HUMUS DETERMINATION PROTOCOL

Frank Chambers

1. Sample sufficient peat (e.g. for very fibrous and/or very wet peat, use 1 x 1 x 0.5 cm; for less fibrous peat, use less) and place in weighing boat. Cut up peat if very stringy; otherwise just leave in boat. Dry samples in boats in an oven (~50ºC) or under infra-red heat lamps.

2. Prepare 1 litre 8% NaOH solution. To do this, dissolve 80g NaOH granules of AnalaR grade in about 500 ml deionised water and make up to 1 litre.

3. Grind up each peat sample separately in an agate pestle and mortar, returning to weighing boats. *Note: clean the pestle and mortar between each grind using dry paper or clean cloth - do not wash.

4. Weigh out 0.2 g of sample accurately on a top-loading balance (to 3 dp) - use a piece of paper and tare weight - then shake into 150 ml beaker.

5. Turn on hotplate to preheat.

6. Add 100 ml of 8% NaOH solution to each beaker using a 100 ml measuring cylinder.

7. Place beaker on hotplate and simmer. Do not cover or boil fiercely.

8. Top up beaker occasionally with deionised water to prevent drying out and to ensure solution does not become too concentrated.

9. Turn hotplate off after 1 hour of slow boiling (simmering).

10. Pour beaker's contents into 200 ml labelled volumetric flask using a funnel, and wash all residue into flask with deionised water.

11. Top up flasks (when slightly cooled) to mark; stopper flask, then shake well.

12. Filter 50 ml of contents of each flask into separate 50 ml labelled volumetric flask, through filter funnel in rack using Whatman No. 1 grade papers (size 15 cm). Use filter paper in a corrugated folding mode, to speed filtration. Ensure only 50 ml filtrate is transferred.

13. Decant filtrate into labelled 100 ml volumetric flask.

14. Turn on spectrophotometer to stabilise. Set to 540 nm.

15. Top 100ml flask up to mark with distilled water and stopper. Then shake well.

16. When all flasks are ready, pipette small volume from first one into each of three* cuvettes and using spectrophotometer (previously allowed to stabilise), measure absorbance (upper scale) and % transmission of first cuvette and record it.

* alternatively, re-use cuvette twice more for the same sample only

17. Repeat for second cuvette (or for a second time) and, if a different reading is obtained, repeat for third cuvette (or time).

N.B. DO THESE READINGS IN NUMERICAL FLASK ORDER!

References
Aaby, B. and Tauber, H. 1975. Rates of peat formation in relation to degree of humification and local environment, as shown by studies of a raised bog in Denmark. Boreas 4, 1-14.

Blackford, J.J. and Chambers, F.M. 1993. Determining the degree of peat decomposition for peat-based palaeoclimatic studies. International Peat Journal 5, 7-24.

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