Skills Handbook Finalv3 - LGC Group
LaboratorySkillsTraining HandbookCharlotte BaileyVicki BarwickAM
Laboratory Skills Training HandbookCharlotte BaileyVicki BarwickMay 2007ISBN 978-0-948926-25-9
1Introduction1.1 The need for a laboratory skills training handbookAlthough analysis of samples is frequently carried out by using sophisticated instruments,the importance of basic laboratory skills cannot be overlooked.The majority ofinstruments require calibration via the analysis of sets of calibration standards. It istherefore essential that analysts are able to prepare such solutions accurately. Theanalyst also needs to be able to handle and prepare samples correctly and be able tomake ‘routine’ measurements, such as measurements of pH, reliably. Laboratories areincreasingly being required to demonstrate the competence of their staff to carry outparticular tasks and to document how competence was assessed. As well as soundpractical skills, analysts should have at least a basic understanding of important qualityissues such as method validation, traceability and quality control.The aim of this handbook is to provide a basic training package in key laboratory skillsand to provide an introduction to important quality topics. Those responsible for traininganalysts can use the handbook to help plan training programmes. Trainees can use thehandbook as a guide to best practice for a range of laboratory skills and to gain a basicunderstanding of quality issues.1.2 Structure of the handbook and how to use itThis handbook is divided into two sections. Part A is aimed at the trainee analyst.Chapter A1 contains the essential health and safety information that analysts should befamiliar with to enable them to work safely in the laboratory. It also covers the selectionof test methods and equipment plus key aspects to consider when planning and carryingout an analysis. Chapter A2 covers sample handling and storage. Chapter A3 covers thekey laboratory skills that analysts need in order to be able to carry out analytical workwith the required level of accuracy. There are questions relating to each ‘skill’ to testunderstanding. Chapter A4 introduces the key topics relating to quality assurance andquality control that analysts should be familiar with. Finally, Chapter A5 addresses datahandling and reporting of results.Part B of the handbook is aimed at those responsible for planning and carrying out thetraining of analysts. For each of the key laboratory skills covered in Chapter A3 there arekey learning points, suggestions for assessing competence and observations which mayindicate that retraining is required.1.3 A note about units of volumeVolumes can be expressed in a number of different ways, e.g. cm3, mL, dm3, L. In thishandbook we have used millilitres (mL) and litres (L) as the units of volume. However,you may encounter cm3 and dm3 in other texts or standard operating procedures.Remember that 1 mL is equivalent to 1 cm3 and that 1 L is equivalent to 1 dm3 (and that1 L contains 1000 mL).1.4 AcknowledgementsProduction of this handbook was supported under contract with the Department of Tradeand Industry as part of the National Measurement System Valid Analytical Measurement(VAM) programme.The authors would like to thank everyone who provided suggestions on the content ofthe handbook, in particular, members of the VAM Clinical, Food, Industrial and LGC Limited 2007
Environment Reference Materials User Groups, and members of the ACB EducationCommittee. We would also like to thank Pete Colwell (LGC), Andy Earls (LGC), MartynEgerton (West Park Hospital, Epsom), Stephen Halloran (Royal Surrey County Hospital)and Elizabeth Prichard (LGC) for reviewing and commenting on drafts of the handbook. LGC Limited 2007
ContentsPART A: INFORMATION FOR THE ANALYST1A11A1.1WORKING IN THE LABORATORYHealth and safety issues1A1.2Method and equipment selectionA1.2.1Method selectionA1.2.2Equipment selection445A1.3The importance of standard operating procedures6A1.4Carrying out an analysis7A2SAMPLE HANDLING AND STORAGE9A2.1Receiving 2.3.2Storage conditions101010A2.411A3Sample trackingKEY LABORATORY SKILLS12A3.1Measuring volumeA3.1.1Types of equipment availableA3.1.2Markings on equipment used for volumetric measurementsA3.1.3Selecting a suitable piece of equipmentA3.1.4Cleaning glasswareA3.1.5Checking the accuracy of the volume of liquid contained in/delivered by the equipmentA3.1.6Checklists for making measurements of volumeA3.1.7Questions1212151618192025A3.2Measuring massA3.2.1Mass versus weightA3.2.2Types of balance availableA3.2.3Selecting a suitable balanceA3.2.4Checking the balance accuracy and set upA3.2.5Correct use of balances for different applicationsA3.2.6Checklist for making measurements of massA3.2.7Questions2727272829313435A3.3Measuring pHA3.3.1What is pH?A3.3.2Equipment for measuring pHA3.3.3Choosing a suitable electrodeA3.3.4Care of electrodesA3.3.5Calibration of pH metersA3.3.6Measuring the pH of the test sampleA3.3.7Checklist for making pH measurements using a pH g solutions of known concentrationA3.4.1When are solutions of known concentration used?A3.4.2Calculating the concentration of solutionsA3.4.3Selecting a suitable material to prepare the solutionA3.4.4Practical steps in preparing solutions of known concentrationA3.4.5Labelling and storage of solutions464647525254 LGC Limited 2007
