Bot

tl
edDr
inki
ngWat
erPr
ocess

Wr
it
en,
Arr
anged&Ver
if
iedby
;

Hay
atKhanAf
ri
di
(
Qual
i
tyCont
rol
Manager
)
AquaNat
uralWat
er,
Peshawar

MuhammadNadeem
(
Dir
ect
or)
I
NDUSTRI
ALENGI
NEERI
NG&CHEMI
CALS(
PVT)LTD.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Gr
oundwat
er.
Pur
eground/
Rawwat ercont
ains11.88%Hydrogenand88.
02%Oxy
genbywei
ght
.Iti
sodor
less,
Col
orl
essandtast
eless.Waterisnecessar
yforhumanbody.

Rawwat erobtai
nedfrom gr
oundmaycont ai
nhighamountofTDS,highAlkali
nit
y,Turbi
dit
y,
mineral
sandMi croor
gani
smswhi chmayaff
ectthequal
i
tyofthefi
nishedproduct.Rawwat eri
s
puri
fi
edandt r
eatedtomakei taccordi
ngt
otherequi
redstandar
ds.

Obj
ect
ivesf
orWat
erpr
ocessi
ngUni
t.
 Auni
for
m gr
oundwat yf
ersuppl oral
ltheseasonsoft
hey
ear
.
 Remov
aloft
hecol
l
oidal
andsuspendedmat
ter
s.
 Remov
alofodorandcol
or.
 React
ioni
nAl
kal
i
nit
ytodecr
easei
t.
 Remov
alofundesi
rabl
eof
f-
tast
emat
ter
s.
 Fr
eef
rom mi
crobi
alhazar
dbychl
ori
nat
iont
ocont
rol
atcont
ami
nat
ion.

Wat
erpr
ocessi
ngst
eps
1.Chl
ori
nat
ion
2.SandFi
l
ter
3.Car
bonPur
if
ier
4.Wat
erPol
i
shi
ngf
il
ter
s
5.ROMembr
anes
6.Addi
ti
onofMi
ner
als
7.Ozonat
ion
8.UVLamp
9.Absol
uteFi
l
trat
ion
10.Pr
ocedur
eforSt
orageTankSani
ti
zat
ion.
11.Wat
erPl
ant&Pl
antFumi
gat
ionPr
ocedur
e.
12.Pr
ocedur
eForbot
tl
esWashi
ng&Fi
l
li
ng
13. Bot
tl
eWat
erQual
i
tyTest
ing.

1.Chl
ori
nat
ionofRawWat
er.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Chl ori
nati
oni sobt
ainedbymi xi
ngchlori
newit
hrawwat er.I
tisappliedi
nwateratthe
rateof04- 12ppm tof orm Calci
um Hypochl
ori
te(CaOCL)inwat erwhichact
sasa
bleachingagentanddi si
nfect
antinit
.Ital
sokil
l
sy east
,Bacter
ia,Algaeandother
mi crobial
lif
e.Ital
soremov eodor,
Taste,ManganeseandHy drogenSulphi
de.Itshoul
d
ber etai
nedi nwaterfortwohour s.
Thedozingandt hepresenceofchl
ori
neinwat
ermaycheckedbyusi
ngOTSol
uti
onas
aChlori
neIndicator
.Ift
hewatercol
orbecomeyel
l
owishbyusingOTSol
uti
oni
tmeans
Chl
orinepresentinwater.

TheDet
ermi
nat
ionofChl
ori
neConcent
rat
ioni
nWat
er

Pur
pose:
 I
nthi
sexper
imenty
ouwi
l
lmeasur
etheamountofdi
ssol
vedchl
ori
nepr
esent
i
n watersamples.Bri
ng a watersample,i
ncludi
ng poolwater,f
rom homei fyou
wish 
(makesur
ei ti
sinanairt
ightcont
ainer
,li
keTupperware,andiskeptascoldas
possi
ble)
.

Backgr
ound:
 Chl
ori
ne i
s of
ten added t
o dr
inki
ng wat
ersuppl
i
es t
o ki
l
lhar
mful
microor ganisms.Chl or ineisnotonl yanef fecti
vedi sinfectant,buti talsor eact swith
ammoni a, i
ronandot hermet al
sandsomeor ganiccompoundst oi mpr oveov er al
lwater
quality.Therei sal i
mi tt otheuseofchl or i
ne,howev er,asnegat i
v er esultsar epossi ble
witht headdi ti
onoft oomuchchl ori
ne.Badt astesorodor sinwat erar eof tenenhanced.
Excessofchl orinecanbehar mfultofishandot heraquat i
cani mal swhent hewat er
contai nsnitrogencompounds.Fi nall
y,t
hef or mat i
onofchl oroform andot hersuspect ed
carcinogensi spossibl e.Itisv er
yimpor tantf orwat ersuppl ierst omoni torcl oselyt
he
l
evelsofchl orinepresenti nthewat erforwhi chtheyar er esponsi ble.Pr actical ly
,water
systemsl ookf oralev elofchl ori
neremai ningi nthewat eraftertreat mentt obe1mg/ L
orlessf ormi nimizi
ngadv erseeffectswhi l
emai ntainingdi si
nfect antpr oper ties.Abov e
1mg/ L,odorandt asteof t
enbecomepr oblemat i
c.

Themet hodofdetermi ni
ngchl or
inei
nthisexperi
mentrel
iesonacolorindi
cator,DPD
(N,N-di
ethyl
-p-
phenylene-diamine)
.Inthepresenceofchlori
ne,DPD react
sr apidl
yto
form ar edcolor,t
hei ntensit
yofwhichisani ndi
cat
orofchlori
neconcentrati
on.The
highertheabsorbance,thehigherthechl
ori
neconcentr
ati
on.

DPD+Chl
ori
ne 
à Redcol
ormeasur
edat515nm

Bycreati
ngaseriesofstandar
dsofknownchlori
neconcentrat
ion,i
tispossi
bleto
const
ructa cal
i
brati
on cur
vet o measur
ethe chl
ori
ne concentr
ati
on i
n unknown
samples.

Pr
ocedur
e: 
The pr
ocedur
e f
or t
his exper
iment i
nvol
ves t
he use of t
he
spect
rophot
omet
erasout
li
nedi
near
li
erexper
iment
s.Thechl
ori
nepr
esenti
nthewat
er
 Bot
tl
edDr
inki
ngWat
erPr
ocess
sampleswi l
lreactwi t
ht heDPDi ndicatorformingar edcoloredcompoundwhichi s
all
owed to dev elop for a set per iod of t i
me and t hen measured withthe
spectr
ophotomet erat515nm.I twil
lbenecessar ytofir
stmixsever
alknownstandar
ds
andmeasur et heirabsorbance.Thesewi l
lbeusedt ogenerateacalibr
ati
oncurve.
Unknownwat ersampl escant henbepr eparedandtheirabsor
banceusedtodetermi
ne
thei
rchlor
ineconcentrati
on.Adetail
edpr ocedurefol
lows.

Pr
epar
ati
onoft
heSt
andar
dCur
ve:

1.Takefourfl
asksorbeaker scapableofholdi
ngatleast100mlandaddtothem
thefol
lowi
ngquant i
ti
esofst andardsol
utionanddeioni
zerwat
er.Usepi
pet
sto
measurethev olumesofst andar
dsol ut
ionineachcase.Thewat ercanbe
measuredwithyourgraduatedcyli
nder
.

Vol
ume St
andar
d Vol
umeDIWat
er(
ml) Chl
ori
neConc.(
mg/
L)
(ml
)

100 0 2.
0

50 50 1.
0

25 75 0.
5

5 95 0.
1

2.To each  ofthesesol uti

onsadd  t
hree 
(3)DPDt abletsandst irt
odi ssolv
e.You
shouldnot i
cet her edcol ordevel
opingasthetabletsdissolv
e.Ifyoudonotsee
thecolornot ifyy ourinstr
uctor.
3.All
ow t he colort o develop f
or10 mi nutes and read the standards i
nt he
spectrophot omet ersetat515nm.
4.Whilet hecol orisdev el
opingyoucanpr eparet
heunknownsampl esorcalibr
ate
thespect rophot omet er.

 
Prepar
ati
onoft
heUnknownSampl
es:

1.Theunknownsampl esmustbepr epar

edexactlyasthestandar
dsweref ort
he
testtobeaccur at
e.100mlofeachunknownneedst obemeasur edintothe
graduatedcyli
nderandtransferr
edt oabeakerorfl
ask.
2.Beforedoingthenextst ep,testtheodorofeachunknownasdemonst r
atedby
yourinstr
uctor.Doy oudetectany" chl
ori
nesmel l
"?Repor
tyourobser
vati
onsin
the 
Results 
secti
on.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
3.Into each unknownsol utionplace t
hr ee DPDtablet
sandst irtodissol ve.
4.Allow t hecol ort odev el
opf or10mi nutesandr eadt heabsor banceont he
spect rophotomet er.
5.Ify oubr oughtawat ersampl ef r
om home,i tispossibl
et hatthewat erisv er
y
highi nchlorine,especi all
ypoolwat er,andwillneedtobedi lut
edf orthetestto
wor k.Ifthecol orofy ourunknowni sdarkerthanthedar kestst andar d,consul
t
youri nstr
uctorf orfur
theri nst
ructi
ons.
6.Allsol uti
onsf rom thisexper i
mentcangodownt hedrainandanyopenedDPD
tablets can be t hrown i nthe trash can.Any  
unopened  DPD t ablets can be
returnedt otheexper i
mentboxf orlateruse.

 
Anal
yzi
ngt
heSol
uti
ons:

1.Tur nt hespectrophotometeronandal l
owi tt
owar m upforatleast15minut
es.
2.Sett hewav elengthto515nm.
3.Wi t
ht hesampl echamberempt y
,adjustthemetertoread0%t r
ansmitt
ancewith
thel eft-
handknob.
4.Inser tacuv ett
ewi t
hdeionizedwater.Closethecoverandsett hemeterto100%
transmi tt
ancewi ththeri
ght-handknob.
5.Measur etheabsor banceofeachoft hefourstandar
dsol ut
ionsandtheunknown
wat ersampl es.Recordtheabsor bancesalongwi t
ht hechlori
neconcentrat
ion
l
istedi nthepr ev
ioustabl
e.Thesev alueswillbeusedt oconstructt
hestandard
curv e.

 
Resul
ts:
 TheResul
tssect
ionf
ort
hisexper
imentshoul
dincl
udeacal
i
brat
ioncur
veof
absorbancev s.mg/lchlor
inebasedontheval
uesobtainedforthefourstandar
ds.Pl
ot
thepointsanddr aw thebeststrai
ghtl
i
nepossibl
ethrought hem.Remembert ousea
ful
lgr aph and use cor rect scal
e.Using the cal
ibrat
ion cur
v e,determi
ne the
concentrat
ionofchlori
neintheunknownsamplesandreportthesevalues.

1.Thesesampl eswer et akenf r

om awat ertreatmentf acil
it
ythathasahi st
oryof
varyi
ngchl orinel evels.Basedont hesesampl es,doesi tappeart heyhav et he
problem undercont rol?Whyorwhynot ?
2.Ifyoubr oughtasampl efrom home,orshar edsomeoneel se’ssample,howdi d
thechlorinel ev elcompar etotheunknownsampl es?
3.Whichoft hesampl eswoul dyoupr efertodr i
nki ftherewerenoknownpr oblems
withmi croor gani sms?Whatabouti ftherewasapasthi stor
yofmi croorganism
contami nationi nt hewat er?
4.Inthesampl e( s)wher et herewasar elati
velyhighchl ori
nelevel
,werey ouablet o
detectanyodor ?Woul dodorbeagoodi ndicatorofl owchlori
nelevels?Whyor
whynot ?
 Bot
tl
edDr
inki
ngWat
erPr
ocess
2.SandFi
lt
er.
SandFilt
rat
ionmaydefi
nedastheprocessofremov i
ngsmallpar
ti
clesoft
he
suspendedmat t
ersf
rom wat
erbypassingi
tthroughaporousmedium.Sandfi
l
terwor
k
fort
hispurpose.
I
tr emovesanyfl
ockpart
icl
esandothersuspendedmatterwhi
chmayhav
ebeencarr
ied
outfrom rawwat
erstor
agesource.Thesandfil
teri
schargedandr
efi
l
lwi
thsandand
gravels.
Thefinestsandparti
cles(Si
l
ica)makeupthetoplay
erwhi
l
ethelar
gestgr
avel
satt
he
bott
om oft hef
il
ter.Rawwat erent
ersf
rom upwar
dandmovesdownwardthr
ought
he
fi
l
terandi mpuri
ti
est r
appedhere.

