ebook img

Basic Concepts in Clinical Biochemistry: A Practical Guide PDF

174 Pages·2018·4.19 MB·English
Save to my drive
Quick download
Download
Most books are stored in the elastic cloud where traffic is expensive. For this reason, we have a limit on daily download.

Preview Basic Concepts in Clinical Biochemistry: A Practical Guide

Vijay Kumar Kiran Dip Gill Basic Concepts in Clinical Biochemistry: A Practical Guide Basic Concepts in Clinical Biochemistry: A Practical Guide Vijay Kumar (cid:129) Kiran Dip Gill Basic Concepts in Clinical Biochemistry: A Practical Guide VijayKumar KiranDipGill DepartmentofBiochemistry DepartmentofBiochemistry MaharshiDayanandUniversity PostgraduateInstituteofMedicalEducation Rohtak,Haryana,India &Research Chandigarh,India ISBN978-981-10-8185-9 ISBN978-981-10-8186-6 (eBook) https://doi.org/10.1007/978-981-10-8186-6 LibraryofCongressControlNumber:2018935904 #SpringerNatureSingaporePteLtd.2018 Thisworkissubjecttocopyright.AllrightsarereservedbythePublisher,whetherthewholeorpart of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation,broadcasting,reproductiononmicrofilmsorinanyotherphysicalway,andtransmissionor informationstorageandretrieval,electronicadaptation,computersoftware,orbysimilarordissimilar methodologynowknownorhereafterdeveloped. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt fromtherelevantprotectivelawsandregulationsandthereforefreeforgeneraluse. Thepublisher,theauthorsandtheeditorsaresafetoassumethattheadviceandinformationinthisbook arebelievedtobetrueandaccurateatthedateofpublication.Neitherthepublishernortheauthorsorthe editorsgiveawarranty,expressorimplied,withrespecttothematerialcontainedhereinorforanyerrors oromissionsthatmayhavebeenmade.Thepublisherremainsneutralwithregardtojurisdictionalclaims inpublishedmapsandinstitutionalaffiliations. Printedonacid-freepaper ThisSpringerimprintispublishedbytheregisteredcompanySpringerNatureSingaporePteLtd. The registered company address is: 152 Beach Road, #21-01/04 Gateway East, Singapore 189721, Singapore Preface Biochemistryisoneofthefundamentalsubjectsoflifescience,andknowledgeofits practical aspects is absolutely essential for students and researchers. Performing a practical in biochemistry requires adequate facilities and training laboratories for organizing meaningful practicals both at undergraduate and postgraduate levels. Clinicalbiochemistryisadiagnosticssubject,whichaimstousestandardmethods, tomonitordiseasedevelopmentandtreatmentbybiochemicalmethods.Laboratory investigations pertaining to clinical biochemistry provide useful information to clinicians, both in the diagnosis of illness and the monitoring of treatment. The necessity for a practical manual on orientation toward various aspects of clinical biochemistry is always desired. The present manual is written in simple language covering various experimental aspects related with clinical biochemistry such as urinary analysis of bioorganic constituents and estimation of proteins, urea, sugar, uricacid,bilirubin,etc.,inbloodwhichisessentialforearlydiagnosisofadisease andalsoassessmentofitstherapyefficacy. The book is specifically helpful to BSc, MLT, MSc, and MSc biochemistry students. Each chapter begins with theoretical aspects of the practical, and the experiments are further well supported by self-explanatory tables and simple calculations that will make the students understand protocols very easily. The manual is also helpful to teachers who are taking clinical biochemistry practicals toguidestudentsinasimpleway. Theauthorshopethatthemanualwillmeettherequirementofundergraduateand postgraduatestudentsstudyingbiochemistryandwillbegladtoacceptconstructive criticisms and suggestions from the faculty, students, and readers to make this manualabetteroneinthefuture. Rohtak,Haryana,India VijayKumar Chandigarh,India KiranDipGill v Reference Biochemical Values in Serum/Plasma Acidphosphatase 1–4KAU/dl Albumin 3.5–5g/dl Totalproteins 3.5–5.5g/dl Globulins 2.5–3.5g/dl Alaninine 3–40IU/L Alkaline 3–13KAU/dl transaminase phosphatase (ALT,SGPT) Amylase 5–45SU/dl Aspartate 5–45IU/L Calcium 9–11mg/dl transaminase (AST,SGOT) Calcium(U) 100–200mg/ day Bilirubin(total) 0.2–1mg/dl Ceruloplasmin 25–50mg/dl Bilirubindirect <0.4mg/dl Cholesterol 150–200mg/dl Bilirubinindirect <0.6mg/dl HDL 30–60mg/dl Creatinine 0.5–1.5mg/dl LDL 60–130mg/dl Creatinine(U) 0.8–2g/day VLDL 20–40mg/dl Chloride 103–105mEq/L Inorganic 3.0–4.5mg/dl phosphorus Creatinekinase, 24–195U/L Phosphorus(U) 0.5–1.5g/day CK-MB Creatinekinase,total 10–400U/L LDH 100–190U/L Fibrinogen 0.2–0.4g/dl Urea 15–40mg/dl Glucose,fasting 70–100mg/dl Urea(U) 15–40g/day Lipase 0.5–160U/L Uricacid 3.5–8mg/dl Transferrin 200–300mg/dl Uricacid(U) 200–500mg/ day Triglyceride,fasting 50–175mg/dl Urobilinogen(U) 1–3.5mg/day Sodium 135–145mEq/L Potassium 3.5–5mEq/L Chloride 103–105mEq/L U-Urine vii Contents 1 CommonClinicalLaboratoryHazardsandWasteDisposal. . . . . . 1 1.1 WasteDisposalinLaboratory. . . . . . . . . . . . . . . . . . . . . . . . 2 2 BloodCollectionandPreservation. . . . . . . . . . . . . . . . . . . . . . . . . 5 2.1 BloodCollection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5 3 QualityControlinLaboratory. . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 3.1 TypesofLaboratoryErrors. . . . . . . . . . . . . . . . . . . . . . . . . . 10 3.2 MethodstoMinimizetheLaboratoryErrors. . . . . . . . . . . . . . 11 4 AutomationinClinicalLaboratory. . . . . . . . . . . . . . . . . . . . . . . . . 13 4.1 TypesofAutoanalyzers. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 5 Photometry:ColorimeterandSpectrophotometer. . . . . . . . . . . . . . 17 5.1 ColorimeterandSpectrophotometer. . . . . . . . . . . . . . . . . . . . 18 6 PreparationofGeneralLaboratorySolutionsandBuffers. . . . . . . 21 6.1 MolarSolutions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 6.1.1 Molarity(M). . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 6.2 NormalSolutions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22 6.2.1 Normality(N). . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23 6.3 Percent(%)Solutions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 6.4 BufferSolutions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 6.5 PhysiologicalBuffersintheHumanBody. . . . . . . . . . . . . . .. 25 6.6 PreparationofCommonLaboratoryBuffers. . . . . . . . . . . . . . 25 6.6.1 0.2MAcetateBuffer(pKa4.86). . . . . . . . . . . . . . . . 25 6.6.2 0.2MSodiumPhosphateBuffer(pKa6.86). . . . . . . . 26 6.6.3 0.2MTris-HClBuffer(pKa8.1). . . . . . . . . . . . . . . . 26 7 ExaminationofUrineforNormalConstituents. . . . . . . . . . . . . . . . 29 7.1 PreservativesUsedforUrineCollection. . . . . . . . . . . . . . . . . 29 7.2 PhysicalExaminationofUrine. . . . . . . . . . . . . . . . . . . . . . . . 30 7.2.1 ColorandOdor. . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 7.2.2 Appearance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 ix x Contents 7.2.3 SpecificGravity. . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 7.2.4 Volume. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 7.2.5 pH. . . .. . . . . . . . . . .. . . . . . . . . .. . . . . . . . . . .. . 31 7.3 ChemicalExaminationofUrine. . . . . . . . . . . . . . . . . . . . . . . 31 7.4 TestsforInorganicConstituentsofUrine. . . . . . . . . . . . . . . . 31 7.5 TestsforOrganicConstituents. . . . . . . . . . . . . . . . . . . . . . . . 32 7.6 DetectionofUrea. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 8 ToPerformQualitativeTestsforUrinaryProteins. . . . . . . . . . . . . 33 8.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33 8.2 TestsforUrinaryProteins. . . . . . . . . . . . . . . . . . . . . . . . . . . 34 8.2.1 DipstickTest. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 8.2.2 TheBoilingTestforCoagulableProteins. . . . . . . . . . 34 8.2.3 SulphosalicylicAcidTest. . . . . . . . . . . . . . . . . . . . . 35 8.2.4 NitricAcidRingTest(Heller’sTest). . . . . . . . . . . . . 35 8.2.5 Bence-JonesProteins. . . . . . . . . . . . . . . . . . . . . . . . 35 9 ToDeterminetheQuantityofProteinsinUrineSample UsingBiuretReaction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 9.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 9.2 SpecimenRequirements.. . . . .. . . . .. . . . . .. . . . .. . . . . .. 39 9.3 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 9.4 Reagents. . . . .. . . . . .. . . . .. . . . . .. . . . . .. . . . .. . . . . .. 40 9.5 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 9.6 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 10 ToEstimatetheAmountofTotalProteinandAlbumininSerum andtoFindA/GRatio. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 10.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 10.2 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 10.3 SpecimenRequirements. .. . . . .. . . . .. . . . .. . . . . .. . . . .. 44 10.4 Reagent. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 10.5 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 10.6 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 10.7 EstimationofAlbumininSerum. . . . . . . . . . . . . . . . . . . . . . 46 10.8 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 10.9 Reagents. . . . . .. . . . .. . . . . .. . . . .. . . . . .. . . . .. . . . . .. 46 10.10 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46 10.11 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 10.12 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 10.13 Precautions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48 Contents xi 11 ToPerformQualitativeTestforReducingSubstancesinUrine. . . 49 11.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 11.2 QualitativeTestforReducingSugars. . . . . . . . . . . . . . . . . . . 49 11.2.1 Benedict’sTest. . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 11.2.2 Benedict’sQualitativeReagent. . . . . . . . . . . . . . . . . 50 11.2.3 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 11.3 CLINISTIX/Uristix. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 11.3.1 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 11.4 Precautions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 11.5 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 12 QuantitativeAnalysisofReducingSugarsinUrine. . . . . . . . . . . . . 53 12.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 12.2 SpecimenRequirement. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 12.3 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53 12.4 Reaction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 12.5 Reagents. . . . . .. . . . .. . . . . .. . . . .. . . . . .. . . . .. . . . . .. 54 12.6 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 12.7 Precautions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54 12.8 Calculation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55 12.9 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55 13 EstimationofBloodGlucoseLevelsbyGlucoseOxidaseMethod. . 57 13.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57 13.2 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57 13.3 SpecimenRequirements. .. . . . .. . . . .. . . . .. . . . . .. . . . .. 58 13.4 Reagents. . . . . .. . . . .. . . . . .. . . . .. . . . . .. . . . .. . . . . .. 58 13.5 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58 13.6 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 13.7 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59 14 ToDeterminetheBloodGlucoseLevelsbyFolin andWuMethod. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 14.1 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 14.2 Reagents. . . . . .. . . . .. . . . . .. . . . .. . . . . .. . . . .. . . . . .. 61 14.3 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62 14.4 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62 15 ToPerformGlucoseToleranceTest. . . . . . . . . . . . . . . . . . . . . . . . 63 15.1 MethodofCarryingGTT. . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 15.2 SpecimenRequirements. .. . . . .. . . . .. . . . .. . . . . .. . . . .. 63 15.3 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 15.4 Reagents. . . . . .. . . . .. . . . . .. . . . .. . . . . .. . . . .. . . . . .. 64 15.5 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64 15.6 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66 15.7 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66 xii Contents 16 EstimationofUreainSerumandUrine. . . . . . . . . . . . . . . . . . . . . 67 16.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 16.2 SpecimenRequirements. .. . . . .. . . . .. . . . .. . . . . .. . . . .. 67 16.3 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 16.4 Reagents. . . . . .. . . . .. . . . . .. . . . .. . . . . .. . . . .. . . . . .. 68 16.5 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 16.6 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 16.7 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70 17 ToDetermineUreaClearance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 17.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 17.2 MaximumUreaClearance. . . . . . . . . . . . . . . . . . . . . . . . . . . 71 17.3 StandardUreaClearance. . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 17.4 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 17.5 Precautions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 17.6 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72 18 ToEstimateCreatinineLevelinSerumandUrine byJaffe’sReaction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75 18.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75 18.2 SpecimenRequirements. .. . . . .. . . . .. . . . .. . . . . .. . . . .. 75 18.3 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76 18.4 Reagents. . . . . .. . . . .. . . . . .. . . . .. . . . . .. . . . .. . . . . .. 77 18.5 PreparationofProtein-FreeFiltrate. . . . . . . . . . . . . . . . . . . . . 77 18.6 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77 18.7 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78 18.8 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78 19 ToDetermineCreatinineClearance. . . . . . . . . . . . . . . . . . . . . . . . 79 19.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79 19.2 SpecimenRequirementandProcedure. . . . . . . . . . . . . . . . . . 79 19.3 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80 20 ToDeterminetheUricAcidConcentrationinSerumandUrine. . . 81 20.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81 20.2 SpecimenRequirements. .. . . . .. . . . .. . . . .. . . . . .. . . . .. 81 20.3 Principle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82 20.4 Reagents. . . . . .. . . . .. . . . . .. . . . .. . . . . .. . . . .. . . . . .. 82 20.5 Procedure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82 20.6 Calculations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83 20.7 ClinicalSignificance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83 20.8 Precautions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84 21 EstimationofTotalCalciuminSerumandUrine. . . . . . . . . . . . . . 85 21.1 Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85 21.2 SpecimenRequirements. .. . . . .. . . . .. . . . .. . . . . .. . . . .. 85

Description:
This book is a practical guidebook in biochemistry, for medical as well as life sciences' students. The book covers reference values, sample collection procedure and detailed protocol to perform experiments. Each experiment starts with a brief introduction of the protocol, followed by specimen requi
See more

The list of books you might like

Most books are stored in the elastic cloud where traffic is expensive. For this reason, we have a limit on daily download.