A3.4.6A3.4.7Checklist for preparing solutions of known concentrationQuestions5557A3.5Preparing reagent solutionsA3.5.1Calculating the concentration of reagent solutionsA3.5.2Preparing reagent solutionsA3.5.3Labelling and storage of reagent solutionsA3.5.4Checklist for preparing reagent solutionsA3.5.5Question595960606161A3.6Carrying out a titrationA3.6.1Principles of titrationA3.6.2Carrying out a standardisation experimentA3.6.3Detecting the end-pointA3.6.4Carrying out a titrationA3.6.5Titration calculationsA3.6.6Checklist for carrying out a fugationA3.7.1What is centrifugation and when is it used?A3.7.2rpm versus gA3.7.3Different types of centrifugeA3.7.4Correct operation of a centrifuge: safety and quality issuesA3.7.5Checklist for using a TY ISSUESDefinition of quality8282A4.2Quality management, quality assurance and quality controlA4.2.1International quality standards8384A4.3Method validationA4.3.1Definition of validationA4.3.2When is method validation required?A4.3.3How much validation is required?A4.3.4Method performance parameters8585868687A4.4Calibration and traceability92A4.5Documenting test methods95A4.6Introduction to control charts97A4.7Proficiency testing and external quality assessmentA4.7.1Scoring systems in PT schemesA4.7.2Evaluation of performance scores9899100A4.8Errors and uncertaintyA4.8.1Random errorsA4.8.2Systematic errorsA4.8.3Evaluating measurement uncertaintyA4.8.4Why is measurement uncertainty important?100101101101104A5A5.1DATA HANDLING AND REPORTING RESULTSEssential statistical terms105105A5.2Units of measurementA5.2.1Concentration expressed as a percentageA5.2.2Concentration expressed as m/m or m/vA5.2.3Concentration expressed as parts per million or parts per billion107107108108A5.3109Reporting results LGC Limited 2007
A5.3.1A5.3.2A5.3.3A5.3.4A5.4Decimal places and significant figuresRounding rulesDeciding the number of significant figures to reportChecking dataQuestions109111112114115PART B: INFORMATION FOR THE SUPERVISOR119B1ACCREDITATION REQUIREMENTS119B2GENERAL APPROACH TO ASSESSING COMPETENCE120B3KEY LABORATORY SKILLS121B3.1Measuring volumeB3.1.1Key learning points for measuring volumeB3.1.2Assessing competence in using volumetric glasswareB3.1.3Observations indicating that retraining may be required121121123124B3.2Measuring massB3.2.1Key learning points for measuring massB3.2.2Assessing competence in making measurement of massB3.2.3Observations indicating that retraining may be required124124125125B3.3Measuring pHB3.3.1Key learning points for measuring pHB3.3.2Assessing competence in making measurement of pHB3.3.3Observations indicating that retraining may be required126126126126B3.4Preparing solutions of known concentrationB3.4.1Key learning points for preparing solutions of known concentrationB3.4.2Assessing competence in preparing solutions of known concentrationB3.4.3Observations indicating that retraining may be required127127127128B3.5Preparing reagent solutionsB3.5.1Key learning points for preparing reagent solutionsB3.5.2Assessing competence in preparing reagent solutionsB3.5.3Observations indicating that retraining may be required128128129129B3.6Carrying out a titrationB3.6.1Key learning points for carrying out a titrationB3.6.2Assessing competence in carrying out a titrationB3.6.3Observations indicating that retraining may be required129129130130B3.7CentrifugationB3.7.1Key learning points for using a centrifugeB3.7.2Assessing competence in using a centrifugeB3.7.3Observations indicating that retraining may be required130130131131APPENDIX 1: GLOSSARY OF TERMS133APPENDIX 2: ANSWERS TO QUESTIONS137APPENDIX 3: TRAINING RECORD149APPENDIX 4: ADDITIONAL RESOURCES152 LGC Limited 2007
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Part AInformation for the analyst LGC Limited 2007
LGC Limited 2007
Part A: Information for the analystA1 Working in the laboratoryThe laboratory is a potentially hazardous working environment. You may well be usingchemicals and/or equipment which, if not handled correctly, could cause you or yourcolleagues harm. However, with the proper procedures in place, work can be carried outsafely. It is essential that you familiarise yourself with the general safety procedures inplace in your laboratory and with any special procedures required to carry out aparticular test method safely. When working in the laboratory you also need to knowhow to select an appropriate test method and equipment, and understand the importanceof following standard operating procedures. This section covers: Health and safety issues (section A1.1); Selecting a test method and equipment (section A1.2); Importance of standard operating procedures (section A1.3).A1.1 Health and safety issuesWhen working in the laboratory you must always: Wear suitable eye protection (safety glasses or goggles); Wear a laboratory coat; Wear suitable footwear (e.g. do not wear open-toed shoes or sandals).In the laboratory you should never: Eat or drink; Smoke; Apply cosmetics.You should know the meaning of common warning and hazard signs used in thelaboratory. You will see different coloured signs: Blue signs are mandatory; Red signs are prohibitive (or relate to fire alarms/fire-fighting equipment); Green signs give safety instructions; Yellow signs give warnings.Some examples of signs you may see in the laboratory are shown in Figure A1.1. LGC Limited 20071
