Klubsy Bear Medtech Review and Tutorials PDF

KLUBSY BEAR MEDTECH REVIEW AND TUTORIALS CLINICAL CHEMISTRY MARCH...

KLUBSY BEAR MEDTECH REVIEW AND TUTORIALS CLINICAL CHEMISTRY MARCH RMT Prepared by: Maria Kristina D. Gaquit, RMT RMT TABLE OF CONTENTS BASICS OF CLINICAL CHEMISTRY................................................................................................ 2 QUALITY ASSESSMENT & QUALITY CONTROL..........................................................................9 LABORATORY SAFETY................................................................................................................... 16 SPECIMEN COLLECTION & HANDLING..................................................................................... 20 ANALYTICAL METHOD.................................................................................................................. 25 CARBOHYDRATES......................................................................................................................... 30 LIPIDS AND LIPOPROTEINS......................................................................................................... 38 PROTEINS........................................................................................................................................ 46 NONPROTEIN NITROGEN.............................................................................................................. 53 ENZYMES......................................................................................................................................... 58 LIVER FUNCTION............................................................................................................................ 66 ELECTROLYTES............................................................................................................................... 71 ACID-BASE BALANCE................................................................................................................... 77 TRACE METALS.............................................................................................................................. 80 ENDOCRINOLOGY........................................................................................................................... 81 THERAPEUTIC DRUG MONITORING........................................................................................... 93 TOXICOLOGY................................................................................................................................... 96 VITAMINS......................................................................................................................................... 99 TUMOR MARKERS........................................................................................................................ 100  COMMON CHEMISTRY ANALYTES BASICS OF CLINICAL CHEMISTRY UNITS OF MEASURE ANALYTE: a biologic solute or constituent contain.it siitnitt'Multiply SI system units (SI) – based on the metric system ANALYTE Conv. unit SI Unit CF Ammonia ug/dL umol/L 0.587 SI system seven basic units Bilirubin 17.1 BASE QUANTITY NAME SYMBOL Creatinine mg/dl umol/L 884 88.4 Length m Iron 0.179 Meter Mass kg Lithium mEq/L umol/L 1 Kilogram Time Seconds s Sodium 1 Electric current Ampere A To Potassium normality molarity 1 Thermodynamic K Chloride mEq/L mmol/L 1 Kelvin If Amount of substance Mol Bicarbonate N.mx valence 1 Mole Luminous intensity Candela cd Magnesium MNvalence 0.5 Total Protein 10 PREFIX USED WITH SI UNITS tf Albumin Globulin g/dL g/L 10 10 FACTOR PREFIX SYMBOL Mnemonics Phospholipid 0.01 1018 exa E mg/dL mg/L 10 1119 1015 peta P Immunoglobulins mg/dL g/L 0.01 1012 tera T Thyroxine ug/dL nmol/L 12.9 109 giga G PCO2 0.133 106 mega M mmHg kPa pO2 0.133 103 kilo k King Triglycerides 0.0113 102 101 hecto deka h da forHector Died Uric acid Phosphorus 881 0.0595 0.323 100 Liter, meter, gram Basic unit By Calcium mg/dL mmol/L 0.25 10-1 deci d Drinking Trinking Cholesterol 0.026 10-2 centi c Chocolate chocolate Glucose 0.0555 10-3 milli m Milk BUN 8855755 0.357 10-6 micro u Monday 10-9 nano n Night BUN to Urea 2.14 10-12 pico p Urea to BUN 0.467 10-15 femto F N to protein 6.54 10-18 atto a 2|Page | Prepared by: Maria Kristina D. Gaquit, RMT REAGENTS Secondary Substance of lower purity secondary standard Its concentration is determined by comparison with a primary standard CHEMICALS standard depends not only on its composition but also on the analytic reference method Labels for 1. State the actual impurities reagents 2. List maximum allowable impurities WATER SPECIFICATIONS 3. Clearly printed with percentage of impurities 4. Initials of its grade of purity (e.g., AR or ACS) Water water Most frequently used reagent in the laboratory OSHA 1. Indicate the lot number Substantially for lab procedures including reagent and standard preparation requirement 2. Physical hazard purified water 3. Biologic health hazard Distilled water water purified by distillation 4. Precautions for safe use Deionized water water purified by ion exchange chromatography 5. Storage Reverse osmosis purified water by reverse osmosis; pumps water across a Safety Data Formerly known as Material Safety Data sheet (RO) water semipermeable membrane Sheet (SDS) A required document provided by the manufacturer which contains Other