2005 AMD7 Reff2018
2005 AMD7 Reff2018
2005 AMD7 Reff2018
IS 13428 : 2005
(Reaffirmed 2014)
(Reaffirmed 2018)
~~~~-~
(Reaffirmed 2013)
( CJ\\lIYI ~8fUT )
(Reaffirmed 2012)
Indian Standard
(Reaffirmed 2011)
PACKAGED NATURAL MINERAL WATER-
SPECIFICATION
(Reaffirmed 2010)
(Second Revision)
First Reprint DECEMBER 2006
(Reaffirmed 2009)
(Reaffirmed 2007)
(Reaffirmed 2006)
(Reaffirmed 2005)
C BlS200S
BUREAU OF INDIAN STANDARDS
MANAK BHAVAN. 9 BAHADUR SHAH ZAFAR MARG
NEW DELHI 11 0002
Price Group II
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FOREWORD
This Indian Standard (Second Revision) was adopted by the Bureau of Indian Standards, after the draft finalized
by the Drinks and Carbonated Beverages Sectional Committee had been approved by the Food and Agriculture
Division Council.
This standard w~s published in 1992 and subsequently revised in 1998 in view of the following:
a) To delete provisions of fortified mineral water which were covered in the earlier version;
b) To align with the revised Codex Standard for natural mineral water, except for the following which are
either not covered or partially covered in the Codex Standard:
1) Organoleptic and physical parameters have been retained. Minimum and maximum values have
been indicated for total dissolved solids in order to ensure presence of minimum minerals content
in the water and at the same time limiting their content for the sake of palatability;
2) Requirements for zinc, silver, chloride, sulphate and alkalinity have been retained and the
requirements ofmagnesium, calcium, sodium, and sulphide have been added as these were considered
relevant characteristics from the point of water quality; and
3) In microbiological parameters, requirements for yeast and mould, salmonella and shigella, vibrio,
cholera and V. parahaemolytlcus have been retained and that of staphylococcus aureus added in
addition to those specified in the Codex standard, to provide additional safeguard. The requirement
of aerobic microbial count has been deleted as the same has not been prescribed in the Codex
Standard.
c) To include hygienic practices in line with Codex (CAC/RCP 33-1985) 'Code of practice for collecting,
processing and marketing of natural mineral waters'.
This revision has been undertaken to incorporate five amendments alongwith the technological developments,
check list for hygienic requirements, and consumer requirements. It is expected that this standard would help in
achieving the above objective.
In the preparation of this standard due consideration has been given to the provisions of the Prevention ofFood
Adulteration Act, 1954 and the Rules framed thereunder. The standard is, however, subject to the restrictions
imposed under this Act and Rules, wherever applicable.
In the preparation of this standard assistance has been derived from the EEC Directive, 80/778IEEC 'Council
directive relating to the quality of water intended for human consumption'.
A separate standard IS 14543 : 2004 'Packaged drinking water (other than packaged natural mineral water)' has
been established.
For the purpose of deciding whether a particular requirement of the 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
.rounded off value should be the same as that of the specified value in this standard.
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IS 13428 : 2005
Indian Standard
PACKAGED NATURAL MINERAL WATER-
SPECIFICATION
( Second Revision)
1 SCOPE 3.1.1 Naturally Carbonated Natural Mineral Water
- Natural mineral water which, after possible
This standard prescribes the requirements,methods of
treatment in accordance with 4.1 and re-incorporation
sampling and test for natural mineral waters offered
of gas fromthe same source and after packagingtaking
for sale in packaged fonn for human consumption.
into consideration usual technical tolerance, has the
NOTE -It does not apply to natural mineral water sold or same content of carbon dioxide spontaneously and
usedfor otherpurposes. visibly given off under normal conditions of
2 REFERENCES temperature and pressure.
The standards listed in Annex A contain provisions, 3.1.1 Non-carbonated Natural Mineral Water -
which through reference in this text, constitute Natural mineral water which, by nature and after
provisions ofthis standard. At the time of publication, possible treatment in accordance with 4.1 and after
the editions indicated were valid. All standards are packaging taking into consideration usual technical
subject to revision and parties to agreements based on tolerance, does not contain free carbon dioxide in
this standard are encouraged to investigate the excess of the amount necessary to keep the hydrogen
possibility of applying the most recent editions of the carbonate salts present in the water dissolved.
standards indicated at Annex A. 3.1.3 Decarbonated Natural Mineral Water -
Natural mineral water which, after possible treatment
3 DEFINITION in accordance with 4.1 and after packaging, has less
For the purpose of this standard the following carbon dioxide content than that at emergence and
definitions shall apply. does not visibly and spontaneously give off carbon
dioxide under normal conditions of temperature and
3.1 Natural Mineral Water - Water clearly pressure.
distinguishable from ordinary drinking water because:
3.1.4 Natural Mineral Water Fortified with Carbon
a) it is obtained directly from natural or drilled Dioxide from the Source - Natural mineral water
sources from underground water-bearing which, after possible treatment in accordance with 4.1
strata for which all possible precautions and after packaging, has more carbon dioxide content
shouldbetakenwithinthe protected perimeters than that at emergence.
to avoidany pollutionof, or externalinfluence
on, the chemical and physical qualities; 3.1.5 Carbonated Natural Mineral Water - Natural
b) it is characterized by its content of certain mineral water which, after possible treatment in
mineralsaltsand their relativeproportions and accordance with 4.1 and after packaging, has been
the presence of trace elements or of other made effervescent by the addition of carbon dioxide
constituents; from another origin.
c) of the constancy of its composition and the NOTE- Mineral watermeans natural mineral wateras defined
stability of its discharge and its temperature, in 3.1.
due accountbeingtakenofthe cyclesof minor 3.2 Packaged Natural Mineral Water - Natural
natural fluctuations; mineralwater filledintohermeticallysealedcontainers
d) it is collectedunderconditions whichguarantee of various compositions, forms and capacities that is,
the original microbiological purity and suitable for direct consumption without further
chemicalcomposition of essential components; treatment.
e) it is packaged close to the point ofemergence
4 TREATMENT AND HANDLING
of the source with particular hygienic
precautions; and 4.1 Treatments permitted include separation from
f) it is not subjected to any treatment other than unstable constituents, such as compounds containing
those permitted by this standard. iron, manganese, sulphur or arsenic, by decantation
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IS 13428 : 2005
and/or simple filtration up to 0.5 microns, if necessary, 6.1.6 Yeast and Mould, shall be absent in 250 ml
accelerated by previous aeration. sample when tested in accordance with the method
given in IS 5403.
4.2 The treatments provided in 3.1.1 to 3.1.5 and 4.1
above may only be carried out on condition that the 6.1.7 Salmonella and·Shigella, shall be absent in any
mineral content of the water is not modified in its 250 ml sample when tested in accordance with the
essentialconstituents, whichgivethewaterits properties. method given in IS 5887 (Part 3)* and IS 5887 (Part 7)
respectively. Salmonella may also be tested by the
4.3 The transport of natural mineral waters in bulk
method specified in IS 15187.
containers for packagingor for any other processbefore
packaging is prohibited, 6.1.8 Vibrio cholera and V. parahaemolyticus, shall
be absent in 250 ml sample when tested in accordance
5 HYGIENIC CONDITIONS with the method given in IS 5887 (Part S).
Natural mineral water shall be collected: processed, 6.1.9 The membrane filtration technique outlined in
handled, packaged and marketed in accordance with IS 15188 may be used to pass the sample of water to
the hygienic practices given in Annex B. A check-list betested through membrane before the microbiological
for good hygienic practices and food safety system for tests specified from 6.1.1 to 6.1.8 are carried out.
packaged natural mineral water processing units given
NOTE - In case of dispute, the method indicated by'·'
at the end of Annex B.
in 6.1.1 to 6.1.3 and 6.1.7 shall be the reference method.
6 REQUIREMENTS 6.2 Natural mineral water shall also comply with the
6.1 Microbiological Requirements
requirements given in Table 1, Table 2, Table 3 and
Table 4.