Laboratory coatmust be wornEye protectionmust be wornNow washyour handsNoadmittance tounauthorisedpersonnelNo smokingNo entry forpersons with apacemaker alhazardEmergencyshowerEmergencyeyewashFirst aidToxic materialElectricalhazardEmergency exitFigure A1.1: Examples of signs displayed in the laboratoryBefore carrying out any laboratory work make sure: You are familiar with the test method; You know how to use all the necessary equipment/apparatus correctly and safely; You know of any hazards associated with the chemicals and reagents that you willbe using and how to handle them correctlyo consult the material safety data sheet for detailed information about particularchemicals or reagents (an example is shown in Figure A1.2). LGC Limited 20072
Southern Chemical Co.Date of issue: 1 September 2006Safety Data Sheet1.Identification of product and companyProduct name:Sodium dodecyl sulfateSynonyms:Dodecyl sulfate, sodium salt; sodium lauryl sulfateCatalogue No.:1234-ABCCompany:Tel (enquiries):Tel (emergency):Southern Chemical Co.South Park Industrial EstateSouthtownEssexES16 0PQUK 44(0)123 987 3210 44(0)123 100 1002. Composition/information on ingredientsProduct name:Sodium dodecyl sulfateCAS Number:151-21-3EINECS:205-788-1Molecular formula:C12H25OSO3Na-1Molecular weight:288.38 g mol3. Hazards identificationHighly flammableHarmful if swallowedIrritating to eyes and skinEye:Causes moderate eye irritation.Skin:Causes severe skin irritation. May be harmful if absorbed through the skin.Ingestion:Harmful if swallowed. May cause irritation of the digestive tract. May cause nausea and vomiting.Inhalation:Causes respiratory tract irritation. May cause allergic respiratory reaction. May cause irritation of therespiratory tract with burning pain in the nose and throat, coughing, wheezing, shortness of breath. May beharmful if inhaled.4.First aid measuresEye contact:Flush thoroughly with water for at least 15 minutes. Obtain medical attention.Inhalation:Remove from exposure and move into fresh air immediately. Obtain medical attention.Ingestion:Wash mouth out thoroughly with water and give plenty of water to drink. Obtain medical attention.Skin contact:Wash skin thoroughly with plenty of water. Remove contaminated clothing and wash before use. Obtainmedical attention.5. Fire fighting measuresFlammable solid, may evolve toxic fumes in a fire.Use water spray, dry chemical, carbon dioxide, or chemical foam extinguisher.6. Accidental release measuresWear appropriate protective clothing.Mix with sand, vacuum or sweep up material and transfer carefully to a suitable container. Wash site of spillage thoroughly.7. Handling and storageAvoid breathing dust. Avoid contact with skin and eyes.Keep away from sources of ignition. Store in a cool dry place in a tightly sealed container.Figure A1.2: Example of information contained in a material safety data sheetTo avoid accidents and to prevent the possible contamination of test samples it isessential to keep the laboratory area as clean and tidy as possible: Make sure you have sufficient space available at the laboratory bench to enableyou to work safelyo make sure that there is space available in a fume cupboard/hood if necessary; Work areas should be kept clean – clean up any spillages immediately. If you areunsure about how to clear up and dispose of spilled material consult yourlaboratory manager/supervisor; LGC Limited 20073
Take precautions to ensure that the samples cannot be contaminated by externalsources or by coming into contact with other sampleso make sure all glassware and other equipment/apparatus used is clean (seesection A3.1.4 for information on cleaning glassware);o do not prepare calibration standards or quality control materials in the samearea as the samples are prepared (this is critical for trace level analysis wherethe samples contain very low concentrations of the analyte);o avoid handling samples containing high concentrations of the analyte in thesame area as samples containing trace levels of the analyte;o if it is possible samples received at the laboratory may contain very differentlevels of the analyte (but you do not know this in advance of carrying out theanalysis), take special care to avoid cross-contamination when handlingsamples and carefully examine the results from quality