इंटरनेट मानक Disclosure to Promote the Right To Information Whereas the Parliament of India has set out to provide a practical regime of right to information for citizens to secure access to information under the control of public authorities, in order to promote transparency and accountability in the working of every public authority, and whereas the attached publication of the Bureau of Indian Standards is of particular interest to the public, particularly disadvantaged communities and those engaged in the pursuit of education and knowledge, the attached public safety standard is made available to promote the timely dissemination of this information in an accurate manner to the public. “जान1 का अ+धकार, जी1 का अ+धकार” “प0रा1 को छोड न’ 5 तरफ” Mazdoor Kisan Shakti Sangathan Jawaharlal Nehru “The Right to Information, The Right to Live” “Step Out From the Old to the New” IS 15697-1 (2006): Chillies and chilli oleoresins - Determination of total capsaicinoid contents, Part 1: Spectrometric method [FAD 9: Spices and Condiments] “!ान $ एक न’ भारत का +नम-ण” Satyanarayan Gangaram Pitroda ““IInnvveenntt aa NNeeww IInnddiiaa UUssiinngg KKnnoowwlleeddggee”” “!ान एक ऐसा खजाना > जो कभी च0राया नहB जा सकता हहहहै””ै” Bhartṛhari—Nītiśatakam “Knowledge is such a treasure which cannot be stolen” ., — IS 15697 (Part 1) :2006 1s0 7543-1:1994 M 3?)-f?a’a – 6TT 3h-#RN&f $Wm;l{ . 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 -Bureau Of Indian Standards BIS is a statutory institution established under the Bureau of hdian Standards Act, 1986 to promote harmonious development of the activities of standardization, marking and quality certification of goods and attending to connected matters in the country. 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