[LAB] ALP & ACP Flashcards
RECOMMENDED NAME OF ALP
ALKALINE PHOSPHATASE
EC CODE OF ALP
3.1.3.1
OTHER TERMS FOR ALP
PHOSPHOMONOESTERASE
ORTHOPHOSPHERIC ESTER MONOHYDROLASE
TWO MAIN TYPES OF PHOSPHATASES
ALKALINE PHOSPHATASE
ACID PHOSPHATASE
OPTIMUM PH OF THE ALKALINE PHOSPHATASE
PH 9.0
WHICH TYPE OF PHOSPHATASE HAS AN OPTIMUM PH OF 9.0
ALKALINE PHOSPHATASE
CLINICAL APPLICATIONS OF SERUM ALP ASSAYS
DIAGNOSIS OF HEPATOBILIARY AND BONE DISEASES
HIGHEST ELEVATION OF ALP IS SEEN IN WHAT DISORDEER
PAGET’S DISEASE
BONE DISORDERS ASSOCIATED WITH ELEVATED ALP
OSTEOMALACIA
RICKETS
OSTEOGENIC SARCOMA
ELEVATION OF ALP IS SIGNIFICANT IN THESE LIVER DISORDERS
BILIARY TREE OBSTRUCTION
MODERATE ELEVATIONS OF ALP ARE SEEN IN WHAT CLINICAL CONDITIONS
LIVER CELL CANCER
INFECTIOUS HEPATITIS
ALCOHOLIC HEPATITS
LIVER CIRRHOSIS
ALP DETERMINATION IS BASED ON WHAT CHEMICAL REACTION
HYDROLYSIS OF VARIOUS PHOSPHATE ESTERS UNDER SPECIFIED CONDITIONS
PHOSPHATE ESTER IN THE ALP REACTION
P-NITROPHENYL PHOSPHATE
WHO INTRODUCED P-NITROPHENYL PHOSPHATE AS A SUBSTRATE
FUJITA IN 1939
WHO PUBLISHED AN ENDPOINT PROCEDURE OF ALP
BESSEY
LOWRY
BROCK
IN 1966
WHO PUBLISHED THE KINETIC PROCEDURE OF ALP ASSAYS
BOWERS & MCCOMG IN 1966
RECOMMENDEDALP ASSAY METHOD
BOWERS-MCCOMB KINETIC METHOD
ACTIVATORS OR COFACTORS IN ALP ASSAYS
MAGNESIUM
COBALT
MANGANESE
ZINC
MCMZ
SERVES AS THE CONSTITUENT METAL IN ALP ASSAYS
MAGNESIUM
COBALT
MANGANESE
ZINC
MCMZ
PHOSPHATE BUFFER ACCEPTOR
AMP (2-amino-2-methyl-1-propanol)
TRIS HYDROXYMETHYL AMINOMETHANE
DIETHANOLAMINE
AMP ACTS AS WHAT
IN THE ALP REACTION
PHOSPHATE BUFFER ACCEPTOR
TRIS HYDROXYMETHYL AMINOMETHANE ACTS AS WHAT IN THE ALP REACTION
PHOSPHATE BUFFER ACCEPTOR
DIETHANOLAMINE ACTS AS WHAT
IN THE ALP REACTION
PHOSPHATE BUFFER ACCEPTOR
WHO RECOMMENDED THE KINETIC METHOD
AACC
AMERICAN ASSOCIATION OF CLINICAL CHEMISTRY
PRINCIPLE OF ALP ASSAYS
P-NITROPHENYL PHOSPHATE + H2O —ALP—> P-NITROPHENOL + PHOSPHORIC ACID (H3PO4)
PH OF THE ALP REACTION
PH 10.3
ACCEPTOR SUBSTRATE IN THE ALP REACTION
AMP
COLOR OF HYDROLYZED P-NPP
YELLOW
RELATIONSHIP OF P-NPP HYDROLYSIS AND ALP ACTIVITY
DIRECTLY PROPORTIONAL
[SUBSTRATE]
SINOWARA-JONES-REINHART
BETA-GLYCEROPHOSPHATE
[SUBSTRATE]
KING-ARMSTRONG
PHENYL PHOSPHATE
[SUBSTRATE]
MOSS
ALPHA NAPTHOL PHOSPHATE
[SUBSTRATE]
KLEIN, BOBSON, AND READ
BUFFERED PHENOLPTHALEIN
ABSORBANCE OF P-NITROPHENOL IN ALP ASSAYS
400 NM
DISADVANTAGE OF THE SINOWARA-JONES REINHART METHOD
LONGER REACTION
REQUIRES A LONGER INCUBATION TIME
[METHOD]
