A method I have used in the past is the Van Soest  method - Goering H. (1970). There are newer methods from NREL.  

Dear Joan,

Thanks for your reply. We have been using the same NREL method. although the problem is that the value of NDF is more than ADF. which should not happen, as the ADF is inclusive of Cellulose, Hemicellulose and Lignin.

Kindly help me with this.

Dear Geeta,

There are some papers comparing some tehcniques to quantify lignin content...

http://goedoc.uni-goettingen.de/bitstream/handle/1/7442/Journal_Chemical_Ecology.pdf?sequence=1

http://www.fpl.fs.fed.us/documnts/pdf1988/kirk88c.pdf

http://bioenergycenter.org/besc/publications/jiang_rapid_determination.pdf

and structural polysaccharides as well....

http://www.nrel.gov/biomass/pdfs/42618.pdf

http://www.sciencedirect.com/science/article/pii/S0981942809002009

I think that Van Soest method is still usefull for large amount of plant material analyses for comparative purposes.

Best rgards,

Gustavo

Dear Gustavo,

Thanks for sharing the literature. Some of these literatures I have been using. The main problem is that  the value of ADF is more than NDF which should not happen, as the NDF is inclusive of Cellulose, Hemicellulose and Lignin while ADF is only cellulose and hemicellulose. 

Kindly help me with this.

Wise’s chlorite for holocellulose and Kurschner-Hoffer nitric acid method for cellulose and Tappi T 222 om-88,  for lignin. I can explain these methods if you want

You can follow the NREL standard protocol to determine the biomass composition (lignin, cellulose, and hemicellulose)

http://www.nrel.gov/biomass/analytical_procedures.html

In addition, use acetyl bromide method for lignin determination

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0110000

Thank you all for your answers.

Hi Geeta,

Standard Methods of AOAC (2000) are useful in determining the chemical composition of lignocellulosic biomass.

As NDF = Cellulose + Hemicellulose + Lignin

ADF = Cellulose + Lignin

So, NDF can never be less than ADF.

I believe you remember that biomass is hygroscopic in nature. The kind of problem you are facing may generally arise due to imprecise weighing practice. Please, make sure:

Other than weighing issues, please confirm that:

Hope this will help you.

Mustafa,

Do you have all the reference papers for determination of cellulose, hemicellulose and lignin? If you care to share them. Thanks.

I have them. Please send me your email

Gd morning,

i also need material related to analysis of cellulose, hemicellulose and lignin in biomass.If possible from your side please. send me on [email protected].

Hi Chirag

you can use acetyl bromide method for lignin determination

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0110000

ADF ad NDF follow 

http://www.johnmorris.com.au/files/product/attachments/4469/68518_opt2.pdf

http://www.johnmorris.com.au/files/product/attachments/4469/68518_opt1.pdf

Mustafa, 

Please send to [email protected]. Really appreaciate it.

I will explain tomorrow. I am busy now 

The cellulose was obtained by refluxing wood meal three times for 1 h with a 1:4 (v/v) mixture of nitric acid and ethanol. Three replicates were done for each experiment, and mean values were used (Browning, 1967; Tutus et al., 2014; Tutus et al., 2015),.

10 ml Nitric Acid, 40 Ml Ethanol (each replicate)

After 1 hour, remove nitric acid and ethanol mixture with 2 por gooch crucible. Then, again add nitric acid and ethanol. For softwood you should four replicates, for hardwood you should three replicates. And dont forget to use refluxing. 

References:

Browning, B.L., 1967: Methods of Wood Chemistry, Vol. 2. Interscience/Wiley, New York.

Tutus, A., Kazaskeroglu, Y., Cicekler, M., 2015: Evaluation of Tea Wastes in Usage Pulp and Paper Production, Bioresources 10(3), 5407-5416.

Tutus, A., Cicekler, M., Ozdemir, A., Altas, A., 2014: Evaluation of Astragalus membranaceus in Pulp and Paper Production, 3rd International Non-wood Forest Products Symposium, 323-331, 8-10 May 2014, Kahramanmaras-Turkey.

