A) Project No.

ARCL/R&D/Agri-001

Project Title: - Humic Acid extraction & application.

Humic Acid is a mixture of complex organic acid molecules which forms during the
decomposition of organic matter. In fact, they are the most active part of organic matter. It is
present in plant remains, Marsh peats, lignite deposits and in soil also.

Fig 1: Possible structure of Humic Acid

1) There are two ways for the preparation of Humic acid as follows:

Humic Acid

Extraction from natural humus Humification through

(Leonardite, Cow dunk) Biodegradation
(Plant & animal residue)

Extraction

2) Extraction of Humic acid by the NAGOYA method:

 The dried samples were crushed and passed through a 2mm mesh sieve.
 The sieved samples were suspended in a 0.1 N NaOH solution in the ratio of 300 ml to the
corresponding amount of the sample containing 1 g organic carbon which had been analysed,
and allowed to stand under N₂ for 48 h at 25°C with intermittent shaking.
 The extracted solution was separated from the residue by centrifugation at 1,500 x g for 15
min after NaCI was added to make 3%.
 The residue was washed with the same volume of 0.1 N NaOH containing 3% NaC1, and
the washing was collected by centrifugation at 1,500Xg for 15 min and combined with the
first extract.
 The extract was filtered with glass-wool, acidified to pH 1.0 with 3 N HCI, allowed to stand
for 10-16 h, and centrifuged at 1,500 x g for 10 min.
 The precipitated HA fraction obtained by the addition of 3 N HC1 to the extract was
neutralized with 2 N NaOH, redissolved in 0.1 N NaOH containing 3% NaC1, and
centrifuged at 6,000 x g for 20 min under N₂ to remove impurities.
 Then the HA solution was diluted with 50 times the volume of 3% NaC1 solution,
reprecipitated with 3 N HC1, and allowed to stand for 10-16 h. The precipitate was collected
by centrifugation at 1,500 x g for 10 min.
 Neutralization, dissolution, dilution and reprecipitation were repeated until the acid
supernatant became almost colourless. The HA precipitate was treated 3 times with HCl-HF
(0.1 M HCl : 0.3 M HF=1 : 1, HA : HCl-HF=1 : 10) by shaking for a day to remove ash, and
dialyzed against distilled water until Cl- was no longer detected with AgNO3, and freeze-
dried to get a powder of HA sample.