6.1.1 'Escherichia coli' (or Thermotolerant bacteria)
6.3 Residues of pesticides for pesticides as given in
shall be absent in any 250 ml sample when tested in
accordance with the method given in IS 5887 (Part 1)* Annex N shall be below the detectable limits. The
or IS 15185. analysisof pesticideshall be conductedby a recognized
laboratory using internationally established test
6.1.2 Coliform, bacteria shall be absent in any 250 ml methods as given in Annex N.
sample when tested in accordance with the method
given in IS 5401 (Part 1)* or IS 15185. 7 PACKING
6.1.3 Faecal streptococci and Staphylococcus aureus, 7.1 Natural mineral water shall be packed in clean,
shall be absent in any 250 ml sample when tested in hygienic, colourless, transparent and tamperproof
accordance with the method given in IS 5887 (Part 2)* bottles/containers, made of polyethylene (PE)
Streptococci (Enterococci) may also be tested by the conforming to IS 10146 or polyvinyl chloride (PVC)
method specified in IS 15186. conforming to IS 10151 or polypropylene conforming
to IS 10910 or polyalkylene terephthalate (PET and
6.1.4 Sulphite reducing anaerobes, shall be absent in PBT) conforming to IS 12252 or polycarbonate
50 ml sample when tested in accordance with the conforming to IS 14971 or polystyrene conforming
method given in Annex C. to IS 10142 or sterile glass bottles suitable for
6.1.5 Pseudomonas aeruginosa, shall be absent in preventing possible adulteration or contamination of
250 ml sample when tested in accordance with the the water. Plastic containers shall be conforming
method given in Annex D. to IS 15410.
,
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IS 13428 : 2005
NOTE - In case of dispute, the method indicatedby '.' shall be the referencemethod.
NOTE - In case of non-eonformity of l'Idio lCtive residues, the source of water shall be abandoned and water shall be recalled
immediately.
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7.2 All packagingmaterialsofplasticorigin shall pass Claims of other beneficial effects related to the health
the overall migration and colour migration limits as of the consumer shall not bemade.
laid down in the relevant IndianStandardsfor products 8.2.2 The name of the locality, hamlet or specified
for respective packaging materials when tested as per place may not form part of the brand name unless
method given in IS 9845. it refers to packaged natural mineral water collected!
8 MARKING processed at the place designated by that brand
name.
8.1 The following particulars shall be marked legibly
8.2.3 The use of any statement or of any pictorial
and indelibly on the label of the bottle/container:
device which may create confusion in the mind ofthe
a) Name ofthe product (that is packagednatural public or in any way mislead the public about the
mineral water); nature, origin, composition and properties of natural
b) Supplementary designations, if any; mineral waters put on sale is prohibited.
c) Name and address of the processor; 8.3 DIS Certification MarkiDg
d) Brand name, if any;
8.3.1 The product may also be marked with the
e) Batch or Code number; Standard Mark.
f) Date of processing/packing;
8.3.2 The use of the Standard Mark is governed by the
g) Best for consumption up to ... (date/month! provisions of Bureau of Indian Standards Act, 1986
year in capital letters); or and the Rulesand Regulationsframed thereunder. The
Best for consumption within days or months details of the conditions under which the licence for
from the date of processing/packing; use of Standard' Mark may begrantedto manufacturers
h) Net volume; or producers may be obtained from the Bureau of
j) Location and name of the source of natural Indian Standards.
mineral water;
9 SAMPLING
k) Direction for storage; and
m) Any other markings required under the Representative samples of natural mineral water shall
Standards of Weights and Measure bedrawnand the criteriafor confonnity to this standard
(Packaged Commodities) Rules, 1977and the shall be established, according to the method given in
Prevention of Food Adulteration Act, 1954 Annex E.
and the Rules framed thereunder.
10 QUALITY OF REAGENTS
8.2 Labelling ProhibitioDs Unless specified otherwise, pure chemicals and
8.2.t No claims concerning medicinal (preventative, distilledwater (see IS 1070)shall be employed in tests.
alleviativeor curative) effects shall be made in respect NOTE - 'Pure chemicals' shall mean chemicals that do not
of the propertiesof the productcoveredby the standard. contain impurities which affect the results of analysis.
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IS 13428 : 2005
ANNEX A
(Clause 2)
LIST OF REFERRED INDIAN STANDARDS
5403 : 1999 Method for yeast and mould count 14194 Radionuclides in environmental
of foodstuffs and animal feeds (first samples - Methods of estimation:
revuion) (Part 1) : 1994 Gross beta activity measurement
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IS 13428 : 2005
ANNEX B
(Clause 5)
HYGIENIC PRACTICES
B-1 FIELD OF APPLICATION conditions and considering the risks of pollution and
physical,chemical and biochemicalreactions, several
The hygienic practices cover appropriate general
perimeterswith separatedimensionsmay be provided
techniques for collecting natural mineral water, its
for.
treatment, bottling, packaging, storage, transport,
distribution and sale for direct consumption, so as to 8-2.1.4 Protective Measures
guarantee a safe, healthy and wholesome product. All possible precautions should be taken within the
B-2 PRESCRIPTIONS OF THE RESOURCES protected perimeters to avoid any pollution of, or
OF NATURAL MINERAL WATER external influence on, the chemical and physical
qualities of natural mineral water.
8-2.1 Protection of Alimentary Reservoirs and
It is recommendedthat regulations be established for
Aquifen
the disposal of liquid, solid or gaseous waste, the use
8-2.1.1 Authorization of substances that might deteriorate natural mineral
water as well as any possibility of accidental
Any spring,well or drilling intendedfor the collection
deterioration of natural mineral water by natural
of natural mineral water should be approved by the
occurrences such as a change in the hydrogeological
Local Health Authority or any other agency having
conditions. Particular consideration should be given
jurisdiction for the region.
to the following potential pollutants such as bacteria,
8-2.1.2 Determination of the Genesis of Natural viruses, fertilizers, hydrocarbons, detergents,
Mineral Water pesticides, phenolic compounds, toxic metals,
radioactive substances and other soluble organic or
As far as it is methodologically possible in each case,
inorganic substances. Even where nature provides
a precise analysis should be carried out on the origin
apparently sufficient protection against surface,
ofnatural mineral waters,the period oftheir residence
pollution, potential hazards arising out of mining,
in the ground beforebeingcollectedandtheirchemical,
hydraulic andengineering facilities etc, shouldbetaken
physical and radiological qualities.
into consideration.
8-2.1.3 Perimeter ofProtection
8-2.2 HYllene Prescriptions for Collection of
Ifpossible areas wherein natural mineral water might Natural Mineral Water
be polluted or its chemical and physical qualities
otherwise deteriorated should be determined by a 8-2.2.1 Extraction
hydrologist. Where indicated by hydrogeological The withdrawal of natural mineral water shall be
6
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IS 13428 : 2005
performed in conformity with the hydrogeological of extraction should be as lowas possible. The storing
conditions in such a manner as to prevent any water should furthermore guarantee protection against
otherthan the natural mineralwater from enteringor, contamination or deterioration.
should there be pumping facilities, prevent any
extraneous water from entering. The natural mineral 8-2.4 Transport of Natural Mineral Water
waterthus collectedor pumpedshould be protectedin 8-2.4.1 Means of Transport, Piping and Reservoirs
such a way that it is not polluted(whether caused by
natural occurrenceor actionsor neglector ill will). Anyvehicle, pipingor reservoir used inthe processing
of natural mineral water from its source to the
8-2.1.1 Materials bottlingfacilities, should comply with the necessary
The pipes, pumps or other possible devices coming requirements and be, made-of inert material such as
into contact with natural mineral water and used for ceramic and stainless steel which prevents any
its collection should be made of such material as to deterioration, be it by water, handling, servicingor by
guarantee that originalqualityof naturalmineral water disinfection; it should allow easy cleaning.
is not changed.
8-2.4.2 Maintenance 0/ Vehicles and Reservoirs
8-1.2.3 Protection ofthe Extraction Area
Any vehicle or reservoir should be properly cleaned
In the immediate surroundings of springs or wells, and if necessary, disinfected and kept in good repair
precautionary measures should be taken to guarantee so as not to present any danger of contamination to
that no pollutant whatsoever can enter the extraction natural mineral water and of deterioration of the
area.Theextraction areashould be inaccessible to non- essential qualities of natural mineral water.
authorizedpeople by providingadequate devices(for
example enclosure). Any use not aiming at the B-3 ESTABLISHMENT FOR PROCESSING
collection of natural mineral watershould be forbidden NATURAL MINERAL WATERS - DESIGN
in this area. AND FACILITIES
8-2.2.4 Exploitation ofNatural Mineral Water 8-3.1 Location
The condition of the extraction facilities, areas of Establishments should be located in areas which are
extraction and perimeter protection as well as the free from objectionable odours; smoke, dust or other
qualityof the natural mineral watershouldperiodically contaminants and are not subjectto flooding.
be checked. To control the stability of the chemical
and physical particulars of the natural mineral water 8-3.2 Roadways and Areas Used by Wheeled
derived, besides the natural variations, automatic Trame
measurements of the typical characteristics of water Such roadways and areas serving the establishment
should be carried out and recorded (for example, which are within its boundaries or, in its immediate
electricalconductance, temperature, contentof carbon vicinityshould have a hard paved surfacesuitable for
dioxide) or frequent partial analysisshould be done. wheeled traffic. There should be adequate drainage and
8-2.3 Maintenance of Extraction Facilities provision should be made for protection of the
extraction area in accordance with 8-2.2 where
8-2.3.1 Technical Aspects appropriate. Adequate road signals may be provided
Methods and procedures for maintaining the extraction to call the attention of road users to the existence of
facilities should be hygienic and not be a potential naturalmineralwater extraction area.
hazard to human health or a source of contamination
8-3.3 Buildings and Facilities
to naturalmineralwater. Fromthe hygiene standpoint,
servicing of the extraction installations should meet 8-3.3.1 Type ofConstruction
the same requirements asthoserequired forthe bottling
Buildings andfacilities should be of soundconstruction
or for treatment.
in accordance with the provisions of 8-2.2 and
8-2.3.1 Equipment and Reservoirs maintained in good repair.
