IS 15699: Ground (powdered) paprika (capsicum) annum linnaeus) - Microscopical examination

2006·1.1 MB·English

by  Bureau of Indian Standards

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IS 15699 (2006): Ground (powdered) paprika (capsicum) annum
linnaeus) - Microscopical examination [FAD 9: Spices and
Condiments]
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IS 15699:2006
ISO 7542:1984
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Indian Standard
GROUND (POWDERED) PAPRIKA (CAPSKWM
ANNUUM LINNAEUS) — MICROSCOPICAL
EXAMINATION
ICS 67.220.10

G BIS 2006
BUREAU OF INDIAN STANDARDS
MANAK BHAVAN, 9 BAHADUR SHAH ZAFAR MARG
NEW DELHI 110002

November 2006
Price Group 3
Spices and Condiments Sectional Committee, FAD 9
NATIONAL FOREWORD
This Indian Standard which is identical with ISO 7542 : 1984 ‘Ground (powdered) paprika
(Caps;cum annum Linnaeus) — Microscopical examination’ issued by the International Organization
for Standardization (ISO) was adopted by the Bureau of Indian Standards on the recommendation of
the Spices and Condiments Sectional Committee and approval of the Food and Agriculture Division
Council.
The text of ISO Standard has been approved as suitable for publication as an Indian Standard without
deviations. Certain conventions are, however, not identical to those used in Indian Standards. Attention
is particularly drawn to the following:
a) Wherever the words ‘International Standard’ appear referring tothis standard, they should
be read as ‘Indian Standard’.
b) Comma (,) has been used as a decimal marker while in Indian Standards, the current
practice is to use a point (.) as the decimal marker.
This standard is subject to the restrictions imposed under the The Prevention of Food Adulteration
Act, 1954 and the Rules framed thereunder, wherever applicable.
For the -purpose of deciding whether a particular requirement of this standard is complied with, the
final value, observed or calculated, expressing the result of a test or analysis, shall be rounded off
in accordance with IS 2 : 196-0 ‘Rules for rounding off numerical values (rev;se~’. The number of
significant places retained in the rounded off value should be the same as that of the specified value
in this standard.
IS 15699:2006
ISO 7542:1964
Indian Standard
GROUND (POWDERED) PAPRIKA (CAPS/CUM
AN/VUUM LINNAEUS) — MICROSCOPICAL
EXAMINATION
1 -Scope and field-of application Tha calyx isattached to the lower part ofthe fruit; itsshape is
characteristic of the variety. In general, it is formed by five
This International Standard deacrib&s the morphological and fused peaked sepals. The calyx isaprolongation ofthe pedun-
anatomical structure of peprika (Capsicum anrruum Linnaeus) cle, ofwhich the shape and the length are characteristic ofthe
and specifies a method for the microscopical examination of variety. The fruit may be upright, hanging orside-ways.
ground (powdered) paprika.
The placenta, the calyx and the peduncle are together called
the “stalk”.
2 Reference
ISO 7540, Ground (powdered) paprika (Capsicum annuum Lin- 4.2 Anatomical structure (see figura 1)
naeus) — Specification.
The anatomical structure of paprika can be more or less
recognized in the ground product. The calls of tha apidermis
3 Definitions (exocarp) of the pericarp sre flat and polygonal when viewad
from above, the walls being thicker atthe‘cornersandslightly
3.1 paprika : The whole fruit of paprika, consisting of the pitted. Their outer surface is covered with a thick cuticula,
pericarp, the seeds, the peduncle, the calyx and the placenta. whera sharp, transverse furrowa canbeobserved, especiallyin
vertical sections.
3.2 ground [powdered] paprika :The product obtained by
Inthe mesocarp, two parts can badistinguished :onthe out-
grinding varieties of dried ripe paprika.
side, the hypodermic adjoining the epidermis, and on the in-
side, the fleshy pericarp.