BackwashPr
ocedur
e.
Backwashi
saprocedureofwashingandcl
eani
ngoutthesandfil
terimpuri
ti
esthat
havebeenr
emovedfrom waterduri
ngnor
malfi
lt
eri
ngoperat
ion.Backwashprocedure
i
saccompli
shedbyreversi
ngthefl
owofwatert
hroughthesandfil
terasfol
lows;
 Waterfl
owrateduringbackwashingshoul dbe4-5timesgreaterasf
lowast he
fl
owduringnormal f
ilt
rat
ion.Thepurposeofthi
sf astfl
owr at
eistoexpandthe
sandusedtoaboutt wiceitscompactv ol
umeandal lowthesandgrainstorol
l
overandscr
ubt ogether,
hereby,againfeedi
ngtheimpur i
ti
esofrawwat er
.
 Cont
inuet
hebackwashpr
ocessunt
ilt
hebackwashi
ngdr
ainwat
erwoul
dn’
tbe
cl
earandt
ranspar
ent
.
 Retur
nthefl
owofwatert
oitsnor
mal di
recti
on,nowrunt
hewat
ert
owast
eunt
il
thewaterwoul
dn’
tbeper
fect
lycl
earandtranspar
ent
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess

I
nst
ruct
ionf
orr
echar
ge/
Ref
il
ltheSandf
il
ter
.
 Sandfi
lt
ershouldberechar
gemaxaf
ter02-
03y
ear
sofwor
kingaccor
dingt
o
thef
oll
owinginstr
uct
ions;
 Cl
oseall
thev
alv
esandopent
hedr
ainv
alv
es,
remov
eal
ltheol
dsandand
gr
avel
.
 Cl
eant
hesandf
il
teri
nter
iort
hor
oughl
y.
 I
nspectthel
i
ningofi
nter
nal
sideoft
hesandf
il
terandr
epai
ranybr
eakagei
n
t
heli
ning.
 Makesur
ethatt
heunderdr
ainpi
pingi
sper
fect
lyi
nlev
el.
 Nowt hesandf
il
terfi
l
lagai
nwiththenewsand,gr
avel
andwat
erbyl
evel
i
ng
car
efull
yandattheendfi
ll
wit
hf i
nestsand(
Sil
i
ca).
 Backwasht
henewlychar
gedsandf
il
terbyf
lowi
ngwat
ersl
owl
yfr
om t
he
bott
om oft
hef
il
ter
.
 St
eri
l
izet
hesandf
il
terwi
thchl
ori
ne.
 Fil
lthesandf
il
tert
othet
opwi
tht
hewat
erandst
irt
omi
xchl
ori
net
hroughout
thefil
ter
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Makeupt
hechl
ori
nesol
uti
onof70%i
n05gal
l
onofwat
er.
 Dr
ainedt
hesandf
il
terunt
ilt
hechl
ori
newoul
dbecompl
etel
yni
l
.
 Al
l
owt
hef
il
tert
ost
andf
oratl
east02hour
sandt
hendr
aincompl
etel
y
 Fil
lthefi
lt
eragainwit
hwat erandbackwasht
hesandfi
lt
erunt
ilt
hest
rong
ster
ili
zi
ngsolut
ionisclear
edandwashedout.Re-
washedtodrai
nandplace
thesandfil
teri
nservi
ce.

Pr
ecaut
ionsf
ort
heSandFi
lt
er.
 Makebackwashpr
ocedur
eaf
terev
ery12hourofwor
king.
 Mai
ntai
nthebackwashpr
ocedur
eti
l
l30mi
nut
es.
 Maint
ainthepr
essur
eofbackwash04t
imesgr
eat
ert
hant
henor
mal
pr
essure.

Fl
owDi
agr
am. SandFi
lt
er.
 Bot
tl
edDr
inki
ngWat
erPr
ocess

3.Car
bonFi
lt
er/Pur
if
ier
.
TheAct i
vatedCar bonpuri
fi
erismorethanafilt
er.Itisdesi
gntor emovechlori
neand
othertasteandodorf r
om rawandt r
eatedwater.Thewat ershouldbecompl etel
y
clear
edandf reefrom suspendedmattersbef
oreent eri
nginthi
suni t
.Theactiveli
feof
thecarbonent i
rel
ydependsupont heamountoft hemat eri
alsremovedfrom thewater
andther ateofcontaminati
ons.However,t
heactivatedcarbonalwaysbechangedat
l
eastonceeachi nyearwithtr
eatmentandtwiceineachy earwi t
houttr
eatment .
Ifproper
lycareforcarbonpuri
fi
er,car
bonusuall
ylastabout01yearsofworki
ng.I
tis
verybett
ertochanget hecarbonbeforet
hechlori
neorforei
gntasteact
ual
lystar
ts
comi ngoutthr
ought hecarbonfi
l
ter.
Aftersandf
il
terr
awwat erent
ersi
ncar
bonpurif
ierwher
eact
ivat
edcar
bonr
emov
est
he
taste,
odorandchlor
ine,
Hence,weobt
ainedcl
earwater
.

I
nst
ruct
ionsf
orBackwashi
ngpr
ocedur
e.
Thebackwashi
ngpr
ocedur
eisi
dent
ical
l
ythesamef
ort
hecar
bonpur
if
ierasf
orsand
 Bot
tl
edDr
inki
ngWat
erPr
ocess
f
il
terbyusi
ngst
ri
ctpr
ecaut
ionswi
thr
espectt
her
ateofbackwashf
low.

I
nst
ruct
ionf
orr
echar
ge/
Ref
il
ltheCar
bonf
il
ter
.
 Cl
oseall
thev
alv
esandopent
hedr
ainv
alv
es,
remov
eal
ltheol
dcar
bonand
gr
avel
.
 Cl
eant
hecar
bonf
il
teri
nter
iort
hor
oughl
y.
 I
nspectthel
i
ningofi
nter
nal
sideoft
heCar
bonf
il
terandr
epai
ranybr
eakagei
n
t
heli
ning.
 Addnewgr
avel
suppor
tbed,
backwashgr
avel
andst
eri
l
izedt
hegr
avel
s.
 Backwashagai
nbef
oreaddi
ngcar
bon.
 Car
bonbeddept
hshoul
dbeappr
oxi
mat
ely24i
nches.
 Byusi
ngtr
eatedwat
er,
wetthecarbont
hor
oughl
yandl
ett
ingt
hewat
erf
lowi
nto
thet
opofthecar
bonfi
l
terunt
ilf
il
l.
 All
owthecar
bont
osoakunderpr
essur
eov
erni
ghtov
erni
ghtorasl
ongas
possi
ble.
 Backwashandr
e-washt
hecar
bonpur
if
ierunt
ilt
hecl
ear
.
 TheAlkal
i
nit
yofwatermaybechangedf
orf
ewday
sunt
ilt
hecar
bonhasbeen
thor
oughl
ywashed.

Met
hodf
orPr
epar
ati
onofAct
ivat
edCar
bon.
 Readytouseacti
vatedcarboni
sopt
ional
whi
l
etheact
ivat
edcar
boncanbe
prepar
edbyfol
lowingprocedur
e,
 Fir
stheattocarbonathight
emperaturethenwashwit
haci
d,mostl
yHCLisused
forwashingpurpose.Af
tert
hiswaterisusedtowashdownt
heaccessamount
acid.Thi
sprocessisdonetothecarbonblackwhi
chisi
npart
icl
esi
zeorgr
anular
form.
 Carbonplatesareact
ivatedbyelect
ri
city.Theseplat
esareneutr
ali
zedaft
er
backwashing.El
ectr
icplatesarechargedpositi
veionsandnegati
veionsby
el
ectri
cit
y.Posit
ivei
mpur it
iesgoestonegativeionspl
ateswhit
ethenegati
ve
i
mpur i
ti
esgoest opositi
veionsplat
es.

Pr
ecaut
ionsf
ort
heCar
bonPur
if
ier
/Fi
lt
er.
 Checkthecar
bonpuri
fi
eraf
terev
ery03mont
hofwor
kingandmai
ntai
nthe
quant
it
yofcarbon.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Mai
ntai
nthebackwashi
ngpr
ocedur
eaf
terev
ery12hourofwor
king.
 Mai
ntai
nbackwashi
ngpr
ocedur
efor30mi
nut
esathi
ghpr
essur
e.
 Checkt
hebackwashwat
erf
lowpr
essur
emustbe04t
imesgr
eat
ert
hant
he
nor
malpressur
e.
 Dr
aint
hewat
ert
hatst
andi
nthef
il
terov
erni
ghtbef
orest
art
ingt
hepr
oduct
ion.

Fl
owDi
agr
am,
Car
bonPur
if
ier
.

4.Wat
erPol
ishi
ngFi
lt
ers.
Thefil
ter
swhi chareconnect edi nt het r
eatedwat erlineafterthecar bonpuri
fi
erare
knownaswat erpolishingfil
ters.Itisimpor tanttohav ea05Mi cronand01Mi cron
Cart
ri
dgef i
lt
ersAssembl yplaceddi rectlyaftertheCar bonpurifier,thiswil
lpr
eventthe
multiMediafi
ltr
ati
onf rom damagi ngdownst ream pumpsandf oul i
ngt heROSy st
em.
Thesefil
ter
sconv ertthetreatedwat erintopur emol eculesofwat er.Thesefi
lt
ers
removethesmal lparti
clesofr ustandt urbi
dmat erialsthatmayhav ebeenpickedup
duri
ngprocessingandal sor emov emi cropar t
icl
esf rom tr
eatedwat er
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Aft
ert
hesepol
i
shi
ngfi
lt
erspur
etr
eat
edwat
erent
ersi
ntot
heROPl
antwher
ewecont
rol
TDSoftr
eat
edwat
er.

5.Rev
erseOsmosi
sPl
ant
.
Rever
seosmosesiscommonl
yrefer
redtoasROisapr
ocesswherey
oudeminer
ali
ze
ordei
onizewat
erbypushi
ngi
tunderpr
essur
ethr
oughasemiper
meabler
ever
se
osmosismembrane.

Osmosi
s.
Osmosisisanaturall
yoccur
ri
ngphenomenonandoneoft hemostimportantprocesses
i
nnaturethi
sisapr ocesswhereaweakersali
nesol
uti
onwi l
ltendtomigratetoa
str
ongsali
nesol
ut i
on.Examplesofosmosisarewhenplantrootsabsor
bwat erfr
om
thesoi
landourkidneysabsorbwaterf
rom ourbl
ood.
ReverseOsmosi sisthepr ocessofOsmosi sinrever
se.Wher easOsmosi soccurs
naturallywit
houtener gyrequir
ed,toreversetheprocessofosmosi sy ouneedtoapply
energyt othemor esal i
nesoluti
on.Ar everseosmosi smembr aneisasemi permeable
membr anethatall
owst hepassageofwat ermoleculesbutnotthemaj or
it
yofdissol
ved
salt
s, organi
cs,bacteriaandpy r
ogens.Howev er
,youneedt o‘push’thewat ert
hrough
therev erseosmosismembr anebyappl yingpressurethati
sgr eat
ert hanthenatural
l
y
occurringosmot i
cpr essureinordertodesal i
nate(demineral
i
zeordei oni
ze)waterin
theprocess, al
l
owi ngpur ewaterthroughwhi l
eholdingbackamaj ori
tyofcontaminants.
HowdoesRev
erseOsmosi
swor
k?
Rev erseosmosi swor ksbyusi ngahi ghpressur epumpt oincreaset hepressur eont he
saltsideoft heROandf or cet hewat eracrosst hesemi permeabl eROmembr ane,
l
eav ingal mostal l(
ar ound95%t o99%)ofdi ssolvedsaltsbehindi nther ejectst ream.
Theamountofpr essur er equi reddependsont hesaltconcentrat i
onoft hef eedwat er.
Themor econcent ratedt hef eedwat er,themor epressureisrequiredt oov er comet he
osmot i
cpr essure.Inv erysi mpl eterms,feedwat erispumpedi ntoaRev er seOsmosi s
(RO)sy stem andy ouendupwi t
ht wotypesof
wat ercomi ngoutoft heROsy stem:goodwat erandbadwat er.Thegoodwat ert hat
comesoutofanROsy stem hast hemajorit
yofcont aminantsr emov edandi scal led
permeat e.Anothert ermf orper meatewat erispr oductwater–t heymeant hesame
thi
ng.Per meat eisthewat ert hatwaspushedt hroughtheROmembr aneandcont ai
ns
veryl i
ttl
econt aminant s.
The‘ bad’ wateristhewat ert hatcont ai
nsall ofthecont aminant sthatwer eunabl eto
passt hr oughtheROmembr aneandi sknownast heconcent r
at e,r
eject,orbr i
ne.Al l
threet erms( concent rate, r ej
ect ,andbrine)ar eusedi nt
erchangeabl yandmeant he
samet hing.Belowi sasi mpl eschemat i
cthatshowshowanROsy stem wor ks.Ast he
feedwat erenterstheROmembr aneunderpr essur e(enoughpr essur etoov ercome
 Bot
tl
edDr
inki
ngWat
erPr
ocess
osmot icpressure)the
wat ermoleculespasst hroughthesemi permeabl
emembr aneandt hesaltsandother
cont aminantsarenotal l
owedt opassandar edi
schargedthroughtheconcent r
ate
stream, whichgoest odrainorcanbef edbackintothefeedwat ersupplyinsome
ci
rcumst ancestober ecycl
edthr ought
heROsy stem t
osav ewat er
.Thewat erthat
makesi tthrought heROmembr aneiscall
edper
meat eorproductwat erandusuall
yhas
around95%t o99%oft hedissolvedsalt
sremovedfrom it
.
Rev
erseOsmosi
sal
soknownasUl
tr
aFil
tr
ati
onbythei
ndust
ryrepr
esentt
hestat
eofart
i
nwatert
reat
mentt
echnol
ogy
.ROPl
anti
softenusedi
nconj
uncti
onwithcar
bonpur
if
ier
.
ROPlantt hatusesasemi permeabl emembr anesf i
lt
ertoseparatet heorgani c
compoundsandcont aminant s.ROPl antishighlyef
fect
iveinremov ingsev eral
i
mpur iti
esf rom wat
ersuchasTDS, Turbi
dit
y,asbestos,
lead,andot herheav ytoxic
metals,radium andmanydi ssolv edorgani
ccompoundsi nwater.Thi sprocessal so
remov eschl or
ineandcanal sor emovenuclearreducti
onsuchasr adioacti
vepl utoni
um
orstr
ont i
um i nthedrinki
ngwat er.Theref
ore;therev
erseosmosi splantseemst obe
mostadv ancedwat erpuri
ficati
onmet hoddev el
opedsof ar
.