PRODUCT IS P-NITROPHENOL, YELLOW
BESSEY, LOWRY, AND BROCK
METHOD THAT REQUIRES A LONGER INCUBATION TIME
SINOWARA-JONES-REINHART
METHOD THAT REQUIRES PROTEIN REMOVAL
KING-ARMSTRONG
DISADVANTAGE OF THE KING-ARMSTRONG METHOD
PROTEINS MAY INTERFERE WITH THE ASSAY
[PRODUCT]
HUGGINS AND TALALAY
PHENOLPTHALEIN
RED
[METHOD]
PRODUCT IS PHENOLPTHALEIN, RED
HUGGINS AND TALALAY
[PRODUCT]
MOSS
ALPHA NAPTHOL
[METHOD]
PRODUCT IS ALPHA NAPTHOL
MOSS
INTERFERENCES THAT CAUSE FALSELY INCREASED RESULTS
- HEMOLYZED SERUM
- UNFRESH SERUM SAMPLE
- SERUM THAT IS IN CONTACT WITH CLOT FOR A LONG TIME
- LIPEMIC OR ICTERIC SAMPLES
- BILIRUBIN
Hb CONCENTRATION THAT CAUSES A FALSE INCREASE
> 100 mg/dL
PERCENTAGE OF INCREASED ALP ACTIVITY WHEN SERUM SAMPLE IS UNSTABLE
3-10%
STORAGE CONDITION OF ALP SAMPLE
2-8C
UP TO 1 WEEK
TIME LIMIT FOR ALP SAMPLE TO BE ASSAYED
4 HOURS
ALP SAMPLE STORAGE CONDITION FOR ALP TO REACH FULL ENZYME REACTIVATION
THAWED
KEPT AT RT FOR 18-24 HOURS
BEFORE MEASUREMENT
PERCENTAGE OF ALP INCREASE HEN SAMPLE IS IN CONTACT WITH CLOT FOR A LONG TIME
20-30%
DUE TO GRADUAL DEVELOPMENT OF MORE BASIC PH AS CO2 IS LOST
REASON WHY PROLONGED CONTACT OF SERUM SAMPLE WITH CLOT FALSELY INCREASES ALP ACTIVITY
GRADUAL DEVELOPMENT OF A MORE BASIC PH IN THE SYSTEM AS CARBON DIOXIDE IS LOST
IS PLASMA USED IN ALP ASSAYS
NO
ANTICOAGULANTS INHIBIT ALP ACTIVITY
ANTICOAGULANTS THAT INHIBIT ALP ACTIVITY
CITRATE
OXALATE
EDTA
EFFECT OF ANTICOAGULANTS ON ALP ACTIVITY
FALSE DECREASE
INHIBITS ACTIVITY
BINDS WITH COFACTORS
WHAT DO THE ANTICOAGULANTS BIND WITH AND WHAT ARE THE EFFECTS
BIND WITH COFACTORS (MAGNESIUM AND ZINC)
CAUSE FALSE DECREASE IN ALP ACTIVITY
EFFECT OF LIPEMIC OR ICTERIC SAMPLES
FALSE INCREASE
BILIRUBIN CONCENTRATION THAT MAY INTERFERE WITH ALP ASSAY RESULTS
> 20 mg/dL
WHY ARE ALP VALUES HIGHER IN CHILDREN THAN ADULTS
DUE TO INCREASED OSTEOBLASTIC ACTIVITY
CAN THE ALP ASSAY DETERMINE THE ISOENZYME
NO
WHY IS THERE ANEED FOR A BUFFER IN THE REACTION
REACTION IS FASTER IF A PHOSPHATE ACCEPTOR IS PRESENT
INCREASES ENZYME ACTIVITY
SHORTENS REACTION TIME
REQUIRES A LESSER VOLUME OF SAMPLE
WHAT HAPPENS TO THE ALP REACTION IF A PHOSPHATE ACCEPTOR IS PRESENT
REACTION SPEEDS UP
INCREASES ENZYME ACTIVITY
SHORTENS REACTION TIME
REQUIRES A LESSER VOLUME OF SAMPLE
LINERAITY OF ALP ASSAY
1000 IU/L
TROUBLESHOOTING SAMPLES WITH VALUES EXCEEDING THE LINEARITY
DILUTE WITH EQUAL VOLUME OF SALINE
REASSAY
MULTIPLY RESULTS BY 2
ABSORBANCE TREND OF ALP ASSAY
INCREASING
