x2 ml 47182 o-Tolualdehyde-2.4-DNPH solution 100 μg/ml ampule of 1 ml 47181 Propionaldehyde-2.4-DNPH solution 1000 μg/ml ampule of 1 ml 47184-U p-Tolualdehyde-2.4-DNPH solution 100 μg/ml ampule of 1 ml 40117-U Pyridine-2-Aldehyde-DNPH Standard in acetonitrile pkg of 3x2 ml 40081-U Pyridine-4-Aldehyde-DNPH solution 1mM in acetonitrile:DMSO, 4:1 pkg of 3x2 ml 47285-U TO11/IP-6A Aldehyde/Ketone-DNPH Mix 15 μg/ml each component in acetonitrile ampule of 1 ml 47185-U Valeraldehyde-2.4-DNPH solutions 100 μg/ml ampule of 1 ml

Table 2 DAIH- and DNPH-standards, in solution and mixes

Cat. No. Product Description Pack Size BCR5512 2.4-Dinitrophenylhydrazones, solutions Set of 4x BCR-551 and 1x BCR-552 (blank) set BCR548 Acrolein-2.4-DNPH 10 mg BCR5534 Formaldehyde-2.4-DNPH on filter Set of 2x BCR-553 and 1x BCR-554 (blank) set BCR550 Glutaraldehyde 2.4-DNPH 10 mg

Table 3 BCR certified reference material

References: [1] Possanzini, M., Di Palo, V. Chromatographia 1997, 46, 5/6, 235. [2] Vogel, M., Büldt, A., Karst, U., Fresenius, J. Analy. Chem. 2000, 366, 781.

New Analytical Standards for the Analysis of Herbal Medicinal Products

Matthias Nold, Product Manager Analytical Standards [email protected]

Herbal medicinal products, pharmaceutical products which Cat. No. Brand Description Pack Size are derived from medicinal plants, are very popular as an 42251 Fluka® Apigenin 10 mg alternative to synthetic drugs. In recent years, regulatory 91466 Fluka Betulinic acid 10 mg pressure for herbal medicinal products has increased, 43412 Fluka (+) Catechin 10 mg resulting in the need for monitoring the concentrations of 95082 Fluka Chrysin 50 mg active ingredients and other characteristic components. 06536 Fluka trans-Cinnamaldehyde 50 mg 97013 Fluka trans-Cinnamic acid 50 mg 38907 Fluka Isorhamnetin 10 mg Sigma-Aldrich off ers analytical standards as well as primary 69545 Fluka Kaempferide 10 mg reference standards of active ingredients and marker com- 96353 Fluka Kaempferol 10 mg pounds of medicinal plants. This portfolio of products con- 49968 Fluka Luteolin-7-glucoside 10 mg Standards tinues to expand at a quick pace. Just as we presented 06279 Fluka Mangiferin 10 mg several new products in the fi rst two issues of Analytix in 28660 Fluka Procyanidin A2 10 mg 2011, we are once again introducing new additions to this 16654 Fluka Quercetin 3-glucoside 10 mg rapidly growing product line. 69686 Fluka Shikimic acid 50 mg Table1 New analytical standards and certified reference materials Our webpage at sigma-aldrich.com/medicinalplants (CRMs) of active ingredients and marker substances for the analysis contains an up-to-date listing of these products catalogued of herbal medicinal drugs not only by alphabetical order, but also according to sub- stance class and by genus for a wide array of medicinal plants.

sigma-aldrich.com/standards 20

Inorganic Custom Standards An Interactive Online Platform

TraceCERT® standards for AAS, ICP and IC are Certifi ed For all TraceCERT Custom Standards we guarantee: Reference Materials fulfi lling the highest-quality standards.* In addition to our comprehensive portfolio of catalogue • Certification under double accreditation following products we also off er you a custom service where you ISO/IEC 17025 and ISO Guide 34 can defi ne your own multi-component standards at the • Highest level of accuracy and reliability same high quality, using our Inorganic Custom Standards • Minimized uncertainties and lot-specific values Online Platform: • Traceability to at least two independent references (i.e. NIST, BAM or SI unit kg) sigma-aldrich.com/csp • Printed certificate according to ISO Guide 31 • Light- and gas-tight aluminium foil bag packaging allowing up to four years’ shelf life

With a few simple mouse clicks you can specify the desired characteristics among the following:

• Element standard • Ion standard • Defining matrix • Analytes • Concentrations

The Custom Standards Platform is a dynamic web page: elements, ions and matrices can only be chosen if the combinations are chemically allowed.