control and blanksamples to help identify any anomalies. Take precautions to ensure that any chemicals or reagents used cannot becontaminatedodo not pipette reagents directly from the bottle – pour a suitable amountinto a beaker or flask;odo not return any unused chemicals or reagents to the original container –dispose of any excess material correctly;Always tidy up after you have finished your worko clean apparatus and return to the correct storage place;o return reagents, chemicals and samples to the correct storage place.A1.2 Method and equipment selectionA1.2.1 Method selectionAnalysis is always carried out for a reason. You will analyse test samples to help answera question or solve a particular problem. For the results of an analysis to be useful asuitable test method must be used. Once you know the reasons for carrying out theanalysis, how do you decide on a suitable test method? The two key aspects to considerare: Method scopeo can the test method be used to measure the analyte(s) of interest in the typesof sample submitted to your laboratory at the concentrations likely to bepresent in the samples?o the method scope can be summarised as the analytes, sample matrices andanalyte concentration range for which the method is applicable; Method performanceo what will be the likely uncertainty in results obtained (see section A4.8) and isthis acceptable?o are the key performance parameters, such as precision, bias and limit ofdetection, fit for purpose (see section A4.3)?o is the method performance acceptable across the required range of sampletypes and analyte concentrations?Other aspects to consider when selecting a test method include: Cost; Time available to complete the analysis (‘sample turnaround time’); LGC Limited 20074
Availability of apparatus/equipment and staff.There are a number of different sources of test methods: Methods nationalstandardso e.g. BSI (UK), CEN (Europe), ISO (international); Methods produced by government bodies or agencieso e.g. British Pharmacopoeia (BP), UnitedEnvironment Protection Agency (EPA); StatesPharmacopoeia(USP),Methods published by professional organisationso e.g. Royal Society of Chemistry (RSC), Association of Official AnalyticalChemists (AOAC); Methods supplied by trade organisationso e.g. Institute of Petroleum (Energy Institute); Methods described in regulationso e.g. The Fertilisers (Sampling and Analysis) Regulations 1991 (S.I. No. 973); Methods published in the scientific literatureo e.g. The Analyst, Journal of AOAC International, Journal of Chromatography,Journal of Analytical Atomic Spectrometry; Methods developed in-house by a laboratory.Usually, in-house method development is a last resort as developing a method to a stagewhere it can be used with confidence for the analysis of test samples can be costly andtime-consuming.In some situations there will not be any choice about which test method is used – it willbe specified in a regulation or requested by the customer to meet a specific regulatoryrequirement.In some cases there may be a method available which does not quite meet all of yourrequirements. For example, the method may not have been validated for the particularrange of analyte concentrations or sample types that you need to analyse. In suchcases, the method must be tested to ensure that it is capable of producing reliableresults for the required sample types (see section A4.3). If the performance is notsatisfactory then further development and revalidation of the method will be required.Remember that the performance of all methods (including standard and regulatorymethods) must be assessed before they are used for the analysis of test samples. This isdiscussed in section A4.3.A1.2.2 Equipment selection‘Equipment’ is everything other than the chemicals and reagents needed to carry out aparticular test. The equipment required to carry out a particular test should be specifiedin the standard operating procedure (see section A1.3). However, before you use anyitem of equipment you will need to make sure that it is in an acceptable condition.Things to check include: Is the equipment clean? (see section A3.1.4 for information on cleaningglassware) Is the equipment in working order?o faulty equipment should be clearly labelled; LGC Limited 20075
o do not use equipment that is damaged (e.g. chipped or cracked glassware) orawaiting repair; Is the equipment set up correctly? Is the equipment correctly calibrated (see section A4.4 for information oncalibration)?o for items calibrated by a third-party check that the calibration is still current;o familiarise yourself with any in-house ca
analysts can use the handbook to help plan training programmes. Trainees can use the handbook as a guide to best practice for a range of laboratory skills and to gain a basic understanding of quality issues. 1.2 Structure of the handbook and how to use it This handbook is divided into two sections. Part A is aimed at the trainee analyst.
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