Equipment and reservoirs usedfor extraction of natural 8-3.3.2 Disposition of Holding Facilities
mineral water should be constructed and maintained
in order to minimize all hazards to human health and Rooms for recreation, for storingor packaging of raw
to avoid contamination. material and areas for cleaning of containers to be
reused should be away from the bottling areas to
8-2.3.3 Storage at the Point 01 Extraction prevent theendproduct from beingcontaminated. Raw
Thequantity of natural mineralwaterstoredat the point materials and packaging materials which come into
7
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IS 13428 : 2005
contact with natural mineral water should be stored 8-3.3.7.6 Stairs, 11ft cages and mal/lory Itrllctllra
apart from other material.
Platforms, ladden, chutes, should be so situated and
8-3.3.3 Adequateworking space should be provided constructed as not to cause contamination. Chutes
to allowfor satisfactory performance of all operations. shouldbe constructed withprovisionof inspection and
8-3.3.4 The design should be such as to permit easy c1eanina batches.
and adequate cleaning and to facilitate proper 8-3.3.7.7 Piping
supervision of hygiene of natural mineral water.
Piping for natural mineral water lines should be
8-3.3.5 Thebuildings and facilities shouldbe designed independent of potable and non-potablewater.
to provide separation by partition, location or other
effective means betweenthose operationswhich may 8-3.3.,8 In natural mineral water handling area all
cause cross contamination. overheadstructures and fittings should be installed in
such a manner as to avoid contamination directly or
8-3.3.6 Buildings and facilities should be designed indirectly of natural mineral water and raw materials
to facilitate hygienic operations by means of a regulated by condensation and drip, and should not hamper
flow in the process from the anival of the natural cleaning operations. They should be insulated where
mineral water at the premisesto the finished product, appropriate and be so designed and finished as to
and should provide for appropriate temperature prevent the accumulation of dirt and to minimize
conditionsfor the process and the product. condensation, mould development and flaking. They
8-3.3.7 Natural Mineral Water Handling, Storing and should be easy to clean.
Bottling Areas
8-3.3.9 Living quarters, toilets and areas where
8-3.3.7.1 Floors animals arekept should be completelyseparatedfrom
andshouldnotopendirectly on to natural mineral water
Where appropriate, floors should be of water-proof, handling areas.
non-absorbent, washable, non-slip and non-toxic
materials, withoutcrevices, and shouldbeeasy to clean 8-3.3.10 Where appropriate, establishments shouldbe
and disinfect. Where appropriate, floors should have so designedthat access can be controlled.
sufficientslope for liquids to drain to trapped outlets. 8-3.3.11 The use of material which cannot be
8-3.3.7.2 Walls adequately cleaned and disinfected, such as wood,
shouldbe avoidedunlessits use wouldnot be asource
Where appropriate, should be of water-proof, Don- of contamination.
absorbent, washable and non-toxic material andshould
be light coloured. Up to a height appropriate for the 8-3.3.12 Canalization, Drainage Lines
operation they should be smooth andwithoutcrevices, Canalization, drainageand used water lines as well u
and should be easy to clean and disinfect. Where any possible waste storage area within the protected
appropriate, angles betweenwalls, betweenwalls and perimeter should be built and maintained in such a
floors, and between wallsandceilingsshould be sealed manner as not to present any risk whatsoever of
and smoothento facilitate cleaning. pollutingaquifers and springs.
B-3.3.7.3 Ceilings 8-3.3.13 Fuel Storage Area
Ceilings should be so designed, constructed and Any storage area or tank for the storing of fuels such
fmished as to prevent the accumulation of dirt and is coalor hydrocarbons shouldbe designed, protected,
minimize condensation, mouldgrowth andflaking, and controlledand maintained in such a manner as not to
should be easy to clean. present a risk of aquifen and springs being polluted
8-3.3.7.4 Windows during the storage and manipulation ofthese fuels.
Windows and other openingsshould be soconstructed 8-3.4 BYllenle Facilities
as to avoid accumulationof dirt and those whichopen
should be fitted with screens. Screens should beeasily 8-3.4.1 Waler Supply
movable for cleaning and kept in good condition. 8-3.4.LI Amplesupplyof potablewaterunderadequate
Internal window sills, if present, should be sloped to pressure and of suitable temperature should be
prevent use as shelves. available with adequatefacilitiesfor its storage,where
necessary, and distribution with adequate protection
B-3.3.7.5 Doors
against contamination. The potable water should
Doorsshould have smooth, non-absorbent surfaces and, conform to the standard for drinking water (,ee
whereappropriate, beself-closing and closefJttina type. IS 10Soo).
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8-3.4.2 EfJluent and Waste Disposal Adequate natural or artificial lighting should be
provided throughout the establishment. Where
Establishments should have an efficient effluent and appropriate, the lighting should not alter colours and
waste disposal system which should at all times be the intensity should not be less than:
maintained in good orderand repair. All effluent lines
(including sewer systems) should be large enough to a) 540 lux (50 foot candles) at all inspection
carry full loadsand should be so constructed as to avoid points,
contamination of potable water supplies. b) 220 lux (20 foot candles) in work rooms, and
c) 110 lux (10 foot candles) in other areas.
8-3.4.3 Changing Facilities and Toilets
Lightbulbsand fixtures suspended over naturalmineral
Adequate, suitable and conveniently locatedchanging
water in any stage of production should be of a safer
facilities and toilets should be provided in all
type and protectedto prevent contaminationof natura.I
establishments. Toilets should be so designed as to
mineral water in case of breakage.
ensure hygienic removal of waste matter. These areas
should be well lighted, ventilated and should not open 8-3.4.7 Ventilation
directly on to natural mineral water handling areas.
Adequate ventilation should be provided to prevent
Hand washing facilities with warm or hot and cold
excessive heat, steam condensation and dust and to
water, a suitable hand-cleaning preparation, and with
removecontaminatedair. The direction of the air flow
suitable hygienic means of drying hands, should be
should never be from a dirty area to a clean area.
provided adjacent to toilets and in such a position that
Ventilationopeningsshould be provided with a screen
the employee will have to use them when returning to
or other protecting enclosure of non-corrodible
the processing area. Where hot and cold water are
material. Screens should be easily removable for
availablemixingtaps should be provided. Where paper
cleaning.
towels are used, a sufficient number of dispensersand
receptacles should be provided near each washing 8-3.4.8 Facilities for Storage of Waste and Inedible
facility. Care should be taken that these receptacles Material
for used paper towels are regularly emptied. Taps of a
Facilities should be provided for the storage of waste
non-handoperatable type are desirable.Noticesshould
and inedible material prior to removal from the
be posted directing personnel to wash their hands after
establishment. These facilities should be designed to
using the toilet. ,
prevent access to waste or inedible material by pests
8-3.4.4 Hand Washing Facilities in Natural Mineral and to avoid contamination of natural mineral water,
Water Processing Areas potable water, equipment, buildings or roadways on
the premises.
Adequate and conveniently located facilities for hand
washing and drying should be provided wherever the 8-3.5 Equipment and Utensils
processdemands.Whereappropriate, facilities for hand
8-3.5.1 Material
disinfection should also be provided. Wann or hot and
cold water should be available and taps for mixing the All equipment and utensils used in natural mineral
two should be provided. There should be suitable waterhandlingareasand whichmay contactthe natural
hygienic means of drying hands. Where paper towels mineral water should be made of material which does
are used, a sufficient number of dispensers and not transmit toxic substances, odour or taste, is non-
receptacles should be provided adjacent to each absorbent, is resistant to corrosion and is capable of
washing facility. Taps ofa non-hand operatable type withstanding repeated cleaning and disinfection.
are desirable. The facilities should be furnished with Surfaces should be smooth and free from pits and
properly trapped waste pipes leading to drains. crevices. The use of wood and other materials which
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IS 13418 : 1005
cannot be adequately cleaned and disinfected should be drawn up for each establishment to ensure that all
be avoidedexceptwhentheiruse wouldnot be a source areas are appropriatelycleaned and that critical areas,
of contamination. The use of different materials is equipment and material are designated for special
exercized in such a way that contactcorrosionthat can attention. An individual, who should preferably be a
occur, should be avoided. permanent member of the staff of the establishment
and whosedutiesshouldbe independent ofproduction,
8-3.5.2 Hygienic Design, Construction andInstallation
should be appointed' to be responsible for the
All equipment and utensils should be so designedand cleanliness of the establishment. He should have a
constructedas to prevent hazardsand permit easy and thorough understanding of; the significance of
thorough cleaning and disinfection. contamination and the hazards involved. All cleaning
personnel should be well-trained in cleaning
8-4 ESTABLISHMENT: HYGIENE techniques.
