IS 15697-1: Chillies and chilli oleoresins - Determination of total capsaicinoid contents, Part 1: Spectrometric method

2006·1.1 MB·English

by  Bureau of Indian Standards

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IS 15697-1 (2006): Chillies and chilli oleoresins -
Determination of total capsaicinoid contents, Part 1:
Spectrometric method [FAD 9: Spices and Condiments]
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., —
IS 15697 (Part 1) :2006
1s0
7543-1:1994

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Indian Standard
CHILLIES AND CHILLI OLEORESINS —
DETERMINATION OF TOTAL CAPSAICINOID
CONTENT
PART 1 SPECTROMETRIC METHOD
ICS 67.220.10

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

November 2006 PriceGroup 2
Spices and Condiments Sectional Committee, FAD 9
NATIONAL FOREWORD
This Indian Standard (Part 1)which is identical with 1S0 7543-1 :1994 ‘Chillies and chilli o]eoresins —
Determination of total capsaicinoid content — Part 1: Spectrometric method’ 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 to this 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.
In this adopted standard, reference appears to the following International Standard for which Indian
Standard also exists. The corresponding Indian Standard which is to be substituted in its place is
listed below along with its degree of equivalence for the edition indicated:
International Standard Corresponding Indian Standard .Degree of
Equivalence
ISO 2825 : 1981 Spices and IS 1797:1985 Method of test for spices Technically
condiments — Preparation of a and condiments Equivalent
ground sample for analysis
For the purpose of deciding w h ether 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 : 1960 ‘Rules for rounding off numerical values (revised)’. The number of
significant places retained in the roundaf off value should be the same as that of the specified value
in this standard.
IS 15697 (Part 1) :2006
1s07543-1 :1994
/ndian Standard
CHILLIES AND CHILLI OLEORESINS —
DETERMINATION OF TOTAL CAPSAICINOID
CONTENT
PART 1 SPECTROMETRIC METHOD
1 Scope 3 Principle
this part of ISO 7543 specifies a method for the de- Determination of the absorption of a methanolic sol-
termination, by a spectrometric method, of the total ution of the extracts of chiliies or chilli oleoresins at
capsaicinoid content of whole or powdered chillies wavelengths of 248 nm and 296 nm.
(usually Capiscm frutescent L.) and their oleoresins.
3,1 Chilliesin powder form
This method of analysis requires discoloration by car-
bon black. In certain cases, this discoloration cannot
Extraction with tetrahydrofuran, then determination
be achieved under the experimental conditions de-
of the capsaicinoids by smectrometrv, in accordance
scribed. Then it is necessaw to use the method of
with the method descr;be~ in this pa~ of ISO 7543.
determination of capsaicinoids by high-performance
liquid chromatography as described in ISO 7543-21).
3.2 Whole chillies

Preparation by grinding the sample, then extraction
with tetrahydrofuran, followed by determination of the
capsaicinoids by spectrometry, in accordance with the
2 Normative reference method described in this part of ISO 7543.
The following standard contains provisions which, 3.3 Qleoresins of-chillies
through reference in this text, constitute provisions
of this part of ISO 7543. At the time of publication, the Dilution of the oleoresin in methanol, then determi-
edition indicated was valid. All starrdards are subject nation of the capsaicinoids by spectromet~, in ac-
to revision, and parties to agreements based on this cordance with the method described in this part of
1s0
part of ISO 7543 are encouraged to investigate the 7543.
possibility of applying the most recent edition of the
standard indicated below. Members of IEC and ISO
maintain registers of currently valid International 4 Reagents
Standards.
Use only reagents of recognized analytical grade, un-
ISO 2825:1981, Spices and condiments — Prepara- less otherwise specified, and distilled or demineral-
tion of a ground sample for analysis. ized water or water of equivalent purity.
1) ISO 7543-2:1993, Chdlies and chilli oleoresins — Determination of total capsaicinoid content — Part 2: Method using
high-performance liquid chromatography.

