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Lecture 6 Sample preparation

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

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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!