FINALS LABORATORY TOPICS Flashcards
Routine or Commonly used total protein method
Biuret Quantitative Colorimetric Method
Principle of total protein?
Biuret Quantitative Colorimetric Method
Biuret Reaction
TOTAL PROTEIN
Volume of biuret reagent: ?
Volume of serum: ?
Volume of biuret reagent: 1 mL
Volume of serum: 10 uL
TOTAL PROTEIN
Total protein after addition of reagents should be left to stand at what temp and fow how many mins?
RT for 5 mins.
TOTAL PROTEIN
End Products and Color
Cuprous Protein Complex (purple/violet)
TOTAL PROTEIN
Absorbance of total protein
550 nm
TOTAL PROTEIN
SI unit of total protein?
g/L
TOTAL PROTEIN
How to convert g/dL to gL for total protein?
mutiply by 10
TOTAL PROTEIN
Reference value of total protein>
6.5 to 8.3 g/dL
TOTAL PROTEIN
more than 8.3 g/L indicates what condition for total protein?
Hyperproteinemia
Increased
TOTAL PROTEIN
less than 6.5 g/L indicates what condition for total protein?
Hypoproteinemia
Decreased
TOTAL PROTEIN
Reference method for total protein?
Kjeldahl Method
TOTAL PROTEIN
Principle of kjedahl method?
Polypeptide bonds + cupric ions in an alkaline medium
ALBUMIN
Most common/Routine method for albumin
Bromcresol green
ALBUMIN
Most sensitive method for albumin detection?
Bromcresol purple
ALBUMIN
Principle for albumin?
Bromcresol Green
Dye Binding
ALBUMIN
Volume of albumin reagent: ?
Volume of serum?
Volume of albumin reagent: 1 mL
Volume of serum: 10 uL
ALBUMIN
After addition of reagents, for what temp and how many minutes should sample be left to stand?
RT for 30 mins
ALBUMIN
End Products and Color of albumin?
Green-Colored Complex
ALBUMIN
Absorbance of albumin?
550 nm
ALBUMIN
How to convert g/dL to g/L for albumin?
mutiply by 10
ALBUMIN
Reference values?
3.5 to 5.5 g/dL
ALBUMIN
more than 5.5 g/dL indicates what clinical significance for albumin?
hyperalbuminemia
GLOBULIN
For getting globulin value, what formula should be used?
G = TP - Albumin
GLOBULIN
Reference value?
2.3 to 3.5 g/dL
What is the Albumin:Globulin Ratio?
1.3 - 3:1
Globulin
Higher globulin value in A:G ratio indicates what clinial signifiance?
with multiple myeloma/ Waldenstrom macroglobulinemia
(increased IgM and IgG)
BILIRUBIN
Method of bilirubin detection?
Modified Evelyn-Malloy Method
BILIRUBIN
Principle of bilirubin detection?
Diazotization
BILIRUBIN
Volume of bilirubin reagent: ?
Volume of serum: ?
Volume of antioxidant: ?
TOTAL BILIRUBIN
Volume of bilirubin reagent: 1 mL
Volume of serum: 50 ul
Volume of antioxidant: 1 drop
BILIRUBIN
In total bilirubin detection, what temp and how many minutes should sample be left to stand?
TOTAL BILIRUBIN
RT for 5 mins
BILIRUBIN
Volume of bilirubin reagent: ?
Volume of serum: ?
Volume of antioxidant: ?
DIRECT BILIRUBIN
Volume of bilirubin reagent: 1 mL
Volume of serum: 10 ul
Volume of antioxidant: 1 drop
BILIRUBIN
In Direct bilirubin detection, what temp and how many minutes should sample be left to stand?
DIRECT BILIRUBIN
RT for 3 mins
BILIRUBIN
End Products and Color
Azobilirubin (pink-purple)
BILIRUBIN
Absorbance
540 nm
BILIRUBIN
SI unit of bilirubin?
umol/L
BILIRUBIN
How to convert mg/dL to umol/L for bilirubin?
multiply by 17.1
BILIRUBIN
Reference value of total bilirubin?
TOTAL BILIRUBIN
0.05 mg/dL (0 to 88.5 μmol/L)
BILIRUBIN
Reference value of direct bilirubin?
DIRECT BILIRUBIN
0 to 1.20 mg/dL (0 to 20.52 μmol/L)
BILIRUBIN
Increased bilirubin indicates what clinical condition?
liver function
BILIRUBIN
T or F
Indirect bilirubin is acquired through computation in which (I = Total bilirubin - Direct bilirubin)
T
BILIRUBIN
Formula of Indirect bilirubin?
Indirect Bilirubin = TB - Direct Bilirubin
BILIRUBIN
sum of the direct + indirect bilirubin
Total bilirubin
BILIRUBIN
aka B2 (water soluble, polar, and
conjugated)
Direct bilirubin
BILIRUBIN
aka B1 (water insoluble, non-polar,
and unconjugated)
Indirect bilirubin
ALT/SGPT
Method for ALT?
