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6 succesive values falling on either side of the mean
6 succesive values falling on one side of the mean or the other each successivly farther away from the mean.
- Carbon Dioxide
- Calcium (Ca)
- Phosphorus (PO4-3)
- Magnesium (Mg)
Increased aldosterone concentration will do what?
- Increase NA concentration
- Decrease K concentration
Increased NA cause
- CUSHING’S SYNDROME
- SEVERE DEHYDRATION
- DIABETIC COMA AFTER INSULIN THERAPY
Decreased NA causes
- METABOLIC ACIDOSIS
- ADDISON’S DISEASE
- RENAL TUBULAR DISEASE
Preferred method and specimen type for Na?
- Ion selective electrode
- Glass ion exchange membrane selective for Na
Increased K causes
- URINARY OBSTRUCTION
- RENAL FAILURE DUE TO SHOCK
- RENAL TUBULAR ACIDOSIS
- ADDISON’S DISEASE (HYPOADRENALISM)
Decreased K causes
- INADEQ INTAKE OF K+
- EXCESSIVE USE OF DIURETICS
- CUSHINGS SYNDROME (HYPERADRENALISM)
K preferred fluid and method
ISE method vanomycin membrane selective for K
Increased Chloride causes
- Any condition causing decreased renal blood flow
- Sever renal tubular disease
Decreased chloride causes
- Salt losing nephritis
- Addison's disease (hypoadrenalism)
- Metabolic Acidosis
- Prolonged Vomiting
Preffered speciman and method for testing Chloride (Cl-)
Serum or heparinized plasma
- Cl sensitive ion exchange membrane that selectively binds Cl
- Tirtration methods
- Mercuric nitrate titration with diphenyl carbazone as indicator end point is excess of mercury.
Coulormetric-Amperometric titration with silver ions to form AgCl; endpoint in excess silver.
Carbon Dioxide dissolves in extracellular fluids to form what?
Carbonic Acid (H2CO3)
Increased CO2 causes
- Cardiac Disease
- Hypoventilation caused by drugs
- General Anesthesia
- CNS Trauma
- Compensation for increased HCO3 concentration
Decreased CO2 causes
- High external temps
- Hysteria anxiety
- Salicylate use of mechanical ventilation
- Compensation for decreased HCO3
Carbon Dioxide speciman collection methods
- Preffered arteriol whole blood
- Anaerobic conditions otherwise decreased CO2, increased O2
- STAT otherwise increased CO2 and Decreeased O2
- On Ice otherwise increased CO2 and Decreased pH
Carbon Dioxide preffered method
- Modified pH electrode
- CO2 diffuses across glass membrane into the chamber with enclosed buffer
pH change in buffer is measured by internal pH electrode
Arterial Blood gasses
Resperitory Opposites Metabolic Equal
Resperitory Opposites Metabolic Equal
Primary HCO3 deficit caused by?
- Uncontrolled Diabetes
- Renal Disease
- Aspirin/ Methanol/ Ethylene glycol poisoning
Lab results for Metabolic acidosis
Decreased pH, CO2, and HCO3
Resporitory alkalosis causes
- Fever and High external temps
- Hysteria, anxiety
- Excessive use of mechanical ventilation
Respiratory alkalosis lab results
- increased pH
- Decreased CO2
- Decreased HCO3
Calcium Ions do what?
- DECREASE MUSCULAR EXCITABILITY
- DECREASE CELL MEMBRANE PERMEABILITY
- TRANSMIT NERVE IMPULSES
- BLOOD COAGULATION
- ENZYME ACTIVATOR
Increased Ca causes?
- PRIMARY HYPERPARATHYROIDISM
- HYPERVITAMINOSIS (VIT D)
- MULTIPLE MYELOMA
- NEOPLASTIC BONE DISEASE
Decreased Ca causes?
- NEPHROSIS; NEPHRITIS
Ca collection anticoagulant?
- Serum or heparinized plasma preffered
- Most anticoagulants not sutable due to the chelation of calcium to prevent clotting
ISE and O-Cresolphthalein complexone
- O-CRESOLPHTHALEIN FORMS A COLORED COMPLEX WITH Ca+2 AT pH of 10 – 12; MEASURED SPECTROPHOTOMETRICALLY
- 8-HYDROXYQUINOLONE IS ADDED TO REMOVE INTERFERING SUBSTANCES LIKE PO4 & Mg
When is phosphorus monitered?
- After infusion of dextrose solutions
- Nutritional recovery syndrome
- Alchoholic withdrawal
- to evaluate kidney function
Methods for Mg measurment.
- Titan Yellow
- Methyl Thymol Blue
Dietary Fe+3 converted to Fe+2 by ______ then absorbed by________?