Fl
owDi
agr
am,
SemiPer
meabl
eMembr
ane.

I
nst
ruct
ionsf
orROPl
ant
.
 Checkthatt
het r
eatedwat
ershoul
dbecl
eanandhav
enocol
orbef
orepassi
ng
thr
oughROPl ant.
 Chl
ori
neshoul
dbecompl
etel
yni
lbef
orepassi
ngt
hroughROPl
ant
.
 Af
terev
eryonehourcheckt
heROWat
ersampl
esf
rom sampl
i
ngpoi
nts.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Bef
oretur
ningONROPl
antcheckt
hatt
her
awwat
ert
anksshoul
dbef
ull
of
wat
er.
 Makesurethatt
hev olumeofAnt
i-
scal
enti
nROPlantisaccurat
ebecause
Membraneseffi
ciencyremai
nsgoodduetothi
sant
i-
scalent
.
 Checkthatthewaterpol
i
shingf
il
ter
sar
ecl
eanandwhi
te,
ift
heyhav
eanycol
or
orexpi
redthenrepl
acedthem.
 Pr
essuredi
ff
erencebetweeni
nletandoutl
etoft
hepoli
shingf
il
tersmustnotbe
morethan05psiandifi
tisgr
eaterthan05psit
henchangeCartr
idgeFi
l
ters.
 Aft
ert
urni
ngONt heROPlant,checkpr
oductf
lowr
ate,
Rej
ect
ionf
lowr
ate,
TDS
andpHValueaf
terever
yhalfanhour.
 Toremovetheminorpossibil
i
tyofchl
ori
neinROMembr anes,useSodium
MetaBi
sul
phate(
NaMS) .Itisafoodgr
adechemicalandvanish/absor
bthe
chl
ori
necomplet
elywhichcanpassesthroughCar
bonpurif
iertoROMembr anes.

Sodi
um Bi
sul
fi
te.
Byaddi
ngsodium bi
sul
fi
te(SBSorSMBS),whichisar
educer,
tothewat
erst
ream bef
ore
anRO,
wit
hproperdoseyoucanremoveresi
dual
chlori
neandchl
oramines.

6.Addi
ti
onofMi
ner
als.
Af
terRever
seOsmosisprocess,wegetpermeat
e/puret
reat
edROwater.I
nthispur
e
waterweaddminer
alswhichareessent
ial
forhumanbodysuchasCal
cium Chl
ori
de,
Magnesi
um Sul
phat
e,Sodium chl
ori
deandSodium Bi
car
bonate.
Af
teraddi
ti
onofmi
ner
als,
final
miner
alwat
erent
ersandst
orei
nast
oraget
ank.

7.Ozonat
ion
Aft
eraddit
ionofminer
als,
final
mi ner
alwaterent
ersandstor
einastoragetankwher
e
anOzonat
erisworki
ngasgerm kil
lerwit
hproperandstandar
dlimi
tsofozonati
on.
I
tsmainpri
nci
pleisbasedont hesucti
onofoxygenthroughinl
etandconv
erti
tint
o
ozonegasasadisinf
ectantathi
ghv ol
tageof45000volt-
55000volt
.

Wor
kingPr
inci
ple.
I
tswor
kingpr
ocedur
eincl
udest
hreest
eps:

 Ozoneisagasproducedfr
om ai
rathi
ghvol
tagewheni
nject
edintowat
er,
ozone
i
sox i
dizessol
ubl
emetalsi
ntofi
l
ter
abl
epart
icl
esandki
l
lsbacter
ia.

 Wat
erpumpedi
ntot
heaer
atorf
orces,
ozoneandot
hergasesoutt
hroughasel
f-
 Bot
tl
edDr
inki
ngWat
erPr
ocess
cl
eani
ngv
alv
e.

 Oxi
dizepar
ti
clesar
efi
l
ter
edt
hroughanat
ural
medi
athatr
equi
resonl
ywat
erf
or
backwashandnoot
hert
reat
ment
s,i
tisf
ull
yaut
omat
ic.

Pr
ecaut
ionsf
orOzonat
er.
 El
ect
ri
csuppl
ypr
oper
lymai
ntai
ned.
 Ef
fi
ciencyofOzonat
orshoul
dbecheckedoncei
nashi
ftt
hroughozonet
estki
t.
 OzoneVal
ueshoul
dbe02-
03ppm.

Fl
owdi
agr
am,
Ozonat
ionpr
ocedur
e.

8.ULTRAVOI
LETLAMP.
Af
terpr
opermi
xi
ngwi
thozonegasf
inal
mi neral
watergoest
ofil
l
erandpassest
hrough
aUVLampwhichi
sal
soworki
ngasgerm kil
lert
hroughUVRays.

I
tspr
inci
plei
sbasedonpenet
rat
ion.
UVi
sthepar
tofel
ect
romagnet
icspect
rum
 Bot
tl
edDr
inki
ngWat
erPr
ocess
betweenv i
sibl
eli
ghtandX- r
ays.Ithasastrongger
mici
dalef
fect.UVkil
l
s
microorgani
smsbypenet rat
ingtheircel
lmembranesanddamagi ngt
heDNAof
Bacteri
a,makingthem unabletoreproduceandeff
ect
ivel
yki
ll
ingthem.

Wor
kingPr
inci
ple.
I
tswor
kingpr
ocedur
eincl
udesf
oll
owi
ngst
eps:

 Thewaterenterst
hroughinl
etvalv
et heUVlampnev ercontactthewater,
iti
s
ei
therhousedinquart
zglasssleevei
nsidet
hewat erchamberormount ed
ext
ernalt
ot hewaterwhi
chflowsthroughUVtransparentTeflontubes.

 Someul t
ra-
purewat
ersy
stem uses185nm UVuni
tforr
educi
ngTOC(
tot
al
organi
ccarbon).

Pr
ecaut
ionsf
orUVLamp.
 UVRodsshoul
dbeofst
andar
dqual
i
ty.
 UVl
ampbodyshoul
dbeofst
ainl
essst
eel
mat
eri
al.
 Ther
eshoul
dbepr
operel
ect
ri
cindi
cat
or.

Fl
owDi
agr
am.UVLampOper
ati
on.
 Bot
tl
edDr
inki
ngWat
erPr
ocess

9.Absol
uteFi
lt
rat
ion/
Micr
ofi
lt
rat
ion.
Mi
crof
iltr
ationiseffecti
v einremov i
ngcol l
oidalandbact eriamat terandhasa
poresizeofonly0. 1t o0.5µm.Membr aneconfigur
at i
oncanv arybet ween
manuf actur
ers,butthe" holl
owf i
ber "t
ypei sthemostcommonl yused.Ty pical
ly,
the
waterispumpedf rom theout si
deoft hef i
bers,andthecl eanwat eriscol l
ectedfrom
theinsideofthef i
bers.Mi cr
ofil
trati
onmembr anesusedi npot ablewat erappl i
cati
ons
usuall
yoper at
ein“ dead-end"f l
ow.I ndead-endflow,allofthewat erfedt ot he
membr anetofi
lterthrought hemembr ane.Afterthi
sf i
nal f
il
tr
at i
on, f
inal product/
Mineral wat
ergoest of i
lli
ngunitf orMineralwaterProduct i
on.

10.Pr
ocedur
eforSt
orageTankSani
ti
zat
ion.
St
oraget
anksani
ti
zat
ioni
sdonet
hroughf
oll
owi
ngst
eps,

 Makef
reshsol
uti
onofchl
ori
neandmakessur
ethatconcent
rat
ionofchl
ori
nei
s
 Bot
tl
edDr
inki
ngWat
erPr
ocess
100ppm.
 Addt hesoluti
oninalltanks; f
ill
withwat erupt othelevel
 Thesol ut
ionremainsinwat erstor
aget ank/tanksfor55mi nute
 Drainouttanksafter55mi nutes
 Addf r
eshwat er& agai ndr aintoassur et herearenot r
acesofsanit
izat
ion
soluti
on
 Testthetankwat erforpresenceofsani t
izat
ionsolution.
 Rinsetanksagainwi t
hcl eanwat erifsanit
izationsol
ut i
onissti
l
lpr
esent
 Recordact i
vi
tyinlogbook

11.Wat
erPl
ant&Pl
antFumi
gat
ionPr
ocedur
e.

Toensurethest
eri
l
izat
ionoft
heworkingenv
ironment
.
Theprocedur
eappli
edtothef
umigat
ionperf
ormedtoall
wat
erpl
antpr
emi
ses.

 Thisacti
vi
tyst ar t
swhi lePlantclosingt wiceamont h.
 Usesgloves&maski frequired
 Puts1gm Pot assium permagnat ei nsomedi sposablegl
ass/beaker
/contai
ner
 Adds5ml f
or mal ininit.
 Keepsthismi xtur eincorner.
 Thesmel l
/fumesemi tt
edf rom mixturewhi chactasaf umigat
ionagent.
 Washeshandsaf tercompl etionofthisact ivi
ty
 Discar
dthismi xtureeverymor ni
ngwi thcar eindrain.
 Washeshandsaf terdiscardingthismi xture.
 Entert
hedet ail offumigationi ndail
yLogBook.

12.Pr
ocedur
eforbot
tl
esWashi
ng&Fi
ll
ing.

Washi
ngandf
il
li
ngpr
ocedur
eisdonet
hroughf
oll
owi
ngst
eps,

 Segregatet he dir
ty,smelly
,damage & bot tl
e contai
ning st
icky thi
ngs of
aesthet
ical
lybadbot tl
e&neverfi
l
lsuchbot t
les/contai
ners.
 Washbot tl
esexternall
ywit
hcleanwater
 Rinsebottl
eat1stst agewithH2O2 addedwat erfor15secondsoraci d-
base
washing(hotpolybri
teat40CandOxoni a).
 Rinse/steri
li
zebottl
eatsecondstagewi t
h45Chotwat erfor15seconds
 Rinse/steri
li
zebottl
eatthir
dstagewithproductwaterfor10seconds
 Bypassessecondst agei
ncaseofnon-avail
abil
i
tyofthefacil
it
y.

 Fil
lthe05gal
lonsbott
lesasperr
equi
rement
.
 Inspectthefi
ll
ed bottl
eswithfl
uor
escentscr
een & ensur
ether
eisnot
hing
suspendedorsett
ledi
nside.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 I
fsomesedi mentpr esentinside,drai
nstheenti
rebottle.
 Cleansthebot t
leandf i
ll
si tagain.Ifst
il
lther
ear esomesedi mentsorparti
cles
i
nsidethebot t
lethennev err efi
l
lsitagai
nunlessitisthoroughl
ycl
eaned.
 I
fsomepar t
icl
escomi ngwi thfil
li
ngwaterthenreplacetheendAbsolutefi
l
ters&
takenecessaryaction.

