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CHEM 211 > Unit 1 > Flashcards

Unit 1 Flashcards

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1
Q

assay

A
  • process of determining amount of analyte in sample
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2
Q

analyte

A
  • chemical substance being measured
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3
Q

signal

A
  • observable change in some property
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4
Q

Advantages of visual detection

A
  • low cost and maintenance
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5
Q

Disadvantages of visual detection

A
  • subjectivity affects accuracy/precision
  • may not be very sensitive
  • may require large sample volumes
  • often time-consuming
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6
Q

voltage

A
  • electrical potential energy between two points
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7
Q

current

A
  • rate of flow of charge past a point in a circuit (usually electrons moving)
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8
Q

transducer

A
  • device that converts input stimulus into electrical output
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9
Q

Advantages of electrical detection

A
  • objective
  • often very sensitive
  • often faster
  • can analyze smaller sample sizes
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10
Q

Disadvantages of electrical detection

A
  • high cost/maintenance

- calibration required

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11
Q

analog signal

A
  • “real world”
  • takes on any value
  • transducer input signal
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12
Q

digital signal

A
  • computer world
  • recorded as bits
  • discrete values
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13
Q

signal

A

-a measured quantity that is correlated to the amount of analyte

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14
Q

noise

A
  • unwanted variation in a measured quantity

- often takes the form of random fluctuations in a measured signal

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15
Q

signal-to-noise ratio (S/N)

A
  • the magnitude of the signal divided by the magnitude of the noise
  • similar term: signal-to-background ratio
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16
Q

detection limit

A

-the amount of analyte that corresponds to a signal just greater than the mean of the background plus three standard deviations of its noise

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17
Q

background

A

-an approximately constant signal, measured in the absence of analyte

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18
Q

How can you increase S/N ratio?

A
  • multiple scans

- signal averaging

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19
Q

Determine how many more scans are required to achieve a S/N ratio of 2?4?9?

A

N=4,16,81

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20
Q

Blank

A
  • a measured sample that lacks the analyte

- contains solvent, reagents, etc. used in the analysis

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21
Q

sample matrix

A
  • all the components of a sample except the analyte

- blank tries to approximate the sample matrix

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22
Q

positive control

A
  • a standard sample that contains a known quantity of the analyte of interest
  • prevents false negative results
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23
Q

negative control

A
  • a standard sample that does not contain any analyte

- prevents false positive results

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24
Q

interference

A

a specific chemical substance in a sample matrix that causes a systematic error in a measured quantity

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25
Q

Which of the following combinations best matches the definition of analyte, sample matrix and blank? Pick all that apply.

(a) Glucose; Blood; Glucose
(b) Glucose; Blood; Synthetic blood
(c) Glucose; Blood; Saliva
(d) Glucose; Sucralose; Blood
(e) None of the above
(f) Lead and Mercury; Blood; Distilled water
(g) Saliva; Lead and Mercury; Saliva
(h) Steroid drugs; Urine; Synthetic Urine

A

(b) glucose; blood; synthetic blood

(h) Steroid drugs; Urine; Synthetic Urine

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26
Q

In a single-molecule fluorescence detection experiment, the average background was measured to be 330 cps ± 50 cps (±1 std. dev.). What must be the minimum magnitude of a signal burst to record detection of a single molecule?

(a) 180 cps
(b) 280 cps
(c) 330 cps
(d) 380 cps
(e) 480 cps

A

e) 480cps

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27
Q

How can interferences affect results?

A
  • act on the analyte (or a measured form thereof)
  • act on a reagent used in the detection method
  • be the source of a large background signal
  • cause negative/positive bias
  • cause absolute/proportional errors
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28
Q

selectivity

A
  • the extent to which other substances interfere with the determination of an analyte
  • typically via reactivity/molecular interactions
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29
Q

good selectivity

A

analysis method has minimal interferences

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30
Q

poor selectivity

A

analysis method prone to certain interferences

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31
Q

masking agent

A

a reagent that prevents one or more components in a sample matrix from interfering with an analyte

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32
Q

What is the analytical signal, and possible interferences that would be found in colourimetric analyses via complexation?