70% of Fe+2 used in what?
Ferrous Iron is what?
Ferric Iron is what?
Normal serum iron range
- 50-150 ug/dl in adults
- lower in adults over 50
TIBC normal range
Normal iron % saturation
20-45 in adults
Serum Iron Ferritin normal ranges
- 90 ug/L in males
- 20-250 ug/L in females
How to calculate percent saturation?
Serum Fe / TIBC x 100 = % Saturation
- Unsaturated Iron Binding Capacity (UIBC)
- Unoccupied binding sites for the Fe on the transferrin molecule.
Blood urea nitrogen is?
- Waste product of protein metabolism
- Made from ammonia (N3) in the liver.
- BUN is filtered by the glomerulus 40% reabsorption by the renal tubules.
BUN decreased causes?
Seen only in instage liver disease.
BUN Increased causes?
Prerenal: Cardiac compensation, dehydration, increased protein catabolism.
Renal: Acute Glomerulonephritis, Chronic Nephritis, Polycystic Kidney, Nephrosclerosis, Tubular Necrosis.
Postrenal: Obstruction of urinary tract. Stones, tumors, prostate disease.
MEASUREMENT OF BUN
- FEARON REACTION REACTION
- HEATED TO 100 C FOR 10 MINS
- DIRECT REACTION OF UREA WITH DIACETYL MONOXIME TO FORM COLORED DIAZINE DERIVATIVE
ENZYMATIC USING UREASE
- 100% filtered by kidneys
- 0% reabsorbed
- Formed by dehydration of creatine phosphate (made in the liver stored in muscle)
- muscle energy source
Renal Disease specifically affecting glomerular function.
- Due to low normal range (0.5-1.4)
- Low creatinine is not significant.
- JAFFE METHOD
- REAGENT: Alkaline picrate solution
Uric acid formation?
Breakdown product of nucleic acid metabolism
Uric acid significance
Indicates renal disease but rises only after Bun and Creatinine are already elevated.
Diagnosis of gout
Measurement of Uric Acid
URIC ACID + PTA + Na2CO3 ------> CO2 + ALLANTOIN + TUNGSTEN BLUE
Urea Clearance tells what?
- Measure of overall kidney function
- 100% filtered with 60% reabsorbed.
Creatinine clearance tells what?
- Measure of glomerular function
- 100% filtered
- 0% reabsorbed
Hormones that increease blood glucose
- Growth Hormone
Diabetes Mellitus Type 1
- AKA Juvenile Diabetes
- 10% of known diabetics
- Prone to ketosis
- Abrupt Onset
- Genetic Inheritance
- Will die without insulin.
- Inslet antibodies present in most
DIABETES MELLITUS: TYPE II
- Adult Onset
- 80% of all Diabetics
- 12% of entire population
- Not prone to Ketosis
- Insulin may or may not be necessary.
- INSULIN INDUCED IN KNOWN DIABETICS
- HEPATIC DISEASE
- IDIOPATHIC HYPOGLYCEMIA IN INFANTS
ENDOCRINE DISORDERS: PANHYPOPITUITARISM, HYPOTHYROIDISM, HYPOADRENALISM
GLUCOSE OXIDASE METHOD
- Glucose oxidase
- Glucose + O2 + H2O ----> Glucuronic acid & H2O2
- H2O2 + O-dianisidine -----> oxidized O-dianisidine + water
GLUCOSE METHODS HEXOKINASE
Glucose + ATP ----------> Glucose-6-phosphate & ADP
Glucose-6-phosphate +NAD -----> 6-phosphogluconate + NADH
O2 consumption Glucose method
O2 depleted after the first step of the glucose oxidase reaction is proportional to glucose concentration.
HA1C determines what?
Average glucose conc. that cells have been exposed to over past 2 months
Increased HA1C means what?
Can detect uncontrolled diabetics who are compliant with treatment only immediatly prior to Dr. visit.
Available HA1C methods
- Cation-exchange chromatography
- Iso-electric focusing
Total protein Functions
- pH Buffers
- Maintain Osmotic pressure
- Synthesis of body tissues
Increased total protein means
- Prolonged Stasis during venipuncture
- Chronic disease state e.g.
- SARCOIDOSIS, CIRRHOSIS, COLLAGEN DISEASES, CHRONIC INFLAMMATORY CONDITIONS
Decreased total protein means?
- Nephrotic Syndrome
- Extensive Burns
- Protein Losing Lesions in GI tract
- Dietary protein deficiency
- Malabsorption syndromes
- Liver disease
Measurement of total protein
3 CONH + ALK CuSO4 --> Chelated Cu complex
*Chelated Cu complex has violet blue color
- Made almost exclusivly by the liver
- Supports osmotic pressure
- Helps control water distribution in the body
- Transports small molecules
- Binds substances: Bilirubin, calcium, free fatty acids, aspirin
Increased Albumin means what?