purification Ultrafiltration safetyData technical data sheets for each chemical manufactured methods Ultraviolet light SheetSDS are only required for hazardous chemicals Sterilization not primarily intended for use by the general consumer Ozone treatment must also contain Sections 12 through 15, to be consistent with the Reagent grade water → Laboratory required water UN Globally Harmonized System of Classification and Labeling of Categories of 1. Clinical laboratory reagent water (CLRW) Chemicals (GHS) reagent grade 2. Special reagent water water 3. Instrument feed water 4. Water supplied by method manufacturer GRADES OF PURITY 5. Autoclave and wash water 1.American ACSchemical standard- society Aka Analytic reagent (AR) 6. Commercially bottled purified water gradeGrade standard reference For Laboratory use CLRW Processes required in preparation of reagent grade water (CLRW): material reference material toiaietlater Suitable for use in most analytic laboratory procedures 1. Prefilters are glass or cotton microfibers that remove 98% of the 2. Ultrapure Have undergone additional purification steps particulate matter. chemicals 2. Activated carbon removes organic matter and chlorine. Used in specific procedures: chromatography, atomic ultrapurechemical 3. A submicron filter removes all particles or microorganisms larger than absorption, immunoassays, molecular diagnostics, elfication the membrane pore size. standardization 4. Reverse osmosis is a process that removes 95-89% of bacteria and 3. United states used to manufacture drugs organic and other particulate matter. an USPUnited states Pharmacopeia 5. lon exchange is a system of resin cartridges or tanks connected in series pharmacopoeia National pure enough to be used in most chemical procedures but may or (USP) & National may not meet all assay requirements that remove cations and anions to make deionized water. Formulary Formulatory (NF) Special May require different preparation than CLRW according to intended 4. CP or pure grade preparation of these chemicals is not uniform (impurity Reagent Water speggepeagent use, such as sterility specification for tissue or organ culture, nucleic ChemicallyPure limitations are not stated) acid content for DNA testing, metal content for trace metal analysis, etc PureGrade not recommended for reagent preparation in clinical laboratories unless further purification or a reagent blank is included. Instrument Used for Internal Instrument rinsing, making dilutions, etc., and instrymettfeed feed water 5. Technical/ primarily in manufacturing needs to meet manufacturer's specifications. Technical commercial Water supplied Label states Intended use; do not substitute for CLRW or SRW unless commercial by methodby NEVER used in Clinical Laboratory watersupply Grade grade reagent methodmanufacturer label indicates it is of such quality. manufacturer Autoclave & REFERENCE MATERIALS wash purified to contain only low levels of organics, inorganics, and Y wash water particulate matter so it does not leave residue on glassware or Primary Highly purified chemical primary Standard measured directly to produce a substance of exact known concentration contaminate solutions and media in autoclaves standard Commercially exercise care because some plastic containers permit and purity bottledLettled 59nF microorganism growth due to air permeability Standard Used instead of ACS primary standard materials purified water Reference Certified reference materials Materials used in clinical chemistry laboratories (SRM) used to verify calibration or accuracy/bias assessments 3|Page | Prepared by: Maria Kristina D. Gaquit, RMT  CLSI- CLRW Guidelines Osmotic pressure pressure that opposes osmosis when a solvent flows through a TYPE 1 Most stringent requirements and generally suitable for routine laboratory osmoticpressure 17104 semipermeable membrane to establish equilibrium between use compartments of differing concentration Required for critical lab applications (HPLC), mobile phase preparation, Note: V & F ↓; B & O ↑ iiiiiiii blanks & sample dilution for analytic techniques preparation of buffers & media for mammalian cell culture & IVF, reagents CLINICAL LABORATORY SUPPLIES for molecular biology applications, preparation of solutions for electrophoresis and blotting THERMOMETERS TYPE 2 used in buffers, pH solutions, and microbiological culture media preparation Thermometers an integral part of an instrument or need to be placed in the device preparation of reagents for chemical analysis. For clinical analyzers, cell culture incubators for temperature maintenance Feed water to type 1 systems Three types: liquid-in-glass (replaces mercury type), electronic thermometer (or thermistor probe), and digital thermometer TYPE 3 Glassware rinsing Calibration Must be calibrated using an NIST-certified thermometer Filling autoclaves, heating baths, & humidifiers Feed water to type 1 systems SI for temp Kelvin C to F F = C (9/5) + 32 or (C x 1.8) + 32 F to C C = (F-32) * 5/9 or (F – 32)/ 1.8 SOLUTION PROPERTIES C to K K = C + 273 Solute Solvent Solution Substance that is dissolved in a liquid iii.int Liquid in which the solute is dissolved GLASSWARES GLASSWARE INSCRIPTIONS I Is Biologic solute + biologic fluid mn Class A Those that satisfy NIST specifications Basic properties of Concentration, saturation, colligative