Colorimetric Determination of Humic Matter with 0.

2N NaOH Extraction
David H. Hardy

Application and Principle

Organic matter OM! i" compo"ed of plant and animal material" in #ariou" "tage" of degradation. Humu" i" the relati#el$ "ta%le fraction of OM that i" &nown to %e a ma'or component of a "oil(" CEC) water holding capacit$) and "tructure. *t" a%undance gi#e" "oil" a dar& %rown to %lac& color. Organic matter i" determined %$ #ariou" method" including lo"" on ignition +O*! and chemical method" "uch a" ,al&le$-.lac& /012!. Chemicall$) humu" i" fractionated into humic and non-humic material" %a"ed on "olu%ilit$ in acid or al&ali "olution". Humic and ful#ic acid" are the ma'or component" of the humic material. 3he Agronomic Di#i"ion- 4oil 3e"ting 4ection- North Carolina Department of Agriculture and Con"umer 4er#ice"( la% determine" humic matter HM! primaril$ to cla"" "oil". *n North Carolina) a "ignificant acreage of Hi"to"ol" and mineral "oil" with hi"tic epipedon" exi"t in northea"tern 3idewater! and "outhern region" of the coa"tal plain". 4ome of the"e "oil" are a""ociated with Carolina .a$". 3he underl$ing rea"on for a cla"" determination i" related to target pH and the lime re5uirement. Acidit$ a""ociated with high le#el" of OM i" not t$picall$ pre"ent a" exchangea%le Al61 "ince it i" in"olu%le due to complexation with OM E#an" and 7amprath) /080!. Hence) a lower target pH of 9.0 to 9.9 can %e utili:ed in ma&ing lime recommendation" if "oil" are cla""ed into group" %a"ed on humic matter content. 3he addition of HM with other "oil te"t data al"o pro#ide" a more comprehen"i#e o#er#iew of "oil fertilit$.

E5uipment and Apparatu"

/. 2-mm "ie#e 2. 4coop) / cm1 #olume and le#eling rod 1. ;ial"< pol$"t$rene) 99 m+ capacit$ 1= x 89 mm! or of "imilar "i:e. 2. 4$ringe pump or pipette> 20-m+ #olume 9. Diluter ? di"pen"er@ 20@20) /0@10 or 9@19 m+ ratio depending on "tandardi:ation. =. A;?;*4 colorimeter) 2-cm light path with a =90 nm filter

Note@ Ea"e of anal$"i" i" made if the colorimeter i" e5uipped with a fi%er-optic pro%e. 3he Agronomic Di#i"ion- 4oil 3e"ting 4ection- North Carolina Department of Agriculture and Con"umer 4er#ice" utili:e" the .rin&man Model 0/0 colorimeter.

Beagent"
/. 4odium h$droxide NaOH!) fw C 20.0 Dunction@ 4olu%ili:e" humic acid" and Na6 act" a" a di"per"ant 2. Dieth$lenetriaiminepentaacetic acid DP3A!) penta"odium "alt 2/E in water) D, C 901.2= Dunction@ Aid" in the di"per"ion of larger Ca-humate compound" 1. Eth$l alcohol) denatured C2H9OH! Dunction@ Aid" in wetting h$dropho%ic OM "urface"

Procedure
/. Extractant@ 0.2N NaOH 6 0.002M D3PA 6 2 E Alcohol A"ing a 90-+ car%o$) add /0 + of water) 200 g NaOH and di""ol#e. Add 200 m+ of D3PA) / + of eth$l alcohol and ma&e to #olume with water and thorough mixing. Once mixed) the pH of the extractant "hould %e a%out /2.0. 3hi" pro#ide" "ufficient #olume for /200 "ample". 2. 4tandardi:ation Dor the "emi-5uantitati#e purpo"e of thi" procedure) "tandardi:ation i" not routinel$ conducted. 3he humic acid reagent u"ed in the original Mehlich /0F2! method i" not a#aila%le toda$.
1. Anal$"i" Mea"ure /.0 cm1 "oil G2 mm! into 99 m+ pol$"t$rene #ial" and add 20 m+ of extractant with enough force to mix with "oil. After /.0 hour) add another 20 m+ of extractant with enough force to mix> allow "ample" to "et o#ernight. 3ran"fer 9.0 m+ of undi"tur%ed "upernatant top half of "olution! from each "ample and add 19 m+ H20 into pol$"t$rene #ial" with enough force to mix. A"ing a .rin&man pro%e fi%er-optic colorimeter with a =00 nm filter) "et in"trument to auto-:ero in a "olution of 9.0 m+ extractant 6 19 m+ of water in"trument "hould read /00 E3!. *mmer"e pro%e into un&nown "ample and read percent tran"mi""ion E3!. Calculation of g HM?/00 cm1 in the "oil i" "hown %elow.