REQUIREMENTS
8-4.4 Storage and Disposal of Waste
8-4.1 Maintenance
Waste material should be handled in such a manner
The buildings, equipment, utensils and all other
as to avoid contamination of natural mineral water or
physical facilities of the establishment, including
potable water. Care should be taken to prevent access
drains, should be maintained in good repair and in an
to waste by pests. Waste should be removed from the
orderly condition. As far as practicable,rooms should
natural mineral water handling and other working
be kept protected from steam, vapour and surplus
areas as often as necessary and at least daily.
water.
Immediately after disposal of the waste, receptacles
8-4.2 Cleaning and Disinfection used for storage and any equipment which has come
into contact with the waste should be cleaned and
8-4.2.1 Cleaning and disinfection should meet the disinfected. The waste storage area should also be
requirements of this standard. cleaned and disinfected.
8-4.1.2 To prevent contaminationof natural mineral
B-4.S Exclusion of Animals
water,all equipmentand utensilsshould becleaned as
frequently as necessary and disinfected, whenever Animalsthat are uncontrolled or that could be a hazard
circumstances demand. to health should be excluded from establishments.
8-4.1.3 Adequate precautions should be taken to 8-4.6 Pest Control
preventnaturalmineralwaterfrombeingcontaminated
during cleaning or disinfection of rooms, equipment 8-4.6.1 There should be an effective and continuous
or utensils,by water and detergentsor by disinfectants programme forthe controlof pests. Establishments and
and their solutions. Detergents and disinfectants should surrounding areas should be regularly examined for
be suitable for the purpose intended and should be evidence of infestation.
acceptable to the official agency having jurisdiction. 8-4.6.2 Shouldpests gain entrance to theestablishment,
Any, residues of these agents on a surface which may eradication measures should be instituted. Control
come in contact with natural mineral water should be measuresinvolvingtreatmentwith chemical, physical
removed by thorough rinsing with water, before the or biological agents should o~ be undertaken by or
area or equipment is again used for handling natural under direct supervision of personnel who have a
mineral water. thorough understanding of the potential hazards to
8-4.2.4 Eitherimmediately aftercessationof workfor healthresultingfromthe use ofthese agents, including
the day or at such other times as may be appropriate, those hazards which may arise from residues retained
floors, includingdrains, auxiliarystructuresand walls inthe naturalmineral water,suchmeasures shouldonly
of natural mineral water handling areas should be be carriedout in accordance withthe recommendations
thoroughly cleaned. of the official agency havingjurisdiction.
8-4.2.5 Changing facilitiesand toilets should be kept 8-4.6.3 Pesticides should only be used, if other
clean at all times. precautionary measures cannot be used effectively.
Before pesticidesare applied, careshould be taken to
8-4.2.6 Roadwaysand yards in the immediate vicinity safeguard natural mineral water, equipment and
of and serving the premises should be kept clean. utensils from contamination. After application,
contaminated equipment and utensils should be
8-4.3 HYIJene Control PrOlramme
thoroughly cleaned to removeresidues prior to beina
A permanent cleaning and disinfection schedule should used again. '
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with all requirements of B-5.1 to 8-5.8 and the contamination. Only packaaina material required for
responsibility should be specifically allocated to immediate use should be kept in the packing or filling
competent supervisory personnel. area.
. B-6 ESTABLISHMENT: HYGIENIC 8-6.4.2 Productcontainersshouldnot have been used
PROCESSING REQUIREMENTS for any purpose that may lead to contaminationof the
product. In case of new containers, if there is a
8-6.1 Raw Material Requirements possibility that they have been contaminated should
To guarantee a good and stable quality of natural be cleaned and disinfected. When chemicals are used
mineral water, certain criteria should be monitored for these purposes, the container should be rinsed as ,
regularly, namely, prescribed under 8-4.2.3. Containers should be well
drained after rinsing. Used and, when necessary,
8-6.1.1 Spring discharge, temperature of the natural unused containers should be inspected immediately
mineral water. before filling.
8-6.1.2 Appearance of the natural mineral water.
8-6.5 Filling and Sealing of Containen
8-6.1.3 Odour and taste of the natural mineral water.
8-6.5.1 Packaging should be done under conditions
8-6.1.4 The conductance of natural mineral water or that precludethe introductionofcontaminantsinto the
any other adequate parameter. product.
8-6.1.5 The microbiological flora. 8-6.5.1 The methods, equipment and material used
for sealing should guarantee a tight and impervious
8-6.2 Should there be a perceptible lack in meeting
sealing and should not damage the containers, nor
the requirements, the necessary corrective measures
deteriorate the physical, chemical,microbiological and
are immediatelyto be taken.
organoleptic qualities of natural mineral water.
8-6.3 Treatment
8-6.6 Packaging of Contalnen
The treatment may include decantation, filtration,
The packaging of containers should protect the latter
airing and where necessary application of off take of
fromcontamination and damageand allowappropriate
carbon dioxide.
handling and storing.
8-6.3.1 Processing should be supervised by
technically competent personnel. 8-6.7 Lot IdentiOcatlon
8-6.3.1 All steps in the productionprocess, including Eachcontainershall be permanently markedwith code
packaging, should be performed without unnecessary to identify the producing factory and the lot. A lot is
delay and under conditions which will prevent quantity of natural mineral water produced under
the possibility of contamination, deterioration or identical conditions, all packagesof whichshould bear
development of pathogenic and, spoilage micro- a lot number that identifies the production during a
organisms. particulartime, interval,and usually from a particular
processing line or other critical processing unit.
8-6.3.3 Rough treatment of containers should be
avoided to prevent the possibilityof contamination of 8-6.8 Processing and Production Records
the processed product.
Permanent, legible and dated records of pertinent
8-6.3.4 Treatment are necessary controls and should processing and production details should be kept
be such as to protect against contamination or concerningeach lot. These records should be retained
development of a public health hazard. It also protect for a period that exceeds the shelf life of the product
against deterioration within the limits of good or longer if required. Records should also be kept for
commercial practice. the initial distribution by lot.
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under such conditions as will preclude contamination of the end-product should take place to ensure that
with and/or proliferation of micro-organisms, and only natural mineral water which is fit for human
protect against deterioration of the product or damage consumption is despatched and that end-product
to the container. During storage, periodic inspection specifications are complied with.
CHECKLIST FOR GOOD HYGIENE PRACTICES AND FOOD SAFETY SYSTEMS FOR
PACKAGED NATURAL MINERAL WATER PROCESSING UNITS
(Clause 5)
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ANNEX C
(Clause 6.1.4)
DETECfION AND ENUMERATION OF THE SPORES OF SULPHITE-REDUCING
ANAEROBES (CLOSTRIDIA)
c-r PRINCIPLE C-3 ENRICHMENT CULTURE
The sporesofsulphite-reducing anaerobes (clostridia) Detection and enumeration of spores of sulphite-
are widespread in the environment. They are present reducing anaerobes by inoculating volumes of the
inhumanandanimalfaecalmatter, inwastewaterand, sample into liquid enrichment media. followed by
in soil. Unlike Escherichia coli and other coliform incubation at 37 % loe for 44 % 4 h in anaerobic
OrLllP;Sm, the sporessurvivein waterfor longperiods conditions.
as they are more resistant than vegetative forms to the
C-4 CULTURE MEDIA AND REAGENTS
actionofchemical andphysical factors. Theymaythus
give an indication of remote or intermittent pollution. C-4.1 Basle Materials
They may even be resistant to chlorination at levels In order to improve the reproducibility of the results,
which are normally used for the treatmentof water. it is recommended that, for the preparation of
C-1 APPARATUSANDGLASSWARE the diluents and culture media, dehydrated basic
components or complete dehydrated media be used.
C-1.1 SCrew-cap bottles or vialsandstoppers of boron Similarly. commen:ially prepared reagents may also
silicateglass of capacities 200. 100 and 25 ml. be used. The manufacturer's instructions shall be
C-2-2 Volumetric Pipettes- ofcapacities 10mland followed.
