SPAD meter is very frequently used as an effective tool for measuring chlorophyll in field .But i agree , its not as accurate compared to destructive means of chlorophyll analysis conventionally used in laboratories...

Hello

Chlorophyll content was traditionally measured in the laboratory by extracting the chlorophyll itself from a leaf sample using acetone before calculating the chlorophyll concentration by spectrophotometrically measuring absorption at 663 nm and 645 nm. This process is time consuming, destructive to the plant material and above all, impractical in applied situations.

The other option is to use a hand-held device such as the Chlorophyll Content Meter which calculates relative chlorophyll content using dual wavelength absorbance measurements through a leaf at 620 nm and 940 nm. The last noninvasive methods are fast, easy to use, and produce reliable estimates of relative leaf chlorophyll, but the results are not much accurate in low light intensity.

So a rapid image processing method has been proposed to get the better results. Using a video camera and personal computer, chlorophyll content of leaves was estimated from the red and blue wavelengths and its accuracy was improved using the normalized difference (red-blue)/(red+blue).

Hi

For measuring chlorophyll content, you can use LICHTENTHALER protocol that uses spectrophotometry method. Also, you can use a spad device for measuring chlorophyll.

Regards

Extract Chlorophyll by Acetone (80%), then read OD in certain wavelength, followed by Arnon's method.

We need to establish these methodologies with certain growth or yield attributing features of crops to arrive at most suitable method..

Sundus Abdulkariem Alabdulla

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The chlorophyll content estimation according to Arnon (1949), tissue pieces macerated with 80% acetone and pinch of calcium carbonate also added. The homogenate is centrifuged at 5000 rpm for 10 min; the supernatant is made upto known volume with 80% acetone. OD of the supernatant is taken at 645 and 663 nm against 80% acetone as blank. Formula for chlorophyll content :

Dr Lie to has picked up very good point. SPAD - based chlorophyll is still quite popular looking at the ease of measurement , cost effectiveness and time consumed in facilitating such analysis. Most important is to standardise the time of sampling in terms crop phenophase so that it related well with crop performance or biomass production., unless this is done , we will continue facing dubicity about SPAD....

Easy way of measuring chloropyll a, b and carotenoids content is homogenization of leaves in 96% ethanol (Lichtenthaler, 1987). Then filtration of homogenate through filter paper to obtain extract of leaves. Optical dencity of pigments in plant extract is determined at 664 nm for Chl a, at 648 nm for Chl b, at 470 nm for car (x+c). The concentration of pigments is calculated according to the formulas:

Chlorophyll a= (13,36 x A664- 5,19 x A648) x 25/dry mass

Chlorophylll b= (27,43 x A648- 8,12 x A664) x 25/dry mass

Carotenoids (x+c)= (4,785 x A470 + 3,657 x A664- 12,76 x A648) x 25/dry mass

A very quick and simple route is to use optical absorption in the red and far red to give a relative chlorophyll content. The CCM200+ is a small, hand-held unit which makes this measurement and stores the results for later download via a USB port.

Another convenient technique is to measure chlorophyll fluorescence at two wavelengths (705nm and 735nm) and the ratio of the result is linearly correlated with chlorophyll content. The CCM300 uses this method. The advantage is that this technique gives absolute units in mg/m2, it has a much larger measurement range, we do not need to completely fill the sample window and because it does not need to pass light through the sample, can be used with thick leaves, pine needles, etc.

If you have questions then contact me at [email protected]

Extract with 96% ethanol and use the "dioxane method". Only chlorophylls are precipitated with dioxane.

Minolta SPAD 502DL plus Chlorophyll Meter

The chlorophyll meter measures leaf absorbance of red light at 650nm and infra-red light at 940nm. This has been calibrated for the chlorophyll content of tissues and for tissue greenness. The information on leaf chlorophyll content (greenness) will be useful for monitoring post-anthesis senescence profiles.

Hello

Spectrophotometrically method is a good and approximately accurate manner to determine chlorophyll content in plants tissue. You can measure Chl a, Chl b and total carotenoids in plant according to Lichtenthaler 1987.

Regards

hello

you can use of porra 2002 and porra 1989 for determine chlorophyll content in plant leaves.

hello

you can use of this article.

I believe for convenience, the non-destructive method using either SPAD or atLeaf chlorophyll meter will do.

of acetone or HPLC

G'day From Perth, Western Australia

There are a few mentions of Arnon (1949) as a means to calculate chlorophyll concentration in extracts of leaf tissue. Additional to Arnon (1949) other common references used for this calculation are Lichtenthaler (1987), Wellburn (1994) and Porra (1989, 2002).

The equations published by Arnon (1949) for aqueous 80% acetone extracts have been shown to be based upon the underestimated extinction coefficients of Mackinney (1941), thereby leading to underestimate of chlorophyll a and b concentrations.

I urge people to read Esteban et al (2018) and Porra & Scheer (2019). No disrespect meant in this communication, as scientists we should all be open to advancements in methodologies, being made aware of such changes does take time.

stay safe!

More suitable method to measure chlorophyll a, b and carotenoids content is a method proposed by Lichtenthaler (1987).

For destructive method SPAD 502 or atLEAF devices

For non-destructive method use spectrophotometer (80% acetone and 100% ethanol).

https://www.adc.co.uk/products/mpm-100-multi-parameter-meter/

measures chlorophyll, flavonol, anthocyanin and nitrogen-flavonol index.

You can use method SPAD or spectrophotometric method.

Here is a link that can help

Article Nondestructive Determination of Total Chlorophyll Content in...

Yes, SPAD method and some other devices are available, No way for obtaining less precise results otherwise the sample should be cut for spectrophotometry, cutting plant parts or individuals reduces the designed plant density resulting in a bias in other measured traits. Analysis by either of the methods depends on which data is preferred by you.

I would recommend the answer of Murat Sabri Sadiklar

This article can help you very well the easiest way of measuring chlorophyll content of leaf.

Thank you!

I would like to recommend this paper:

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