properties, redox potential, 20oC Temp of calibration. Temp of glassware & solutions for maximum accuracy solution conductivity, density, pH, and ionic strength TC To contain. TD To deliver. Vessel calibrated to deliver specific volume (e.g. graduated cylinder) CONCENTRATION Percent solution Equal parts per hundred or the amount of solute per 100 total units TYPES OF GLASS ii Percent solution w/w, v/v, w/v (most common) ii.itiiiiiiiiN Molarity Borosilicate Most common type of glassware in volume measurements number of moles per 1L of solution (Pyrex/ Kimax) High degree of thermal resistance Molality MY amount of solute per 1Kg solvent Borosilicate it lntity Normality number of gram equivalent weights per 1L of solution Low alkali content gew L Can be heated or autoclaved Pyrex (corning, corning, N.Y.), Kimax (Kimble, Vineland, N.J.) SOLUTION SATURATION Aluminosilicate Strengthened chemically rather than thermally Dilute relatively little solute Aluminosilicate (Corex) Six times stronger than borosilicate glass Intentrated Concentrated large quantity of solute in solution Resists clouding & scratching better Saturated excess of undissolved solute particles Corex (Corning N.Y.) supefactated Supersaturated greater concentration of undissolved solute Boron free Used for highly alkaline solutions BoronFree Alkali resistant Glass Poor heat resistance COLLIGATIVE PROPERTIES High Silica Heat, chemical, & electrical tolerance Vapor pressure pressure at which the liquid solvent is in equilibrium with the water HighSilica Excellent optical properties vaporpressureto3mmHg vapor Used for high precision analytic work optical reflectors, mirrors Freezing point temperature at which the vapor pressures of the solid and liquid Flint glass Soda-lime glass containing oxides of sodium, silicon & Calcium Freezingpoint v1.86c phases are the same FlintGlass Least expensive but poor resistance to high temp and sudden most commonly used & preferred; uses wheatstone bridge changes of temp Boiling point temperature at which the vapor pressure of the solvent reaches one Used for some disposable glasswares Boilingpoint to.ae atmosphere Low-actinic High thermal resistance w/ amber or red color Low Actinic glassware Maximum protection to light-sensitive materials iiii.EE Glassware Hiiiii.fiure Used to store control material and reagents (ex: bilirubin standards) 4|Page | Prepared by: Maria Kristina D. Gaquit, RMT LABORATORY VESSEL GLASSWARE According to Drainage Characteristics Beaker Wide-mouthed, straight-sided jar with pouring spout. Not accurate Blow-out raining Self-draining Beaker Blowout selfDNo III enough for critical measurements Has a continuous etched ring etched ring or markings Last drop of liquid should be expelled by Contents drain by gravity eII Volumetric flask Used to prepare standards for quantitative procedures volumetric blowing out Ex: Mohr, Volumetric, Pasteur Pear shaped, long neck with single calibration mark. Manufactured to strict standards Ex: Ostwald Folin, Serologic spshoutout MVP Erlenmeyer flasks Sloping sides, graduated markings. Used to hold liquids, mix solutions, According to type ErlenmeyerFlask 1111 1,11 measure noncritical volumes transfer Transfer Graduated/Measuring Graduatedmeasuring Graduated cylinder Upright, straight-sided tube with flared base. Used for noncritical Dispense one volume without further Dispenses several different volumes Graduated measurements subdivisions cylinde.gg 1. Volumetric 1. Serologic 11 Shouldn’t be used to measure ±2s Random Warning rule controls >1s rule from mean. Initiates from mean on Warning flag of testing of same side possible change other rules. 125 in accuracy or If 415 Systematic precision. no violation of other rules, run is considered in control 13s 1 control >±3s Random Rejection 10X 10 consecutive Systematic Rejection from mean rule controls on rule same side of 135 Random mean 10x Systematic 13 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT  COMPARISON OF METHODS (COM) EXPERIMENT RANDOM ERRORS SYSTEMATIC ERRORS COM experiment – involves measuring patient specimens by both an existing (reference) 13S, R4S 22s, 41s, 10x method and a new (test) method o Values obtained by the reference method are plotted on the x-axis and the values obtained by the test method are plotted on the y-axis o agreement between the two methods is estimated from the straight line that best fits the points o Linear Regression analysis: provides objective measures of the location and dispersion for the line ▪ ▪ ▪ Three factors: slope (m), y-intercept, correlation coefficient ® Defined by the equation: y=mx+b Perfect correlation generates a slope of 1 and a y-intercept of 0 itin liitiident Slope 1 Proportional Systematic error SPICY y-intercept 0 Constant systematic error o Bland-Altman plot/ Difference plot: alternate approach to visualizing paired data; indicates either the percent or absolute bias (difference) between the reference and test method values over the average range of values; useful tool to visualize concentration-dependent error o COM experiments measures two kinds of error: random error and systematic error Standard – is a solution of a known concentration often used when calibrating instruments METHOD EVALUATION Delta check – most commonly used patient based-QC technique; comparison of current results with