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Cell And Molecular Biology > Bio Analysis > Flashcards

Bio Analysis Flashcards

1
Q

What is quantitative analysis

A

X

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2
Q

Disease

A

• Dysregulation of the cell cycle is common during tumorigenesis (cancer display uncontrolled growth)

• Increased levels of CDK molecules and cyclins are sometimes found in human tumours, such breast cancer and brain tumours

• Inhibition of certain CDKs has been shown to inhibit tumour cell growth, induce apoptosis and cause tumour regressions in animal models

• CDKs are potential targets for new anticancer therapy

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3
Q

Analytical chemistry

A

a branch of chemistry that deals with the identification of compounds and mixtures (qualitative analysis) or the determination of the proportions of the constituents (quantitative analysis): techniques commonly used are titration, precipitation, spectroscopy, chromatography, etc. 1

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4
Q

Biochemical analysis:

A

characterisation of biological components within a sample using appropriate laboratory techniques

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5
Q

Qualitative Analysis

A

indicate whether a particular substance (analyte) is present above a threshold level

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6
Q

Marquis test

A

presence of alkaloids

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7
Q

Trinder spot test

A

presence of salicylates (aspirin)

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8
Q

hCG

A

human chorionic gonadotrophin

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9
Q

Quantitative Analysis:

A

determines the amount of a particular analyte present in the sample (e.g. the concentration of a drug in blood serum).

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10
Q

Accuracy

A

Closeness of a measured or derived data value to its true value

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11
Q

Precision

A

Closeness of values with repeated measurements

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12
Q

Sensitivity

A

• ability to detect small amounts of analyte in a sample
• OR the percentage of patients with the disease that will be correctly
identified as disease positive

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13
Q

Specificity:

A

• ability to detect only the analyte of interest in a sample
• OR the percentage of patients without the disease that receive a negative result

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14
Q

Why measure biological molecules?

A

Medicine:
• Blood hormone levels • Enzyme activity
• Drug levels
• Metabolites
• Clinical trials

Toxicology:
• Xenobiotics
• Drugs / metabolites
• Pharmacokinetics
• Anti-doping testing in sports

Forensics:
• Xenobiotics
• Drugs / metabolites
• Environmental toxins
• Other toxic compounds

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15
Q

Typical analytical scheme

A

Sample
Sub-sample
Extraction ; the test substance or remove the interfering substance before analysis can proceed
Separation; Separation procedures depend on physical or chemical characteristic of the compound
Detection
Result
Interpretation

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16
Q

Separation technique for polar molecules that are volatile,soluble,and absorbable

A

Gas-liquid chromatography Liquid-liquid chromatography
Liquid-gas chromatography

17
Q

Separation technique for ionic molecules that are charged

A

Ion-exchange chromatography
Electrophoresis

18
Q

Separating technique for molecules with a mass that have the physical property are diffusion and sedimentation

A

Gel filtration chromatography
Dialysis Ultracentrifugation

19
Q

Separating technique for molecules with a shape that have the physical property of ligand binding

A

Affinity chromatography

20
Q

Chromatography

A

-developed by a Russian botanist

-Technique that allows the resolution of a mixture of compounds as a consequence of the different rates at which they move through a stationary phase, under the influence of a mobile phase

21
Q

Planar chromatography

A

The stationary phase is supported on a flat plate or in the fibres of a paper.
The mobile phase moves through the stationary phase by capillary action or by gravity.
• Paper chromatography
• Thin layer chromatography (TLC)

22
Q

Column Chromatography:

A

The stationary phase is held in a tube through which the mobile phase is forced either by pressure or by gravity.
• Simple column chromatography
• High pressure liquid chromatography (HPLC) • Gas chromatography (GC)

23
Q

Thin layer chromatography

A

-TLC plate (stationary phase): a sheet of glass, metal, or plastic coated with a thin layer of a solid adsorbent (usually silica or alumina)
-Samples are “spotted” at the base of the sheet and dried.
-Sheet is placed in the tank containing a shallow layer of solvent (mobile phase)

-Substances reacting to the stationary phase more strongly will have retarded migration.

24
Q

Rf

A

distance travelled by substance /distance travelled by solvent

25
Q

Gel Filtration (Column) Chromatography

A

-Separates proteins, peptides and oligonucleotides on the basis of size
-The stationary phase (gel) consists of beads (e.g. Sephadex)
-The mobile phase (buffer) is used to elute the analytes from the gel matrix based on their size

26
Q

Gel Filtration Chromatography: the Column

A

Gel matrix: Sephadex (separation Pharmacia dextran) a cross-linked dextran gel

27
Q

Parts of the gel filtration column in chromatography

A

Vo: volume required to elute molecules bigger than the pore size of the column gel
Vt=Vs+VI +Vo
VI= volume of solvent in the pores
Ve: elution volume of a particular solute

Partition coefficient: Kav =(Ve-Vo)/(Vt-Vo)

28
Q

Partition coefficient:

A

Kav =(Ve-Vo)/(Vt-Vo)

29
Q

Gel Filtration Chromatography

A

Relative molecular weight (Mr) of analyte can be estimated by plotting the partition coefficient Kav against the logarithm of the molecular weight of standard protein samples

30
Q

Typical analytical scheme 2

A

Sample
Sub-sample
Extraction
Separation
Detection ; Liquid chromatography detectors: • UV / visible
• Fluorescence
• Electrochemical
Result Interpretation

31
Q

Detection and Analysis
Spectroscopy

A

Measures the absorption and emission of electromagnetic radiation by atoms and molecules in solution
Isaac Newton (1666):
uses a glass prism to split sunlight into a monochrome spectrum

32
Q

Io

A

intensity of light striking a substance

33
Q

I

A

intensity of light transmitted through the substance

34
Q

Spectroscopy: Beer-Lambert law

A

-Describes the absorbance of monochromatic light as it passes through a solution

-log10(io/I)=A=ECL

-I/io=10 -ecl.

l = the distance travelled by the light through the substance [in m]
ε = molar extinction coefficient of the substance c = concentration of the substance [mol/l]
A= absorbance

A=ecl Absorbance is linearly related to the concentration of the analyte
With known ε and l the concentration of a substance can be estimated using a standard plot – Abs against concentration

35
Q

How do spectrophotometers work

A

-light source
-collimator(lens)
-monochromator(prism or grating|)
-wavelength selector
-sample collation in a cuvette
-detector (photocell)
-digital display or meter

Cell And Molecular Biology (20 decks)

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