1
15697 (Part 1) :2006
1s07543-1 :1994
4.1 Carbonblack,for analysis Sampling is not part cf the method specified in this
International Standard. A recommended sampling
4.2 Methanol, spectrometric grade. method is given in lSO 9482).
4.3 Methanol solution, obtained by mixing 70 parts 7 Preparation of test sample
by vo!ume of methanol (4.2)with 30 parts by volume
of water.
7.1 Chilliesin powder form
4.4 Hydrochloricacidsolution, c(HCI) = 1 nTO1/1.
7.1.1 Check that all of the powder passes through
the 500 pm sieve (5.8). H this is not the case, grind
4.5 Sodium hydroxide solution, freshly prepared, the powder in accordance with ISO 2825 until the re-
c(NaOH) = 1 mol/1. quired particle size is obtained, then homogenize.
4.6 Tetrahydmfuran, freshly distilled or spectro- 7.1.2 Weigh, to the nearest 0,01 g, about 10 g of
metric grade. homogenized powder (7.1,1 ), and transfer it
quantitatively to the continuous-extraction apparatus
(5.5).
5 Apparatus
7.1.3 Extract for 8 h using 100 ml of tetrahydrofuran
Usual laboratory equipment and, in particular, the fol-
(4.6).
lowing.
Evaporate the solvent to the maximum extent poss-
5.1 One-mark volumetric flasks, of 25 ml, 100 ml ible in the rotaty vacuum evaporator (5,6) under re-
and 250 ml capacity. duced pressure in a 250 ml round-bottomed flask on
the water bath (5.7).
5.2 Spectrometer, single-beam or double-beam,
suitable for measurements of absorption at wave- 7.2 Whole chillies
lengths between 220 nm and 370 nm, and fitted wi~h
quartzcellsof 1cm optical path length.
7.2.1 Grind the chillies in accordance with ISO 2825
until the powder obtained passes entirely through the
5.3 Magneticstirrer. 500 km sieve (5.8). Homogenize the powder after
sieving.
5.4 Membrane filter,with aperture size 0,45 Vm.

7.2.2 Weigh, to the nearest 0,01 g, about 10 g of
5.5 Continuous-extraction appa r atus, Soxhlet- the homogenized powder (7.2.1), and transfer it
type, with aflat-bottomed flask of 250 ml capacity and quantitatively to the continuous-extraction apparatus
a ground neck. (5,5).
5.6 Vacuum evaporator, rotary. 7.2.3 Extract for 8 h using 100 ml of tetrahydrofuran
(4.6).
5.7 Water bath,
Evaporate the solvent to the maximum extent poss-
ible in the rotary vacuum evaporator (5.6) under re-
5,8 Testsieve,with aperture size 500 vm. duced pressure in a 250 ml round-bottomed flask on
the water bath (5.7).
5,9 Analyticalbalance, capable of weighing to an
accuracy of ~ 0,00Q 1 g. 7.3 Oleoresinsof chillies
6 Sampling 7.3.1 Thoroughly homogenize the oleoresin.
Itis important that the Iabmatory receive a sample 7.3.2 Weigh, to the nearest 0,0001 g, 0,5 g to 1 g
which is truly representative and has not been dam- of the homogenized oleoresin (7.3.1) in a 250 ml
aged or changed during transport or storage. volumetric flask (5.1)with a ground glass stopper.
2) ISO948:1980, Spices and condiments — Sampling.

2
IS 15697 (Part 1) :2006
1s07543-1 :1994
8 Procedure — 2 ml of sodium hydroxide solution (4.5)
Dilute to the mark with methanol (4.2).
8.1 Preparation of test solutions
This solution is the “blank alkali solution” (B)
8.1.1 Chillies,whole or in powder form
8.2.3 Take three 25 ml volumetric flasks (5.1) and
mark them al, a2 and a3 respectively. Transfer to
To the extract obtained m 7.1.3 or in 7.2.3, add
each flask:
0,05 g to 0,1 gof carbon black (4.1) so as to maintain
a ratio of the order of 10 between the extract and
— 1 ml of the filtrate obtained in 8.1.1 or 8.1.2,
carbon black.
— 2,7 ml of water,
Add about 90 ml of methanol solution (4.3).
Agitate on the magnetic stirrer (5.3) for 30 min. Allow — 2 ml of hydrochloric acid (4.4).
the solutlon to stand for 5 min. Filter through the
Dilute each flask to the mark with methanol (4.2).
membrane filter (5.4) into a 100 ml volumetric flask
(5.1). Dilute to the mark with methanol solution (4.3).
8.2.4 Take three 25 ml volumetric flasks (5.1) and
The filtrate shall be clear. Experience has shown that mark them bl, b2 and b3 respectively. Transfer to
a light yellow colour does not affect the determi- each flask:
nation.
— 1 ml of the filtrate obtained in 8.1.1 or 8.1.2,
8.1.2 Oleoresins of chillies
— 2,7 ml of water,
To the test portion (7.3.2), add 0,05 g to 0,1 g of car-
— 2 ml of sodium hydroxide solution (4.5).
bon black (4.1) so as to maintain a ratio of the order
of 10 between the oleoresin and carbon black. Dilute each flask to the mark with methanol (4.2)
Add about 90 ml of methanol solution (4.3).
8.3 Spectrometric measurements
Agitate on the magnetic stirrer (5.3) for 30 min. Allow
the solution to stand for 5 min. Filter through the 8.3.1 Foradouble-beam spectrometer
membrane filter (5.4) into a 100 ml volumetric flask
(5.1). Dilute to the mark with methanol solution (4.3). Adjust the zero and the 100 % absorption with
methanol solution (4.3).
The filtrate shall be clear. Experience ha s shown that

