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Analytix 5/2007 Analytix Issue 5 • 2007 New European Pharmacopoeia Regulations for Karl Fischer Titration • Ketone, Aldehyde and Wine Analysis Standards • APPI Dopants, HPLC Buffers and Vials • Pseudomonas & Yersinia Media and Tests • Ultra Pure Maldi Matrices • Titration overview ™ ATX_5.indd 1 23.10.2007 18:54:02 2 Isotec® Stable Isotopes Your Stable Isotope Source Dear Colleague, • NMR solvents If you’ve read recent issues of Analytix, • NMR reference standards you’ll likely know that analytical standards • NMR tubes and accessories Editorial are a very important product area for us. In • NMR equipment and software this issue, I’d like to discuss another import- ant group of standards that you may not be This is just a small portion of our complete offering, aware that we supply: stable isotopes. which ranges from isotopically-enriched gases to la- beled amino acids to complex biomolecules. Isotec Scientists use stable isotope labeled com- routinely incorporates D, 13C, 15N, and 18O into these pounds to probe metabolic pathways and compounds to meet our customers’ needs. as internal standards for reliable quantita- tive analysis. Stable isotopes are routinely If you cannot fi nd a labeled compound to meet your used in: requirements, Isotec’s R&D Group will work with you to Lisa Roth • Metabolic research design and synthesize a custom molecule. Our exten- Technical Marketing Manager • Quantitative proteomics sive experience, combined with our inventory of basic Stable Isotopes • Protein structure determinations starting materials, enables us to deliver unique, custom • Breath test kits and research stable isotope labeled compounds. • Mass spectrometry • Positron emission tomography (PET) Along with the other Sigma-Aldrich brands, our com- • Magnetic resonance imaging (MRI) mitment to quality and service is found throughout • Agricultural research Isotec. Our on-site synthesis, purifi cation and isotopic enrichment facilities and expert Quality Control team Isotec®, the Stable Isotope Group of Sigma- and Quality Assurance department provide you with Aldrich, is a leading producer of stable the highest quality products available. Isotec also pro- isotope labeled compounds and enriched vides a wide variety of packaging options. We also pro- stable isotopes, such as 13C. The most rec- vide custom packaging on request whenever possible. ognizable stable isotope products are NMR Our knowledgeable and well-trained Customer Service deuterated solvents. Other NMR products team handles your requests and answers your toughest we offer include: questions. We look forward to serving your needs. Kind regards, The Analytix is published 5 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: Dave R. van Staveren, PhD www.sigma-aldrich.com/analytix ATX_5.indd 2 23.10.2007 18:54:11 3 Feature Article Microbiology 04 New European Pharmacopoeia Regulations 14 Pseudomonas Media and Tests for Karl Fischer Titration Detection, identification and cultivation of Our pyridine-free HYDRANAL® reagents Pseudomonas species comply with Ph. Eur. Regulation 5.7 for water Contents of Table 16 Yersinia Diagnostic determination Detection, identification, differentiation and cultivation of Yersinia species Standards 06 Wine Quality Analysis Spectroscopy Wine analysis stock solutions for the 17 Ultra-pure MALDI Matrices determination of wine components in Absence of organic and inorganic impurities sweet and dry wines and improved solubility make them ideal for 08 Ketone and Aldehyde Analysis the most demanding MALDI applications Our portfolio of DNPH-labeled standards for US and European regulations Titration 20 Titration Reagents, Solutions and Chromatography Indicators 09 APPI Dopants An overview of the evolving Sigma-Aldrich Solvents and post-column additives for photo- portfolio ionization 11 Selecting the Right Reversed-phase HPLC New Product Corner Buffers 23 Ionophore for Quick and Reliable An overview of our high-purity HPLC buffers Determination of Benzoate as solutions, solids or concentrates 23 Natural Compounds Analytical Standards 13 New High-quality Vials and Accessories for Analytical Applications Sigma-Aldrich expands its range of vials and accessories www.sigma-aldrich.com/analytix ATX_5.indd 3 23.10.2007 18:54:13 4 New European Pharmacopoeia Regulations for Karl Fischer Titration Freedom of choice of pyridine-free reagents and other improvements Helga Hoffmann, Technical Service HYDRANAL® Manager [email protected] Andrea Felgner, Product Manager Analytical Reagents [email protected] 2. In our earlier comparison tests, we often recommended the add- ition of a suitable solubilizing agent for the sample. To accommo- date this improvement, the new pharmacopoeia