Fl
owDi
agr
am,
Bot
tl
edr
inki
ngwat
erPl
ant
.

13.Bot
tl
eWat
erQual
it
yTest
ing.
 Ensur
esaf
ebot
tl
edr
inki
ngwat
er.
 I
dent
if
ypr
obl
emsandAdoptpr
ecaut
ionar
ymeasur
es.
 Rai
seawar
enessandSel
ectanappr
opr
iat
ewat
ersour
ce.
 Det
ermi
net
heef
fect
ivenessofwat
ert
reat
mentt
echnol
ogi
es.
 I
nfl
uencepol
i
ciest
osuppl
ysaf
ebot
tl
edr
inki
ngwat
er.
 Mai
ntai
nthephy
sical
andchemi
cal
test
s.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Cont
rol
measur
ement
sforMi
crobi
ologi
cal
hazar
ds.
 Prov
idethestr
icthy
gieni
ccondi
ti
onwhi
chi
sfr
eef
rom t
hechanceofhazar
dof
cont
aminati
ons.

Phy
sicalandChemi
calt
est
sofBot
tl
edr
inki
ngWat
er.
Tomaintai
nthequal
i
tyofthebott
ledri
nki
ngwatermaint
ainthephy
sicalandchemi
cal
test
saccor
dingt
othest
andardsandspeci
fi
cat
ioncondi
ti
ons.
Theset
est
sar
egi
venbel
ow,
1.Sensor
yEv
aluat
ionTest
.(TOA)
.
2.PHTest
.
3.TDS(
Tot
aldi
ssol
vedsol
i
ds)
.
4.Tot
alHar
dness.
5.Cal
cium Test
.
6.Magnesi
um Test
7.Chl
ori
deTest
.
8.M.
Alkal
i
nit
y.
9.Sul
phat
esandNi
tr
it
est
est
.
10.
Sodi
um andPot
assi
um Test
.
11.
Arseni
cTest
.
12.
OzoneTest
.
13.
FreeChl
ori
neTest
.

Mi
crobi
alTest
sofBot
tl
edr
inki
ngwat
er.
Basi
cmi
crobi
alt
est
sforbot
tl
edr
inki
ngwat
erar
egi
venbel
ow,
1.Tot
alpl
atecountBact
eri
atest
.
2.Tot
alCol
i
for
mtest
.
3.Escher
ichi
acol
itest
.
4.Sal
monel
l
atest
.
5.Fecal
Str
ept
ococcust
est
 Bot
tl
edDr
inki
ngWat
erPr
ocess
6.pseudomonast
est

1.Det
ermi
nat
ionofPH.
Thephofadr inki
ngwaterdefinethel
ogarit
hmicacidi
tyofhy
drogeni
onsconcent
rat
ion
whichi
sani ndicat
orofAcidicorBasiccharacterofwat
erandusedformonit
ori
ngfor
thebot
tl
ingprocessandwat ertr
eatmentcarbondioxi
demeasur
ements.
Thebasi cpri
ncipleofphmeasur ementisthedetermi nationofacidit
yofhydr
ogeni ons
bypot enti
all
yusi ngast andardhy drogenel ect
rodeandar ef
erenceelect
rode.The
potenti
aldif
ferencebet weent hesetwoel ect
rodesbot hi mmersedi nasamesolutionis
l
inearfuncti
onoft hi
ssolutionphv alue.Thisli
nearrelationshi
pisdescri
besbypl otti
ng
themeasur ed pot enti
alproduced i nt heglassel ectrodeagainstt heknown ph of
dif
ferentbuff
ersol ut
ions.

Pr
epar
ati
onofBuf
ferSol
uti
ons.
Reagent
s.
1.Pot
assi
um Hy
drogenpht
hal
ate(
KHC8H4O4)
.
2.Pot
assi
um Di
hydr
ogenphosphat
e(KH2PO4)
.
3.Pot
assi
um Di
hydr
ogenphosphat
e(KH2PO4)
.
4.Sodi
um Di
hydr
ogenphosphat
e(Na2HPO4)
.
5.Ammoni
um Chl
ori
de(
NH4CL)
.
6.Ammoni
um Hy
droxi
de(
NH4OH)
.
Buf
ferSol
uti
onPH4.
00.
Di
ssolv
edry10.12gm ofPotassi
um Hy
drogenpht
hal
ateat103 0 C i
nonel
i
terof
di
sti
l
ledwat
ert
omakebuffersol
uti
onPH4.0.
Buf
ferSol
uti
onPH7.
00.
Takedry1.179gm ofPot assium Dihy
drogenphosphat
eand4.303gm ofSodi
um
Dihy
drogenphosphat
eanddissolv
einonelit
erofdi
sti
ll
edwat ween1100Ct
erbet o130
0
CTemperatur
etomakebuffersolut
ionPH7.00.
Buf
ferSol
uti
onPH10.
00.
Take5.
7gm ofAmmoni um Chl
ori
deand58mlofAmmonium Hy
droxi
det
hendi
ssol
ve
i
noneli
terofdi
sti
l
ledwatert
omakebuff
ersol
uti
onPH10.
00.

2.Det
ermi
nat
ionofTot
aldi
ssol
vedSol
ids(
TDS)
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Thedeter
minati
onofTot
aldi
ssol
vedSol
i
d/Mi
ner
alsofbot
tl
edr
inki
ngwat
eri
spr
oceed
thr
oughadigi
talTDSMet
er.

3.Tot
alHar
dness.
Def i
niti
on:Wat erhar dnessi sthetraditionalmeasur eoft hecapacityofwat ertoreact
withsoap, hardwat err equiringconsider ablymor esoapt oproduceal ather.Hardwat er
oftenpr oducesanot iceabl edeposi tofpr ecipitate(e.g.insolublemetals,soapsorsal ts)
i
ncont ainers,i
ncludi ng“ bat htubri
ng” .Itisnotcausedbyasi nglesubstancebutbya
variet
yofdi ssolvedpol yvalentmet all
ici ons,predomi nantlycalci
um andmagnesi um
cations,althoughot hercat ions( e.
g.al umi num, bari
um, iron,manganese, stronti
um and
zinc)alsocont r
ibute.Har dnessi smostcommonl yexpressedasmi l
li
gramsofcal ci
um
carbonat eequi valentperl i
ter .Watercont ainingcal ci
um car bonateatconcent r
ati
ons
below60mg/ li
sgener all
yconsi deredassof t ,
60–120mg/ l
,moderatelyhar d,120–180
mg/ l,hard,andmor et han180mg/ l
,veryhar d.Al thoughhar dnessiscausedbycat i
ons,
i
tmayal sobedi scussedi nt ermsofcar bonat e( t
empor ary)andnon- carbonate
(permanent )hardness.
Det
ermi
nat
ionofTot
alHar
dness.
Appar
atus.

 Aut
oBur
ett
e.50ml

 Ti
tr
ati
onf
lask.100ml

 Measur
ingcy
li
nder
.50ml

 Cal
cul
ator
.
Reagent
s.

 Buf
fersol
uti
onPH10.
00

 Har
dnessI
ndi
cat
or

 EDTASol
uti
on0.
01M.
Pr
epar
ati
onofReagent
s.
Har
dnessI
ndi
cat
or.
Dissol
ve01gm ofEr
ichr
omeBlack-
T(EBT)in75mlTr
iet
hanol
eami
net
henadd25ml
ofEthanol
tomake100ml ofHar
dnessIndi
cat
or.
0.
01M EDTATi
tr
antSol
uti
on.
Di
ssol
ve3.
78gm ofEt
hyl
endi
ami
net
etr
aacet
icaci
d(EDTA)i
nonel
i
terofdi
sti
l
led
 Bot
tl
edDr
inki
ngWat
erPr
ocess
wat
ert
omake0.
01M EDTASol
uti
on.
Pr
ocedur
e.

 Take50ml
ofsampl
einTi
tr
ati
onf
laskbyusi
ngmeasur
ingcy
li
nder
.

 Addf
ewdr
opsofhar
dnessi
ndi
cat
orasr
equi
red.

 Addf
ewdr
opsofBuf
fersol
uti
onPH10.
00andst
irt
omi
xit
.

 Nowt
it
rat
eagai
nst0.
01M EDTASol
uti
on.

 I
nit
ial
col
orwi
l
lbepi
nkwhi
l
eendpoi
ntwi
l
lbet
heskybl
ue.
Har
dnessCal
cul
ati
on=Bur
ett
eReadi
ngxHar
dnessFact
or(
20)
.

Har
dnessFact
orDer
ivat
ion.

For
mul
a
=
1000xRequi
redchemi
cal
atomicmassxTi
tr
antConcent
rat
ion
sampleVol
ume
Requi
redchemi
cal
=T.
Har
dness=CaHCO3=At
omi
cmass=101
Ti
tr
antConcent
rat
ion=0.
01
Sampl
evol
ume=50ml

1000x101x0.
01
Har or =
dnessFact =20
50
4.Cal
cium.
Definiti
on:Cal cium i
suniqueamongnut ri
ent
s,inthatthebody ’
sreserveisal so
functional;
i
ncr easingbonemassi sli
near
lyrel
atedtoreducti
oni nfr
actureri
sk.Thet ot albody
storesar eont heorderof1200g,withabout99%i nbonesandt eeth.Al argebodyof
primar yevi
dencef rom r
andomizedcontrol
ledtri
alsshowst hati
ncreasingcal cium
i
ntake, especiall
yinthosewhohav ehadhabitual
lylowcalcium i
ntakes,increases
bonemassdur inggrowthandreducesbonelossandf r
actureri
sklateinlife.
Osteopor osisisoneoft hemostpreval
entofage-relat
eddiseases.Calcium and
 Bot
tl
edDr
inki
ngWat
erPr
ocess
v
itami
nDar
ejoi
ntl
ybenef
ici
ali
nincr
easi
ngbonemass.
Inadequat ei
ntakesofcal ci
um hav ebeenassoci atedwithincreasedrisksof
osteopor osi
s,nephrol
ithi
asis(ki
dneyst ones),colorect
alcancer,hypertensionand
stroke,coronaryarter
ydisease,insuli
nresistanceandobesi ty.Mostoft hesedisorder
s
hav etreatments,butnotcures.Owi ngtoal ackofcompel li
ngev idencef ortherol
eof
calcium asacont ri
butoryelementinr el
ati
ont othesediseases, esti
mat esofcalci
um
requirementhav ebeenmadeont hebasisofboneheal thoutcomes, witht hegoalof
optimizingbonemi neraldensity
.

Det
ermi
nat
ionofCal
cium.
Appar
atus.

 Aut
oBur
ett
e.50ml

 Ti
tr
ati
onf
lask.100ml

 Measur
ingcy
li
nder
.50ml

 Cal
cul
ator
.
Reagent
s.

 4NNaOH

 Cal
cium I
ndi
cat
or

 EDTASol
uti
on0.
01M.
Pr
epar
ati
onofReagent
s.
Cal
cium I
ndi
cat
or.
Di
ssol
ve02gm ofCal
concar
boxl
icAci
din100mlofdisti
ll
edwatert
omakeCal
cium
I
ndi
catorandMur
oxi
dePowderisopti
onal
asCal
cium Indi
cat
or.
4NSodi
um Hy
droxi
deSol
uti
on.
Di
ssol
ve40gm ofSodi
um Hy
droxi
dei
n250ml
ofdi
sti
l
ledwat
erandst
irt
omi
xit
.
0.
01M EDTATi
tr
antSol
uti
on.
Di
ssol
ve3.
78gm ofEt hy
lendi
aminetet
raacet
icaci
d(EDTA)i
nonel
i
terofdi
sti
l
led
wat
ertomake0.
01M EDTASoluti
on.
Pr
ocedur
e.

 Take25ml
ofsampl
einTi
tr
ati
onf
laskbyusi
ngmeasur
ingcy
li
nder
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Addf
ewdr
opsofCal
cium i
ndi
cat
orasr
equi
red.

 Addf
ewdr
opsof4NNaOHandst
irt
omi
xit
.

 Nowt
it
rat
eagai
nst0.
01M EDTASol
uti
on.

 I
nit
ial
col
orwi
l
lbepi
nkwhi
l
eendpoi
ntwi
l
lbet
heskybl
ue.
Cal
cium Cal
cul
ati
on=Bur
ett
eReadi
ngxCal
cium Fact
or(
16.
04)
.

Cal
cium Fact
orDer
ivat
ion.