I ml. The chemical products used for the preparation of the
culturemediaand the reagents shall be of recognized
C-2-3 Water Batbs - thermostatically controlled.
analytical quality.
C-2.4 Test Tubes - 150mm x 13mm. . The water used shall be distilled or deionized water,
C-2.S Iron Wire free from substances that might' inhibitthe growth of
micro-organisms underthe test conditions.
C-2.6 Incubator - capable of being maintained at
37~ 1°C. Measurement of pH shall be madeusing apH-meter,
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measurements being referred to a temperature C-4.4 IroD (III) Cltnte(C,H.,O, Fe), 7 percent(mIm)
of 2SoC. solution.
If the preparedculturemediaare notused immediately, Dissolve 7 g of iron(nl) citrate in 100 ml of water.
they shall unlessotherwisestated,be stored in the dark Sterilize by filtration.
at approximately4°C, for no longer than 1 month.
Store at between2 and SoC.
C-4.2 Differential Reinforced Clostridial Medium It is advisable to prepafea fresh solutionevery 14 days.
(DRCM)
C-4.! Complete Medium
C-4.2.1 Single Strength Basal Medium
C-4.S.1 On the day of analysis,mix equal volumesof
Composition the solutions of sodium sulphite (see C-4.3) and iron
Peptone tryptic digest of meat 10 g (III) citrate (see C-4.4).
Meat extract 109 C-4.!.2 Add 0.5 ml of the mixture (see C-4.S.I) to
Yeast extract 1.S g each bottle of single strength medium (see C-4.1.1)
Starch 1g which has been freshly heated and cooled.
Hydrated sodium acetate Sg C-4.S.3 Add 0.4 ml of the mixture (see C-4.S.1) to
Glucose 1g each 10ml, and 2 ml to each SO mit of double strength
L-Cysteine-hydrochloride O.S g medium (see C-4.2.2) similarlytreated.
Water 1 000 ml
C-4.6 SelectioD of Spores (Tecbnlque)
Mixthe peptone,meatextract, sodiumacetate andyeast
Before the test," the sample of water should be heated
extract with 800 ml of water.
in a water bath at 75 :i: SoC for 15min from the time it
With the remaining 200 ml of distilled water, prepare reaches that temperature. A similar bottle containing
a starch solution by mixing the starch in a little cold the samevolumeof water as the test sample should be
water to fonn a paste. Heat the rest of the water to used periodically as a control in order to check the
boiling point and slowly add it to the paste with heatingtime required. The temperatureof the water in
constant stirring. the control bottle can be constantly recorded by
thermometer.
Then add this starch solution to the first mixture and
heat to boiling point until it dissolves. C-4.7 Inoculation and Incubation
Finally,add the glucoseand L~ysteine-hydrochloride Add SO ml of sample (lee C-4.6) to a 100 ml screw-
and dissolve. cap bottle containing SO ml of the double strength
Adjust the pH between 7.1 and 7.2 with sodium complete medium (see C-4.S.3).
hydroxide. Add 10 ml of sample (see C-4.~) to a series of five
Transfer 25 ml aliquots of the medium into screw- 2S ml screw-cap bottles containing 10 ml of double
capped bottles of capacity 25 ml. Sterilize in the strength completemedium, (see C-4.5.3),
autoclave at 121 ± 1°C for 1S min. Add 1 ml of sample(see C-4.6) to a seriesoffive25 ml
C-4.2.2 Double Strength Basal Medium
screw-cap bottles containing25 ml of single strength
completemedium (lee C-4.5.2).
Preparethe double strength medium as in C-4.1.1 but
Ifnecessarytadd I mlof. I to 10dilutionof thesample
reduce the volume of water by half.
(see C-4.6) to a series of five 2S ml screw-cap bottles
Transfer 10 ml and 50 ml aliquotsofthe medium into containing25 ml ofsingle strength complete medium
screw-capped bottles of capacities25 ml and 100 ml (see C-4.S.2).
respectively.
In order to carry out a qualitative examination of
C-4.3 Sodium Sulphite (N~SOJ, 4 percent (mlIII) 100 ml of mineral water or packaged water without
solution. makina an MPN count, usea 200 ml vial filled with a
mixture of 100ml of doublestrenathcompletemedium
Dissolve 4 g of anhydroussodium sulphite in 100 ml
(lee C-4.5.3) and 100 ml of sample (lee C-4.6).
of water. Sterilize by filtration.
If necessary, top up all the bottles with the single
Store at between 2 and SoC.
strenath complete medium (lee C-4.5.2) to brina the
It is advisableto prepare a freshsolution every 14days. volumeofiiquid levelwith the neck and to ensure that
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only a very small volume of air remains, then seal the addition of iron wire, heated to redness and placed
bottles hermetically, or incubate under anaerobic into the medium before inoculation, may aid
conditions. anaerobiosis.
Incubatethe inoculated bottles at 37 ± I°Cfor 44 ± 4 h. Bottles in which blackeningis observed, as a result of
Large volumes ofculture in hermetically sealed glass the reduction of sulphite and the precipitation of iron
bottles may explode due to gas production. The (II) sulphide, shall be regarded as positive.
•
ANNEX D
(Clause 6.1.5)
DETECTION AND ENVMERATION OF PSEUDOMONAS AERUGINOSA
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ANNEX 2D
(Clauses D-2 andD-IO)
FURTHER INFORMATION ABOUT PSEUDOMONAS AERUGINOSA
Pseudomonas aeruginosa is the type specie of the Leifson test. It generally reduces nitrate beyond the
genuS Pseudomonas. stage of nitrite and produces ammonia from the
breakdown of acetamide.
It is a gram negative, non-sporing rod which is oxidase
and catalase positive. It is capable of growth at 42°C Gelatin is liquefied, casein is hydrolysed, but
but not at 4°C; it usually produces a water soluble starch is not hydrolyzed. The pigment pyocyanine
fluorescentpigment (98 percentofstrains)and exhibits (blue-green) is produced by more than 90 percent of
oxidative metabolism as indicated by the Hugh and strains.
ANNEX E
(Clause 9)
SAMPLING PLAN FOR PACKAGE NATURAL MINERAL WATER
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the number of bottles in the sample shall be taken at randomnessof selection, procedure given in IS 4905
random from these cartons and then the required shall be followed.
number of bottles picked up according to E-l.3.1.
Table 7 Scale 01 SaDlpliBI
E-I.3.3 The sample bottles selected as in E-l.3.1
or E-l.3.2 shall be divided at random into three equal (Clause E-2.2.3)
sets and labelled with all the particulars of sampling. SINo. No. 01CoataAnen i. tile Lot N•• ber olS.aapaa
One of these sets of sample bottles shall be for the (I) (2) (3)
purchaser, anotherfor vendorand the third for referee.
i) 0-500 5
E-l.3.4 Referee Sample iI) 501 to 1 200 8
iii) 1 201 to 3 200 13
Refereesample shall consist of a set of sample bottles iv) 3 201 and above 20
marked for this purpose and shall bear the seals of
the purchaser and the supplier. These shall be kept at E-2.2.4 These containers shall first be examined
a place agreed to between the purchaser and the visually for the condition of packing, external
supplierso as to be used in a case ofa dispute between appearance and the fill. The lot shall be considered
the two. satisfactory for inspection of othercharacteristics given
E-l.4 Criteria for Conformity in the specification, if all the containers are found
satisfactory for these characteristics.
The tot shall be declared as conforming to the
requirements of the specification, if allthe requirements E-2.2.S In case any defective container is found
are complied with. according to E-2.2.4, twice the number of containers
shall be examined for these characteristic(s). If no
E-2 GENERALREQUIREMENTS OF SAMPLING defectivecontaineris found,the lot shall beconsidered
FOR ABOVE 2 LITRES CONTAINERS satisfactory for inspection of othercharacteristics given
in the specification.
E-2.1 In drawing, preparing, storing and handling
samples,the following precautions and directions shall E-2.3 Preparation of Test Samples
be observed as far as possible:
E-2.3.1 Out of the containers selected according
a) Sample shall be drawn in original sealed to E-2.2.3, any three containers shall be selected at
container and kept in protected place not random and stored separately.
exposed to damp air, dust or soot; and
b) Each container in originalshall be sealedand E-2.3.2 Each of the sample containers selected as
marked with full details of sampling. in E-2.3.1 shall be divided at random into three equal
sets and labelled with all the particulars of sampling.
£-2.2 Scale or Sampling One of these sets of sample containersshall be for the
purchaser, anotherfor vendor and the third for referee.