now states in ref- erence to the solvent “…or recommended by the supplier of the Feature ArticleFeature titrant.” An example is gentamicin sulphate, for which the Ph. Eur. pre- scribes methanol to be used in the titration vessel. During suitability tests, we found that gentamicin did not fully dissolve during titra- tion, which resulted in erroneously low water content, and after- wards a too-high recovery of added water. The test results thus did not fulfi ll the requirements of the suitability test. However, when we used a mixture of HYDRANAL® Methanol dry and HYDRANAL® The European Pharmacopoeia (Ph. Eur.) specifi es Karl Formamide dry in the titration vessel, the gentamicin dissolved well Fischer titration to measure the water content of many and the results fulfi lled the requirements of the suitability test. Both solvents, chemicals and other substances. Historically, documents are available from us upon request. and even until very recently, pyridine-containing KF re- agents were specifi ed. This means that for a very long 3. In addition to the freedom in the choice of titrant, the new time, the Ph. Eur. did not meet the state-of-the-art in KF Ph. Eur. also permits choices in the medium to be used in the titra- technology, since pyridine-free KF reagents have been tion vessel, specifi cally “…methanol R, or the solvent indicated in available and used successfully since 1980. If one wanted the monograph or recommended by the supplier of the titrant.” to take advantage of the improved safety and other Acetone provides a perfect example. The Monograph for ac- benefi ts of pyridine-free reagents, tedious compari- etone states that pyridine, which is malodorous and toxic, be in- son tests had to be carried out with both types of re- troduced into the titration vessel. We have performed successfully agents. Sigma-Aldrich, an innovator in pyridine-free KF the requisite suitability test using both pyridine and odorless reagents, supported its customers by conducting over HYDRANAL® KetoSolver, which requires no hazardous declaration, 100 of these comparison tests in the HYDRANAL® in the titration vessel. These test results are also available on request. laboratory. However, the newly released Ph. Eur. 5.7 contains an Method B important change: The use of a pyridine-based KF re- This has basically the same requirements as Method A, except after agent is no longer compulsory. Chapter 2.5.12 “Water: addition of the sample an excess of titrant has to be added. The Semi-Micro Determination” describes in Method A the titrant, which is not consumed by the water in the sample, has to be direct titration of water and in Method B the indirect back titrated using a standard with a known amount of water, like method. In practice, the direct method, described HYDRANAL® Water in methanol Standard 5.00. below, is easier to carry out. Suitability Method A The accuracy of the determination with the chosen titrant must Introduce into the titration vessel methanol R, or the be verifi ed for each substance to be examined. The following solvent indicated in the monograph or recommended procedure, given as an example, is suitable for samples containing by the supplier of the titrant. Where applicable for the 2.5–25 mg water. apparatus used, eliminate residual water from the mea- The water content of the substance to be examined is deter- surement cell or carry out a pre-titration. Introduce the mined using the reagent/solvent system chosen. Thereafter, sequen- substance to be examined rapidly and carry out the tial known amounts of water R are added in an appropriate form titration, stirring for the necessary extraction time. (at least 5 additions) and the cumulative water content determined after each addition. Several points are important to elucidate: 1. The titration agent is a matter of choice. If methanol 1. Calculate the percentage recovery is used in the titration vessel, the titrant of choice is the 2. Calculate the regression line of the cumulative water ® one-component reagent, HYDRANAL Composite. 3. Calculate the percentage mean recovery www.sigma-aldrich.com/hydranal ATX_5.indd 4 23.10.2007 18:54:13 5 The reagent/solvent system is considered to be acceptable if: The German edition of Ph. Eur. 5, Chapter 4.1.2 • The mean recovery is between 97.5% and 102.5% reference solutions for limit tests contains following • The slope b is between 0.975 and 1.025 (deviation ±2.5%) information: • The error e1 and e2 are not greater than 2.5% Reference solution for Micro 1147300 The suitability test with all necessary calculations is not easy to carry determination of Water R out; neither is the preparation of the necessary documentation. To support our HYDRANAL®
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