past results Method Evaluation – used to verify the acceptability of new methods prior to reporting Outlier – Control result outside established limits; occurs more than once in 20 runs, patient results investigation must be carried out preliminary evaluation: analysis of a series of standards to verify the linear range and then replicate analysis (at least eight measurements) of two controls to obtain estimates of short- term imprecision. REFERENCE INTERVAL STUDIES General CLIA regulations of method validation Reference intervals include all the data points that define the range of observations. A pair of Nonwaived 1. Demonstrate test performance comparable to that established by medical decision points that span the limited of results expected for a defined healthy FDA approved manufacturer. population tests a. Accuracy b. Precision Erroneously called "normal ranges." c. Reportable range o All normal ranges are reference intervals, but not all reference intervals are normal ranges) 2. Verify reference (normal) values appropriate for patient population Therapeutic range: Reference interval applied to a therapeutic drug Nonwaived 1. Determine Three main categories of the application of reference intervals FDA approved a. Accuracy 1. Diagnosis of a disease or condition tests modified b. Precision 2. Monitoring of a physiologic condition or developed c. Analytic sensitivity 3. Monitoring therapeutic drugs by laboratory d. Analytic specificity (including interfering substances) Establishing a reference interval: A new reference interval is established when there is no e. Reportable range of test results existing analyte or methodology in the clinical or reference laboratory with which to conduct f. Reference/ normal ranges comparative studies, It is a costly and labor-intensive study that will Involve laboratory g. Other performance characteristic resources at all levels and may require from 12 0 to as many as 700 h. Calibration and control procedures 120 700 study Individual, verifying a reference interval (transference): This is done to confirm the validity of an existing reference interval for an analyte using the same (identical) type of analytic system (method and/or instrument). These are the most common reference Interval studies performed In the clinical laboratory and can require as few as 20___ study Individual. 20 14 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT  SINGLE VALUE CRITERIA OF WESTGARD TYPE OF ERROR TEST USED TO DETERMINE Random error Replication experiment Proportional error Recovery experiment Constant error Interference experiment Systemic error Comparison of methods (COM) Total error Replication and comparison DIAGNOSTIC EFFICIENCY Determine how good a given test is at detecting and predicting the presence of disease (or a physiologic condition) o True positive: Assay correctly identifies a disease or condition in those who have it o False positive: Assay incorrectly identifies disease when none is present First thing to do: Determine imprecision and inaccuracy. Both of these errors (total error) are o True negative: assay correctly excludes a disease or condition in those without it compared with the maximum allowable error o False negative: assay incorrectly excludes a disease when it is present o Total error > maximum allowable error (unacceptable and must be modified and reevaluated or rejected) Measures of Diagnostic efficiency o MEASUREMENT OF IMPRECISION/ RANDOM ANALYTIC ERROR Description Formula Other notes ▪ Imprecision: dispersion of repeated measurement around a mean (true level) Diagnostic Proportion with the Sensitivity (%) Analytical sensitivity ▪ Estimated from studies in which multiple aliquots of the same specimen (with Diagnostic sensitivity disease who have a 𝑇𝑃 - is a measure of the constant concentration) are analyzed repetitively sensitivity 𝑥 100 positive test result 𝑇𝑃 + 𝐹𝑁 smallest increment of o ESTIMATION OF INACCURACY/ SYSTEMATIC ANALYTIC ERROR the analyte that can ▪ Inaccuracy: difference between a measured value and its actual value due to the truepositive be distinguished by presence of a systematic error the assay ▪ Estimated from studies in which multiple aliquots of the same specimen (with Diagnostic Proportion without the Specificity Analytical specificity is constant concentration) are analyzed repetitively Diggitiity specificity disease who have a 𝑇𝑁 the ability of the assay to 𝑥 100 negative test result Tegative 𝑇𝑁 + 𝐹𝑃 distinguish TYPES OF STUDIES TO ESTIMATE ACCURACY Positive Proportion with a disease PPV % PPV predicts the Recovery Study determines how much of the analyte can be identified in the Predictive who have a positive test 𝑇𝑃 probability that an sample Eititive Value result compared with all 𝑇𝑃 + 𝐹𝑃 𝑥 100 individual with a positive Recovery A small aliquot of concentrated analyte is added (spiked) into a patient value individuals who have a assay result has he sample (matrix) and then measured by the method being evaluated positive test result disease or condition Amount recovered is the difference between the spiked sample and the Negative Proportion without a disease NPV % NPV the probability that Negative Predictive who have a negative test 𝑇𝑁 an individual with a patient sample (unmodified) 𝑥 100 Predictive Value result compared with all 𝑇𝑁 + 𝐹𝑁 negative assay result