a light yellow colour does not affect the determi- Measure the blank absorbance at wavelengths of
nation. 248 nm and 296 nm by placing first the blank alkali
solution (B)in the measuring cell and the blank acid
solution (A) in the reference cell.
8.2 Preparation of dilutions for
spectrometricmeasurement Measure the absorbance of each sample solution at
wavelengths of 248 nm and 296 nm by placing the
8.2.1 Transfer the following to a 25 ml volumetric solution from flask bl in the measuring cell and the
flask (5.1): solution from flask al in the reference cell. Then
measure the absorbance with solutions from flasks
— 3 ml of water, b2and a2,and flasks b3 and a3,respectively.
— 2 ml of hydrochloric acid (4,4). 8.3.2 Forasingle-beam spactrometar
Dilute to the mark with methanol (4.2). Adjust the zero and the 100 ‘X. absorption with
methanol solution (4.3).
This solution is the “blank acid solution” (A),
Re-adjust the zero of the apparatus by placing solution
8.2.2 Transfer the following to a 25 ml volumetric B in the measuring cell and then measure the
flask (5.1): absorbance of the solutions from flasks al, a2 and
a3, respectively, at wavelengths of 248 nm and
— 3 ml of water, 296 nm.

3
IS 15697 (Part 1) :2006
1s07543-1 :1994
Then measure the absorbance at wavelengths of A’b k the absorbance of the blank solution;
248nm and 296nm by placing solution A in the
measuring cell, and then the solutions from flasks d is the dilution factor (equal to 25 x 100
bl, b2andb3, respectively, under the operating conditions described
inthis part of ISO7543);
9 Calculation
m is the mass, in grams, of the test portion.
9.1 Calculate the total capsaicinoid content, W248, as Carry out an additional dilution when the absorbance
a percentage by mass, at a wavelength of 248 nm, is greater than 0,8.
using the following formula:
9.3 The difference between the results of the
W24B = [‘A=-_– A,-)’ X d measurements obtained at wavelengths of 248 nm
314xm and 296 nm shall not be more than 10-Yo.
where
If this is not the case, repeat the analysis.
A, is the absorbance of the sample solution;
10 Test report
Ab is the absorbance of the blank solution;
The test report shall specify
d is the -dilution factor (equal to 25 x 100
under the operating conditions described
— -the method in accordance with which sampling
in this pan of ISO 7543);
was carried out, if known,
m is the mass, in grams, of the test portion.
— the method used,
Carry out an additional dilution when the absorbance
— the test result(s) obtained, and
is greater than 0,8.
— if the repeatability has been checked, the final
9.2 Calculate the total capsaicinoid content, wz~~,as
quoted result obtained.
a percentage by mass, at a wavelength of 296 nm,
using the following formula: It shall also mention all operating details not specified
in this part of ISO 7543, or regarded as optional, to-
(A’, - A’b) x d
w*ga = gether with details of any incidents which may have
127xm
influenced the test result(s).
where
The test report shall include all information necessary

A’, is the absorbance of the s ample solution; for the complete identification of the sample.

4
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Amendments Issued Since Publication
Amend No. Date of Issue Text Affected
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