metabolics

Question 1

what is metabolics?

Answer 1

what is happening now, it allows the study and characterisation phenotype within living organism

Question 2

what elements are metabolites made from?

Answer 2

H, C, N, O and S

Question 3

what is MS2

Answer 3

if know elements and their weights and atomic mass it is a direct refection of make up

Question 4

which AA are associated with detoxification reactions

Answer 4

cysteine and methionine- sulphur containing AA

Question 5

what are metabolites

Answer 5

anything that can be metabolised

Question 6

what is the citric acid cycle

Answer 6

where you make ATP, ADP and AMP implication in cancer and aging

Question 7

what does it mean if see genomic expression but no metabolimics?

Answer 7

the event isn’t stable enough

Question 8

what is primary metabolites

Answer 8

biomolecules that are synthesised by the cell because they are essential for growth and maianataince of a biological system

Question 9

what are secondary metabolites

Answer 9

organic compounds produced by bacteria, fungi, or plants which aren’t directly involved in normal growth and mainatance of a biological system

Question 10

what are examples of primary metabolism

Answer 10

amino acids, carbohydrate and alchohols and vitamins B2 and B12

Question 11

what are examples of secondary metabolites

Answer 11

Phenolic, steroids, essential oils and alkaloid metabolites

Question 12

what is 2HG

Answer 12

muatations in IDG result in the conversion of isocitrate to 2-HG instead of alpha ketoglutarate. This leads to altered methylation in cell and promotes tumour growth

Question 13

how is tyoe 2 diabetes caused

Answer 13

icreased level of TAG which leads to saturation of acetyl-coA leading to desythestization

Question 14

what is the Warburg effect?

Answer 14

cancer and tumourgenesis are due to tumour cells generating energy through the non oxidative breakdown of pyruvate.

Question 15

why do plants have largest metabolome

Answer 15

because they can’t. move

Question 15

why do plants have largest metabolome

Answer 15

because they can’t. move

Question 16

what are all lipids made from

Answer 16

C, O, H double bond

Question 17

what is metabolite target analysis s

Answer 17

quailitive and quantitive anaylsis of one, or several metabolites related to a specific matabolic reaction

Question 18

what is metabolic foot printing

Answer 18

analysis of the metabolites screted or excreted by an organism; if the organism is growing in culture it would include its envioemnt

Question 19

what is metabolic profiling

Answer 19

Identification and quantification of a selective number of predefined metabolites, which are related to a specific pathway

Question 20

metabolic finger printing

Answer 20

global, high throughput, rapid analysis to provide sample classification. Also utilised as a screening tool to discriminate between samples from different biological status or orgin.

Question 21

what are the main steps of untergetting metabolomics work flow

Answer 21

• experimental design
  -sample prep
  -sample anaylis by MS and NMR
  -pre processing data analysis
  -metabolite identification
  -hypothesis
  -ecperimental validation

Question 22

what should you do in discovery work

Answer 22

identify a panel of biomarkers taht can refine disovery to validation

Question 23

what is a spectroscopic technique

Answer 23

a non destructive technique taht uses radiowae laser so can profile samples at a decent enough reoliutio

Question 24

what are the main spectroscopic techniques that are used?

Answer 24

NMR and FTIR

Question 25

what is FTIR

Answer 25

it gives out a vibartional signature but is sensitive to water so needs to be processed properly

Question 26

what is the gold standard in metabolic?

Answer 26

LC/MS

Question 27

Which mass analyses are good for discovery work

Answer 27

QTOF and orbi trap

Question 28

what is the advantages if quadrouple mass analyser

Answer 28

Robust, simple, low maintaince, relatively inexpensive

Question 29

what are the disadvantages of the quadruple mass analysers

Answer 29

low mass resolution, low sensitivity, slow scan speed

Question 30

what is the disadvantage of having a too long. TOF

Answer 30

the ions can diffuse

Question 31

how does orbital work

Answer 31

the heavier the ion the longer it takes to orbit, ms2 analyses the results

Question 32

what is the simplist mass spec analyser

Answer 32

quadropole

Question 33

how does quadropole Give you the mass

Answer 33

the ratio of voltage is directly proptional to the mass

Question 34

what is MS3 used for

Answer 34

when you know the fragments after further smashing up fragments after using low res mass spec

Question 35

how do we know that the sample is fragmenting in MS2?

Answer 35

precursor intensity decreases with higher eV as more % is converted into fragments

Question 36

what does MS1 do

Answer 36

quick ID based on hi res ion mass measurement

Question 37

what does MS2 do

Answer 37

structural fragments for confirmation

Question 38

what does electro spray ionisation do

Answer 38

converts liquid samples to gas phase ions

Question 39

what is the mobile phase in GC

Answer 39

a coil

Question 40

what is the mobile phase in LC

Answer 40

pac column

Question 41

how does chromotogrpahy work

Answer 41

through plate theory and van deemter

Question 42

what does the van deemter equation state

Answer 42

the plates height can be decreased or enlarged by temp flow

Question 43

what is the most basic type of LC

Answer 43

TLC

Question 44

When would you use GC

Answer 44

if you sample was a gas, anything with a smell

Question 45

how does gas chormotgrpahy work

Answer 45

uses volatility go the gases. The most volatile are eluted quicker. It is then fragmented by election impact

Question 46

what is the major cost of gas chromoatgrpahy

Answer 46

electricity and gas

Question 47

what would not be detected in NMR

Answer 47

low abundance metabolites

Question 48

which chromatography is the gold standard

Answer 48

LC

Question 49

which types of chromatography would you use to separate complex liquid mixtures

Answer 49

PR, HILIC and lipidomics

Question 50

what analysis is required after LC

Answer 50

DDA or DIA

Question 51

What are the disadvantages of LC

Answer 51

- easily broken -long downtime -high operational skill needed -sophisticated downstream analysis needed

Question 52

what makes the high pressure in LC

Answer 52

solvent, column, flow rate and gradient composition

Question 53

what would you use GC for

Answer 53

sweat analysis, breath analysis, pesticides

Question 54

what disadvantage of GC

Answer 54

low coverage can only look at low molecular weigh metabolites

Question 55

what are the 3 different phases of LC metabolimics

Answer 55

reverse pahse- non polar. helix phase- polar and ion chomrotgarphy- sugar

Question 56

what are the advantages and disadvantages of polarity switching

Answer 56

reduced the time but reduces quality of data as one mode will be surpassed and other will be over expressed

Question 57

why is plastic a contimanation an issue and how is it over come

Answer 57

use plastic tools, have to use low bais to stop plastic leaching from the solvent. Would show as a bleed across the results. At proeitn would see much less as would be absorbed

Question 58

what are QCs

Answer 58

10% of samples go into a master pool. It is themes reproducible data point. Inject aliquots of QC at regular intervals to demonstrate stability and quality of aquisition

Question 59

what is unsupervised anaylisis

Answer 59

clustering algortm which doesn't now you labelling or class structures. Allows you to see outliers

Question 60

what is the most commonly used data analysis in unsupervised

Answer 60

pca, which removes noise from data set. Anything after PC1,2, and 3 is noise