WHY IS THE ABSORBANCE TREND INCREASING
INCREASED SUBSTRATE HYDROLYZED = INCREASED ABSORBANCE
NORMAL VALUES FOR ALP
32-123 IU/L
TRUE OR FALSE
NON SPECIFIC ACP IS WIDELY DISTRIBUTED THROUGHOUT THE BODY
TRUE
EC CODE OF ACP
3.1.3.2
MORE IMPORTANT ACP FRACTION
PROSTATIC ACP
OTHER TISSUE SOURCES OF ACP
BONE
LIVER
SPLEEN
KIDNEYS
ELEVATED ACP LEVELS ARE OBSERVED IN WHAT CLINICAL CONDITIONS
PAGET’S DISEASE
HYPERPARATHYROIDISM WITH SKELETAL INVOLVEMNT
BONE CANCERS
SUBSTRATES IN THE ACP REACTION
PHENYL PHOSPHATE
P-NITROPHENYL PHOSPHATE
THYMOPTHALEIN PHOSPHATE
SUBSTRATE PROPOSED BY BABSON
ALPHA-NAPTHYL PHOSPHATE
SUBSTRATE PROPOSED BY HILLMAN
DIAZOTIZED 2-AMINO-5-CHLOROTOLUENE (FAST RED TR)
FAST RED TR
DIAZOTIZED 2 AMINO-5-CHLOROTOLUENE
SAID THAT ALPHA-NAPTHYL PHOSPHATE CAN BEHYDROLYZED BY OTHER ENZYMES
AMADOR
ABSORBASE OF THE DIAZO DYE IN ACP ASSAYS
405 NM
SPECIFIC INHIBITOR IN ACP ASSAYS
L-TARTRATE
PRINCIPLE OF ACP ASSAY
Alpha-napthylphosphate + H2O —ACP—> Alpha-napthol + inorganic phosphate
Alpha-napthol = Fast Red TR —> Diazo Dye
PHOSPHATE BUFFER ACCEPTOR IN ACP
1,5-NAPTHALEIN DISULFONATE
1,5-NAPTHALEIN DISULFONATE ACTS AS WHAT
PHOSPHATE BUFFER ACCEPTOR IN ACP
RELATIONSHIP OF THE FORMATION OF RED COLORED COMPLEX WITH ACP ACTIVITY
DIRECTLY PROPORTIONAL
INHIBITOR IN THE ACP REACTION
L-TARTRATE
WHAT DOES L-TARTRATE DO
INHIBIT PROSTATIC ACP
BUT DOES NOT INTERFERE WITH THE REACTION MECHANISM
TRUE OR FALSE
L-TARTRATE INTERFERES WITH THE ACP REACTION
FALSE
IT ONLY INHIBITS PROSTATIC ACP
SUBSTRATE OF CHOICE FOR MOST ENDPOINT REACTIONS
THYMOLPTHALEIN MONOPHOSPHATE
THYMOLPTHALEIN MONOPHOSPHATE IS THE SUBSTRATE OF CHOICE FOR WHAT KIND OF ACP ASSAYS
ENDPOINT ASSAYS
ALPHA-NAPTHYLPHOSPHATE IS THE SUBSTRATE OF CHOICE FOR WHAT KIND OF ACP ASSAYS
CONTINUOUS MONITORING ASSAYS
SUBSTRATE OF CHOICE FOR CONTINUOUS MONITORING ASSAYS
ALPHA-NAPTHYLPHOSOHATE
IMMUNOCHEMICHAL TECHNIQUES FOR ACP
RADIOIMMUNOASSAY
IMMUNOPRECIPITATION
[SUBSTRATE AND PRODUCT]
BODANSKY
S: BETA-GLYCEROPHOSPHATE
P: GLYCEROL
[SUBSTRATE AND PRODUCT]
GUTMAN, KING, ARMSTRONG
S: PHENYLPHOSPHATE
P: PHENOL
[SUBSTRATE AND PRODUCT]
HUDSON
S: P-NITROPHENYLPHOSPHATE
P: P-NITROPHENOL
[SUBSTRATE AND PRODUCT]
BABSON AND REED
S: ALPHA-NAPTHYLPHOSPHATE
P: ALPHA-NAPTHOL
[SUBSTRATE AND PRODUCT]
ROY
S: THYMOPTHALEIN MONOPHOSPHATE
P: THYMOPTHALEIN
— Abs 590 nm
[SUBSTRATE AND PRODUCT]
RIETZ, GUILBAULT
S: 4-METHYLUMBERLLIFERONEPHOSPHATE
P: METHYLUMBERLLIFERONE
[OTHER CHARACTERISTICS]
NON SPECIFIC TO PROSTATIC ACP
BODANSKY
GUTMAN, KING, ARMSTRONG