Seen only when there is a loss of plasma water (dehydration) or excessive IV albumin therepy
Decreased Albumin means?
- Low protein intake
- liver failure
- Increased protein loss
- reduced synthesis of proteins
- increased protein catabolism
- inadequate digestion or absorption of injested protein
Dye binding assays for the measurement of Albumin
- Binding of albumin to dye causes color change
- Methyl Orange
- 2,4-Hydroxy, Azobenzene-Benzoic Acid (HABA)
- Bromcresol Green
- Bromcresol Purple
Dye binding reaction color changes for Albumin
ALBUMIN + INDICATOR DYE -----> ALBUMIN-DYE COMPLEX
- BCG Yellow--->Green
- BCP Yellow---->Purple
Globulin formula from total protein
TP Value - Albumin Value= Globulin
Albumin globuin ration
Albumin / Globulin
Protein Electrophoresis migration order
(+) Albumin, Alpha 1, Alpha 2, Beta, Gamma (-)
Bilirubin is formed how?
- Hemoglobin Metabolism
- Hemoglobin Breakdown
Billirubin forms from broken down heme then excreted through the liver.
What happens to the iron from broken down HGB
Recycled into new HGB
What happnes to protein leftover from the breakdown of HGB
Recycled into the amino acid pool
Progression of Hemoglobin breakdown.
- Free protein bound Bilirubin--->
- Conjugated Bilirubin (Liver-->bile duct)--->
- Urobilinogen by bacterial intestinal flora--->
- Feces and kidney/urine
- processed through liver and conjugated with glucuronic acid
- can be excreted through kidneys; non toxic
- Most likely cause of Jaundice
- Water Insoluble
- Bound to albumin for transport
Increased Total Bilirubin
Prehepatic Conditions- Hemolysis
Hepatic Conditions-anything that causes damage to the liver
Post hepatic conditions- Gallstones, neoplasms, bile duct obstructions.
Decreased total bilirubin means
Not a damn thing
Total Bilirubin methods
- Based Van Den Bergh Reactions
- Jendrassik-Grof Method
- Evelyn- Malloy Method
Based Van Den Bergh Reaction
Measurement of bilirubin
BILIRUBIN + SULFANILIC ACID + HCl + NaNO2---> AZOBILIRUBIN
Jendrassik - Grof Method
For measurement of Bilirubin
USES CAFFEINE BENZOATE TO MAKE DIRECT & INDIRECT BILI WATER SOLUBLE; ALKALINE TARTRATE SHIFTS pH TO ALKALINE (BLUE COLORED COMPLEX)
USES METHYL ALCOHOL TO MAKE ALL TYPES OF BILI WATER SOLUBLE; END PRODUCT READ AT ACID pH
TOTAL BILI REACTION
TBILI + DIAZO RGT + ALCOHOL OR CAFFEINE ----> AZOBILIRUBIN
- IF pH ACIDIC AZOBILIRUBIN IS RED
- IF pH ALKALINE AZOBILIRUBIN IS BLUE
DIRECT BILI REACTION
TBILI + DIAZO RGT ----> AZOBILIRUBIN
- IF pH ACIDIC AZOBILIRUBIN IS RED
- IF pH ALKALINE AZOBILIRUBIN IS BLUE
Formation of ammonia
Formed in the liver from removal of NH2 amino acids.
Detoxified by conversion to urea.
Normal Ammonia range
Critical range and meaning
- >400 ug/dl
- reyes's syndrome & impending coma
UV KINETIC (DECREASE ABS) method
Measurement of Ammonia (NH3)
- NH3 + ALPHA KG + NADH -----> NAD + H2O + GLUTAMATE
- Glutamate Dehydrogenase= GDH
AST is what?
- AST catalyzes transfer of an amino group
- Found primarily in liver, heart & skeletal, and kidney
Markedly increased AST mean what?
AMI (24-36 hours after onset of pain)
- Viral Hepatitis
- Toxic liver necrosis
Moderatly elevated AST means what?
- Skeletal Muscle Disease
- After Trauma or Surgery
- Severe Hemolytic Anemia
Not commonly seen
Uremia and Vit B12 deficiencies may cause low values
AST measurement methods
Coupled enzymatic reaction
L-aspartate + alpha-oxoglutarate -- Oxaloacetate + L-glutamate
Oxaloacetate + NADH + H+ -- Malate + NAD
ALP is what?