4 Structure of paprika The hypodermic is uni- or multi-seriate : its cells, like the
epidermis, arethicker at the corners andareslightlypitted, the
4.1 Description and morphological structure walls beingsuberized and yellow. Insidethe hypodermic, there
isa layer of wider pare’nchyma cellswhose walls become-pro-
Paprika isan inflated berry. Several varieties of paprika having gressivelythinner, butwhi hare notsuberizedandinwhich, in
different-shapes, are cultivated. Ground (powdered) paprika is varieties of paprika which% e red whan mature, contain red
produced from long orshort cylindro-conical varieties. chromatoplasts. When not quite mature, globoid starch par-
ticles may be seen.
The pericarp isrelatively thin, slightly juicy, coriaceous onthe
outside, and with a smooth red or dark red surface when Dispersed throughout the fleshy pericarp, fibro-vascular
mature. bundles of ringed, spiral and spotted vesselscan beseen.
The fruit is Ioculated by a septum into two, three or, On the inner side of the mesocarp, socalled “giant” cells, of
sometimes, fourparts which are well developed in the lower diameter 1to 2 mm, can be seen, separated by layersof thin
partofthefruit and flatten towards the apex, sothat the upper walled cellswhich cannot berecognized inthegroundproduct.
partof the fruit isunilocular.
The endocarp, composed of a single layer of cells, is im-.
The semi-spherical shaped central placenta isinthe lower part mediately adjacent to thegiant cells.Thisconsistsoftwo kinds
ofthefruit where the majority offhe seedsdevelop. Seeds ap- of cells :
pear to a lesser extent on the septum. The seeds are 2,8 to
4,5 mminlength, flat ovoidorkidney shaped andthicker atthe a) thick walled, yallow “bead string” sclerouscells,which
edges. Theopen-edged hilum isatthevantral sideoftheseed. aremore or less elongated and parallel to the axis of the
The seedsare ofayellowish-white or yellowish-brown colour, ‘fruit, and which appear undulated when viewed from
with a fine, glittering surface, slightly darker towards the above; they are slightly iignif~d and grouped under the
edges. giant cells;
1
IS 15699:2006
ISO 7542:1964
b) elongated, undignified,thin walled cells separating the 5 Microscopical examination
grouped cellsmentioned ina), forming a continuous net-
work which canbeeasilylorn. Only their fragments canbe 5.1 Principle
detected inthe ground product.
Clarificationofapinchofground paprikaona-microscopeslida
Thecellsoftheseptumaresimilartothoseofotherpartsofthe and examination of pe~icies under appropriate magnification.
fleshy pericarp. They do not contain chromatoplasts, the
vascularbundlesarethicker andthere aremore spiralelements
5.2 Reagents
with wider cavities. The pungent principle of paprika, cap-
saicin, develops inthe glandular cellslocated ingroups inthe
All reagents shall,be of recognized analytical grade and the
epidermal tissueoftheseptum. Glandular cellscontaining cap-
water usedshallbedistilledwater orwater ofatleastequivalent
$.aicinare not identifiable in the ground product. Neither the
purity.
seedsnorthe pericarp contain capsaicin.
5.2.1 Glycerol, solution.
The central placenta consistsmainly of colorless thin walled
parenchyma cellssimilartothoseofthefleshypericarp andthe
KXluteone part byvolumwof glycerol with onepartbyvolume
septum. The vascularbundlesarestrongly developed, Insome
ofwater.
places, very characteristic stone cells with slightly Iignified
walls can beseenamong the thin walled parenchyma cells.
5.2.2 Sodium hypochiorite, solution.
The surface of the seedsiscovered by a hard yellowish seed
Mix five voiumes of sodium hypochiorite soiution with two
coat, theoutward layerofwhich istheepidermis, consistingof
volumes of hotwater.
large greenish-yellow cells, The radial walls of the cells, as
viewed from above, are strongly undulated (mesentery cells)
and, together with“theinnerwalls, are considerably thickened 5.2.3 Iodine-potessium iodide soiution (Lugoi solution).
and highlylignifiedwith “slit-shapedpits.
Dissolve2 g of potassium iodide and 1g of iodine in a little
water, and diluteto 100 ml.
The thicknessandheight ofthe cellwalla incraaeetowards the
edgesoftheseedanddecrease towards theflat surfaceof the
5.2.4 iodine-sodium hypochiorite soiution.
seed. The outer cell waif is thin, more or less convex,
undignified,anditisonly inthe innermost part that averythin
Add iodineto the sodium hypochlorite soiutitm (5.2.2) untila
layer of cells with Iignified walls can be found. Under the
brown colour isobtained.
epidermis (exocarp) arethe parenchyma cells, nexttowhich is
thethickerendocarpwhich consistsofcellswith Iignifiedwalls.
The seedprotein tissueconsistsof comparatively thick walled 5.3 Apparatus
polygonalcellsofuniform diameter (isodiametric cells)contain-
ingoiland aleurone. 5.3.1 Binocuier microscope. with magnification of X 50to