For
mul
a
=
1000xRequi
redchemi
cal
atomicmassxTi
tr
antConcent
rat
ion
sampleVol
ume

Requi
redchemi
cal
=Cal
cium =Ca=At
omi
cmass=40.
10
Ti
tr
antConcent
rat
ion=0.
01
Sampl
evol
ume=25ml

1000x40.
10x0.
01
Cal
ci or =
um Fact =16.
04
25

5.Magnesi
um.
Defini
tion:Magnesium isthefourthmostabundantcat i
oninthebodyandt hesecond
mostabundantcationi nintr
acel
lul
arfl
uid.Itisacofact
orforsome350cel lular
enzymes, manyofwhi chareinvolv
edinener gymetaboli
sm.Itisalsoinvolvedinpr otei
n
andnucl ei
caci
dsynt hesi
sandi sneededf ornormalvascul
artoneandi nsulinsensiti
v i
ty.
Totalbodystor
esareont heorderof25g, withabout60%inbone.Tot albodybur denis
dif
fi
culttoquant
ify
,becauseonl yasmal lportioni
sinbloodorflui
ds,andi tcanbe
vari
able.Lowmagnesi um level
sareassoci at
edwithendothel
ialdysf
unction, i
ncreased
 Bot
tl
edDr
inki
ngWat
erPr
ocess
vascularreacti
ons,el
ev atedcir
culati
nglevelsofC-r
eacti
veprotei
n(aproinf
lammatory
mar kerthati
sar i
skf actorforcoronaryheartdi
sease)anddecreasedinsul
insensit
ivi
ty.
Lowmagnesi um st
atushasbeeni mpl i
catedinhyper
tensi
on,coronar
yhear tdi
sease,
type2di abetesmelli
tusandmet aboli
csyndrome.
Magnesium def iciencyhasbeeni mpli
catedint hepat
hogenesisofhy pertensi
on, wi t
h
someepi demi ol
ogi calandexperiment
alstudiesdemonst r
atinganegat ivecorrelati
on
betweenbl oodpr essureandser um magnesium level
s.Howev er,datafrom cli
nical
studi
eshav ebeenl essconvinci
ng.Cardi
acar rhythmi
asofv entri
cularandat r
ialorigin
havebeenr eport edinpati
entswi t
hhypomagnesaemi aandinpost menopausal women
i
ncont r
olleddietst udi
es.Indeed,aseri
ouscar diacar
rhyt
hmi a,TorsadedePoi ntes,is
tr
eatedwi thintr
av enousmagnesi um t
herapy.

Det
ermi
nat
ionofMagnesi
um.
Thevalueofmagnesi
um isderi
vedfr
om t
hediff
erenceofbur
ett
ereadi
ngofTot
al
Hardnessandt
heburett
er eadi
ngofCal
cium.
Magnesi
um Cal
cul
ati
on=(
B.Readi
ngofTot
alHar
dness–B.
Readi
ngofCal
cium)xMg
Fact
or
Magnesi
um Fact
orDer
ivat
ion.

For
mul
a
=
1000xRequi
redchemi
cal
atomicmassxTi
tr
antConcent
rat
ion
sampleVol
ume

Requi
redchemi
cal
=Magnesi
um =Mg=At
omi
cmass=24
Ti
tr
antConcent
rat
ion=0.
01
Sampl
evol
ume=50ml

1000x24x0.
01
Cal
ci or =
um Fact =4.
08
50
6.Det
ermi
nat
ionofChl
ori
des.
Appar
atus.
 Aut
oBur
ett
e.50ml
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Ti
tr
ati
onf
lask.100ml

 Measur
ingcy
li
nder
.50ml

 Cal
cul
ator
.
Reagent
.

 Pot
assi
um chr
omat
eIndi
cat
or(
K2Cr
O4)
.

 0.
0282NSi
l
verNi
tr
ateSol
uti
on(
AgNO3)
Pr
epar
ati
onofReagent
s.
Chl
ori
deI
ndi
cat
or.
Di
ssol
ve05gm ofPot
assi
um chr
omat
ein100mlofdi
sti
ll
edwat
ert
omakeChl
ori
de
I
ndi
cator
.
0.
0282NSi
lv
erNi
tr
ateSol
uti
on.
Di
ssol
ve4.79gm ofSi
lv
erNitr
ategranul
esin1000mlofdi
sti
ll
edwat
erandst
irt
omi
x
i
tandstor
einacolor
edglassbott
lebecausei
tabsor
bli
ght.

Pr
ocedur
e.

 Take50ml
ofsampl
einTi
tr
ati
onf
laskbyusi
ngmeasur
ingcy
li
nder
.

 Addf
ewdr
opsofChl
ori
dei
ndi
cat
orasr
equi
red.

 Nowt
it
rat
eagai
nst0.
0282AgNO3Sol
uti
on.

 I
nit
ial
col
orwi
l
lbey
ell
owi
shwhi
l
eendpoi
ntwi
l
lbet
hebr
ickRed.
Chl
ori
deCal
cul
ati
on=Bur
ett
eReadi
ngxChl
ori
deFact
or(
20)
.

Chl
ori
deFact
orDer
ivat
ion.

For
mul
a
=
1000xRequi
redchemi
cal
atomicmassxTi
tr
antConcent
rat
ion
sampleVol
ume
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Requi
redchemi
cal
=Chl
ori
de=Ca=At
omi
cmass=35.
5
Ti
tr
antConcent
rat
ion=0.
01
Sampl
evol
ume=50ml

1000x35.
5x0.
0282
Chl
ori or =
deFact =20
50
7.M.Al
kal
ini
ty.
Defi
niti
on: Al
kali
nit
ymeanst hebuffer
ingcapacit
yofwat er;
thecapaci
tyofwaterto
neutr
alizeit
sel
f.Itprevent
sthewat erpHlevelsfrom becomi ngtoobasi
coracid.I
talso
addscar bontowat er.Al
kal
init
ystabil
i
zewat eratpHl evelsaround07.However,
when
theacidit
yishighinwat er
,Alkali
nit
ydecreases,whichcancausehar mfulcondi
ti
onsf or
aquati
cl i
fe.
Inwaterchemist
ryAlkal
init
yisexpressedinppm ormg/Lfr
om equi
val
entt
ocalci
um
carbonat
e.Total
Alkali
nit
yofwat eristhesum ofal
lthr
eesor
tsofAl
kal
ini
ty;
car
bonates,
bicar
bonatesandhydroxideAlkal
ini
ty.

Det
ermi
nat
ionofM.Al
kal
ini
ty.
Appar
atus.
 Aut
oBur
ett
e.50ml

 Ti
tr
ati
onf
lask.100ml

 Measur
ingcy
li
nder
.50ml

 Cal
cul
ator
.

 Al
kal
i
net
estki
topt
ional
.
Reagent
.

 Met
hyl
Pur
pleI
ndi
cat
or.

 0.
02NSul
fur
icAci
dSol
uti
on(
H2SO4)
Pr
epar
ati
onofReagent
s.
Met
hylPur
pleI
ndi
cat
or.
Dissol
ve0.
66gm ofMet
hyl
Redand0.34gm Br
omocr
esol
greeni
n100ml
ofEt
hanol
andsti
rtomixt
omakeMethyl
Purpl
eIndi
cat
or.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
0.
02NSul
fur
icAci
dSol
uti
on.
Di
ssol
ve0.
54mLofSul
fur
icAci
din1000ml
ofdi
sti
l
ledwat
erandst
irt
omi
xit
.
Pr
ocedur
e.

 Take25ml
ofsampl
einTi
tr
ati
onf
laskbyusi
ngmeasur
ingcy
li
nder
.

 Addf
ewdr
opsofMet
hyl
Pur
plei
ndi
cat
orasr
equi
red.

 Nowt
it
rat
eagai
nst0.
02NH2SO4Sol
uti
on.

 I
nit
ial
col
orwi
l
lbeGr
eeni
shwhi
l
eendpoi
ntwi
l
lbet
heReddi
sh.
M.
Alkal
i
nit
yCal
cul
ati
on=Bur
ett
eReadi
ngxAl
kal
i
nit
yFact
or(
40)
.

8.Det
ermi
nat
ionofSul
phat
es(
SO4)bySpect
rophot
omet
ry.
Appar
atus
 Spect
rophot
omet
er.
 Sampl
i
ngcov
ets/cel
lwi
thl
i
d10mm.
Chemi
cal
s
 Readyt
ousesul
phat
ereagent
-01
 Readyt
ousesul
phat
ereagent
-02
Pr
ocedur
e.
 Fi
rstwashanddr
ytheCov
etwi
tht
issuepaper
.
 Thesamplecovetshoul
dbecompl
etel
ycl
earandt
ranspar
entandf
reef
rom any
fi
ngerpr
intoni
t.
 Takesampl
ewat
eri
ncov
ett
obet
est
.
 Addtwodr
opsofsul
phat
ereagent
-01andt
wodr
opsofsul
phat
ereagent
-02t
hen
st
irt
omixi
t.
 Nowgi
vest
ayt
imet
osampl
ecov
etf
or10mi
nut
es.
 Tur
nONt
heSpect
rophot
omet
erandkeepi
tonabsor
banceMode.
 Setwav
elengthat420nm andadj
ustSamplecovet,
Disti
l
ledwat
ercov
etand
bl
ankCelli
nthespeci
fi
ccompartmentofspect
rophotometer
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Runt
hebl
ankcel
ltomakespect
rophot
omet
erbl
ankf
or15mi
nut
es.
 Rundi
sti
l
ledwat
erf
or10mi
nut
esandnot
eit
sreadi
ng.
 Agai
nrunt
hebl
ankcel
ltomakespect
rophot
omet
erbl
ankf
or10mi
nut
es.
 Nowr
unt
hesampl
efor15mi
nut
esandnot
ether
eadi
ng.
 Aft
ertaki
ngsulphat
ereadi
ngbl
ankt
hespect
rophot
omet
eragai
nforf
urt
heruse
andnextreadi
ng.

Readi
ngofdi
sti
l
ledwat
erabsor
bance
Sul
phat
eCal
cul
aton =
i x10
Readi
ngofsamplewat
erabsor
bance

9.Det
ermi
nat
ionofNi
tr
it
es(
NO2)bySpect
rophot
omet
ry.
Appar
atus
 Spect
rophot
omet
er.
 Sampl
i
ngcov
ets/cel
lwi
thl
i
d10mm.
Chemi
cal
s
 Readyt
ouseNi
tr
it
esr
eagent
-01
 Readyt
ouseNi
tr
it
esr
eagent
-02
Pr
ocedur
e.
 Fi
rstwashanddr
ytheCov
etwi
tht
issuepaper
.
 Thesamplecovetshoul
dbecompl
etel
ycl
earandt
ranspar
entandf
reef
rom any
fi
ngerpr
intoni
t.
 Takesampl
ewat
eri
ncov
ett
obet
est
.
 Addtwodr
opsofNi
tr
it
esr
eagent
-01andt
wodr
opsofNi
tr
it
esr
eagent
-02t
hen
st
irt
omixi
t.
 Nowgi
vest
ayt
imet
osampl
ecov
etf
or10mi
nut
es.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Tur
nONt
heSpect
rophot
omet
erandkeepi
tonabsor
banceMode.
 Setwav
elengthat510nm andadj
ustSamplecovet,
Disti
l
ledwatercov
etand
bl
ankCelli
nthespeci
fi
ccompartmentsofspect
rophotometer
.
 Runt
hebl
ankcel
ltomakespect
rophot
omet
erbl
ankf
or15mi
nut
es.
 Rundi
sti
l
ledwat
erf
or10mi
nut
esandnot
eit
sreadi
ng.
 Agai
nrunt
hebl
ankcel
ltomakespect
rophot
omet
erbl
ankf
or10mi
nut
es.
 Nowrunthesampl
efor15mi
nut
esandnot
ether
eadi
ngwhi
chwi
l
lbef
inal
Ni
tr
it
ereadi
ng.
 Aft
ertaki
ngNitri
tesreadi
ngbl
ankthespect
rophot
ometeragainf
orf
urt
heruse
andnextreadi
ng,ifnoti
nusethenShutOFFthespect
rophot
ometer
.

10.Det
ermi
nat
ionofSodi
um (
Na)byFl
amephot
omet
ery
.
Definiti
on:Sodi um (Na)isthemaj orext racel
lularcaut i
onandi tplaysar oleinbodyf l
uid
di
stributi
on.Concent rat
ionofsodi um ionsi nsidet heplasma( extracell
ular)is130- 145
mmol /l
.Higherandl owerconcent rat
ionsar er eferredtoashy pernatremiaand
hypernatremi arespecti
vely
.Whenasol utioncont aini
ngcat i
onsofsodi um and
potassium isspay edintofl
ame, t
hesolv entev apor at
esandi onsar econv ertedinto
atomi cstate.Intheheatoft heflame( t
emper atureabout1800º C),small f
ractionoft he
atomsi sexci t
ed.Relaxati
onoft heexci tedatomst othelowerener gylevel i
s
accompani edbyemi ssi
onofl ight(photons)wi t
hchar acter
isti
cwav el
ength( Na: 589nm,
K:766nm) .Intensit
yoftheemi t
tedli
ghtdependsont heconcent rati
onofpar t
icular
atomsi nf l
ame.