After your request has been submitted online, we will

Standards evaluate the production costs and respond with a quota- tion by e-mail.

It’s easy, try it out!

* At sigma-aldrich.com/tracecert you can fi nd the extensive TraceCERT portfolio of certifi ed single- and multi-element standards.

sigma-aldrich.com/csp 21

New TraceCERT® 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]

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, traceability can be established between two dif- ferent compounds, in contrast to most other analytical techniques.

With a small set of NIST traceable internal standards, we are able to certify basically any organic compound by measur- ing a gravimetrically produced mixture of the analyte and the internal standard. The accurately determined mass to mass ratio then allows for the calculation of the analyte content with a very high level of confi dence. Typical uncer- tainties range from 0.5% down to 0.1%.

For analytical testing labs, ISO/IEC 17025 accreditation is New organic TraceCERT products gaining increased importance for demonstrating compe- The portfolio of organic TraceCERT CRMs intended for use tence in testing and calibration. Essential to this accredita- as chromatography standards is comprised of over 50 prod- tion is the use of suitable calibration standards, with the ucts and is continually growing. The products include recommendation that these standards are traceable to amino acids, PAHs, fatty acids /FAME, pesticides and phar- internationally recognized reference materials. maceuticals, as published in previous articles [1].

While the availability of inorganic certifi ed reference materials The most recent product additions comprise several natu- (CRMs) is usually high (see our corresponding TraceCERT ral products as listed below. portfolio at sigma-aldrich.com/inorganiccrm), there has been a lack of sources for traceable organic CRMs. Please visit our website (sigma-aldrich.com/organiccrm) where you can find a complete and current listing of all In the last year, Sigma-Aldrich has introduced the innova- products and a library of technical articles and examples of tive new product line of organic TraceCERT standards [1]. certifi cates that can be downloaded. These premium CRMs are certifi ed in a double accredited laboratory, fulfi lling both ISO/IEC 17025 and ISO Guide 34, Cat. No. Brand Description Pack size using high-performance quantitative NMR (HP-qNMR®) 93485 Fluka® AMOZ 50 mg for content determination with direct traceability to NIST and 94664 Fluka Anhydrotetracyclin HCl 50 mg SI. 51868 Fluka trans-Caffeic acid 50 mg 56396 Fluka Caffeine 100 mg 52229 Fluka trans-Ferulic acid 50 mg The organic TraceCERT reference materials are character- 76635 Fluka Fumaric acid 100 mg ized by: 91215 Fluka Gallic acid 100 mg 52341 Fluka Salicylic acid 100 mg • Certified Content by HP-qNMR 68654 Fluka Vanillic acid 50 mg • Superior level of accuracy, calculated uncertainties and Table 1 NEW organic neat CRMs TraceCERT

lot-specific values Product Corner New • Traceability to NIST References: • [1] TraceCERT Organic Certified Reference Materials, Analytix, Comprehensive documentation delivered with the Vol 2 and 3, 2010; Vol 1, 2011. product (certification according to ISO Guide 31)

sigma-aldrich.com/organiccrm 22

Volumetric Titration VOLPAC® Solution Containers with new adapter

Andrea Felgner, Product Manager Analytical Reagents [email protected]

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 fl exible 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 diff erent-sized adapters that allow direct connection of the titrating tube to the VOLPAC container.

Cat. No. Description Acid solutions 35335-5L-VOLPAC, 35335-10L-VP Hydrochloric acid solution Reag. Ph. Eur. 0.1 mol/L 35329-5L-VP Hydrochloric acid solution Reag. Ph. Eur. 0.5 mol/L 35329-10L-VP 35328-5L-VOLPAC Hydrochloric acid solution Reag. Ph. Eur. 1 mol/L 35328-10L-VP 35327-10L-VOLPAC Hydrochloric acid solution 2 mol/L 35358-5L-VP Sulfuric acid solution Reag. Ph. Eur. 0.05 mol/L 35358-10L-VP 35357-5L-VP Sulfuric acid solution Reag. Ph. Eur. 0.1 mol/L 35357-10L-VP 35355-10L-VP Sulfuric acid solution Reag. Ph. Eur. 0.25 mol/L

