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CCHM 2 > LAB4 - LIPASE and AMYLASE > Flashcards

LAB4 - LIPASE and AMYLASE Flashcards

1
Q

a turbidimetry assay used for lipase determination

A

tietz-fiereck turbidimetry assay

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2
Q

the kit for tietz-fiereck turbidimetry assay conists of

A

lipase buffer
olive oil substrate
lipase activity standard

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3
Q

the pH level of lipase buffer

A

pH 7

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4
Q

amount of lipase activity standard used

A

60 U/L

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5
Q

principle of tietz-fiereck turbidimetry assay

A

the lipolytic action of lipase in the serum hydrolyzes the olive oil emulsion to fatty acids, mono and di-glycerides and glycerol. The decrease in turbidity at 550 nm is proportional to the lipase activity of the serum

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6
Q

Give the values or the amount of reagent needed for cuvette 1 (STANDARD)

A

lipase buffer- 1200 ul
olive oil - 250 ul
standard - 25 ul
serum - no serum needed

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7
Q

Give the values or the amount of reagent needed for cuvette 2 (UNKNOWN/TEST SERA)

A

lipase buffer- 1200 ul
olive oil - 250 ul
standard - NO STANDARD
serum - 25 ul

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8
Q

after preparing the solution needed, how many minutes we do need to stand after shaking vigorously/

A

10 mns standing at room temp

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9
Q

wavelength needed to read lipase

A

550 nm

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10
Q

read blank tube needed for the method

A

distilled water

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11
Q

formula for lipase

A

abs of sample/ abs of standard

X 60 u/l

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12
Q

ref range of lipase

A

30 - 200 u/l by clsi, nccls

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13
Q

increased level of lipase are associated to what conditions

A

acute pancreatitis
carcinoma of pancreas
intestinal obstruction
perforated peptic ulcer
acute peritonitis

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14
Q

it is generally agreed that lipase values have about the same significance as ____ values in the diagnosis of acute pancreatitis

A

amylase

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15
Q

during pancreatic disturbances, the serum lipase activity may rise slowly that the serum amylase, but it may remain elevated for a longer time so that it may be useful in the diagnosis and follow up of acute pancreatitis

true or false?

A

true

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16
Q

what should we do if the chemistry analyzer minimum’s readability is 2.5 ml

A

multiply by 2 the volume of the standard/test/ and the control and reagent

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17
Q
A
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18
Q

the amylase reagent sit we used and studied is for the (quantitative, qualitative) kinetic determination of amylase activity

A

quantitative

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19
Q

the amylase reagent sit we used and studied is for the quantitative kinetic determination of amylase activity.

intended to use for (manual, automated, both) procedures in human serum and urine

A

both

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20
Q

amylase was first measured QUANTITATIVELY by an ___

A

iodometric method

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21
Q

amylase was first measured QUANTITATIVELY by an iodometric method by who?

A

Wohlegemuth in 1908

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22
Q

In 1938, he then introduced a procedure that STANDARDIZED the amounts of starch and iodin

A

somogyi

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23
Q

His work became the basis for the widely used amyloclastic and saccharogenic methods

A

Somogyi

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24
Q

Again, what is the method of somogyi that is widely used and became the basis for lipase determination?

A

amyloclastic 1956 and saccharogenic methods 1960

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25
Q

what is the disadvanatage of amyloclastic and saccharogenic methods of somogyi

A

long incubation time
endogenous glucose interference
unstable reaction colors resulting in poor reproducibility and reliability

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26
Q

several procedures have been suggested such as the ______, however they are still subjected to long incubation time,
endogenous glucose interference and interference in the formation of NADH

A

use of defined substrate maltoteraose

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27
Q

he introduce a substrate that will eliminate the interference from endogenous glucose and pyruvate

A

wallenfells et.al.

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28
Q

wallenfells et.al

he introduce a substrate that will eliminate the interference from endogenous glucose and pyruvate

what substrate is it?

A

p-Nitrophenylglycosides

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29
Q

the present procedure is based on the modification of __

A

wallenfells et.al

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30
Q

the present procedure we are doing right now is based on the modification of wallenfells et.al’s method.

What could be the modification?

A

p-Nitrophenyl-D-Maltoheptaoside with the terminal glucose blocked to reduce spontaneous degradation of substrate by glucosidase and glucoamylase

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31
Q

the modified method of wallenfells is performed in what mode

A

kinetic mode with very short time lag and offers much greater stability than previous amylase methodologies.