X 1300, fitted with a friarmicrometer eyepiece and a buiit-in

The upperepidermis(onthesideclosestto thefruit) ofthefive lamp.
pointed-sepaled calyx consistsof ceilswith slightly undulated
walls. The upper epidermis carries short glandular hairs con- 5.3.2 Microscope siides.
sisting of a dark red plurilocular ovoid gland supponed on a
short peduncle.
5.3.3 Cover siides.
The cellsof the lower epidermis are larger, octagonal and are
5.3.4 Watch-glass.
interspersedwith stomata buthave noglandular hairs. Between
the two epidermes, there isaspongy parenchyma, with spiral
vascular bundles,green chromatoplasts and calcium oxalate 5.3,5 Mounted needle.
crystals.
5.3.6 Scaipei.
The peduncle of the paprika iscovered by an epidermis con-
taining agreencolouring material which turnstobrown atma-
5.4 Preparation of the sample
turity. Under the epidermis, vascular tissue can beseeninthe
parenchyma cellswith thinwalls. The elements ofthexylem(of
Prepareseveraiaempiesfor microscopical examination. Piecea
the~ascular bundles)areIignified, with bordered pitsshowing
pinch of ground (powdered) paprika on a microscope slide
reticular thickening. The elongated thick walled xylem fibres
(5.3.2), mix carefuily with the clarifying solutions and cover
with slit-likepitsandthe fibreeofthe phloem serveassuppor-
with acover siide(5.3.3).
tingtissues.
Glyceroisolution (5.2.1) makes tha preparation iightandmore
Parenchyma cells are found next to the long fibres in the translucent; it siightiy dissolves coiouring matter, and
phloem. colourleasparts become translucent.
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IS 15699:2006
ISO 7542:1964
Sodium hypochlorite hasa bleaching effect making coloured by comparison of relative dimensions. (See figure 1 for the
particlesbecome pale and itdissolvaa agreat deal of the con- transverse sectionof the pericarp of paprika.)
temsof the cell, rendering the preparation more translucent.
If foreign elements are detected, a reference preparation shall
Thisbleaching effect can be accelerated by carefully heating beexamined.
theslide i-mtilthe sodium hypochlorite solution begins to boil,
butreplacingthe water lostfhrough evaporation. The main microscopical characteristics whtch distinguish
paprika are asfollows :
The iodine-potassium iodide solution isused most usuallyfor
the detection of foreign starch. The pericarp of ripe paprika a) exocarp (epidermis) of the paricarp (seefigure 2);
doesnotcontain starch.
b) “bead string” sclerous cells of the paprika andocarp
The iodine-sodium hypochlorite solution isusedfor the detec- (seefigure 3);
tion offine starch particles from unripe pericarp. The reagent
rendersthe material clear{ swelling starch particles andcolour- c) epidermis of the paprika seed (mesenfery cells)Isea
ingthem blue. figure 4);
d) epidermis of the calyx (see figure 5);
5.5 Procedure e) stone cellsof the placenta (see figure 6);
The examination isusuallycarried out under amagnif~ation of f) elements of the peduncle (seefigure 7);
X 200 toX 600. Insome cases, the origin of aparticular tissue
can only bedetermined by seeking small structural details, or g) large numbara of red oildrops.

3
IS 15699:2006
ISO 7542:1984
[email protected]._-
—Omomoao0000 Epidermis(exocarp)
&&g:i
~Hypodermis
la

\
Lw
Unicallulatlayerofendocarp
~Thin-wallad CSIIS
Figure 1 – Transverse section of paprika pertcarp
IS 15699:2006
ISO 7542:1984
Figure2 – Exocarp of thepericarp of paprika
Figure3 — “Bead string” cells of the andocarpof the
paricarp of paprika, seen from above

F}gure4 – Pericarp of the paprika sead (mesentary
cells), scan from above
5