Practical No.

11, 12 Time (180 minutes)

A. Title: Determination of total potassium content in plant tissues

B. Aims and Objective:

Learn procedure to estimate total potassium level in different tissue (leaves, fruits etc.)
of horticultural plants.

C. Material required:

1. Heating plate
2. Spectrophotometer
3. 100 ml beaker
4. Watch glass
5. 100 ml volumetric flask
6. 100 plastic bottles (for storage of sample)
7. Filter paper
8. Nitric acid (HNO3)
9. Per chloric acid (HClO4)
10. Sulphuric acid (H2SO4)
11. Distilled water
12. Potassium dihydrogen phosphate (KH2PO4)
D. Procedure/ activity:

1. This practical requires two practical classes. In first practical class, students will be
briefed about the underlying principles of total potassium determination in plant
tissues and analytical procedures involved in it. In next practical class, students will
conduct the practical work.
2. Students will wash, dry, grind leaf samples.
3. Information about digestion of organic matter and use of flame photometer will be
given with practical demonstration.
4. On-site determination of total potassium contents in leaf samples (add step wise
detail of whole procedure).
5. For estimation of K, digestion of plant material is done as per method described by
Yoshida et al. (1976).
6. For wet digestion, one-gram oven dried, powdered sample and 10 ml of tri-acid
mixture (HNO3, HClO4 and H2SO4; 5:2:1) is taken in 100 ml beaker.
7. The beaker is covered with watch glass and allowed to stand for about 4 h for initial
reaction to subside.
8. The mixture is heated slowly till solid material gets disappeared, then vigorously until
clear colourless solution is obtained.
9. When the volume of mixture is reduced to 1.5 ml it is removed and allowed to cool.
10. On cooling the solution is shifted to 100 ml volumetric flask and volume is made up to
mark by adding distilled water.
11. Potassium is determined by using flame photometer as per method described by
Chapman and Parker (1961).
12. Quantity of element is assessed in ppm by comparing the emission of flame
photometer with that of standard curve which is then converted to percentage by
using the formula:
ppm on graph × dilution
K (%) = × 100
106

Flow Chart
1 g oven dried sample + 10 ml tri-acid mixture (HNO3: HClO4: H2SO4 @ 5:2:1)
↓
Covered with watch glass and allowed to stand for four hours
↓
Heated slowly until solid disappear
↓
Heated vigorously till colourless
↓
Remove when volume reduced to 1.5 ml
↓
Cool, volume made up to 100 ml
↓
Stored in bottles
↓
Feed samples into flame photometers
 E. Activity to be conducted by the students:

1. Students in groups will perform hands on training of titration to determine ascorbic

acid contents in different fruit juices.
2. Students will write down and note the reading of various samples in their practical
note book.
3. At the end, groups will compare their readings to conclude the overall observation
taken by whole class.
Key Messages
1. Principle of flame photometry and its application in determination of potassium contents.
Flame photometry is a process in which emission of radiation by neutral atoms is measured.
Sample is introduced into the flame to obtain neutral atoms. When a solution containing
cations of potassium is spayed into flame, the solvent evaporates and ions are converted into
atomic state. In the heat of the flame, small fraction of the atoms is excited. Relaxation of the
excited atoms to the lower energy level is accompanied by emission of light (photons) with
characteristic wavelength i.e. Na at 589 nm, K at 766 nm. Intensity of the emitted light
depends on the concentration of understudy atom in flame. Since radiation is emitted it is
also called as flame emission spectroscopy.