Appar
atus
Fl
amephot
omet
erwi
thSodi
um Fi
l
ter
Sampl
i
ngBeaker
s100ml
.
Chemi
cal
s
Readyt
ouseSodi
um st
andar
dsol
uti
on1000ppm.
Hy
drochl
ori
cAci
dforgl
asswar
eswashi
ng
Pr
epar
ati
onofChemi
cal
.
Herewehave1000ppm sol
uti
onofSodi
um andwehav et
omakether
equir
ed
mini
mum ppm sol
uti
onaccor
dingt
otheSodium v
aluebyusi
ngf
oll
owi
ngformul
a,
 Bot
tl
edDr
inki
ngWat
erPr
ocess
C1V1=C2V2
Chemi
cal
avai
l
abl
exr
eq.v
olume(
1000ppm sol
)=Req.ppm Sol
uti
onxReq.Sol
uti
on
Volume
1000ppm xV1=40ppm x250ml

40X250
V1 = =10ml
1000

10mlofSol
uti
onf
rom 1000ppm Sol
uti
onr
equi
redt
omake40ppm Sol
uti
oni
n250ml
vol
ume.
Pr
ocedur
e.
 Swi
tchONt
heFl
amephot
omet
er.
 Sel
ectpr
ogr
am f
orSodi
um Fi
l
termode.
 St
artI
gni
ti
onandadj
ustf
lamewi
thr
egul
atorknob.
 Nowstandardi
zetheflamephotomet
erwi
thZeromodebyr
unni
ngdi
sti
l
ledwat
er
i
nitbysucti
onfrom capi
ll
aryt
ubein100mlbeaker
.
 RunSodi
um st
andar
dsol
uti
onandf
inei
tandst
abi
l
izet
her
eadi
ng.
 St
andardi
zet
hef
lamephot
omet
erwi
thZer
omodeagai
nbyr
unni
ngdi
sti
l
led
wateri
nit
.
 Repeatt
hispr
ocessf
ort
hreet
imes.
 Nowr
unt
hesampl
ewat
erf
orSodi
um Readi
ng.
 Keepupthecapi
ll
arytube10mm f
rom thebot
tom ofSampl
ebeakeranddon’
t
toucht
hecapi
ll
arytubewit
hbot
tom sur
faceofbeaker
.
 Runt
hesampl
efort
hreet
imesandnot
ethef
inal
aver
ager
eadi
ngofSodi
um.
 St
andardi
zethefl
amephotomet erwi
thZeromodeagai
nbyr
unni
ngdi
sti
l
led
wateri
nitf
orfur
theruseornextsample.

11.Det
ermi
nat
ionofPot
assi
um (
K)byFl
amephot
omet
ery
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Defini
tion:Potassium (K)i
sthemajorcautionfoundi
nsideofcell
s.Thepr
operlevelof
potassium isessenti
alfornormal
cellfuncti
on.Anabnormalincr
easeofpotassium
(hy
per kalemia)ordecreaseofpot
assium (hypokal
emia)canprof
oundlyaf
fectthe
nervoussy stem andheart,
andwhenext reme,canbefatal
.
Thenor
mal
bloodpot
assi
um l
evel
is3.
5-5.
0mi
l
li
mol
es/
li
ter(
mmol
/l
).
Appar
atus
Fl
amephot
omet
erwi
thPot
assi
um Fi
l
ter
Sampl
i
ngBeaker
s100ml
.
Chemi
cal
s
Readyt
ousePot
assi
um st
andar
dsol
uti
onof1000ppm.
Hy
drochl
ori
cAci
dforgl
asswar
eswashi
ng
Pr
epar
ati
onofChemi
cal
.
Herewehave1000ppm sol
uti
onofPot
assi
um andwehavetomaket
herequi
red
mini
mum ppm sol
uti
onaccor
dingt
otheSodi
um val
uebyusi
ngfol
l
owingf
ormula,

C1V1=C2V2
Chemi
cal
avai
l
abl
exr
eq.v
olume(
1000ppm sol
)=Req.ppm Sol
uti
onxReq.Sol
uti
on
Volume
1000ppm xV1=40ppm x250ml

40X250
V1 = =10ml
1000

10mlofSol
uti
onf
rom 1000ppm Sol
uti
onr
equi
redt
omake40ppm Sol
uti
oni
n250ml
vol
ume.
Pr
ocedur
e.
 Swi
tchONt
heFl
amephot
omet
er.
 Sel
ectpr
ogr
am f
orPot
assi
um Fi
l
termode.
 St
artI
gni
ti
onandadj
ustf
lamewi
thr
egul
atorknob.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Nowstandardi
zetheflamephotomet
erwi
thZeromodebyr
unni
ngdi
sti
l
ledwat
er
i
nitbysucti
onfrom capi
ll
aryt
ubein100mlbeaker
.
 RunPot
assi
um st
andar
dsol
uti
onandf
inei
tandst
abi
l
izet
her
eadi
ng.
 St
andardi
zet
hef
lamephot
omet
erwi
thZer
omodeagai
nbyr
unni
ngdi
sti
l
led
wateri
nit
.
 Repeatt
hispr
ocessf
ort
hreet
imes.
 Nowr
unt
hesampl
ewat
erf
orPot
assi
um Readi
ng.
 Keepupthecapi
ll
arytube10mm f
rom thebot
tom ofSampl
ebeakeranddon’
t
toucht
hecapi
ll
arytubewit
hbot
tom sur
faceofbeaker
.
 Runt
hesampl
efort
hreet
imesandnot
ethef
inal
aver
ager
eadi
ngofPot
assi
um.
 St
andardi
zethefl
amephotomet erwi
thZeromodeagai
nbyr
unni
ngdi
sti
l
led
wateri
nitf
orfur
theruseornextsample.
 I
fnotinusef
ornextsampl
ingst
opigni
ti
onandi
gni
ti
onsour
cef
ir
standt
hen
SWITCHOFFtheFlamephotomet
er.

I
nst
rumentSaf
ety

 Pl
easer
eadt
hei
sinf
ormat
ioncar
eful
l
ypr
iort
oinst
all
i
ngorusi
ngt
hisequi
pment
.
 Theuni
tdescr
ibedi
nthi
smanual
isdesi
gnedt
obeoper
ateonl
ybyt
rai
ned
per
son.
 Anyadj
ust
ments,mai
ntenanceandrepai
rmustbecarri
edoutasdef
inedi
nthi
s
manual
byaper
sonquali
fi
edt
obewar eofthehazar
dsinv
olved.
 Iti
sessent
ial
thatbot
hoper
ati
ngandser
viceper
sonnel
empl
oy,
topr
ovi
deasaf
e
syst
em ofwork.
 Thecov er
soft heunitshoul
donlyberemov
edbypersonall
ywhohavebeen
tr
ainedto
avoidtheriskofshock.Atthi
spointt
heuni
tmustbedisconnect
edf
rom al
l
servi
cesi.e.
gasandel ectr
ic.
 Refer
encesshoul
dalwaysbemadet othehealt
handsafet
ydatasuppl
iedwit
h
anychemical
sused.Gener
all
yaccept
edlaborator
ypr
oceduresf
orsafehandl
ing
ofchemical
sshoul
dbeempl oyed.
 I
fiti
ssuspect
edt
hatsaf
etypr
otect
ionhasbeenimpair
edinanyway,theuni
t
mustbemadeinoper
ati
veandsecuredagai
nstanyi
ntendedoper
ati
on.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Thef
aul
tcondi
ti
onshoul
dber
epor
tedi
mmedi
atel
ytot
heappr
opr
iat
eser
vici
ng
aut
hor
it
y.

At
tent
ion:
 Fl
amephot
omet
erusesf
lammabl
egaseswhi
chcancauseexpl
osi
onsi
fused
I
mpr
oper
ly…!
.
 Swi
tcht
hei
nst
rumentonandof
fundersuper
visi
on….
!.

Washi
ngofgl
asswar
esf
orSodi
um andPot
assi
um Test
.
Washt heglasswarestoremoveal
l t
het r
acesoforgani
cmatter
,suspended
par
ti
clesandmaket hem f
reef
rom scali
ngandtranspar
entbywashingwithan
aci
dicmixturerat
io1:1thr
oughfol
l
owi ngsteps,

 Makeanaci
dicmi
xtur
eby1:
1rat
iowi
thdi
sti
l
led/ROWat
er.
 Washanddi
pthegl
asswar
esov
erni
ghti
nthi
ssol
uti
onf
ordecont
ami
nat
ion.
 Ri
nset
hegl
asswar
est
hor
oughl
ywi
thDi
sti
l
led/ROWat
er.
 Dr
yinacl
eandr enat800Ct
yOv emper
atur
e.
 Byusi
ngthi
smet hodofwashi
ng,wecanr
emov
eal
lthet
racesofSodi
um and
Pot
assi
um avai
l
ableingl
asswares.

Det
ermi
nat
ionof
 Ar
seni
c,
 I
ron,
 Ozone,
 Fr
eechl
ori
ne,
Inbott
ledri
nkingwatercanbedet
ectandcalcul
atethr
ought
hepr
ocedur
eofTest
kitMethodsaccor
dingtothemanufact
ureri
nstr
ucti
ons.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Bot
tl
edWat
erPar
amet
er
 
Sr
. Per
missi
bleLi
mit
s BLUWat er
Par
amet
er Uni
ts  
# Standar
ds
WHO P.
F.A PSQCA
Phy
sicalPar
amet
ers
Un- Un-
1
Col
or
(
TCU)
15
obj
ecti
onabl
e obj
ecti
onabl
e
Un-
obj
ect
ionabl
e
2 Conduct
ivi
ty (
µS/
cm) NGVS NGVS NGVS Un-
obj
ect
ionabl
e
Odor Un- Un- Odorless
Odor
less
3 - obj
ectionabl
e obj
ectionabl
e
4 pH - 6.5-8.5 6.
5~8. 5 6.5-8.5 6.5-
8.5
Un- Un- Un-
5
Tast
e
- obj
ectionabl
e obj
ectionabl
e obj
ectionabl
e
Un-
Objectionabl
e
6 TDS (
mg/
l) 500 500 500 130-270mg/ L
Chemi
calPar
amet
ers
1 Cal
ci um (
mg/
l) 100 100 100 25- 50mg/ L
2 Chloride (
mg/
l) 250 250 250 45- 110mg/ L
3 Hardness (
mg/
l) 500 250 250 70-150
4 I
ron (
mg/
l) 0.3 0.3 0.3 <0.2
5 Magnesi um (
mg/
l) 50 50 50 5-20mg/ L
6 Nit
rat e-
N (
mg/
l) 1.0 1.0 1.0 Lesst han0.1
7 Potassium (
mg/
l) 10 10 10 0. 1-1.
5mg/ L
8 Sodium (
mg/
l) 50 50 50 05- 35mg/ L
9 Sul
f ate (
mg/
l) 250 250 250 10- 40mg/ L
Lesst han0.01
10 Ar
seni
c ppb 0.
01 0.
01 0.
01 ppb
Mi
croBi
ologi
calPar
amet
ers
1 TotalColif
orm CFU Ni
l Ni
l Ni
l Ni
l
2 E-Coli CFU Ni
l Ni
l Ni
l Ni
l
3 Salmonella CFU Ni
l Ni
l Ni
l Ni
l
Totalvi
able
4 countat37C CFU Ni
l Ni
l Ni
l Ni
l
TotalViable
countat20
5 c~22c CFU Ni
l Ni
l Ni
l Ni
l

