LYCOPODIUM SPORE METHOD: (QUANTITATIVE MICROSCOPY)

● It is an important analytical technique for powdered drugs especially

when chemical & other methods of evaluation of crude drugs fail as
accurate measures of quality.
● Lycopodium is the spores of the species of club moss, Lycopodium
clavatum (fam: Lycopodiaceae).
● Lycopodium spores are very characteristic in shape and appearance and
exceptionally uniform in size(25µm).
● On an average 94,000 spores/mg of powdered lycopodium are present.
● A powdered drug is evaluated by this technique if it contains:
✔ Well defined particles which may be counted. E.g.: Starch
grains/pollen grains
✔ Single layered cells/tissues, the area of which may be traced under
suitable magnification and actual area calculated.
✔ Objects of uniform thickness, the length of which can be measured
under suitable magnification and actual area calculated.

Procedure:

✔ Dry the powdered drug (ginger) at 105oC and determine its steady
weight at room temperature.
✔ Weigh accurately powdered drug material and lycopodium spores
and mix them in the Proportion of 2:1. Mix them on a glass plate
with flexible spatula.
✔ Make a thin smooth paste by adding a suspending medium
(oil/glycerine: tragacanth: water2:1:2)
✔ Transfer the paste into a stoppered tube by washing with more
quantity of suspending media.
✔ Adjust the final volume by suspending fluid such that about 10-20
spores may be present in a field of 4mm objective.
 ✔ Oscillate the tube gently to get uniform suspension.
✔ Place a drop on slide, spread with a needle, put coverslip and count
the characteristic particles of the organic matter as well as
lycopodium spores in the field.
✔ % purity of an authentic powdered crude drug is calculated using
the equation:
NxWx94,000x100
SxMxP
N=No. of characteristic structures (starch grains) in 25 fields
W=Weight in mg of Lycopodium taken
S=No. of lycopodium spores in the same 25 fields
M=Wt. in mg of the sample calculated on basis of sample dried at 105o
C.
P=2,86,000in case of ginger starch grain powder