bioanalytik at Universität Tübingen

Flashcards and summaries for bioanalytik at the Universität Tübingen

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Database repertoires

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Electrochemical detection (EC)

Exemplary flashcards for bioanalytik at the Universität Tübingen on StudySmarter:

Clean up methods

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Diode array detector (DAD)

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Gases

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Organs/tissues

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Hydrophobic interaction chromatography (HIC)

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Exemplary flashcards for bioanalytik at the Universität Tübingen on StudySmarter:

Ion Exchange (IEX) Chromatography

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Particle material

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UV/Vis Detector

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FSC

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Exemplary flashcards for bioanalytik at the Universität Tübingen on StudySmarter:

Column types

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Exemplary flashcards for bioanalytik at the Universität Tübingen on StudySmarter:

bioanalytik

Database repertoires

  1. Sequence data base (ex. NCBI)
    • DB for DNA/RNA sequences
  2. Database for other primary data:
    • expression data, metabolome data, protein crystal structure
  3. secondary DB (ex. EXPASY):
    • contain info derived from primary DB
  4. cumulative DB (ex. BRENDA):
    • collect data bout specific subject/organism

bioanalytik

Electrochemical detection (EC)

1. Pulsed Amperometry: 

- Voltage between two electrodes.

- Voltage changes if analyte is oxidized at anode or reduced at cathode

-Destructive and electrodes might degrade 


2. Condictivity Detection:

- Constant potential between two electrodes

- Passing analytes change the conductivity

- Non-destructive but less sensitivity

bioanalytik

Clean up methods

  • precipitation
  • extraction (liquid-liquid, SPE)
  • Filtration:
    • material: cellulose (native proteins), polymer (denatured), glass/ceramic (metabolites)
    • consider: pore size, stability of membrane, sticking of analyte to filter, denaturing of proteins, dead volumes

bioanalytik

Diode array detector (DAD)

  • similar to UV/Vis
  • 1st entire light goes through sample then split by prism
  • can measure at any given time point entire UV/Vis spectra 
  • able to check purity of peak by checking spectra at multiple points (if similar peak is pure)
  • Different spectra in 1 peak: impurity 

bioanalytik

Gases

  • mostly easy sample taking & relatively clean sample
  • Gas mouse 
  • liquids that absorb gases
  • integrated sampling with solid absorbing materials
  • small gas molecules can diffuse trough container !

bioanalytik

Organs/tissues

  • very difficult and complex 
  • mostly inhomogeneous (min sample weight 100 mg)
  • removal of different tissues prior analysis
  • Tissue already influenced by sampling procedure (must be quick and shock freeze as soon as possible)
  • sample prep prior storage
  • document: 
    • Plants: developmental stage, daytime, temp, humidity, illumination, treatment, genotype 
    • Animals: sex, race, age, nutrition, health, treatment
  • different parts of organism should analyzed separate to exclude integrated results because of inhomogeneous distribution

bioanalytik

Hydrophobic interaction chromatography (HIC)

  • SP: non polar (aromatic surface)
  • MP: buffer; inverse salt gradient from strong to weak 
  • Hydrophobic & aromatic interactions
  • For when RP/ NP fail PROTEINS; NUCLEIC ACIDS

bioanalytik

Ion Exchange (IEX) Chromatography

Stationary phase: 

  • Anion exchanger: positively charged sp binds anions
  • Cation exchanger: neg charged sp binds cations

Mobile phase:

  • Buffer 
  • Consider: exchanger & analyte MUST be charged; select good buffer for right pH
  • High capacity but low separation power 
  • Used for proteins & DNA
  • EXCELS at Amino acid separation

bioanalytik

Particle material

Responsible for stability of column (solvents; pH; pressure)

  • Silica: stable at acidic pH not at basic; pressure stable
  • Polystyrol-, Acrylate-polymers: wide pH stability; low pressure stability
  • Sugar polymers: low pH and low pressure stable; biocompatible (hydrophilic); mainly for SEC

bioanalytik

UV/Vis Detector

  • measures light absorption at predefined wavelength (190-1000 nm)
  • Prism splits light from source 
  • slit blocks all but selected wavelength
  • versatile
  • non-destructive
  • photomultiplier detector

bioanalytik

FSC

  • Forward scattered light
  • info on size of particle 

bioanalytik

Column types

  • Packed: column filled with small particles that have pores
  • monolithic: no individual particles inside but 1 block with many gaps & pores

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