32
Q

the determination of amylase activity in what sample is most commonly performed for the diagnosis of acute pancreatitis

A

serum and urine

33
Q

in acute pancreatitis, amylase levels are elevated for longer periods of time in what preferred sample

A

urine than in serum

34
Q

since urine amylase levels are elevated for longer periods of time in urine, determining the ratio of amylase and ____ is important in followng the course of the pancreatitis

A

creatinine clearances

35
Q

give the first reaction of amylase determination

A

PNPG7 —– amylase ——- PNPG3+Maltotetraose

36
Q

2nd reaction in amylase determination

A

PNPG3 —- glucoamylase — PNPG1 + glucose

37
Q

3rd reaction of amylase determination

A

PNPG1— glucosidase —-p-Nitrophenol+glucose

38
Q

amylase hydrolyze p-nitrophenyl-D-maltoheptaoside to form

A

p-nitrophenylmaltotriose

39
Q

p-nitrophenylmaltotriose will be hydrolyze glucosidase to form

A

p-nitrophenylglucoside

40
Q

p-nitrophenylglucoside will be hydrolyze again by glucosidase to form

A

p-nitrophenol + glucose

41
Q

the end product, p-nitrophenol + glucose, will form a result color of

A

yellow

42
Q

the rate of ____ in absorbance is measured in 405 nm and is proportional to the amylase activity in the sample

A

increase

43
Q

nm use for amylase

A

405 nm

44
Q

reagent compositions

A

p-nitrophenyl-D-maltoheptaoside
glucosidase
glucoamylase
sodium chloride
calcium chloride
buffer 50 ml

45
Q

the reagents are intended to use only in ____

A

in vitro

46
Q

we must store the dry reagent in what temp

A

2-8 * C refrigerated

47
Q
A
48
Q

reagent are stable until when

A

expiration label

49
Q

reconstituted reagent is stable for at least how many days at room temp 15-30*C

A

10 days

50
Q

reconstituted reagent is stable for at least how many days at refrigerated temp 2-8

A

30 days

51
Q

the reagent should be discarded if

A

turbidity has occurred; turbidity may be a sign of contamination
moisture has penetrated the vial and caking has occurred
reagent does not dissolve completely upon reconstitution
the reconstituted reagent has an absorbance of 0.70 or greater when measured against water at 405 nm

52
Q

if the reconstituted reagent has an absorbance of 0.90 upon measuring against water blank at 405 nm, what is the next course of action?

A

discard the reagent, the reconstituted reagent must have an absorbance of lower than 0.70

53
Q

the ideal specimen for amylase

A

serum

54
Q

can we use plasma in amylase?

A

yes, only heparinized plasma

55
Q

why do other anticoagulant such as citrate and EDTA are not used for amylase determination

A

such anticoagulants bind with calcium which is the ion needed for amylase activity, so only heparin is allowed for plasma use

56
Q

urine specimen must be collected in ____ period

A

24 hr period

57
Q

urine specimen must be collected in 24 hr period and the pH must be adjusted to

A

pH 7 with either 0.1 N NaOH or 0.1 N HCl and keep refrigerated

58
Q

do we need to add preservative to the urine sample?

A

no need for that

59
Q

amylase in serum and urine is reported stable for how many days or weeks in room temp
18-25*C

A

1 week

60
Q

amylase in serum and urine is reported stable for how many days or weeks in refrigerated temp and PROTECTED AGAINST EVAPORATION AND BACTERIAL CONTAMINATION

A

several months

61
Q

macroamylasemia ___ activity of pancreatic amylase in serum

A

increases

62
Q

a number of diseases can affect the determination of amylase

true or false

A

true the fire

63
Q

who listed drugs and other substances that can interfere in amylase activity

A

young et.al

64
Q

lipemia, hemolysis, will ____amylase values

A

increase

65
Q

can insulin affect amylase determination?

A

yes

66
Q

how many minutes we need to prewarm the reagent

A

3 minutes

67
Q

how many minutes do we need to read the sample for amylase

A

read first at 15 seconds then read for every 30 sconds

68
Q

the formula for amylase

A

abs/min X 1.025 X 1000

divide them by

8.5 X 0.025 X 1

69
Q

shortcut for amylase

A

abs/min X 4824

70
Q

how do you get the abs/min then?

A

minus minus plus then divide by 4

71
Q

to convert to SI units (nkat/L) the amylase

A

multiply to 16.67

72
Q

linearity for amylase

A

1500 IU/L

73
Q

if the linearity exceed for amylase, what should we do

A

dilute with an equal volume of saline and reru the multiply the result by 2

74
Q

ref range for amylase in serum

A

up yo 96 IU/L

75
Q

ref range for amylase in URINE

A

18 to 330 IU/L

76
Q
A

CCHM 2 (22 decks)

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