Titration 35354-5L-VOLPAC Sulfuric acid solution Reag. Ph. Eur. 0.5 mol/L 35354-10L-VP 35348-5L-VP Sulfuric acid solution 2.5 mol/L 35348-10L-VP

sigma-aldrich.com/volpac 23

Cat. No. Description Base solutions 35116-5L-VP Potassium hydroxide solution 0.5 mol/L 35125-10L-VOLPAC Potassium hydroxide solution 0.1 mol/L 35262-10L-VOLPAC Sodium hydroxide solution 0.01 mol/L 35263-5L-VOLPAC Sodium hydroxide solution Reag. Ph. Eur. 0.1 mol/L 35263-10L-VP 35256-5L-VOLPAC Sodium hydroxide solution Reag. Ph. Eur. 1 mol/L 35256-10L-VP 35261-10L-VP Sodium hydroxide solution 0.2 mol/L 31086-5L-VP Sodium hydroxide solution 0.25 mol/L 31086-10L-VP 35257-5L-VOLPAC Sodium hydroxide solution 0.5 mol/L 35257-10L-VP pH Buffer solutions 33665-5L-VP Buffer solution pH 4.0 (20 °C) with fungicide (citric acid / sodium hydroxide / sodium pH 4.0 33665-10L-VP chloride) red colored 33643-5L-VP Buffer solution pH 4.0 (20 °C) with fungicide (citric acid / sodium hydroxide / sodium pH 4.0 33643-10L-VP chloride) 33666-5L-VP Buffer solution pH 7.0 (20 °C) with fungicide (potassium dihydrogen phosphate / pH 7.0 33666-10L-VP disodium hydrogen phosphate) green colored 33646-5L-VP Buffer solution pH 7.0 (20 °C) with fungicide (potassium dihydrogen phosphate / pH 7.0 33646-10L-VP disodium hydrogen phosphate) 33667-5L-VP Buffer solution pH 9.0 (20 °C) (borax / hydrochloric acid) pH 9.0 33667-10L-VP blue colored 33648-5L-VP Buffer solution pH 9.0 (20 °C) (borax / hydrochloric acid) pH 9.0 33648-10L-VP 33668-10L-VP Buffer solution pH 10.0 (20 °C) (borax / sodium hydroxide) violet colored pH 10.0 Complexometric solutions 34544-5L-VP Ethylenediaminetetraacetic acid disodium salt solution with zinc complex added (solution B) 1 mL solution = 1 German degree of hardness in 100 mL of water

35102-5L-VP IDRANAL® III solution (EDTA-Na2)0.2 mol/L 35102-10L-VP

34550-5L-VP-R IDRANAL III solution (EDTA-Na2), Reag. Ph. Eur. 0.1 mol/L 34550-10L-VP-R

35103-5L-VP IDRANAL IV solution (DCTA-Na4), Reag. Ph. Eur. 0.1 mol/L 35103-10L-VP Salt solutions 34631-5L-VP Potassium dichromate solution for COD determination according to DIN 38409 0.02 mol/L 34277-5L-VP-R Sodium carbonate solution 0.5 mol/L 34247-10L-VP-R Sodium fluoride solution 40 g/L 34246-10L-VP-R Sodium oxalate solution 25 g/L 35224-5L-VP Sodium thiosulfate solution, for determination of iodine adsorption number according to ASTM D 1510 34449-10L-VP-R Sodium thiosulfate solution 0.05 mol/L 35233-5L-VP Sodium thiosulfate solution 0.2 mol/L 35245-5L-VP Sodium thiosulfate solution Reag. Ph. Eur. 0.1 mol/L 35245-10L-VP

34400-5L-VP Luff-Schoorl-Reagent (CuSO4*5H2O) ~25 g/L

Table 1 Complete list of all Fluka brand products that are available in VOLPAC containers (These products can be easily recognized since they are tagged with a ‘-VOLPAC’ or ‘-VP’ in their product number.)

Special offer on all VOLPAC solution containers!

Receive 45% off your first order of any VOLPAC container. Please quote Titration promotion code 979 when placing your order. Offer valid until July 31, 2011.

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