E-2.2.1 Lot
E-2.3.3 Referee Sample
The quantity of packagednatural mineral water of the
sametype belongingto the same batchof manufacture Referee sample shall consist of a set of sample
and packed in a day, shall constitute a lot. containers marked for this purpose and shall bear the
seals of the purchaserand the supplier. These shall be
E-2.2.2 For ascertaining the confonnityof the material
kept at a place agreed to between the purchaser and
to the requirements ofthe specification, samplesshall
the supplierso as to be used in case of dispute between
be tested from each lot separately.
the two.
E-2.2.3 The number of containersto be selected from
E-2.4 Criteria for Conformity
a lot shall depend on the size of the lot and shall be
drawn at random, according to Table 7. Separate The lot shall be declared as conforming to the
container(s) shall be drawn for testing for the requirements of the specification, if allthe requirements
microbiological requirements. In order to ensure the are complied with.
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ANNEX F
[Table 2, Sl No. (viii)]
DETERMINATION OF BARIUM CONTENT
ANNEX G
[Table 2, SfNo. (ix)]
DETERMINATION OF ANTIMONY BY SPECTROPHOTOMETRIC METHOD
G-3.1 Hydrocblorlc Acid Solution - 6 N. Cool reagents G-3.1, G-3.2, G-3.3 and approximately
100ml benzene, andeight 125 ml separators with teflon
G-3.2 Dilute Phospborlc Acid - 3 N. stopcocks in refrigerator before use. Maintain
Dilute 70 ml phosphoric acid (85 percent) to 1 litre temperature at SoC to 10°C during extraction and
with distilled water. colour development
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ANNEX H
[Table 2, Sl No. (x)]
DETERMINATION OF BORATE
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H-2.6 Boron, standard solution 2 corresponding to aliquots to a volume of just less than 25 rnl and adjust
1.0 mg of B per litre. their volumes to precisely 25 ml by the addition of a
little extra distilled water, as necessary. Carry out the
Dilute 10 ml of borate solution (see 8-2.5) to 100 ml
procedure given in 8-4.1 on these aliquots.
with water.
Carry out a blank determination with each of the
1 ml ofthis standard solution contains 1.0 ug of borate,
aliquots. Ifborate is present in the distilled water, the
expressed as 8.
borate found increases in proportion to the volume of
H-2.7 Calclumhydroslde ICa(OH)2) the aliquot taken. Erratic results indicate external borate
contamination. Relatively high but constant results
8-3 APPARA TUS indicate impure reagents.
8-3.1 Ordinary laboratory apparatus made of 8-4.3 Prevention of Contamination
polypropylene, polyethylene or polytetrafluoroethylene,
where applicable. As borate is widespread in the environment, significant
contamination may occur during trace determinations.
8-3.2 Spectrometer, for use in the wavelength range
of 410 om to 420 nm, with cells of an optical path The following sources ofcontamination, and remedies,
length between 10 mm and SO mm. should be considered.
Laboratory glassware is usually made from borosilicate
8-4 PROCEDURE
glass. Special borate-free thermally resistant glass is
8-4.1 Determination obtainable, but for routine purposes, old borosilicate
glass, well rinsed in hydrochloric acid, may be used
Transfer 25.0 ml ofthe sample, or a smaller amount of
for acidic solutions, but should never be used for
the sample diluted to 25 ml with distilled water, into a
neutral or alkaline solutions, or for prolonged storage
100 ml polyethylene flask. Add 10ml ofazomethine-H
at any pH value. (Borosilicate glassware previously
(see H-2.1). Mix and allow to stand in the dark for 2 h
used with alkaline solutions shall not be used without
at 20 ± 1°C, then measure the absorbance at the
very thorough acid rinsing.) Polyethylene flasks and
absorption maximum in the range of410 nm to 420 nm
plastics pipettes are preferable.
against distilled water in a cell of optical path length
10 mm, using the spectrometer set up according to the Detergents and soaps used for glassware and labcoats
manufacturer's instructions and after setting the zero should be borate free, and the use oftowels and tissues,
with distilled water in the cell. Alternatively use a for drying shall be avoided.
cell of SO mm optical path length for low boron Toiletries, talcum powder and cosmetics used by
concentrations of up to about 0.2 mg ofboron per litre. technicians often contain borate and should be avoided
Check the wavelength of the absorption maximum or removed, especially prior to undertaking accurate
whenever a new batch of this reagent is used. low-level determinations.
NOTE- The reactionIime may be shortened by keeping the
Water and reagents may contain borate and blanks
treatedsampleat a temperature of300C. Inthiscasethe sample,
the blank and the calibration samples should be treated should be carried out at least in duplicate and should
accordingly, because the intensity of colour is temperature agree.
dependent.
"-4.4 Calibration
8-4.2 Blank Test
8-4.4.1 Zero mg/I to 0.20 mg/I ofBoron Calibration
Carry out a blank test by treating 25 ml of water as Graph
described in 8-4.1. Ensure that the blank value is in the
To a series of six 25 ml one mark plastics flasks add
range of 0.1 absorption units to 0.17 absorption units
respectively 0 ml, 1 ml,2 ml, 3 ml, 4 ml and 5 ml of
per 10 mm; if the absorption is higher then check the
boron standard solution .2 (see 8-1.6), dilute to the
reagents and the distilled water for their borate content.
mark with distilled water and mix. This gives
NOTE- The following procedure may be used to check the concentrations of 0 mg; 0.04 mg; 0.08 mg; 0.12 mg;
quality of reagentsand the distilled water.
0.16 mg and 0.20 mg of boron per litre respectively.
Measure into three separate borate-free beakers Analyze each standard solution as described in H-4.1t
(preferably polytetraf1uore~ylene) 2S ml, 100 ml and measuring the absorbance values in a SO mm optical
2S0 ml aliquots of the distilled water. Make each path length cell compared against distilled water.
slightly alkaline by the addition ofthe same small (for Prepare a calibration graph by plotting the absorbance
example 200 mg) amount of calcium hydroxide values against the known concentrations in milligrams
(see B-2. 7) t~ each. Evaporate the 100 ml and 250 ml of boron per litre for each standard.
2S
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ANNEXJ
[Table 2, Sl No. (xi) and Table 3, Sf No. (iv)]
DETERMINATION OF SILVER AND CHROMIUM CONTENTS
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Table 9 Preparation of Metal Staadard SolutioDS the residue. Wash the watch-glass and beaker with
(Clauses J-3.4.1 and J-3.4.2) water and tilter. Wash the filter and discard. Dilute
the filtratewith waterto such a concentrationthat it is
51 Metal Wel.llt Compo.ad DluoIvlDi MedlulD within the range of the instrument.
No. (a) (I litre Total)
(1) (2) (3) (4) (5) J-4.3 Total Metal
i) Silver 1.575 Silver nitrate Water + 10ml Transferan aliquotof well mixedsampleto the beaker
(AgNO) redistilled nibic acid
and add 3 ml nitricacid. Heatandevaporate to dryness
ii) Chromium 1.923 Chromium Water+ 10ml (do not boil). Cool and add 3 ml nitric acid and heat
oxide (Cr103) redistilled nitric acid
untildigestion iscomplete, generally indicated by light
coloured residue. Add2 ml hydrochloric acid( I:I, vlv)
J-3.S Lanthanum Stock Solution and heatgentlyto dissolve the residue. Wash the watch-
Slowly add 250 ml hydrochloric acid to 58.65 g glass and beaker with water and filter. Washthe filter
I~thanum oxide (L~03' purity99.9 percentby mass), and discard. Dilute the filtrate with water to such a
dissolve and dilute to 1 000 ml. concentration that it is within the range of the
instrument.
J-3.6 Ammonium Pyrrolidine Ditbiocarbamate
Solution J-S DETERMINATION
Dissolve 1 g ammonium pyrrolidine dithiocarbamate Transfer an aliquot of the sample to a 250 ml beaker
in 100 ml water. Prepare fresh daily. and dilute to 100 ml with water. Prepare blank and
standard solution in the same manner. Adjust the pH
J-4 PREPARATION OF SAMPLE of the sample and standard solutions to 2.5 with
hydrochloric acid using a pH-meter. Transfer
J-4.1 Dissolved Metals
quantitatively to a 200 ml volumetric flask,add 2.5 ml
As soon as practicable after collection, filter known of ammonium pyrrolidine dithiocarbamate solution and
volume of sample through 0.45 J.1m membrane. Use mix. Add 10 ml methyl isobutyl ketone and shake
firstly50-100 ml to rinse the flaskand discard. Collect vigorously for I min. Let the layers be separated and
the filtrate and preserve the solution by adding 3 ml then add water until the ketone layer is in the neck of
nitric acid (I: I, vlv) per litre. the flask. Centrifuge, ifnecessary. Aspiratethe ketone
layer and record readings of standards and samples
J-4.2 Suspended Metals againstblank. The fuel-to-air ratio should be adjusted
Transfer the residue and membrane from J-4.1 to a to as blue a flame as possible since organic solvents
250 ml beaker and add 3 ml nitric acid. Cover with add to fuel supply. Preparethe calibration curve from
watch-glassand heat gently to dissolvethe membrane. the average of each standard and read the sample
Increase heating and evaporate to dryness. Cool and concentration.
add 3 ml nitricacidand heatuntildigestion iscomplete,
J-6 CALCULATION
generallyindicatedby lightcolouredresidue. Add2 ml
hydrochloricacid (1:1, vlv) and heat gentlyto dissolve Metal content, mg/l = Metal, in mg, in the aliquot/I.
