Select a Method Select a method Sample Preparation Sampling Eliminate Interferences Calculate the Results and Report Writing Perform the Measurement DEFINITION OF THE PROBLEM To determine the best method of analysis the analyst should ask the following questions. SELECT A METHOD Factors to consider include: Accuracy Detection limit Time Cost/number of analyses Complexity of sample - selectivity Equipment Technical expertise LIST OF JOURNALS Analytical Chimica Acta Analytical Abstracts Journal of Analytical Chemistry Analytical Communications Analyst Journal of the Association of Official Analytical Chemists Journal of Chromatography Journal of Chromatographic Science Talanta Trends in Analytical Chemistry LIST OF BOOKS SELECTING AN ANALYTICAL METHOD Performance Characteristics Criteria used to compare which of several instrumental methods would be the best for a particular analysis. e.g. Limit of Detection. Figures of Merit Quantitative (numerical) measures of performance characteristics. e.g. LOD of Pb (AAS) = 15 ppb LOD COMPARISON (g/L) / ppb VALIDATION OF ANALYTICAL METHOD (METHOD VALIDATION) Analysis of Standard Samples (SRM) Analysis by Other Methods Standard Addition to the Sample VALIDATION OF ANALYTICAL METHOD Analysis of Standard Samples - A sample whose analyte concentration is known. - The standard reference material (SRM) can be obtained from The National Institute Of Standard and Technology (NIST). - The analyte concentration in the SRM has been certified by the institute. - Compare the data obtained from the method with the certified value. Analysis by Other Methods - The result of the analytical method can be evaluated by comparison with data obtained from a different method. Standard Addition to the Sample - The known amount of the analyte is added to the sample and then analyzed by the proposed method. The effectiveness of the method can be established by evaluating the recovery of the added quantity. - The standard addition method will reveal errors arising from the way the sample was treated or from the presence of the other compounds in the matrix. SAMPLING Sampling is the process to get a representative and homogeneous sample. Representative means that content of analytical sample reflects content of bulk sample. Homogeneous means that the analytical sample has the same content throughout. OBTAIN A SAMPLE DEPEND ON The size of the bulk to be sampled. The physical state of the fraction to be analyzed (solid, liquid, gas) The chemistry of the material to be assayed. (Nothing can be done that would destroy or alter the identity or quantity of the analyte) SAMPLING BULK MATERIALS Identify the population from which the sample is to be obtained. Collect a gross sample that is truly representative of the population being sampled. Reduce the gross sample to a laboratory sample that is suitable for analysis. SAMPLING METHODS Methods of sampling are given in standard reference books. ASTM (American Society for Testing and Material) APHA (American Public Health Association) AOAC (Association of Official Analytical Chemists International) Homogeneous samples present no problem, a simple grab sample approach taken at random and assumed to be representative. Several samples have to be taken if parent sample is heterogeneous. SAMPLING SOLID Inhomogeneity of the material, make sampling of solids more difficult. The easiest way to sample a material is grab sample, the sample taken at random and assumed to be representative. For reliable results, it is best to take 1/50 to 1/100 of the total bulk. The larger the particle size, the larger the gross sample should be. The gross sample must be reduced in size to obtain a laboratory sample. EXAMPLES Stockpile of cereals: take increment from surface and interior. Compact solids (metals and alloys): obtained by random drilling or by sawing across the metal at random intervals and collecting the `sawdust as the sample. Obtaining a random sample from a bulky material (ore, grain, coal) can be achieved while the material in motion (conveyor belt). Periodically transfer portion into a sample container. SAMPLING SOLID CONE AND QUARTER SAMPLING LIQUIDS Liquid samples are homogeneous and are much easier to sample. The gross sample can be relatively small. If liquid samples are not homogeneous, and have only small quantity, they can be shaken and sampled immediately. Sampling techniques will depend on the types of liquid. EXAMPLES Large volume of liquids (impossible to mix) - Sampled after transfer (during discharge) - If in a pipe, sampled after passing through a pump or at different points in pipe system. Large stationary liquids (lakes, rivers) - Sampled at different depths using a sample thief - Sample thief is a bottle that can be opened and filled at any desired location in the solution. Biological fluids - The timing of sampling is very important e. g. before meal or after meal. SAMPLING GASES Tend to be homogeneous. Large volume of samples is required because of their low density. Examples: - Air analysis: Use a `Hi-Vol sampler that is containing filters to collect particulates. - Liquid displacement method: The sample must has little solubility in