Calculation"

3he humic matter content of a "oil i" determined %$ extrapolating from 3a%le / which contain" humic matter #alue" a" related to tran"mi""ion. 3a%le /. Humic matter g humic matter ? /00 cm1 "oil! a" related to tran"mi""ion.H 0 / 2 1 2 9 = 8 F 0 / /0.00 0.90 0.2/ F.F= F.92 F.2/ 8.0= 8.80 8.29 8.2/ 2 =.00 =.8F =.9F =.1F =.20 =.02 9.F9 9.=0 9.91 9.1F 1 9.21 9.00 2.09 2.F/ 2.=0 2.9= 2.22 2.12 2.20 2.00 2 1.0F 1.F8 1.88 1.=8 1.98 1.28 1.18 1.2F 1./0 1./0 9 1.0/ 2.02 2.F2 2.8= 2.=F 2.=0 2.92 2.22 2.18 2.20 = 2.22 2./9 2.0F 2.00 /.02 /.F8 /.F0 /.82 /.=8 /.=/ 8 /.99 /.20 /.21 /.18 /.1/ /.29 /./0 /./2 /.0F /.02 F 0.08 0.02 0.F= 0.F/ 0.8= 0.8/ 0.== 0.=0 0.9= 0.9/ 0 0.2= 0.2/ 0.1= 0.12 0.28 0.22 0./F 0./1 0.0F 0.02 /00 0.00 H A two-digit tran"mi""ion #alue i" repre"ented %$ com%ining a num%er in the left hand column with a num%er in the upper row. 3he humic matter #alue i" contained within the cell at the point where the"e two tran"mi""ion #alue" inter"ect in the ta%le. Dor example) a tran"mi""ion #alue of 89 corre"pond" with a humic #alue of /.29 g HM ? /00 cm1 "oil. 3he data repre"ented in 3a%le / can %e expre""ed mathematicall$ %$ the e5uation@ $ C /00.0/ e -0.2102x where IxJ and I$J repre"ent g HM ? /00 cm1 "oil and tran"mi""ion) re"pecti#el$.

Anal$tical Performance
Bange and 4en"iti#it$ ,ithout dilution) a le#el of HME of approximatel$ /0 g HM?/00 cm1 "oil can %e attained. 3he "en"iti#it$ depend" on the concentration of humic acid". Preci"ion and Accurac$ No data are a#aila%le a" to the actual accurac$ of thi" mea"urement or method. .a"ed on the anal$"i" of three chec& "ample" in North Carolina(" la%orator$) the following data were attained. Chec& 4oil A E , n C 9= Mean 3ran"mi""ion --E-0=.2 /2./ F=./ 4td. De#iation --E-/./ /.1 2.0 Mean HM 4td. De#iation g HM?/00 cm1 0./= 0.09 0.20 0.22 0.=9 0./0

*nterference" None are t$picall$ encountered. Prior to dilution) an$ "u"pended material ma$ potentiall$ interfere with light mea"urement" although the"e are not normall$ encountered. *f dilution occur" with "uch force that air %u%%le" are created in the "olution to %e read) "ufficient time to allow air %u%%le" to diffu"e through the "ample" i" re5uired.

*nterpretation
North Carolina(" "oil te"ting la% u"e" HM to determine "oil cla""< mineral M*N!) mineral-organic M-O!) and organic OBK! a" follow". M*N L HM M 1.18 g ? /00 cm1 "oil. M-OL HM N 1.18 g ? /00 cm1 "oil and HM M 9.21 g ? /00 cm1 "oil OBK L HM N 9.21 g ? /00 cm1 "oil

4oil" in portion" of North Carolina(" coa"tal plain" region and to a more limited extent in the mountain" contain con"idera%le amount" of organic matter that limit" aluminum "olu%ilit$ at lower pH. Con"e5uentl$) the cla"" "$"tem i" utili:ed to "et target pH for which lime rate i" determined. 3arget pH i" "et at 9.0) 9.9 and =.0 to =.9 for OBK) M-O and M*N cla""e") re"pecti#el$. 3he pH range gi#en for the M*N i" dependent upon the crop to %e grown> for example corn ha" a target pH of =.0 wherea" alfalfa ha" a target pH of =.9.