ANNEX K
[Table 2, S/ No. (xxi)]
METHOD OF TEST FOR ANIONIC SURFACE ACTIVE AGENT
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K-2.2 Spectrometer with Selectors for K-3.1 Ethyl Aeetate. freshly distilled.
Discontinuous Variation. capible of measurement at
650 nm, equipped with cells of optical pathlengths of C••tIoII- EthyllCCUlte Is tl.....ableand toxic.
10 mm and SO mm. K-3.J Chloroform
K-2.3 Gas-Stripplnl Apparatus - One litre C••t10D - Chloroform Is• suspected carclnoaen. Ifnecessary.
capacity (see Fig. 1). purify the chloroform by flItratlon throuah AIZOJ (neutral
...... W200). If it alves rise to hiah results in blankteat.
K-) REAGENTS K-3.4 Ethanol. 95% (vlv).
K-).l Sodium Chloride K·3.S Metha.ol. freshly distilled.
ETHYL
ACETATE
, Itl
Q
TEST Itl
SAMPLE
SINTEREO
Gl.ASS
FIllER GI
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K-3.6 Sulphuric Acid Solution - 0.5 mill. joint with about 30 ml of ethanol and add the rinsings
to the contentsof the flask. Neutralize the solutionwith
K-3.7 Ethanollc Sodium Hydroxide - 0.1 mol/I.
sulphuric acidagainstphenolphthalein until it becomes
Dissolve 4 g of sodium hydroxide pellets in ethanol colourless. Transfer the solution to a I 000 ml
and dilute to 1 000 ml with the same ethanol. volumetric flask, dilute to the mark with water and
mix. This standard solution is stable for 6 months.
K..3.8 Metbylene Blue, Neutral Solution
K-4 PROCEDURE
Dissolve0.350 g ofmethyleneblue in waterand dilute
to I 000 ml. Prepare the solution at least 24 h before K-4.1 Separation of the Surfactant
use.
Non-surfactant methylene blue active substances can
NOTE- Theabsorbance of the chloroform phaseof the blank cause errors in the test of methylene blue index.
test, measured against chloroform, shall not exceed 0.2 per
10 mm of optical pathlength at 6S0 nm. In the case of high
Stripping is recommended for concentrating small
absorbances during the blank test, use other batches of amount of surfactants from water samples. Separate
methylene blue or purify the methylene blue solution by suspended matter by centrifugation, but note that
extractionas under: adsorbed surfactants on suspended matter will not be
Placethe methylene bluesolutionin a suitablylargeseparating determined,
funnel, For each 100 ml of methylene blue solution,add 200
ml of butTer solutionand 200 ml of chloroform. Shakefor 30s Place a measured quantity of the test sample, up to
and allow to separate. Run off the chloroform layer as
completely as possible and rinse the aqueous layer without
1 000 ml in the gas-stripping apparatus. Install the
shakinawith60 mlof chloroform foreach 100mlof methylene stripping apparatus in well ventilated hood to carry
bluesolution. Repeatthe extraction andrinseas before. Discard off ethyl acetate vapour. Separation is improved by
the chloroform extracts; collect for reuse after treatment. additionof sodium chloride. If samplevolumeexceeds
K-3.9 Methylene Blue, Acidic Solution 500 ml, add 100 g of sodiumchloride and dissolve by
passing nitrogen gas or air through it. I f a smaller test
Dissolve 0.350 g ofmethyleneblue in 500 ml of water sample volume is used, dissolve 109 g of sodium
and add 6.50 ml of sulphuricacid (p-1.84 g/ml). Dilute chloride in 400 ml of water and add this solution, to
with water to 1 000 ml after mixing. Prepare the the test sample.
solution at least 24 h before use.
Ifnecessary, add water to bring the sample surface up
The absorbance of the chloroform phase of the blank to the level of the upper stopcock. Add 100 ml ethyl
test, measured against chloroform, shall not exceed acetate.Fill the wash bottle in the gas line(nitrogen or
0.02 per 10 mm of optical path length at 650 nm. In air) two-thirdfull with ethyl acetate. Pass a gas stream
the case of higher blank absorbances, either wash the of20 Vb to so Vb through the gas-strippingapparatus.
methylene blue solution twice with chloroform for Adjustthe gas flow in sucha waythatthe phasesremain
purification, as in K-3.8, or use other batches of separateand no turbulence is producedat the interface.
methylene blue. The significant mixing of the phases and consequent
solution of ethyl acetate in the water is avoided. Stop
K-3.10 Burrer Solution,pH 10
the gas flow after 5 min.
Dissolve 24 g of sodium hydrogen carbonate If a loss of more than 20 percent (wv) of the organic
(NaHC0 3) and 27 g of anhydrous sodium carbonate phase has occurreddue to solution in the water phase,
(N~C03) in water and dilute to I 000 ml.
discard the test sample.
K-J.ll Phenolpbthalein Indicator Solution Runoff the organic phasecompletelyinto a separating
funnel. Return any water in the separating funnel to
Dissolve 1.0 g of phenolphthalein in SO ml of ethanol
the gas-strippingapparatus.
and add, while stirring continuously, 50 ml of water.
Filter off any precipitate that forms, Filterthoethylacetate solution through a dry qualitative
gas-filter paper intoa 250ml flask. Adda further 100ml
K-3.11 Dodecylbenzene Sulpbonic Acid Methyl
of ethylacetateto the gas-stripping apparatus and again
Ester (Tetrapropylene Type), stockstandard solution.
pass nitrogen or air through it for S min. Separate the
Weigh to the nearest 0.1 mg, 400 mg to 450 mg of organic layer as described above, using the same
dodecylbenzene sulphonic acid methyl ester, into a separating funnel, filter, and add it to the first portion.
round-bottomed flask, and add SO ml of ethanol- Rinse the filter paper and funnel with 25 ml of ethyl
sodium hydroxide solution and some anti-bumping acetate. Remove alltheethylacetate solution on a water-
granules. Attach the reflux condenserand boil for I h. bath under a hood. To speed up the process, direct a
Aftercooling,rinsethe condenserandthe ground-glass gentleair stream over the surface of the.solution.
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Dissolve the residue in about 5 ml of methanol and completelyas possibleand swirl the funnel to dislodge
50 ml of water. Transfer the solution quantitativelyto droplets from the sides of the funnel.
a 100 ml volumetric flask and dilute to the mark with
Allow to settle for 2 min, then run as much as possible
water.
of the chloroform layerintoa secondseparatingfunnel,
K-4.2 Blank Test containing 110 ml of water and 5.0 ml of acidic
methylene blue solution. Shake uniformly but not too
Carry out a blank test at 650 nm and subtract the vigorously for 1 min as previously described. Filter
interpolated absorbance, Ao' from the absorbance Ai the chloroform layer through a cotton or glasswool
of the test sample. Under the given conditions, the filter wetted with chloroform into a 50 ml volumetric
absorbance Ao of the blank test shall not exceed 0.02 flask.
per 10 mm optical path length, otherwise equipment
and the reagents shall be checked carefully for any Repeatthe extraction of the alkalineand acidicsolution
contamination. using a 10 ml portion of chloroform for the extraction.
Separatethe chloroform layer and filter it, through the
K-4.3 Test with the Sample same filter, into the volumetric flask. Repeat the
extraction using a further 10 ml portion ofchlorofonn
Transfer a measured volume of the test sample into a and filter that into a SO ml volumetric flask. Dilute to
separating funnel. This test portion should contain the mark with chloroform and mix.
20 ~g to 200 Ilg of MBAS (methylene blue active
substances). In the lower MBAS range, a test portion For each test sample carry out the complete extraction
up to 100 ml may be used. If the volume of the test for a blank determination on 100 ml water.
portion is lessthan 100 ml, dilute with waterto 100 mJ. Measurethe absorbancesfor the test sample as well as
Add 5.0 ml of neutral methylene blue solution, 10 ml for the blank test at 650 nm in cells of optical path
of butTer solution and IS ml chloroform, Shakeevenly length 10 mm to 50 mm against chloroform. The
and gently about twice a second for 1 min, preferably absorbanceof the test sample should not be more than
in a horizontal plane. Allow the layers to separate as that of the blank (see K-4.2).