Interference Determines if specific compounds affect the laboratory tests like interference value individuals who have a does not have the disease hemolysis, turbidity, and icteric Interferents often affect tests by absorbing or scattering light; but they negative test result or condition can also react with reagents or affect creation rates used to measure a given analyte Sample Used to assess presence of error (inaccuracy) in actual patient sample Eifarison comparison Examines patient samples by the method being evaluated (test method) study with a reference method study Westgard et al. & CLIA recommended that 40 to 100 specimens be run each method on the same day over 80 to 20 days (preferably within 4 hours), with specimens spanning the clinical range and representing a diversity of pathologic conditions 15 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT  Elimination Phys inatiiiiin T.IE iiiiiiiiiiiii.gg ar Class I BSC Class II BSC LABORATORY SAFETY SAFETY EQUIPMENT The employer is required by law to have designated safety equipment available, but it is also the responsibility of the employee to comply with all safety rules and to use safety equipment. All laboratories are required to have safety showers, eyewash stations, and fire extinguishers and to periodically test and inspect the equipment for proper operation It is recommended that safety showers deliver 30 to 50 gallons of water per minute at 20 to 50 pounds per square inch (psi) and be located in areas where corrosive liquids are stored or used. Eyewash stations must be accessible (i.e., within 100 feet or 10s travel) in laboratory areas presenting chemical or biological exposure hazards. Fire blankets, spill kits, first-aid supplies BIOSAFETY CABINETS (BSCs) remove particles that may be harmful to the employee who is working with potentially infectious biologic specimens BSCs are designed to offer various levels of protection, depending on the biosafety level of the specific laboratory class o CDC and the National Institutes of Health have described four levels of biosafety. class Class III BSC class 111 CHEMICAL FUME HOODS Fume hoods are required to contain and expel noxious and hazardous fumes from chemical reagents the hood should never be operated with the sash fully opened, and a maximum operating sash height should be established and conspicuously marked. Periodically evaluate ventilation by measuring face velocity with a calibrated velocity meter 111 Class velocity at the face of the hood should be 100 to 120 feet per minute and fairly uniform across the entire opening. 16 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT PERSONNEL PROTECTIVE EQUIPMENT (PPE) Bloodborne pathogens Safety glasses, goggles, visors, or work shields → protect the eyes and face from splashes and o Exposure Control plan – to minimize bloodborne pathogen exposure; The plan must be impact. available to all employees whose duties may result in reasonably anticipated occupational o Contact lenses do not offer eye protection; it is strongly recommended that they not be exposure to blood or other potentially infectious materials. worn in the clinical chemistry laboratory, unless additional protective eyewear is also Airborne pathogens utilized. o A TB exposure control program must be established, and risks to laboratory workers must o If any solution is accidentally splashed into the eye(s), thorough irrigation is required. be assessed. Gloves and rubberized sleeves → protect the hands and arms when using caustic chemicals. o The CDC guidelines require the development of a tuberculosis infection control program o Gloves are required for routine laboratory use by any facility involved in the diagnosis or treatment of cases of confirmed infectious TB. Laboratory coats, preferably with knit-cuffed sleeves, should be full length and buttoned and o TB isolation areas with specific ventilation controls must be established in health-care made of liquid-resistant material. facilities. Proper footwear is required o Those workers in high-risk areas may be required to wear a respirator for protection. o shoes constructed of porous materials, open-toed shoes, and sandals are considered o All health-care workers considered to be at risk must be screened for TB infection ineffective against spilled hazardous liquids FIRE SAFETY Respirators may be required for various procedures in the clinical laboratory. CLASS COMBUSTIBLE MATERIAL TYPE OF COMMENTS o Respirators with high-efficiency particulate air (HEPA) filters must be worn when OF EXTINGUISHER engineering controls are not feasible (ex: working directly with patients with tuberculosis FIRE TB) A Ordinary combustible solid Pressurized water, Dry ordinarycombustibles All contaminated PPE must be removed and properly cleaned or disposed of before leaving the Ardinart materials:: wood, paper, fabric/ chemicals & loaded laboratory cloth, plastic steam DONNING & DOFFING B Dry chemicals, CO2, Do not use water to DONNING Gown → masks/ respirator → goggles/ face shield → gloves Flammable/Combustible mammableliquid gases liquids halon extinguish Basa & gases:: petroleum products, DOFFING Gloves → Goggles/ face shield → gown → mask or respirator → perform hand washing gasoline, thinners, greases C LiveElectric Electrical equipment (Ex: Dry chemicals, CO2, Never use water, Dry HANDWASHING