Materials and Reagents

1. Chloroform (Carl Roth, catalog number: 7331.2 )
   
2. Heptatriacontane (Fluka, catalog number: 51848 )
   
3. Dotriacontane (Fluka, catalog number: 44253 )
   
4. N,O-bis-trimethylsilyl-trifluoroacetamide (BSTFA) (Macherey-Nagel, catalog number: 701220.201 )
   
5. Pyridine (Merck, catalog number: 1074630500 )
   
6. 1.25 M methanol-HCl (Fluka, catalog number: 17935 )
   
7. Sodium chloride-saturated aqueous solution (Applichem, catalog number: A2824 )
   
8. Anhydrous sodium sulfate salt (Applichem, catalog number: A1048 )
   
9. Pectinase (Trenolin Super D) (Erbsloh, catalog number: 20312 )
   
10. Cellulase (Cellulast) (Novo Nordisk AIS, catalog number: CA451088 )
    
11. Citric acid monohydrate (Sigma-Aldrich, catalog number: C1909 )
    
12. Liquid nitrogen
    
13. 20 mM citrate buffer, pH 3.0 supplemented with 1 mM sodium azide (see Recipes)
    

Equipment

1. Freeze-drier
   
2. Nitrogen cylinder and blow-down system
   
3. Block heater with supports for glass scintillation vials
   
4. Glass scintillation vials (for volume 15-20 ml)
   
5. Glass vials with conic bottom (for volume 1-1.5 ml)
   
6. Glass vials (with volume 1.5 ml) and glass inserts for them (with volume 0.2 ml)
   
7. Glass funnels and glass Pasteur pipette
   
8. Screw caps with Teflon or PTFE liners for the glass vials
   
9. Glass syringes (for volumes 1,000, 100, 50 and 10 μl)
   
10. Paper filters
    
11. Nylon filters (with pores 41 μm) (EMD Millipore, model: NY4102500 )
    
12. Glass filtration apparatus
    
13. GC-MS: Temperature controlled capillary gas chromatograph (Agilent Technologies, model: 6890N ) with on-column injection (J&W Scientific; 30 m DB-1, 320 μm i.d., d_f = 1 mm) and a mass spectrometric detector (Agilent Technologies, model: 5973N ; 70 eV; m/z 50–750)
    
14. GC-FID: Capillary gas chromatography (Hewlett-Packard, model: 5890 II ) and flame ionization detection
    

Procedure