Modified IFCC and Bergmeyer Method
ALT/SGPT
Principle of ALT?
transfer of an amino group between alanine and keto-acids to glutamate and pyruvate
ALT/SGPT
What 2 reagents are needed for ALT which make up the working reagent?
- ALT buffer reagent
- ALT enzyme reagent
5 parts buffer to 1 part enzyme reagent
ALT/SGPT
Volume of working reagent: ?
Volume of sample: ?
Volume of working reagent: 1 mL
Volume of sample: 50 uL
ALT/SGPT
After adding the working reagent, what temp and for how many minutes should be the sample be in?
note: this is before adding serum
37-42 c for 3 minutes
ALT/SGPT
How is the reading for ALT ?
Repeat reading twice every 60 seconds
ALT/SGPT
End Products and Color
Lactate and NAD
ALT/SGPT
Absorbance?
340 nm
ALT/SGPT
SI unit of ALT?
U/L
ALT/SGPT
How to convert mg/dL to U/L for ALT?
mutiply by 3,376
ALP/SGOT
Mthod for ALP?
Bower’s and McComb Method
ALP/SGOT
Principle of ALP?
Liberation of inorganic phosphate from organic phosphate ester
ALP/SGOT
What 2 reagents are needed in ALP which makes up the working reagent?
- ALP buffer reagent (R1)
- ALP substrate (R2)
5 parts buffer to 1 part enzyme reagent
ALP/SGOT
Volume of working reagent: ?
Volume of sample: ?
Volume of working reagent: 1 mL
Volume of sample: 20 uL
ALP/SGOT
After adding the working reagent, what temp and for how many minutes should be the sample be in?
37 c for 3 mins
ALP/SGOT
How is reading for ALP?
Repeat reading twice every 60 seconds
ALP/SGOT
End Products and Color
P-nitrophenol (yellow) and a phosphate ion
ALP/SGOT
Absorbance?
405 nm
ALP/SGOT
SI Unit of ALP?
U/L
ALP/SGOT
How to convert mg/dL to U/L for ALP?
multiply by 2,764
ALP/SGOT
Reference value for ALP?
34 to 114 U/L
ALT/SGPT
more thhan 35 U/L indicates what clinical significance?
Liver abnormalities (not always hepatitis)
ALP/SGOT
more than 34 U/L indicates what clinical condition?
Biliary obstruction and bone disease
What specimen is preferred for detection of urine protein and creatinine?
24 hour specimen
URINE PROTEIN
Method of urine protein?
Watanabe (Dye-Binding) Colorimetric Method
URINE PROTEIN
Principle of urine protein?
Pyrogallol Red-Molybdate + Protein
URINE PROTEIN
T or F
You can also use CSF sample for urine creatinine
T
URINE PROTEIN
Volume of total protein reagent: ?
Volume of sample: ?
Volume of total protein reagent: 3 mL
Volume of sample: 50 uL
URINE PROTEIN
In urine protein detection, what temp and how many minutes should sample be left to stand?
RT for 10 mins
OR
37 c for 5 mins
URINE PROTEIN
End Products and Color?
Blue-Purple Complex
URINE PROTEIN
Absorbance?
600 nm
URINE PROTEIN
Reference value for random specimen
1 to 14 mg/dL
URINE PROTEIN
Reference value for 24 hour specimen
22 to 120 mg/24hrs
URINE PROTEIN
more than 14 (for random) and more than 120 (for 24 hour) indicates what clinical significance?
Proteinuria
URINE CREATININE
Method of urine creatinine?
Fabinay and Eringhausen Kinetic Jaffe Method
URINE CREATININE
Principle of urine creatinine?
Creatine + Picric Acid in 10% NaOH
URINE CREATININE
What are the reagents needed for urine creatinine which makes up the base reagent?
- Creatinine base reagent
- Creatinine acid reagent
URINE CREATININE
Volume of base reagent: ?
Volume of sample: ?
Volume of base reagent: 0.5 mL
Volume of sample: 50 uL
URINE CREATININE
After adding base reagent (before sample), sample should be let it what temp and how many minutes?
37 c for 3 minutes
URINE CREATININE
How is reading done for urine crea?
Read absorbance after 20 secs. then repeat after 60 secs.
URINE CREATININE
End Products and Color?
Creatinine Picrate
(yellow/orange- red tautomer)
URINE CREATININE
Absorbance?
510 nm
URINE CREATININE
Conversion:
mg/L → mg/dL
multiply by 10
→ the most widely used marker of GFR
→ is produced constantly, freely filtered by the glomerulus, not reabsorbed, but minimally secreted
→ a good measure of the completeness of a 24H urine specimen
CREATININE CLEARANCE
CREATININE CLEARANCE
Reference Value FOR MALE
85-125 mL/min
CREATININE CLEARANCE
Reference Value FOR MALE
75-112 mL/min