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CC RMT AUG2024 > Spectrophotometry > Flashcards

Spectrophotometry Flashcards

1
Q

Ultraviolet

A

<400nm

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2
Q

Visible light

A

400-700 nm

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3
Q

Infrared

A

<700 nm

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4
Q

Planck’s Formula

A

E=hv

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5
Q

relationship between wavelength and energy

A

inversely proportional to energy and frequency

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6
Q

shortest wavelength = __ energy frequency

A

↑ energy frequency

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7
Q

longest wavelength = __ energy frequency

A

↓ energy frequency

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8
Q

the concentration of a substance is directly proportional to the amount of light absorbed and is inversely proportional to the logarithm of transmitted light

A

Beer-Lambert’s Law

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9
Q

A=abc

what is ‘a’?

A

molar absorptivity

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10
Q

A=abc

what is ‘b’?

A

light path

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11
Q

A=abc

what is ‘c’?

A

concentration

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12
Q

Beer’s formula:

A

A= 2-log%T

(2 log Tayo after beer)

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13
Q

formula to determine the concentration of an unknown analyte

A

Cu= AuCs/As

(AUq CaSe AmpangetS)

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14
Q

distilled water, rgt, sample; sets the spectrophotometer to 0 absorbance

A

blank

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15
Q
  • corrects absorbance caused by the color of REAGENTS
  • used to 0 the instrument before measuring test samples and other blanks
A

reagent blank

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16
Q

used to subtract the intrinsic absorbance caused by hemolysis, icterus, turbidity, or drug interference

A

sample blank

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17
Q

substance of KNOWN purity and concentration; used to determine the concentration of the unknown analyte

a. standard
b. control

A

a. standard

18
Q
  • soln containing various analytes with known target values;
  • analyzed with patient samples to monitor analytical performance (accuracy, precision)
  • aka QC sample

a. standard
b. control

A

b. control

19
Q

values provided by the manufacturer

a. assayed
b. unassayed

A

a. assayed

20
Q

values determined by the laboratory

a. assayed
b. unassayed

A

b. unassayed

21
Q

provides polychromatic light

A

light source

22
Q

deuterium/hydrogen

a. UV, non-colorimetric
b. UV, visible light, colorimetric
c. visible, near IR

A

a. UV, non-colorimetric

23
Q

Xenon/Mercury

a. UV, non-colorimetric
b. UV, visible light, colorimetric
c. visible, near IR

A

b. UV, visible light, colorimetric

24
Q

tungsten, LASER

a. UV, non-colorimetric
b. UV, visible light, colorimetric
c. visible, near IR

A

c. visible, near IR

25
Q

meaning of LASER

A

light amplification by Stimulated Emission of Radiation

26
Q

prevents stray light

A

entrance slit

27
Q

wavelength selector/isolator

A

monochromator

28
Q

continuous, non-linear spectrum, better separation for high-frequency light

a. prism
b. diffraction gratings

A

a. prism

29
Q

continuous, linear, uniform separation of wavelengths; most commonly used

a. prism
b. diffraction gratings

A

b. diffraction gratings

30
Q

controls bandpass;
narrow fraction of the spectrum to reach the cuvette

A

exit slit

31
Q

sample cell; may be round or square

A

cuvette

32
Q

quartz/fused silica cuvette

a. UV-IR
b. UV-visible
c. visible; not for UV work

A

a. UV-IR

33
Q

plastic cuvette

a. UV-IR
b. UV-visible
c. visible; not for UV work

A

b. UV-visible

34
Q

glass cuvette

a. UV-IR
b. UV-visible
c. visible; not for UV work

A

c. visible; not for UV work

35
Q

converts the transmitted light energy into an equivalent amount of electrical energy

A

photodetector

36
Q

processes the electrical signal, performs mathematical operations, and displays the output

A

read-out device

37
Q

Spectrophotometer QA:

if the WAVELENGTH is set to the actual one selected by the monochromator; checked using didymium glass or holmium oxide

A

wavelength accuracy

38
Q

Spectrophotometer QA:

done using glass filters and solutions that have known ABSORBANCE values

A

absorbance check

39
Q

Spectrophotometer QA:

A change in concentration results in a STRAIGHT LINE calibration curve (Beer’s law)
determined using neutral density filters and dichromate solution

A

Linearity

40
Q

Spectrophotometer QA:

any wavelength outside the band of interest; causes absorbance error and loss of linearity; detected using sharp cut off filters

A

stray light

CC RMT AUG2024 (29 decks)

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