Live Equipment Computers, electrical wiring) halon chemicals may damage o Crucial component of infection control and chemical hygiene curyente electrical equipment o Use soap and warm water CO2 leaves no residue o Time: at least 20 seconds and is a good choice for o Important step: mechanical friction computers & analyzers D Sand or ceramic Leave to professional BIOLOGIC SAFETY Combustible/reactive Combustiblemetals metals: barrier; Metal X fire fighters; only try & All blood samples and other body fluids should be collected, transported, handled, and DElements Magnesium, sodium, potassium, isolate burning metal processed using universal precautions (i.e., presumed to be infectious). Aluminum from combustible Finely dispersed aerosols from centrifugation of biologic specimens are high-risk source of surfaces with sand or infection ceramic barrier o ideally, specimens should remain capped during centrifugation, or several minutes should E Arsenal fire Arsenal None (do not attempt Allowed to burn out and Ersena cannot be put out or is liable to to extinguish) nearby materials are be allowed to elapse after centrifugation is complete before opening the lid result in detonation protected o As a preferred option, the use of a sealed-cup centrifuge is recommended K Cooking media: cooking oil, Liquid designed to SPILLS vegetable, animal oil, fats prevent splashing and 1. Alert others in area of the spill. Kusina cool the fire 2. Wear appropriate protective equipment. MULTIPURPOSE DRY CHEMICAL FIRE EXTINGUISHER – CLASS ABC 3. Use mechanical devices to pick up broken glass or other sharp objects. ABC CO2 FIRE EXTINGUISHER – CLASS BC 4. Absorb the spill with paper towels, gauze pads, or tissue. Halogenated hydrocarbon extinguishers – recommended for use with computer equipment 5. Clean the spill site using a common aqueous detergent. Inspect fire extinguishers monthly to ensure they are mounted, visible, accessible and 6. Disinfect the spill site using approved disinfectant or 10% bleach, using appropriate contact charged time. 7. Rinse the spill site with water. 8. Dispose of all materials in appropriate biohazard containers 17 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT  NFPA HAZARD DIAMOND Developed a standard hazard identification system (diamond-shaped, color-coded symbol) RMT Be YOU will REMEMBER: You Were Born Ready!!! Hazard Color 0 1 2 3 4 HEALTH Blue No can cause Can cause can cause Can be Health (left) hazard significant temporary serious or lethal irritation incapacitation permanent or residual injury when a fire is discovered, all employees are Operation of the fire extinguisher “PASS” injury expected to take the action in the acronym FLAMMABILITY Red Will not Must be must be can be Will RACE Flamability (top) burn preheated heated or in ignited vaporize RESCUE – rescue anyone in immediate danger PULL pin for ignition high ambient under & burn ALARM – activate the institutional fire alarm AIM nozzle to occur temp to burn almost all at system SQUEEZE trigger ambient normal CONTAIN – close all doors to potentially SWEEP nozzle side to side temps temp affected areas INSTABILITY Yellow Stable High temp Violent May may Reactivity EXTINGUISH/ EXIT- if possible attempt to (right) makes chemical explode explode extinguish the fire; exit the area unstable change at from high at high temp or temp or normal pressures shock temp & pressure SPECIAL White OXY – oxidizer; ACID- Acid; ALK – Alkaline; COR- Corrosive; Hazard special HAZARDS (bottom) Radiation, W=use no water 0 No hazard 1 Slight hazard 2 Moderate hazard Remember: No SMS Ex 3 Serious hazard (extremely danger) 4 Extreme HAZARD (deadly) 18 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT CHEMICAL SAFETY Other hazards: SDS/ MSDS – Safety Data Sheet o a major source of safety information for employees who may use hazardous materials in Other hazard their occupation ELECTRICAL potential hazards associated with the use of electrical appliances and o employers are responsible for obtaining the SDS from the chemical manufacturer HAZARDS equipment Chemical Hygiene plan Electrical direct hazards: death, shock, or burns o A plan that provides procedures and work practices for regulating and reducing exposure Indirect hazards: fire or explosion Hazard Precautionary procedures: of lab personnel to hazardous chemicals o Use only explosion-rated (intrinsically wired) equipment in hazardous Chemical hygiene officer atmospheres. o Must be designated for any lab using hazardous chemicals o Be particularly careful when operating high-voltage equipment, such Storage & Handling Chemicals as electrophoresis apparatus. o Develop respect for all chemicals and have complete knowledge of their properties o Use only properly grounded equipment (three-prong plug). o Store according to their chemical properties and classification; arrangement depends on o Check for frayed electrical cords. quantities of chemicals needed and the nature or type of chemicals o Promptly report any malfunctions or equipment producing a “tingle” o Storage should NOT be based solely on alphabetical order for repair. o The storeroom should be organized so that each class of chemicals is isolated in an area o Do not work on “live” electrical equipment that is not used for routine work COMPRESSED Combination of hazards: danger of