- Alkaline Phosphotase
- Plays role in membrane transport of molecules
- Group of several enzymes that catalyzes transfer of PO4 (phosphate) group in an alkaline medium.
- Primarily related to liver and bone disease.
10 X increase in ALP means what?
Obstructive Hepatobiliary tract disorders
moderate 1-3 x increase in ALP means what?
Increase in ALP values not caused by Hepatobiliary tract disorders
- Osteoblastic cells of bones:
- Paget's Disease, Ricketts
- In children, adolescents, healing bone fractures
Decreased ALP (Alkaline Phosphotase)
- Severe anemia
ALP does what at room temp?
Based on Bessey, Lowery, Brock method
P-Nitrophenylphosphate changes to p-nitrophenol a yellow color deveopes and is measured at 405 um
ALT is what and does what?
- Alanine Transferase
- ALT catalyzes transfer of amino group
What is the ALT co-enzyme?
- needed for full enzyme activity
What liver enzyme is more specific than AST?
ALT (Alanine Transferase)
ALT (alanine transferase) methods
Coupled enzymatic reaction
Alanine, pyruvate, L glutamate, lactate,
measured at 340 nm
What is CK?
- Creatine Kinase
- CK is a cytoplasmic & mitochondrial enzyme that catalyzes phosphorylation
- Mg required for full enzyme activity
CK is elevated when?
- Predominatly in diseases of skeletal and cardiac muscle.
- Used to diagnose AMI
CK levels after AMI?
- Begin to rise 4-12 hours
- Peak levels at 24 hours
- Returns to normal after 3 days
- Not specific for MI but rises with other cardiac conditions
- Typical pattern
- 4-8 hours
- peak 15-24
- normal 48-72
- Coupled Enzymatic reaction
- End product Phosphogluconate
Troponin is what?
- Regulates muscle contractions
- Specific for cardiac necrosis and is helpful when CK-MB has already returned to normal.
Prognosis of elevated Troponin
Elevated Troponin levels in time
- 4 hours within onset of pain
- remains elevated for 10-24 days
Myglobin useful for what?
- Highly sensitive marker for AMI
- Rises 2-5 hours after Infarc
- peaks at 12 hours
- Cleared by kidneys
- Not specific for cardiac muscle
Amylase main function?
- Aids in digestion of carbohydrates
- First salivary amylase
- then pancreatic
Amylase meaning and activity time frame
Used for the diagnosis of acute pancreatitis
- Rises 2-12 hours after onset of pain
- PEaks at 24 hours
Lipase useful for what?
- Diagnosis of acute pancreatitis when amylase levels have returned to normal
- Remains elevated for up to 14 days
GGT is what and indicates what?
- Gamma-glutamyl Transferase
- Serves as an early indicator of liver disease.
- especially due to alcohol damage
- Also obsturctive liver disease in children or during pregnancy
Increased Lactic dehydrogenase (LD) can mean
- Hemolyzed sample
- Hematological disorders
- Hapatitis/ Liver Disease
- Acute Renal disease
Cardiac Enzyme Profile Tests
Liver Enzyme Profile Tests
Pancreatic enzyme profile
- packaged for removal by the liver
- Leiberman-Burchardt (Fat Burchardt)
- Sulfuric Acidd
- Acetic acid
- Green end product
What are triglycerides?
Fatty acids are stored in cells as tryglycerides
Lipid Electrophoresis order
- Anode to cathode
- HDL, VLDL, LDL, Chylomicrons
- Acidification release to change Fe+3 to Fe+2
- Complexes with color reagent
UIBC is what?
- Unsaturated Iron Binding Capacity
- Part of anemia work-up
- Asses iron binding sites not already bound with iron
UIBC calculated how?
Difference between Serum Iron concentration and TIBC
Thyroid Hormones regulate what?
- Carbohydrate, Lypid and protein metabolism
- CNS development
- Bone & Tissue Growth development
- GI function
- Sexual Maturation
What are the thyroid hormones and what are they used for?
- T3= triiodothyronine
- T4= Thyroxin
- Used to determine the extent of hyperthyroidism or hypo...
- t4 found in higher concentration than T3
Increased T3 & 4 means what?
- Graves disease
- toxic multinodular goiter
- Pituitary tumors
Decreased T3 and 4 means
- Hashimoto's thyroiditis
- Pituitary or Hypothalmic disease
- Spontaneus hypothyroidism
First and best test for thyroid abnormalities?
- uses 3 seperate monocolonal antibodies to give test high specificity
Excess of cortisol?
Addison's disease lab results
- Lack of cortisol
- LAck of aldosterone
- Decreased NA
- Increased K
- Increased BUN
- Decreased Glucose