the liquid and does not react with the liquid - Breath sample: The subject could blow into evacuated bag. SAMPLING GASES Air Samplung Pump Dust Sampler Air Sampling Filters Hi-Vol Hi-Vol SAMPLE STORAGE AND PRESERVATION Samples are preserved to prevent from: Decomposition of biological samples through the action of bacteria. Refrigerated after collection until the time of analysis. Precipitation of metals from water samples. Acidified (10% HNO 3 ) immediately upon collection. Loss of water from hygroscopic material. Loss of volatile analytes from water samples. SAMPLE PREPARATION Sample preparation or sample treatment is a step in chemical analysis where the sample is brought into the correct size form for analysis. Sample preparation: Most laborious Time-consuming Error-prone steps Distribution of Time Analysts Spend on Sample Analysis GENERAL PRINCIPLES FOR SAMPLE PREPARATION PREPARING A LAB SAMPLE Converting the sample to a useful form: Solids are usually ground to a suitable particulate size to get a homogeneous sample. Dry the samples to get rid of absorption water. (Drying at 110 to 120C for 1 hour and cooled in dessicator before weighing). Solid samples must be dissolved in solution. DEFINING REPLICATE SAMPLES Replicate samples are always performed unless the quantity of the analyte, expense or other factors prohibit. Replicate samples are portion of a material of approximately the same size that is carried through an analytical procedure at the same time and the same way. PREPARING SOLUTION Most analyses are performed on solutions. A solvent is chosen that dissolves the whole sample without decomposing the analyte. Digestion should be performed in Teflon or quartz containers to reduce contamination. Several sources of error are encountered in the sample dissolution step: - Incomplete dissolution of the analyte. - Losses of analyte by the volatilization. - Introduction of analyte as a solvent contamination. - Contamination from the reaction of the solvent with vessel walls. DECOMPOSITION AND DISSOLUTION Simple Dissolution Acid Treatment/Wet Digestion Fusion Techniques Organic Solid Dry Ashing Wet Digestion Inorganic Solid SIMPLE DISSOLUTION Dissolution by water. WET DIGESTION Dissolution in various strong mineral acids. HCl Carbonates, phosphates, oxides H 2 SO 4 Organic material at 300C HNO 3 Any metals not dissolve by HCl HClO 4 Steel HF Silica Aqua Regia (HCl:HNO 3 , 3:1) Not stable HNO 3 :HCl:HF (5:15:3) Alloys GRADE OF ACIDS Very High Purity Chemicals - ARISTAR TM (BDH), Ultra-Pure (NBS) Analytical Reagents - Certified AR TM (Fisher) and Puriss TM (Fluka) Chemically Pure (CP) - CP TM (Sigma), GPR TM (BDH) Practical Grade - Purified TM (Sigma), LR TM (BDH) Commercial or Technical Grade WET DIGESTION MICROWAVE DECOMPOSITION FUSION TECHNIQUES Contamination by flux material High salt content High temparature, loss of analyte through evaporation Sample container may react with flux material DISADVANTAGES OF FUSION TECHNIQUES DESTRUCTION OF ORGANIC MATERIAL FOR INORGANIC ANALYSIS For organic materials (animal and plant tissue, biological fluid ) containing inorganic analytes such as trace metals, the organic matrix is destroyed via : Dry Ashing Involves slow combustion at 400-700C , which leaves behind the inorganic residue which is soluble in dilute acid. Wet Digestion Heat organic with oxidising acids (HNO 3 /H 2 SO 4
mixture), inorganic residue left behind. ELIMINATING INTERFERENCES Interferences are substances that prevent direct measurement of the analyte and must be removed. Techniques - Separation - Standard Addition MEASUREMENT The analytical measurement is often the simplest stage of the analytical process. All reagents used must be of high purity (reagent grade). For trace analysis a blank measurement must be performed. Analytical measurements are divided into two types: classical (gravimetric, volumetric) and instrumental. The physical or chemical property proportional to the analyte concentration is measured. Suitable standards must be measured to determine the relationship between analyte quantity and the physical/ chemical property being measured (i.e., calibration). Classical Methods (Gravimetric, Volumetric) Accurate and precise to approx. 0.1% but require large amount of analyte (mmol, mg). Instrumental Methods (Spectroscopy, Chromatography) More selective and sensitive than classical methods but less precise. Accurate to ! 1% Measure of physical/chemical property of an analyte related to concentration. Rapid/may be automated/may be used for determination of multiple analytes at a time. CALCULATING RESULTS Determine the concentration of the analyte in the sample solution. Use results to calculate the amount of analyte in the original (bulk) sample. Evaluation of results Requires appropriate use of statistics Must be reasonable, reliable and related to the problem as originally stated Careful thought should be given to data presentation, so that it is readily digested and conclusions are clearly demonstrated. REPORT Report results with limitation/accuracy information. A professional chemist/charted chemist should verify the report. Test Report an example Professional Chemist