Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose...

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Lecture 6 Sample preparation
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Transcript of Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose...

Page 1: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Lecture 6Sample preparation

Page 2: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.
Page 3: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Component Percent

Water ~92

Proteins 6–8

Salts 0.8

Lipids 0.6

Glucose (blood sugar)

0.1

Composition of blood plasma

Blood serum is blood plasma without fibrinogen or the other clotting factors (i.e., whole blood minus both the cells and the clotting factors)

Blood Samples:Blood: arterial

capillaryvenous

Page 4: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Blood Samples:

Anticoagulants: lithium heparinsodium heparinNaF sodium citrateEDTA

Serum: no anticoagulant

Plasma: centrifugation with anticoagulant present

Blood cells: erythrocytes

Page 5: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Urine 20-50 mL

Stomach content 25 -50 mL

Tissue Hair, nails 2-5 g if possible

Many of options in postmortem work

Page 6: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.
Page 7: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Sampling

LiquidOptional protein precipitation

SolidOptional protein precipitation

Homogenize in buffer

Isolation-Single step liquid-liquid extraction- Multiple step liquid-liquid extraction- Solid phase extraction- (pre)concentration

DATA COLLECTION

Data processing and interpretation

Organic Compounds

Page 8: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Air/gas sampling

canistersbags

Cleaned Polyurethane Foam (PUF) Cartridges and Resins

Page 9: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Solid Phase Extraction cartridges

Page 10: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Headspace vial

Caps & septa

Page 11: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Microdiffusion (Conway) cell

Preconcentration of volatile compounds

Page 12: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

A simple device for headspace LPME. Stirring and thermostating are optional.

A 1 mL drop at the end of a syringe needle.

Liqiud phase microextraction (LPME)

Page 13: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

SPME

Page 14: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Solvent extraction techniques

Page 15: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Filters:Can be also used for Solid Phase Extraction

Page 16: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Solid Phase Extraction

Page 17: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.
Page 18: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Inside the SPE cartridge…

Page 19: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Various sorption techniques:

centrifuge

positive pressure

Page 20: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

Large volumes:

Page 21: Lecture 6 Sample preparation. ComponentPercent Water~92 Proteins6–8 Salts0.8 Lipids0.6 Glucose (blood sugar) 0.1 Composition of blood plasma Blood serum.

SPE extraction:Lots of junk removed!