ANNEX L
[Table 3, Sl No. (vii)]
DETERMINATION OF NICKEL BY FLAME ATOMIC ABSORPTION
SEPCTROMETRIC METHOD
L-I PRINCIPLE Add 100 ml ofnitric acid to 600 ml of water and dilute
to I 000 ml.
Formation of a complex between the metals being
determined and ammonium l-pyrrolidinedithio- L-3.3 Nickel Standard Solution Corresponding
carbamate (APDC) and extraction at pH 2.5 with to 1 000 gil
metbyl-isobutylketone (MIBK). Weigh 1.000 g of pure metal and dissolve it in cone,
Determination of the metals in this organic phase by nitric acid, heating to effect complete dissolution.
flame atomic absorption spectrometry. Allowto cool and transfer each solution quantitatively
to a 1 000 ml volumetric flask, dilute to the mark with
L-2 APPARATUS water and mix.
L-2.1 Atomic absorption spectrometer fitted with For preparing standard solution, it is also permissible
hollow cathode lamps for appropriate metals or to use metal salts of accurately known composition.
electrodeless discharge lamps, and with a suitable
device for allowing for the correction of the non- One ml of the standard solution contain 1.00 mg of
specific absorbance and with a nebulizer-burnerwith nickel.
an acetylene-air flame. L-3.4 Sodium Hydroxide - 2.S moVI.
L-3 REAGENTS Dissolve 100g of sodium hydroxide in wateranddilute
L-3.1 Concentrated Nitric Acid (p-I.4 alml) to 1 litre.
L-3.2 Nitric Acid, 1.5 molll L-3.5 HydrOeblorie Aeld - 0.3 molll.
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L-4.3 Add 4 ml of 15 mol/l nitric acid to 100 ml of L-5.4 Preparation of the Sets of Calibration
the sample and heat until the volume is reduced to Solutions
SOmt.
Dilute with water immediately "before use, the nickel
Place the treated sample in a boiling flask. Add 12 ml standard solution in order to obtain diluted solutions
ofhydrochloric acid (p-l.19 g/ml), Connect the boiling containing 10 mg of nickel per litre.
flask to a condenser and reflux the solution for 2.5 h.
In a SOO ml volumetric flask, place
Cool and filter to remove insoluble materials that could
clog the nebulizer. Collect the filtrate in a 100 ml a) 20 ml of nickel solution that contain 10 mg/l
volumetric flask. of nickel; and
b) O.S ml of nitric acid.
Wash the condenser and filter several times with water,
add this to the contents of the volumetric flask and Make up to the mark with water. This is solution S.
make up to the mark with water. Prepare at least four calibration solutions by diluting
solution S with water so as to cover the ranges of
L-5 PROCEDURE concentrations of nickel from 0 to 200 ug/l.
L-S.l Test Portion Acidify each calibration solution by adding the same
Place in a 100 mIone-mark volumetric flask a test nitric acid which has been added to preserve the
portion of the acidified sample containing S to 20 J.1g samples. The volume added shall be such that the
of nickel being determined. Make up to the mark with concentrations of nitric acid are the same in the sample
water. and in the calibration solutions.
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ANNEXM
[Table 3, Sl No. (viii)]
TEST FOR POLYCHLOROBIPHENYLES (PCB)
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M-3.~ Acetic Acid, glacial, redistilled. correspondence between peak retention times which
M-3.7 Chromium Trioxide, re-crystallised. give indication of the presence of PCB in the sample.
M-4 SEPARA TION OF PCB FROM THE MORE M-5 OXIDATION OF pp'-DOE IN THE PCB
POLAR PESTICIDE RESIDUES FRACTION
Weigh s.o g of the prepared gel and rapidly transfer M-S.1 Adjust the volume of fraction I to 2 ml,
it, by using small amounts of hexane, to the concentrating, if necessary, as described above. Add
chromatographic column, in which a plug of cotton 2 ml of acetic acid and, after replacing the micro-
wool has been placed just above the stopcock. The Snyder column, heat the tube cautiously in the steam-
stopcock may be moistened with solvent but must not bath until all of the hexane has evaporated, as judged
be lubricated with grease. Allow the gel to settle in the by the reduction in volume. Introduce 100 mg of
column and remove and trapped air bubbles by stirring chromium trioxide and place the tube in boiling water
with a glass rod. Drain the surplus hexane from the for 15-20 min. Cool the mixture and shake it vigorously
column until its meniscus just touches the surface of with 2.0 ml of hexane (accurately measured) in the
the gel. Introduce the cleaned-up sample extract as a stoppered tube. Neutralize the acid with 6-7 ml of 5 N
solution in 1 ml of hexane, and allow the hexane to sodium hydroxide, solution. Shake the tube again and
drain until the meniscus again just touches the surface then set it aside until the two layers separate. Keep the
of the gel. Wash the vessel that contained the extract tube well stoppered to prevent loss of hexane.
with two I-ml portions ofhexane, adding each washing
M-6 DETECTION OF PCB
separately to the column' and allowing it to run just
into the gel. Place a receiver, preferably a Kudema- Inject 5 J.11 of the upper hexane layer of the oxidized
Danish evaporator, beneath the column and pass 42 ml mixture on to at least two GLC columns with different
of hexane through the column at a rate not exceeding liquid phases, one of which should be Apiezon L.
0.7 mllmin. Collect the eluate, stopping the elution Compare the retention times of the peaks so obtained
when the meniscus reaches the top ofthe gel, and label with those given by the PCB reference material.
this fraction I. It should contain all the PCB. Agreement between the retention times of the peaks
Hexachlorobenzene, aldrin, O.p-DDT and pp' -DOE, so obtained now indicates the presence of PCB
if present, are also eluted in this fraction. Change the compounds in the sample with a greater degree of
receiver and pass 50 ml of a 10 percent solution of certainty. Any pp'-DDE, which was present in
diethyl ether in hexane through the column. Collect fraction 1, will have been converted into 4, 4'-
the eluate and label it fraction 2. This contains the dichlorobenzophenone. Under the GLC conditions
remainder of the organochlorine pesticide residues, quoted, this compound has a retention time similar to
including usually a small amount ofpp' -DOE, and can that of heptachlor epoxide and so gives a peak on the
be used for the determination of these. Concentrate chromatogram before most of the PCB isomers
fraction I to 5 ml in the Kudema-Danish evaporator encountered in practice. A comparison of the traces
and examine it by GLe with electron capture detection, before and after oxidation of fraction 1 will show how
on at least two stationary phases of different polarities. much pp' -DOE was originally present. Adjust the
Iffurther concentration is necessary, reduce the volume volume of the hexane solution so that the individual
of the solution with a gentle stream of dry air heights ofthe PCB peaks are within the linear response
or nitrogen at room temperature. Compare the range of the electron capture detector. Under the
chromatograms from fraction 1 with zhose given by prescribed GLC conditions for the Apiezon L. column,
solutions of commercial PCB preparations when the last ofthe PCB isomers normally found in wildlife
injected on the same columns as well as the tissues emerges in about 120 min after injection.
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ANNEX N
(Clause 6.3)
STANDARDS ON METHODS or REsmUE ANALYSIS
Sl Name ofPesticide Method01Test, &fto
No. r ...... ,
USEPA AOACIISO
(1) (2) (3) (4)
1. DDT (0, P &, p, p-isomers of DDT, DOE &, DOD) 508 AOAC990.06
2. y-HCH (Lindane) S08 AOAC990.06
3. a,p&5-HCH 508 AOAC990.06
4. Endosulfan (a, Pand Sulphate) 508 AOAC990.06
s. Monocrotophos 8141A
6. Ethion 16S7A
7. Chlorpyrifos S2S.2, 8141A
8. Phorate (Phorate and its oxygen analogue that is, 8141A
phorate sulphoxide and phorate sulpbone)
9. 2,4-D S15.1
10. Butachlor 525.2, 8141A
II. Isoproturon 532
12. Alachor 525.2, S07
13. Atrazine 525.2, 8141A
14. Methyl Parathion(Methyl Parathion and its oxygen 8141A ISO 10695
analogue that is, methyl-paraoxon)
IS. Malathion(Malathionand its oxygen analoguethat 8141A
is, malaoxon)
16. Aldrin and dieldrin 525.2 AOAC990.06
NOTE- Test methods R for Iuidance and reference for testina 11boratory. In case of two methods, USEPA method shill be the
reference method.
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