 Good morning Mustafa,

i also need material related to analysis of cellulose, hemicellulose and lignin in biomass.If possible from your side please. send me on [email protected]

Thanks a lot :)

Can anyone suggest a simple method for determining cellulose, hemicellulose and lignin content of dried leaves? - ResearchGate. Available from: https://www.researchgate.net/post/Can_anyone_suggest_a_simple_method_for_determining_cellulose_hemicellulose_and_lignin_content_of_dried_leaves [accessed Sep 6, 2016].

I recommend Joan's answer

Best Regards

i also need cellulose, hemicellulose and lignin content measurement in %. please send me at [email protected]

The below mentioned researcher has many articles on

determining cellulose, hemicellulose, and lignin content of many types of biomass (including leaves of rice straw etc). Happy new year.

https://www.researchgate.net/profile/Yehia_Fahmy

the 4:1 ethanol:HNO3 method (Kürschner Hoffer) works very well for wood is very simple non toxic - it has beeen originally invented for wood as far as I know in 1929, it works well especially for hardwood; for softoowd digestion it is not so precise.... But with my experience there can be touble for other material (e.g. fossile wood). I have no experience with leaves though. If you need complex assessment of extractives, holocellulose, lignin, then there is a method by Rowell et al 2012, which is easy to follow and at the end you have all parameters. IF you need it I can send the method description to you.

Dear Hofmann, I am interested in your method

ok!

Dear Geeta Yadav , Nasir Ali et al.,

a similar discussion in ResearchGate can also be found here:

https://www.researchgate.net/post/Is_there_an_easy_procedure_to_determine_the_amount_of_lignin_hemicellulose_and_cellulose_present_in_a_biomass_sample

FTIR may be used to quantify lignin and cellulose, for example:

Article Evaluation of morphological and chemical aspects of differen...

https://www.astm.org/Standards/E168.htm

Article ChemInform Abstract: Qualitative and Quantitative Analysis o...

Article Practical uses of FTIR spectroscopy in wood science and technology

Popescu et al. (2011), for example, wrote: "FT-IR spectroscopy can offer the possibility of a quantitative lignin determination in wood. The intensity of the band of aromatic skeletal vibrations around 1510 cm-1 is a measure for the lignin content. Faix [24] indicate that the following non-linear equation describes well the relation between the lignin content and the height of the 1510 cm-1 band x:

% Lignin = -1.23 + 193.4 x + 279:8 x2

The ASTM E168 method may be used to construct calibration curves with biomass constituents (xylan, cellulose, kraft lignin; commercially available from Sigma Aldrich and other suppliers). With the help of the linear equations derived from the calibration curve one can then quantify the amount of the constituent in the sample. Knowing the total amount of sample used for FTIR one can then determine the relative content.

Good luck, Rob.

In order to determine cellulose, you must first determine the neutral detergent fibre (NDF), acid detergent fibre (ADF), and acid detergent lignin (ADL) in your samples, then use the following equations:

- Cellulose = ADF - ADL

- Hemicellulose = NDF - ADF

Using the Van Soest et al , (1991) method is the rignt method. You should first determine the NDF, ADF and ADL content of your samples, then you have the cellulose, hemicellulose by a simple soustraction

Cellulose = ADF-ADL and, Hemicellulose= NDF.

The should determin first ADF NDF ADL.

follow this

https://www.scielo.br/scielo.php?pid=S1517-83822002000300005&script=sci_arttext

YOU CAN USE ASTM D1103-60 for cellulose

Please refer https://shodhganga.inflibnet.ac.in/handle/10603/141185

Use the AOAC methods.

Check https://www.aidic.it/cet/19/72/014.pdf

Use the Nrel method.

for me , i use the Van Soest Detergent Fiber analysis .

Salam Alaikoum

I'm so sorry for the delay. yes, it's clear ..i 've used fibertec for this purpose.

please sent me your email..i will transmit to you all detail.

If you want, you can do this analysis in our center, it's available and free

Sorry again

This is my email: [email protected]

Thank you In advance Amokrane Serine .

Actually, I have used another method, but I found some difficulties especially in the separation.