HUDSON
[OTHER CHARACTERISTICS]
LENGTHY ASSAY
BODANSKY
[OTHER CHARACTERISTICS]
RAPID ASSAY
HUDSON
[OTHER CHARACTERISTICS]
LESS SENSITIVE TO PROSTATIC ACP
BABSON AND REED
[OTHER CHARACTERISTICS]
MOST SPECIFIC FOR PROSTATIC ACP
ROY
[OTHER CHARACTERISTICS]
LESS INTERFERENCES
ROY
[OTHER CHARACTERISTICS]
FLUORESCENCE METHOD
RIETZ, GUILBAULT
TRUE OR FALSE
ONLY NON HEMOLYZED SERUM SHOULD BE USED BECAUSE ACP IS CONTAINED IN RED BLOOD CELLS
TRUE
TIME LIMIT FOR THE SEPARATION OF THE SERUM FROM CLOT IN ACP ASSAY
WITHIN 2 HOURS
WHY SHOULD THE SERUM BE SEPARATED FROM THE CLOT IN ACP
PREVENT LEAKAGE OF ERYTHROCYTIC AND PLATELET ACP INTO THE SAMPLE
STABILIZATION CONDITIONS OF ACP
ACIDIFY WITH ACETATE BUFFER
20 UL OF BUFFER PER 1ML SERUM
TRUE OR FALSE
ACID PHOSPHATASE IS STABLE AT ROOM TEMPERATURE
FALSE
EXTREMELY LABILE AT ROOM TEMPERATURE
STABILITY OF ACP
7 DAYS AT 2-8C
ANTICOAGULANTS THAT INHIBIT ACP ACTIVITY
FLUORIDE
HEPARIN
OXALATE
INTERFERENCES IN ACP THAT CAUSE FALSE DECREASE
ANTICOAGULANTS (FLUORIDE, HEPARIN, OXALATE)
BILIRUBIN
[SUBSTRATE AND PRODUCT]
BESSEY, LOWRY, BROCK
S: P-NITROPHENYL PHOSPHATE
P: P-NITROPHENOL (YELLOW)
[SUBSTRATE AND PRODUCT]
HUGGINS AND TALALAY
S: PHENOLPTHALEIN DIPHOSPHATE
P: PHENOLPTHALEIN (RED)
[SUBSTRATE AND PRODUCT]
MOSS
S: ALPHA NAPTHOL PHOSPHATE
S: ALPHA NAPTHOL
PURPOSE OF ADDING L-LACTATE WHEN MEASURING NON PROSTATIC ACP
INHIBIT PROSTATIC
SUBTRACT NON PROS FROM TOTAL ACP
NORMAL VALUE OF ACP
TOTAL ACP: 0.9 IU/L
PROS ACP: 0-3 IU/L
NON PROS: 0-6 IU/L
WHAT TO DO IF NON LINEARITY IS OBSERVED
CAN DISREGARD THE FIRST OR LAST READING
DUE TO LAG PHASE AND SUBSTRATE DEPLETION PHASE
ONLY ELIMINATE ONE ABSORBANCE READING
WAVELENGTH USED FOR ALP AND ACP
405 NM
SI CONVERSION OF ALP AND ACP
16.67
MULTIPLIER OF ACP
TOTAL: 853
NON PROS: 860
REFERENCE METHOD OF ACP
KINETIC METHOD BY HILLMAN
REFERENCE METHOD OF ALP
BOWERS’MCCOMB KINETIC METHOD
LINEARITY OF ACP
60 U/L AT 37C
STABILITY OF RECONSTITUTED ACP REAGENT
1 DAY AT RT
7 DAYS AT 2-8C
STABILITY OF RECONSTITUTED L-TARTRATE REAGENT
UNTIL EXPIRY DATE AT 2-8C
PH OF AMP BUFFER
PH 10.45
[ALP ASSAY]
VOLUME OF ALP REAGENT
1 ML
[ALP ASSAY]
INCUBATION TIME ADN TEMPERATURE OF THE REAGENT
5 MINUTES
37C
[ALP ASSAY]
VOLUME OF PATIENT SAMPLE
0.025 ML
[ALP ASSAY]
INCUBATION CONDITION FOR SAMPLE
1 MINUTE
37C
IN BETWEEN READINGS
[ACP ASSAY]
VOLUME OF L-TARTRATE REAGENT
1 ML
[ACP ASSAY]
VOLUME OF ACETATE BUFFER
20 UL
[ACP ASSAY]
INCUBATION TIME AND TEMPERATURE IN BETWEEN READINGS
1 MINUTE
37C