fire, explosion, asphyxiation, or GASES compressed mechanical injuries Storage requirements HAZARDS Requirements for safety handling: Gases o Know the gas that you will use. SUBSTANCE STORED SEPARATELY Flammable liquids Flammable solids Hazard o Store tanks in a vertical position. Mineral acids Organic acids o Keep cylinders secured at all times. o Never store flammable liquids & compressed gases in the same area. Caustics Oxidizers o Use the proper regulator, tubing, & fittings for the type of gas in use. Perchloric acid Water-reactive substances o Do not attempt to control or shut off gas flow with the pressure relief Air-reactive substances Others regulator. Heat-reactive substances requiring CRYOGENIC Liquid nitrogen - one of the most widely used cryogenic fluids (liquefied refrigeration cryogenic MATERIALS gases) in the laboratory Unstable substances (shock-sensitive HAZARDS material Hazards: fire or explosion, asphyxiation, pressure buildup, embrittlement of explosives) materials, and tissue damage similar to that of thermal burns. ERGONOMIC physical actions can, over time, contribute to repetitive strain disorders 1. Flammable/ Among the most hazardous materials in the clin chem lab Ergonomic HAZARDS such as tenosynovitis, bursitis, and ganglion cysts. Combustible Classified according to flash point: the temp at which sufficient vapor is Hazard factors associated with repetitive strain disorders: position/posture, chemicals given off to form ignitable mixture with air applied force, and frequency of repetition Flammable liquid – has a flash point below 37.8oC (100oF) Combustible liquid – flash point at or above 37.8oC (100oF 2. Corrosive injurious to the skin or eyes by direct contact or to the tissue of the chemicals respiratory and gastrointestinal tracts if inhaled or ingested Ex: acetic acid, sulfuric acid, nitric & hydrochloric acids; ammonium hydroxide, potassium hydroxide, sodium hydroxide external exposures to concentrated corrosives can cause severe burns and require immediate flushing with COPIOUS amounts of CLEAN WATER 3. Reactive substances that, under certain conditions, can spontaneously explode or chemicals ignite or that evolve heat or flammable or explosive gases. 4. Carcinogenic substances that have been determined to be cancer-causing agents pressurized containers chemicals if a CHEMICAL SPILL occurs: 1. assist/ evacuate personnel 2. confinement and cleanup of the spill 19 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT  Environmental factors associated with evacuated Blood collection tubes SPECIMEN COLLECTION & HANDLING o Ambient temperature, altitude, humidity, light BLOOD COLLECTION EQUIPMENT Ambient Inverse______ inverse relationship Nonsterile, disposable latex, nitrile, neoprene, polyethylene, and vinyl examination gloves are Temperature low temp = increase draw vol high temp = reduced draw vol acceptable for phlebotomy procedures Altitude Inverse______ relationship ANTISEPTICS inverse High altitude = lower draw vol 70% isopropyl alcohol Most commonly used antiseptic used for routine blood collection Humidity Diff. humidity conditions can affect only plastic evacuated Benzalkonium chloride Used for skin cleansing for ethanol testing iiii.fiiiiiiiiiie tubes due to greater permeability to water vapor relative to (zephiran) glass 70% Alcohol followed by Iodophor s CtAD Most commonly used before drawing blood for blood culture Light CTA inmettidamone is a special additive mixture for coagulation testing that is sensitive to light and found only in glass Chlorhexidine Recommended skin disinfectant by CLSI for blood culture, infants evacuated tubes gluconate Gluconate Chlorhexidine 2 months and older and patients with iodine sensitivity EDTA AC of choice for hematology cell counts and cell morphology TOURNIQUET 3to4 inches the above site MOA: Chelates (binds) calcium recommended test purpose Restrict venous flow to distend or inflate the veins, making them larger Lavender-top (KEDTA: plastic,by spray-dried, KEDTA: Liquid form in glass and easier to find, and it stretches the vein walls so they are thinner tubes) and easier to pierce with a needle Pink tubes: used In Immunohematology for ABO grouping. Rh typing, and Time Should not be longer than 1 minute (60 seconds) antibody screening Prolonged Hemoconcentration = ↑ enzymes, proteins, protein-bound substances White-top tubes: contain EDTA and gel; most often used for molecular application (cholesterol, triglycerides, calcium), potassium blood a nalytes diagnostics PCR DNA Note Incorrect application and fist exercise can result in erroneous test Sodium citrate MOA: Chelates (binds) calcium results Light blue-top tube containing 0.105 M orAmmon1.29 M (3.2% or 3.8% Use of tourniquet in lactate determination may result in falsely ↑ results respectively) sodium citrate Blood pressure cuff as tourniquet: 40 to 60 mmHg o used for coagulation testing because it preserves the labile Standard: 60 mmHg coagulation factors Obese patient: 40 mmHg o Blood: anticoagulant ratio = 9:1 Black-top: contain buffered sodium citrate NEEDLES o used for Westergren ESR PARTS Bevel: end that pierces the vein; allows the needle to easily slip into the o Blood: anticoagulant ratio = 4:1 skin and vein without coring (removing a portion of the skin or vein) Heparin Effective anticoagulant in small quantities without