Mi
crobi
ologi
calTestofBot
tl
edr
inki
ngwat
er.
Mi
crobi
ology
.Thest
udyofmi
croor
gani
sm i
sknownasMi
crobi
ology
.
Themai
nmi
crobi
alt
estofbot
tl
edr
inki
ngwat
er,
Jui
cesandbev
eragesar
egi
venbel
ow,
1.Tot
alpl
atecountBact
eri
a
 Bot
tl
edDr
inki
ngWat
erPr
ocess
2.Tot
alcol
i
for
m Bact
eri
a
3.Escher
ichi
acol
i
4.Faecal
Str
ept
ococcus
5.PseudomonasTest
6.Sal
monel
l
aShi
j
ell
aTest
.
7.YeastandMol
d
Mat
ri
x.
I
nmicr
obiol
ogyl
anguagethesampl
ewhichi
susedf
ormi
crobi
ologi
cal
testi
sknownas
mat
ri
x.Ther
earetwotypesofMat
ri
x/sampl
es.
1.Li
qui
dMat
ri
x
 Wat
er
 Bev
erages
 Jui
ces
 Mi
l
k
 Sugarsy
rup
2.Sol
idMat
ri
x
 Al
lty
pesoff
oodpr
oduct
s
 Baker
yandsweetpr
oduct
s
 RawMeat
 Poul
tr
y
 Conf
ect
ionar
y
Mi
crobi
ologi
calTestMet
hod
Accor
dingtotheseli
qui
dsandsoli
dmat
ri
xther
ear
ethr
eemet
hodsofsampl
i
ngar
e
usedi
nmi cr
obiologyi
nindust
ri
es.
1.Di
rectsampl
i
ngmet
hod
2.Membr
anesf
il
tr
ati
ont
echni
que
3.Di
l
uti
onmet
hod/Mul
ti
pleFer
ment
ati
onTubet
echni
que
Di
rectsampl
ingMet
hod.
Thi
smet
hodofsampl
i
ngi
nmi
crobi
ologyi
sof
tenusedf
orl
i
qui
dmat
ri
xinwhi
ch01ml
 Bot
tl
edDr
inki
ngWat
erPr
ocess
sampl
edi
rect
lyi
ncubat
einpet
ridi
sh.
Appar
atus
 St
eri
l
izedpet
ripl
ates
 Cul
tur
ebot
tl
es
 Mi
cropi
pet
tewi
thst
eri
l
izedext
rat
ips.
 Cover
edTher
most
ati
cwat
erbat
htomai
ntai
nthet
emper
atur 500
eofAgarat45-
C.
 Spr
itLamp
 Ver
ti
cal
airf
lowcabi
netwi
thUVLamp.
 Magni
fyi
ngLens
 I
ncubat
orf
orbev
eragesat250CTemper
atur
e.
 I
ncubat
orf
ordr
inki erat370CTemper
ngwat atur
e.
 Dr
yov
erf
ort
hest
eri
l
izat
ionofgl
asswar
es.
 Aut
ocl
avef
orChemi
cal
sst
eri
l
izat
ion.
Chemi
cal
.
 Desi
recul
tur
emedi
a
 Absol
uteEt
hanol
 Spr
it
 70%Et
hanol
spr
ayasDi
sinf
ect
ant
.
Pr
ocedur
e.
 Al
lappar
atusandchemi
cal
sshoul
dbest
eri
l
ized.
 Di
sinf
ectt
hesampl
i
ngar
eawi
th70%et
hanol
spr
ay.
 Col
l
ectt
hedesi
resamplesincul
tur
ebot
tl
esaf
terst
eri
l
izi
ngt
hesampl
i
ngpoi
nts
byf
lami
ngorby70%ethanolspr
ay.
 Bur
nthespri
tlampandkeepar
oundt
hei
ncubat
ionar
eat
ost
opandcont
rol
the
cr
osscont
aminati
on.
 Take01ml
requi
redsampl
einapet
ripl
atebyusi
ngmi
cropi
pet
te.
 Feedtherequi
redagari
npet
ripl
ateandmi
xitandshakei
tcl
ockwi
seandant
i-
cl
ockwiseslowly
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Lett
hepet
ridi
shesf
orsol
i
dif
y.
 Nowi
nvertthepet
ripl
atesandi
ncubat
einani
ncubat
orundert
her
equi
red
t
emper
ature.
 Takei
ncubat
ionr
esul
tsaf
ter24hour
sofObser
vat
ion.
2.Membr
aneFi
lt
rat
ionTechni
que.
Thephar maceut i
cal andcosmet i
csindustri
estypicall
yfocusonmoni toringthei
r
processwat erforPseudomonasspeci es.Theelectronicsindust rymoni t
orsforanyand
al
l microorganismsbecauset heymustkeept hei
rpr ocesswat erfreef r
om ev enthe
smallestor ganisms.Mi crobialmonitoringinthefoodandbev eragei ndust r
ytypi
cal
ly
employ ssev eraltypesoft echniquesbecauseoft hev arietyofsampl est hatare
encount ered.Bev eragesampl escant ypical
lybemoni toredf ormi croor ganismsbythe
MFTechni que, butwhensol i
dsampl escannotbel i
quefied,alternati
v emet hodsmust
beused.

Concl
usi
on
TheMFTechni queisaneff
ecti
ve,acceptedt echni
quefortesti
ngflui
dsamplesf
or
micr
obiol
ogicalcont
aminat
ion.I
tinvol
v eslesspreparat
ionthanmanyt r
adi
ti
onal
methods,andisoneofafewmet hodst hatwi l
lall
owtheisolat
ionandenumerat
ionof
micr
oorganisms.TheMFTechniqueal sopr ovi
despresenceorabsenceinf
ormation
wit
hin24hours.

Appar
atus
 St
eri
l
izedpet
ripl
ates
 Cul
tur
ebot
tl
es/Sampl
i
ngzi
pperbag,
Bev
eragemoni
tor
s(100ml
)
 Cover
edTher
most
ati
cwat
erbat
htomai
ntai
nthet
emper
atur 500
eofAgarat45-
C.
 Spr
itLamp
 Ver
ti
cal
airf
lowcabi
netwi
thUVLamp.
 Magni
fyi
ngLens
 I
ncubat
orf
orbev
eragesat250CTemper
atur
e.
 I
ncubat
orf
ordr
inki erat370CTemper
ngwat atur
e.
 Dr
yov
erf
ort
hest
eri
l
izat
ionofgl
asswar
es.
 Aut
ocl
avef
orChemi
cal
sst
eri
l
izat
ion.
 Fi
l
trat
ionAssembl
ywi
thhol
der
,fi
l
terbaseandf
unnel
.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Vacuum sour
ce
 Membranesfi
l
ters,
whit
e,gridded,
47mm and.
45mi
cronsasspeci
fi
edbyt
he
manuf
actur
erforenumerat
ionofBact
eri
a.
 For
cepst
otr
ansf
ert
hemembr
anef
il
ter
swi
thoutt
ouchi
nganddamagi
ng.

Pr
ocedur
e. 

1.Col
l
ectt
hesampl
e.

2.Selectt
heappropr
iat
enutr
ientorcul
tur
emedi
um and
Dispenseiti
nthepet
ridi
sh.

3.Fl
amet hefor
ceps,
andr
emov
ethemembr
anef
rom t
he
ster
il
epackage.

4.Pl
acet
hemembr
anef
il
teri
ntot
hef
unnel
assembl
y.

7.Ri
nsefunnel
wit
hster
ilebuf
fer
edwater.Turnonvacuum
andal
lowtheli
qui
dtodrawcomplet
elythroughthef
il
ter
.

8.Fl
amethef
orcepsandr
emov
ethemembr
anef
il
terf
rom
thef
unnel
.

 
 Bot
tl
edDr
inki
ngWat
erPr
ocess
9.Pl
acet
hemembr
anef
il
teri
ntot
hepr
epar
edPet
ridi
sh.

10.I
ncubateatt
hepr
opert
emper
atur
eandf
ort
heappr
opr
iat
e
ti
meperiod.

11.Countt
hecol
oni
esunder10-15Xmagni
fi
cat
ion.

12.Conf
ir
mthecol
oni
esandr
epor
tther
esul
ts.

Not
e: 
Obser
veasept
ict
echni
quet
hroughoutt
hepr
ocedur
e. 

Adv
ant
agesofMFTechni
que.

 Per
mit
stest
ingofl
argesampl
evol
umes.

 Reducespr
epar
ati
ont
imeascompar
edt
omanyt
radi
ti
onal
met
hods.

 Al
l
owsi
sol
ati
onandenumer
ati
onofdi
scr
etecol
oni
esofbact
eri
a.

 Pr
ovi
despr
esenceorabsencei
nfor
mat
ionwi
thi
n24hour
s.

 Eff
ecti
veandacceptabl
etechni
que.Usedt
omoni
tordr
inki
ngwat
eri
n
gover
nmentlabor
atori
es.

 Usef
ulforbact
eri
almonit
ori
nginthephar
maceut
ical
,cosmet
ics,
elect
roni
cs,
andfoodandbeveragei
ndustr
ies.

 All
owsforr
emoval
ofbact
eri
ost
ati
corcidal
agentsthatwoul
dnotbe
remov
edinPourPl
ate,
Spr
eadPlat
e,orMPNt echni
ques.
Gener
alasept
ict
echni
ques

 Washhands
 Cl
eanbenchar
eawi
thswabsoaked/
Spr
ayeri
nmet
hyl
atedspi
ri
t.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Don’ttouchanypar tofthecont ainer
,pipet t
e,etc.,cominginCont actwiththe
sampl eorcult
ure.
 Don’tremov eli
dofapet ridishorcapofat esttubel ongerthanabsolutel
y
necessar y
.
 Li
ght l
yflametopoft esttubes, endsofpi pettes,necksandSt oppersofbot tl
es
beforeandaf t
eraddingsampl esorinocul ati
on.
 Transfercult
urefrom onet ubet oafresht ube( subcul t
uri
ng)usingwi r
eloop
ster
il
izedbyheat i
ngtor ednessandcool ed.
Al
way
sf ol
lowt hesemet hodst oav oi
dcont aminat i
on…!

3.TheMostPr
obabl
eNumberMet
hod/Mul
ti
pleFer
ment
ati
onTube
Techni
que.
Thebasi cassumpt ionsoftheMPNmet hodar ethatbact eri
af ol
l
owPoi ssonst at
ist
ics,
andthatasi ngleviablecellwithresul tint ur
bidit
yoft het estmedi aundert heconditions
used.Nut r
ientbr othwi l
lsuppor tgrowt hofor gani smsandt urnturbid.Thebasi cpattern
ofgrowthv s.no- growthcanpr ovideinf ormation,par ticul
ar l
yatlownumber sof
organismswi thlar
genumber sofr eplicates.Howev er, t
hisaccur acycanbegr eat
ly
i
ncreasedbydi luti
ngt heinoculum andt hencompar ingt her ecoveri
esofal ltubesint he
dil
uti
onser ies.Thist hen,i
st hebasi soft heMPN( al
soknownandmul ti
pletube,dil
ut i
on
tube,ordil
ut iontubemet hods)met hod.
Themostpr obablenumber(MPN)met hodisauseful,i
funderuti
li
zed,toolforthe
microbiol
ogist
.Foodhomogenateseasil
yclogthefil
teri
nthefilt
rati
onmet hod, sothat
themostpr obabl
emethodisoftenusedforfoodsoli
dmat ri
x.Thismet hodofmi cr
obial
samplingmaybeusedf orsoli
dfoodmat r
ixthatcontai
nshighlevelofpart
iculate
mattersandhelpustominimizethesamplequantit
yandt ocalculat
ethetoonumer ous
tot
alcount(TNTC)ofthefermentsampleeasily
.
3
I
ft hecounti sexpectedt obemor ethan2.5x10 permlorgr am, pr
eparedecimal
di
lutions,shakeeachdi luti
on25timesin30cm ar c.Foreachdi lut
ionusefreshpi
pette.
Pipette01ml offoodhomogenat efrom 1stdi
lut
ionandtransf er01ml ofsampleto2nd
di
lutiontubecont ai
ning09ml ofdil
uentr ng3rd,
epeatusi 4thandmor etubesunti
lthe
desiredil
utionisobtained.
Appar
atus
 St
eri
l
izedpet
ripl
ates
 Cul
tur
ebot
tl
es/Di
l
uti
onBot
tl
es
 Cover
edTher
most
ati
cwat
erbat
htomai
ntai
nthet
emper
atur 500
eofAgarat45-
C.
 Spr
itLamp
 Bot
tl
edDr
inki
ngWat
erPr
ocess
 Ver
ti
cal
airf
lowcabi
netwi
thUVLamp.
 Magni
fyi
ngLens
0
 I
ncubat
orf
orbev
eragesat25 CTemper
atur
e.
 I
ncubat
orf
ordr
inki erat370CTemper
ngwat atur
e.
 Dr
yov
erf
ort
hest
eri
l
izat
ionofgl
asswar
es.
 Aut
ocl
avef
orChemi
cal
sst
eri
l
izat
ion.
 Wei
ghi
ngbal
ance
 Testt
ubeswi
thscr
ewl
i
ds
 Mi
cropi
pet
tewi
thst
eri
l
izedr
emov
abl
eti
ps
 St
eri
l
izedgr
aduat
edgl
asspi
pet
tes(
01and10ml
)
Chemi
cal
s
 Desi
reCul
tur
eMedi
as.
 Absol
uteEt
hanol
.
 Spr
it
.
 Pept
onesbuf
fer
edwat
er.

