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Biochemistry Practical > Spectrophotometer > Flashcards

Spectrophotometer Flashcards

1
Q

Spectrophotometry or Colorimetry It is a technique that is used to

A

determine the concentration of a colored solution by the application of the Beer—Lambert law

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2
Q

Beer—Lambert law

A

The concentration of a solute is directly proportional to the absorbance (Light absorbed) (optical density) and inversely proportional to the transmittance

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3
Q

A + T =

A

1

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4
Q

Spectrophotometer or Colorimeter

A

It Is a device used to measure the Intensity of light (شدة الضوء) after it has passed through an absorbing medium (sample)

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5
Q

Spectrophotometer or Colorimeter Consists of

A

Sample Compartment – Mode - Zero control (0 A and 0 T)

Readout (display) - Wavelength selection - print

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6
Q

Mechanism of Spectrophotometer action

A
  1. Light source (Infrared / UV / Visible) sent light to the Condenser lens
  2. Lens collect it on Prism which reflect color into 7 spectral colors
  3. By wavelength selector the specific color goes from slit which select transmits the desired wavelength and absorbs all other wavelength and direct light to the sample solution (in cuvette)
  4. Photoelectric result will appear by photocell
  5. Display result in digital display
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7
Q

The sample unknown solution is compared with a

A

known standard to obtain the exact concentration of the unknown sample

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8
Q

Sample Conc.

A

(A Sample)/(A Standard ) X Standard Conc.

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9
Q

If the substance by itself is colorless, it can ..

A

chemically converted to a colored substance

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10
Q

Before using the Spectrophotometer device

A

The zero point of the Colorimeter should be first adjusted with a solution known the blank

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11
Q

The types of blanks

A

Water , Air or Reagent blank this depends upon the procedure used (Kits)

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12
Q

Steps in spectrophotometer

A
  1. Adjust wavelength
  2. Zero control by placing blank
  3. Remove the blank
  4. Place the slandered
  5. Place the sample
  6. Take the reading
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13
Q

Photocell function

A
  • Measures the intensity of the light transmitted through the solution after absorption
  • It converts the radiation to electrical energy
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14
Q

The spectrophotometer needs … Tubes

A

3

Blank – Sample – Slandered

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15
Q

R1

A

Standard known conc.

Present in Standard tube only

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16
Q

R2

A

Coloring reagent

Present in 3 tubes

17
Q

Blood analysis may be carried out on

A

whole blood, plasma or serum depending on the distribution of the substance which is being studied

18
Q

Blood collected in

A
  1. Wasserman tube (Blue cap)
  2. Vacutainer (no anticoagulant) – (Red cap)
19
Q

Lab equipment

A
  1. Tourniquet
  2. Alcohol Swab
  3. Syringe (With beveled end up to direct blood to Syringe)
20
Q

Whole blood

A

Diluted by water to hemolysis RBCs

Detect : Hb , lactate, pyruvate and ammonia

21
Q

Plasma Detect

A

Clotting factors as fibrinogen , prothrombin

22
Q

Serum Detect

A

Glucose Cholesterol , albumin and enzymes

23
Q

Collecting Plasma or serum by using

A

Micropipette

24
Q

What is deference between plasma and serum

A

Plasma : Centrifuge uncoagulated blood / presence of fibrinogen

Serum : Centrifuge coagulated blood

25
Q

Centrifuge blood … to have serum or plasma

A

15 RPM to 10 Min

26
Q

Na or K citrate Mechanism

A

Inhibits blood coagulation by combining with Ca

27
Q

Na or K citrate Cap Color

A

Black → ESR
Light Blue → Prothrombin time

28
Q

Na Fluoride Mechanism

A

Inhibits enolase enzyme so stops the consumption of glucose

29
Q

Na Fluoride detect

A

Glucose

30
Q

Na Fluoride Cap Color

A

Gray

31
Q

EDTA Mechanism

A

Inhibits blood coagulation by chelation of Ca and Mg

32
Q

EDTA detect

A

CBC and blood film

33
Q

EDTA Cap Color

A

Violet

34
Q

EDTA Note

A

Not used in measuring ALP enzyme (no Mg) and Ca and Mg)

35
Q

Heparin Mechanism

A

Anti-thrombin

36
Q

Heparin detect

A

Blood gases (O2 – CO2)

37
Q

Heparin Cap color

A

Green

38
Q

Heparin note

A

Not used in blood film (black film - aggregation of cells) and PCR

39
Q

K oxalate

A

Not used anymore (aggregation of cells)

Biochemistry Practical (1 decks)

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