significant effect on Shaft: Long cylindrical portion many determination (does not affect levels of lons such as calcium) most commonly Hub: end that attaches to the blood collection device usedanticoagulant MOA: Accelerates the action of antithrombin III, neutralizing thrombin Lumen: internal space of the needle inchemistry and preventing the formation of fibrin Green-top tube Needle Indicated by a number that is related to the diameter of the lumen gauge (inverse relationship) Lithium heparin = used for most chemistry tests except lithium and 21-gauge needle: standard for most routine adult antecubital folate venipuncture control Sodium heparin = not used for sodium assays, but recommended for Length: 1-for better and 1.5-inch long needles are most commonly used trace elements, leads, and toxicology (Injectable form) Sodium Gray-top: used for glucose measurements because of It antiglycolytic fluoride properties preventing glycolysis for 3 days EVACUATED TUBE SYSTEM (ETS): most common and efficient system preferred by the CLSI In bacterial septicemia, fluoride inhibition of glycolysis is nether adequate for collecting blood samples nor effective in preserving glucose Parts Multiple sample needles: ETS needles/Special blood-drawing needle; Longer The anticoagulant is potassium oxalate or Na2EDTA bevel pierces the vein, shorter end penetrates the tube stopper Clot tubes Red-top: used for most chemistry, blood bank, and Immunology assay Tube holder: clear, plastic, disposable cylinder with a small threaded opening Red/gray- and gold-top (SSTs): contains a clot activator and separation at one end (AKA hub) where the needle is screwed into it and a large opening gel: most often used for chemistry tests Evacuated tubes o Has thixotropic gel that can absorb certain drugs cuisine falsely Factors Shelf-life is defined by the stability of the additive and vacuum retention lowered results (Ex: Phenytoin, phenobarbital, lidocaine, quinidine, most evacuated tubes have at least 12-month shelf-life and carbamazepine) 20 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT  o Tubes containing gels are not used in BB or for immunologic testing Cephalic vein is second choice if the median cubital is unsuitable. Basilic vein is the = may Interfere with immunologic reactions third choice. It should not be chosen unless no other vein is more prominent due to its Time to clot close proximity to the brachial artery and nerves. o Clotting time of tubes using gel separators: clot approx 30 minutes o Tubes with clot activators: clot in 5 minutes CISI o Plain red-stoppered tubes with no additives: take about 60 minutes ORDER OF DRAW to clot completely ETS & EVACUATED TUBE DERMAL PUNCTURE 1. Blood culture tube (yellow) 1. Capillary blood gas Proper patient Identification – first step in sample collection 2. Coagulation sodium citrate tube (light blue) 2. Slide/ Blood smear Types of Specimens: 3. Serum tubes with or w/o clot activator or gel 3. EDTA tubes o Whole blood - plasma + cells separator 4. Other anticoagulated tubes o Plasma – liquid portion of anticoagulated blood 4. Heparin tubes with or without gel (green 5. Serum tubes o Serum – liquid portion of clotted blood stopper) “BSEON” 5. EDTA tubes (lavender/ pink) o Urine – usually 24 hours 6. Glycolytic inhibitor tubes (gray) o CSF – for glucose and total protein analysis, a blood sample should be analyzed concurrently o Other body fluids: serous fluid, amniotic fluid GENERAL METHODS OF BLOOD COLLECTION 1. ARTERIAL PUNCTURE (arterial blood) A process by which blood is obtained from a patient’s artery Arterial blood – is the oxygenated blood with a bright red color mottled Use: for blood gas analysis (pO2, pCO2), and pH measurement Heparin: preferred anticoagulant Modified Allen A tTest llenest– done before blood is collected from radial artery to determine whether non a nticoag Modified the ulnar cc Site: Radial arter RadialArtery y (most preferred), brachial, femoral, scalp, umbilical - Indwelling umbilical artery: best site for blood gas analysis wanticoag 2. SKIN PUNCTURE (capillary blood) Simple method by which to collect blood samples in pediatric patients (for routine assays requiring small amounts of blood) In the neonate: skin puncture of the heel is the preferred site to collect a blood sample In older children: finger is the preferred site Skin puncture is useful in adults with extreme obesity, severe burns, and thrombotic tendencies, with POCT or with patients performing tests at home (blood glucose) Often preferred in geriatric patients because the skin is thinner and less elastic (venipuncture will be more prone to hematoma) In newborns: frequently used to collect sample for newborn screening tests for Inherited metabolic disorders shouldnotexceed2mm - Deep heel prick is made at the distal edge of the calcanea protuberance following a 5- to 10- minute exposure period to prewired water 3. VENIPUNCTURE (venous blood) a process by which blood is obtained from a patient’s vein Venous blood is the deoxygenated blood with dark red color Sites for venipuncture: antecubital fossa region, veins on the wrist and dorsal aspect of the hands, veins on the ankle Median median ccubital ubital vein: best site for venipuncture because it is the largest and the best anchored vein 21 | P a g e | Prepared by: Maria Kristina D. Gaquit, RMT