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Musculoskeletal Ospe > Biochemistry > Flashcards

Biochemistry Flashcards

1
Q

Difine spectrophotometer?

A

Is used to measure the amount of light that the chemical substances absorbs by measuring the intensity of light as a beam of light passes through sample solution

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2
Q

What is the basic principle of spectrophotometer?

A

Is that each compound absorbs or transmits light over a certain range of wavelength

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3
Q

F of light source in spectrophotometer?

A

Emits radiant E that contains many wavelength

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4
Q

F of monochromator ?

A

Prism , to beak the polychromatic radiation into component of wavelength or bands of wavelength .

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5
Q

F of adjustable aperture?

A

Exit slit that allows only one color to pass to the sample .

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6
Q

F of sample ?

A

Absorbs a portion of the light and transmits the rest through the sample .

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7
Q

F of output?

A

Detector , that convert the radiant E into electrical E .

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8
Q

F of transport vessels ( cuvettes )?

A

To hold the sample .

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9
Q

What is the components of spectrophotometer?

A
  1. A stable and cheap radiation energy source .
  2. Monochromator .
  3. Transport vessels * cuvettes* .
  4. A photosensitive detector and an associated readout system .
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10
Q

How the simple beam spectrophotometer work ?

A

To measure the intensity of the incident light , the sample must be removed , so that the reference can be placed each time .

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11
Q

What is the advantages of simple spectrophotometer?

A

Less expensive and less complicated.

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12
Q

How the double beam spectrophotometer work ?

A

Before it reaches the sample , the light source is split into tow separate beams , from this one passes through the sample and the second one is used for the reference.

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13
Q

What is the advantages of double beam spectrophotometer?

A

The reference reading and the sample reading can take place at the same time .

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14
Q

What is the type of spectrophotometer?

A

Simple beam spectrophotometer.

Double beam spectrophotometer.

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15
Q

Types of spectrophotometer based to the wavelength of light and write the range of each type ?

A
  1. Visible spectrometer ( 400 _ 700 nm) of electromagnetic radiation spectrum .
  2. UV spectrometer (180 _400nm) of electromagnetic radiation spectrum .
  3. IM spectrometer (700_1500 nm) of electromagnetic radiation spectrum .
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16
Q

What is the realation between the amount of light absorbs and the concentration of a substance.?

A

Directly propotional .

17
Q

What tube do we need in our spectrophotometer experiment?

A

Blank tube , reagent and water
Standard tube , reagent and standard
Test tube , reagent and test

18
Q

What’s the beer’s law ?

A 
The concentration of the substances is directly proportional to the light absorbed or inversely proportional to the logarithm of the transmitted light.
A = abc 
A for absorbance 
a for absorptivity 
b for light path in centimetres 
C for concentration of the substances.
19
Q

What is the absorptivity ?

A

It is a constant for a particular substance that means the amount of light absorbed per unit concentration.

20
Q

What is the equation of law ?

A

Ct =( At / Ast )× C st .

21
Q

Where do we get a specimen from the total calcium ?

A

Serum and heparinized plasma .

Serum is prefered.

22
Q

How do we get a sample from the free calcium ?

A
  1. should be collected and handled anaerobically .

2. Take care of the heparinized in the tube .

23
Q

What are the precautionary steps for taking a calcium sample?

A
  1. Without tourniquet.
  2. In sitting position .
  3. Use wide bore needle . ابرة ذات تجويف واسع
  4. Fasting .
24
Q

What is the analytical method for assessing calcium in the serum?

A

By using calcium _binding reagent which exhibit high affinity for calcium and forming a coloured reaction product .

25
Q

List the calcium binding reagent ?

A
  1. O_cresolphathalein complexone .

2. Arsenazolll .

26
Q

What is the OCPC method?

A

Ca+OCPC_____red complex (570_580 nm )

At alkaline pH 10_12 .

27
Q

What is the arsenazolll method ?

A
  1. Measured at acidic pH 6\has much affinity for Ca than other.
  2. Utility of the OCPC assay is limited by the fact that absorption of carbon dioxide by the alkaline reagent causes considerable calibration drift.
  3. Arsenazo III is highly sensitive and specific for calcium, generating a sensitive absorbance peak at (600-660 nm).Because the Arsenazo III reacts under slightly acidic conditions.
  4. Measured at wavelength 650nm.
28
Q

How calculate the ionized calcium ?

A 
iCa= 0.9+(0.55×tCa_ 0.3 × albumin ).
iCa= (6×tCa _TP/3)/(6+TP).
29
Q

How calculate the corrected serum calcium ?

A
  1. Serum calcium / (0.6 + ( total protein/ 8.5)).
  2. Cacium + 0.8 ( normal albumin _ patient albumin )
    Normal albumin = 4.0
30
Q

What is the optimal ph for the enzyme reaction ?

A

9 _ 10.5

31
Q

What is the activators of ALP enzyme ?

A

Mg +2 / zn +2

32
Q

Where is the highest concentration of ALP found?

A 
Intestine 
Live 
Kidney 
Placenta 
Bone
33
Q

Where the ALP found mainly ?

A

In the cells lining the biliary tract .

In the osteoblasts .

34
Q

List the ALP isozymes?

A

ALPL Intestinal.
ALPL tissue non specific live / bone / kidney .
ALPP_ placental.

35
Q

When does ALP increase ?

A
  1. Gut ALP rises after meals .
  2. Bone ALP increase at the time of physiological growth.
  3. Placental ALP normally increase at the third trimester.
36
Q

What is the method and the substrate that use to assay enzyme activity?

A

Kinetic method

37
Q

What is the principle of assay enzyme activity by ALP ?

A

ALP catalyzes the hydrolysis of colourless organic phosphate ester substrate ( p_Nitrophenylphosphate) to the yellow coloured product ( p_Nitrophenol) and phosphate .

38
Q

What is the suitable environment to assay enzyme activity by ALP ?

A

Alkaline ph at 37 ⁰C

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