A
  • analytical signal: colour change upon complexation between analyte and reagent
  • interference with analyte: matrix component complexing with analyte
  • interference with reagent: matrix component complexing with reagent
  • background interference: matrix component that adsorbs same wavelength of light as analytical complex
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33
Q

Accuracy

A

-closeness of an experimental value, xi (or mean value of a set of measurements, x̅) to the true value, μ

34
Q

Precision

A
  • agreement among results, s

- reproducibility between replicate measurements

35
Q

Absolute error

A

-the difference between the measured and true value
-value will be positive or negative
E=xi-μ

36
Q

Relative error

A

-error in measurement
-expressed as a %
E=(xi-μ)/μ

37
Q

Replicates

A
  • samples from the same source, run using the same method, under the same conditions
  • expected to give the same result in the absence of error
38
Q

Random/Indeterminate errors

A
  • introduce uncertainty
  • symmetric about the true value(μ)
  • treat with statistics
39
Q

Systematic/Determinate errors

A
  • introduce bias
  • measured value (xi) always higher/lower than true value (μ)
  • can be proportional or constant
40
Q

Proportional error

A

-effect is independent of the magnitude of the measurement

41
Q

Constant error

A

-effect dependent on measurement

42
Q

Given the data, determine if the errors are constant or proportional:

Measured True Relative Error

  1. 004V 0.004V error too small
  2. 022V 0.020V 10.0%
  3. 389V 0.354V 10.0%
  4. 200V 2.000V 10.0%
A

Proportional error

43
Q

Given the data, determine if the errors are constant or proportional:

Measured True Relative Error

  1. 002V 0.004V 50.0%
  2. 018V 0.020V 10.0%
  3. 352V 0.354V 0.6%
  4. 998V 2.000V 0.1%
A

Constant error

44
Q

What are 3 types of systemic errors?

A
  1. Instrument errors
  2. Method errors
  3. Personal errors
45
Q

Instrument errors

A
  • minimize with careful, regular calibration
  • voltage fluctuations/drift
  • can be corrected
46
Q

Method errors

A
  • chemistry doesn’t behave as expected
  • Difficult to identify and correct
  • incomplete reactions
  • interference from non-analytes
  • false positive/negative results
47
Q

Personal errors

A
  • poor lab technique
  • incorrect recording of data
  • deviations from an established method
48
Q

Which of the following would not lead to a systematic error?

(a) An interfering species in the sample matrix that made some of the analyte unavailable
(b) An interfering species in the sample matrix generates a high background signal
(c) Fluctuating intensity of the lamp during absorbance measurement
(d) Decomposition of the sample during your sample preparation
(e) More than one of the above

A

(c) Fluctuating intensity of the lamp during absorbance measurement

49
Q

Binomial distribution

A

-discrete probability distribution (50/50)

50
Q

Gaussian distribution

A

-known as normal distribution
-used to describe data clustered about a mean value,μ
-continuous and symmetric distribution
-“bell curve” shape
-peak position determined by the mean
-width and height determined by the standard deviation, s
-total area under the curve is 1
-probability of measuring z is the area of that range under the curve
z=(x-x̅)/s

51
Q

At least how many measurements must be made until the sample standard deviation approaches the population value?

A

N > 20

52
Q

Population

A

all possible measurements of interest

53
Q

Sample

A

a limited number of measurements that are representative of the population

54
Q

How are replicate measurements evaluated?

A

Replicate measurements are evaluated using the mean and standard deviation

55
Q

Deviation

A

Difference between a measured value and the mean value of all measurements

56
Q

Standard deviation and Relative standard deviation(RSD)

A

a measure of the uncertainty and precision associated with a measurement

57
Q

Degree of Freedom

A

number of independent measurements

58
Q

Purpose of the t-statistic

A
  • permits use of sample data to test hypotheses about unknown population means without knowledge of the population standard deviation
  • accounts for limited sample size to better reflect population values
  • as N increases, t value decreases
  • as confidence interval increases, t value increases
59
Q

What is the purpose of significance testing?