Effect" of 4torage
Air-dr$ "oil can %e "tored indefinitel$ without affecting thi" mea"urement. 3he "olution &eep" indefinitel$ a" long a" co#ered from out"ide air.

4afet$ and Di"po"al

,hen di"pen"ing the extractant) "afet$ gla""e" and a full face "hield are re5uired gi#en the cau"tic nature of the %a"ic "olution. Other proper precaution and per"onal protection e5uipment re5uired at glo#e" and a la% coat "ince the "olution i" cau"tic. ,a"te "olution and "oil! can %e di"carded without concern.

Beference"
E#an") C. E. and E. O. 7amprath. /080. +ime re"pon"e a" related to percent Al "aturation) "olution Al) and organic matter content. 4oil 4ci. 4oc. Proc. 20@F01-F0=.

Mehlich) A. /0F2. Photometric determination of humic matter in "oil") A propo"ed method. . Comm. 4oil 4ci. Plant Anal. /9 /2!@ /2/8-/222. ,al&le$) A. and *. A. .lac&. /012. An examination of the Degt'areff method for determining "oil organic matter and a propo"ed modification of the chromic acid titration method. 4oil 4ci. 18@ 20-1F.

HUMUS DETERMINATION PROTOCOL Frank Chambers 1. Sample sufficient peat (e.g. for very fibrous and/or very wet peat, use 1 1 !." cm# for less fibrous peat, use less$ and place in weighing boat. Cut up peat if very stringy# otherwise %ust leave in boat. &ry samples in boats in an oven ('"!(C$ or under infra)red heat lamps. *. +repare 1 litre ,- .a/0 solution. 1o do this, dissolve ,!g .a/0 granules of 2nala3 grade in about "!! ml deionised water and make up to 1 litre. 4. 5rind up each peat sample separately in an agate pestle and mortar, returning to weighing boats. 6.ote7 clean the pestle and mortar between each grind using dry paper or clean cloth ) do not wash. 8. 9eigh out !.* g of sample accurately on a top)loading balance (to 4 dp$ ) use a piece of paper and tare weight ) then shake into 1"! ml beaker. ". 1urn on hotplate to preheat. :. 2dd 1!! ml of ,- .a/0 solution to each beaker using a 1!! ml measuring cylinder. ;. +lace beaker on hotplate and simmer. &o not cover or boil fiercely. ,. 1op up beaker occasionally with deionised water to prevent drying out and to ensure solution does not become too concentrated. <. 1urn hotplate off after 1 hour of slow boiling (simmering$. 1!. +our beaker=s contents into *!! ml labelled volumetric flask using a funnel, and wash all residue into flask with deionised water.

11. 1op up flasks (when slightly cooled$ to mark# stopper flask, then shake well. 1*. Filter "! ml of contents of each flask into separate "! ml labelled volumetric flask, through filter funnel in rack using 9hatman .o. 1 grade papers (si>e 1" cm$. ?se filter paper in a corrugated folding mode, to speed filtration. @nsure only "! ml filtrate is transferred. 14. &ecant filtrate into labelled 1!! ml volumetric flask. 18. 1urn on spectrophotometer to stabilise. Set to "8! nm. 1". 1op 1!!ml flask up to mark with distilled water and stopper. 1hen shake well. 1:. 9hen all flasks are ready, pipette small volume from first one into each of three6 cuvettes and using spectrophotometer (previously allowed to stabilise$, measure absorbance (upper scale$ and - transmission of first cuvette and record it. 6 alternatively, re)use cuvette twice more for the same sample only 1;. 3epeat for second cuvette (or for a second time$ and, if a different reading is obtained, repeat for third cuvette (or time$. ..A. &/ 10@S@ 3@2&B.5S B. .?C@3BC2D FD2SE /3&@3F References Aaby, B. and Tauber, H. 1<;". 3ates of peat formation in relation to degree of humification and local environment, as shown by studies of a raised bog in &enmark. Boreas 4, 1)18. Blackfor , !.!. and C"a#ber$, %.M. 1<<4. &etermining the degree of peat decomposition for peat)based palaeoclimatic studies. International Peat Journal &, ;)*8.