A

-to determine whether the difference between two or more values is too large to be explained by indeterminate error

60
Q

Null hypothesis

A
  • used to test experimental results
  • postulates that two observed quantities are the same
  • assume two results are the same, then apply a test to see if the null hypothesis can be statistically rejected
61
Q

case 1 t-test

A
  • statistical test that compares the average experimental data of a sample to actual value
  • t exp < t table =no significant difference
  • t exp > t table =significant difference
62
Q

case 2 t-test

A

-statistical test that compares the means of two different analyses of replicate measurements

63
Q

F-test

A

-statistical test that compares the precision (standard deviation) of two sets of measurements

64
Q

G-test

A

-statistical test used to exclude an outlier from a data set

65
Q

Type 1 error

A

rejection of the null hypothesis when it is actually true

66
Q

Type 2 error

A

acceptance of the null hypothesis when it is actually false

67
Q

case 3 t-test

A

statistical test that compares the methods of single measurements on several different samples

68
Q

Absolute calibration method

A

-based on evaluation with use of fundamental physical constants and/or universal quantities only

69
Q

Empirical calibration method

A

-result of empirical analysis of unknown sample is only as good as the calibration

70
Q

Calibration curve

A
  • plot of measured signal vs known quantity

- used to interpolate unknown x values from measured y values

71
Q

Sensitivity

A

the slope (m) of the calibration curve

72
Q

Dynamic range

A

concentration range over which the calibration curve is analytically useful

73
Q

Least Squares Method of Analysis

A
  • determines line/curve of best fit to experimental data
  • minimizes the residuals between the data points and the line of best fit
  • assumes error only in y data
74
Q

Selectivity of calibration curves

A

-for linear calibration plots, selectivity for method for compound 1 vs compound 2 is reflected by the ratio of their slopes m1/m2

75
Q

Matrix Effect

A

-combined effect of all non-analyte components in a sample on the quantitative measurement of the analyte

76
Q

What are 3 examples of matrix effects?

A
  1. Some component in the sample generates a signal similar to the analyte.
  2. Some component in the sample has a chemical interaction with the analyte.
  3. Some component in the sample is co-isolated with the analyte.
77
Q

Standard Addition Concept

A
  • means of accounting for matrix effects
  • add increasing amounts of a standard analyte to aliquots of the original sample
  • result: amount of analyte to be “removed” from the sample to get signal=0
78
Q

Limitations of Standard Addition

A
  • most precise results obtained when the amount of standard added is comparable in magnitude to the original quantity of analyte
  • time consuming
  • opportunity for dilution error
  • added standard should not overwhelm any interferences
  • cannot account for shifts in baseline
  • dilution of a sample decreases the ability to detect low concentrations
  • may require large quantities of sample
  • common variation is to add an increasing amount of standard in increments to the same volume of sample
79
Q

Internal Standard

A
  • intentionally added substance of known quantity that is not expected to be found in the sample, but is expected to behave similarly
  • accounts for losses during sample processing or fluctuations in instrument signals
  • reference the analyte signal to the internal standard signal
80
Q

Sampling

A
  • process by which a sample population is reduced to a size suitable for laboratory analysis
  • composition of sample must be representative of the population
81
Q

Sampling Process

A
  1. Identify population
  2. Develop sampling strategy
  3. Collect and preserve a gross sample(s)
  4. Reduce the gross sample(s) to a lab sample
  5. Replicat analyses of the sample(s)
82
Q

What are some challenges with real samples?

A
  1. defining the problem (how will the experiment be conducted? i.e., when where how many samples to take? what size sample? is accuracy and precision required? what is the expected concentration range of analyte?)
  2. storing and preserving sample prior to analysis (i.e., protection from light, temperature, and pH changes)
  3. homogenization (i.e., may require crushing, grinding, drying, digesting, decomposing, etc.)
  4. overwhelming matrix effects (i.e., requires separation of analyte from interferences see unit 5)
  5. analyte too dilute for reliable analysis

CHEM 211 (5 decks)

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