Pr
ocedur
e
Al
ltheappar
atusandchemi
cal
sshoul
dbest
eri
l
ized.
Takesampl
ebywai
t01gm or01ml
andmakenecessar
ydi
l
uti
onsbyr
ati
o01:
09.
Addanddissol
vethesamplei
nt esttubecont
aini
ng09ml
ofpept
onesbuf
fer
edwat
er
st
di
luentandswi
rlt
omi xi
tfor1 dilut
ion.
Pipett
e01ml off
oodhomogenat
efom 1stdi
r l
uti
onandtransf o2nddi
ert lut
iont
ube
contai
ning09mlofdi
l omake2nddi
uentt l
utionofthesampleandswi
rlt
omi xi
t.
Repeatbyusi
ngt
hi hodmake3rd,
smet 4thandmor
etubesunt
ilt
hedesi
redi
l
uti
oni
s
obtai
ned.
Shakeeachdi
l
uti
on25t
imesi
n30cm ar
c.
Foreachdi
l
uti
onusef
reshpi
pet
te.
Take01ml
ofsampl
efr
om eachdi
l
uti
oni
ntopet
ripl
atesbyusi
ngmi
cropi
pet
te.
Nowf
eedt
hedesi
recul
tur
emedi
ainpet
ripl
atesandmi
xitbyshaki
ngcl
ockwi
seand
 Bot
tl
edDr
inki
ngWat
erPr
ocess
ant
i-
clockwi
se.
Thet
emper
atur
eoft
hecul
tur asmustbeunder500C.
eMedi
Nowt
hel
ett
hepet
ripl
atesf
orsol
i
dif
y.
I
nvertt
hepetr
i pl
atesandi
ncubat
einani
ncubat
orundert
her
equi
redt
emper
atur
efor
i
ncubati
onpurpose.
Observ
eandcountt hecoloni
esafter24hour
soff i
rstobser
vat
ionandf
inal
l
yaf
ter48
hourofincubat
ionandtherepor
ttheresul
tcaref
ull
y.

101 102 103

t
h
1stD. 2ndD. 3r
dD. 4 Di
l
uti
on......
Sampl
e

Cal
cul
ati
on.
01gorml
sampl
eint
o09ml
ofpept
oneswat
er.
Supposet
hatt
her
ear
e05col
oni
esar
eobser
v n3rddi
edi l
uti
oni
ncubat
edsampl
eit
means,

. . .. .
3RD Di
lut
ion Pet
ripl
ate.

5cf
u
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Tot
alcounti
n01ml e= 5x103
ofsampl
= 5x10x10x10
= 5000cf
u(col
onyf
ormi
nguni
t)
Byusi
ngtheMPNMet
hodofmi
crobi
ologyt
est
ingwecanmaker
esul
tcount
abl
efr
om a
TNTCResul
t.
0
Note:fr
ozensamplecanbesoft
enbystor
ageitfor18hour
sat02-
05 Coftemper
atur
e
butdon’tt
hawthesampleformaki
ngthedi
lut
ions.Thaw:mel
tbygiv
ingt
emperat
ureor
heati
ng.
Pr
ecaut
ion
 Thegener
al asepti
ctechniquesforgoodworki
ngandf
orgoodr
esul
twhi
ch
shoul
dami crobiol
ogistf
ollow,ar
egivenas,
 Washhandsandwearmask,
glov
esandf
ootcov
ers.
 Cl
eananddi
sinf
ectal
lthei
ncubat
ionar
eawi
thswabsoakedi
nmet
hyl
atedspr
it
.
 Useet
hanol
spr
ayi
ntheenv
ironmentofi
ncubat
ion.
 St
eri
l
izeal
lthegl
asswar
esandchemi
cal
s.
 Nocr
oss-
cont
ami
nat
ionoft
hesampl
esandcul
tur
eMedi
aswi
thmi
croor
gani
sm.
 Don’ttouchanypar
toft
hegl
asswar
escomi
ngouti
ncont
actwi
tht
hesampl
eor
cul
ture.
 Don’tr
emovel
i
dofapet
ridi
shorcapofat
estt
ubel
ongert
hanabsol
utel
y
necessar
y.
 Light
lyf
lametopofthetestt
ubes,endsofpi
pet
tes,neckofcul
tur
ebot
tl
esand
thepetr
ipl
atesbef
oreandafteraddi
ngthesamples.
 Transf
ert
hecultur
efrom onet
ubetoanotherfr
eshtubeforsub-
cul
tur
ingal
way
s
useaster
il
izedwir
eloopbyheati
ngtorednessandcooled.
Al
waysfoll
owt
hesepr
ecaut
ionst
oav
oidt
hecr
oss-
cont
ami
nat
iondur
ingi
ncubat
ion
pr
ocess.

Compar
isonbet
weenMPNMet
hodandMFTechni
que
Mul
ti
plef
erment
ati
ont
ubet
echni
que/MPN. Membr
anef
il
tert
echni
que
MFTechni
que.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Sl
ower
:requi
res48hour
sforaposi
ti
ve. Mor
erapi
d:quant
it
ati
ver
esul
tsi
nor
Pr
esumpt
iveposi
ti
veabout18hour
s.

Mor
elabor
-i
ntensi
ve. Lessl
abor
-i
ntensi
ve.

Requi
resmor
ecul
tur
emedi
um. Requi
resl
esscul
tur
emedi
um.

Requi
resmor
egl
asswar
e. Requi
resl
essgl
asswar
e.

Mor
esensi
ti
ve. Lesssensi
ti
ve.

Resultobt
ainedi
ndi
rect
lybystati
sti
cal Resul
tsobt
aineddi
rect
lybycol
onycount
.
approxi
mation.(
Lowpreci
sion)
(
highpr
eci
sion)

Notr
eadi
lyadapt
abl
eforusei
nthef
iel
d. Readi
lyadapt
edf
orusei
nthef
iel
d.

Appli
cabl
etoal
lty
pesofwat
er,
foodand Notappl
icabl
etot
urbi
dwat
ers,
jui
cesand
bever
ages. f
oods.
Maygi
vebet
terr
ecov
eryofstr
essedordamaged
or
gani
smsinsomecir
cumstances

AgarFeedi
ngMet
hod
Ther
ear
etwot
ypesofmet
hodsf
oragarf
eedi
ngi
nmi
crobi
alt
est
ing.
1.Spr
eadPl
ateMet
hod
2.PourPl
ateMet
hod
1.Spr
eadPl
ateMet
hod.
Spr
eadpl
atemethodi
sanagarf
eedi
ngmethodinwhichrequi
redagari
stakeni
npet
ri
pl
ate,
sol
i
dif
yitt
hensampl
eistobespr
eadonitfori
ncubati
on.
2.PourPl
ateMet
hod.
Pourplatemethodisanagarfeedi
ngmet hodi
nwhi chr
equir
edsampl
eistakenina
petr
iplatet
henAgarpourintothesampli
ngplateandswir
ltomixi
tcl
ockwiseandant
i
-cl
ockwiseandthenleti
ttosoli
dif
yfori
ncubati
on.

Compar
isonbet
weenAgarPourPl
ateandAgarSpr
eadPl
ateMet
hod
 Bot
tl
edDr
inki
ngWat
erPr
ocess
PourPl
ateMet
hod Spr
eadPl
ateMet
hod
1.Growthofor
gani
sm thatneedloweroxygen 1.Growthofinj
ur edormicr
oEurophi
l
ic
tensi
oncel
lmaybefavoredsuchasmi cro mayber educedbecausetheAgar
Europhi
l
icor
gani
sm orinj
uredcell
s. surf
acehashi gheroxy
gentensi
on.

2.Volumeplated(0.
1or01ml samples)canbe 2.Onlysmal
l volumes(0.
1-0.
5ml)can
vari
ed;Lar
gev ol
umes01ml maybemor e beplat
ed,thismaybelessaccur
ate
accurat
eindetect
ingLowcellnumbers. fordet
ect
ingl evel
sofBact
eri
a.

3.Colonymor
phologyisnotusual
lyobser
ved, 3.Col
onymor
phol
ogyandPi
gment
ati
on
becausemostcoloni
esareimbeddedinAgar
. ar
eeasi
l
yobser
ved.

4.I
mbeddedcoloni
esdon’
ttendt
ospr
ead
t
hroughoutt
hepl
ate. 4.Spr
eader
”canbeapr
obl
em.

5.Insomecase, hi
ghercount
smay
5.Warm Agarusemayinhi
bitt
hegr
owt
hofhi
ghl
y occurbecauseorganismsarenot
heatsensi
ti
veor
gani
sms. exposedtowar m Agar.

6.Agarmustbest
eamedandt edat450C
emper 6.Agarpl
atescanbepr
epar
edanddr
ied
bef
oreuse. aheadofti
me.

7.Tr
ansl
ucentorOpaquemedi
acanbe
7.Agarmedi
um mustbet
ransl
ucent
.
used.

Bot
tl
edr
inki
ngwat
er
‘
’Waterthatisint
endedforhumanconsumpt i
onandthati
sseal
edinbott
leswit
hno
addedingredi
entsexceptt
hatitmaycontai
nsafeandsuit
abl
eanti
-mi
crobial
agent
wit
hinthelimit
sandmi ner
alsandfl
uori
des.‘
’
Det
ect
ion/Enumer
ati
onofMi
crobesi
nBot
tl
edr
inki
ngwat
er.
1.Tot
alpl
atecountBact
eri
a
2.Tot
alcol
i
for
m Bact
eri
a
3.Escher
ichi
acol
i
4.Faecal
Str
ept
ococcus
5.PseudomonasTest
 Bot
tl
edDr
inki
ngWat
erPr
ocess
6.Sal
monel
l
aShi
j
ell
aTest
.
7.YeastandMol
d

1.Tot
alPl
ateCountBact
eri
a
Theaerobi cpl
atecount(APC)i si
ntendedtoindicat
ethelev el ofmicroor
ganism ina
product.Detai
ledproceduresfordetermi
ningtheAPCoff oodsandbot tl
edr i
nki
ngwater
havebeendev elopedbyt heAssociati
onofOf f
ici
alAnalyt
ical Chemists(AOAC)andt he
Amer i
canPubl i
cHealthAssociati
on( APHA).ThisMicrobi
al testforthedrinki
ngwater
andbev eragescanbedet ectbyusingfol
lowingAgarMedi as.
 Nut
ri
entAgar
 Nut
ri
entBr
oth
 Pl
atecountAgar
 R2AAgar
 Tr
ypt
icSoyAgar

 Tr
ypt
oneGl
ucoseExt
ractAgar

2.Escher
ichi
aCol
i.
Escher ichiacol i,originalyknownasBact
l eri
um col icommune, andi swi delydist ri
but ed
i
nt hei ntest i
neofhumansandwar m-bl oodedani malsandi st hepr edomi nant
facultativeanaer obei nt hebowel andpar toft heessent ial i
ntestinalflorathatmai ntains
thephy si
ol ogyoft heheal t
hyhost .E.col iisamemberoft hefami lyEnt erobacter iaceae
whichi ncl udesmanygener a, i
ncludingknownpat hogenssuchasSal monel la,
Shi gella,
andYer sinia.Al t
houghmostst r
ainsofE.col iar enotregar dedaspat hogens, t
heycan
beoppor tuni sti
cpat hogenst hatcausei nfectionsinimmunocompr omi sedhost s.Ther e
areal sopat hogeni cst rainsofE.col ithatwheni ngested, causesgast rointesti
nal i
llness
i
nheal thyhumans. E.col iasani ndi catoroff ecalcontami nat i
on.Thi swasbasedont he
premi set hatE.col iisabundanti nhumanandani mal fecesandnotusual l
yfoundi n
otherni ches.Fur ther mor e,sinceE.col icoul dbeeasi lydet ectedbyi t
sabi l
i
tytof erment
glucose( l
at erchangedt ol actose),itwaseasi ertoisolatet hanknowngast roi
nt est i
nal
pathogens.He nce, t
hepr esenceofE.col iinfoodorwat erbecameaccept edas
i
ndi cativeofr ecentf ecal cont aminat i
onandt hepossi blepr esenceoff rankpat hogens.
E.coli(commonl yknownaspat hogenicE.coli
),ar
eagr oupofE.coliwhi
chcause
di
ar r
heal di
seaseinhumans.Ther earefourgroupsofdiarr
heagenicE.coliwhi
chhave
previ
ouslybeenassociatedwithfoodborneil
lness;ent
erotoxi cE.col
geni i(ETEC),
enteropat cE.col
hogeni i(EPEC),ent
erhemor r
hagicE.coli(EHEC)andenteroi
nvasi
veE.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
col
i(EI
EC).Thesegroupsar
ecl
assi
fi
edbyt
hei
rvi
rul
encef
act
orsandcanbei
dent
if
ied
bythei
rvi
rul
encetrai
ts.
Thesepathogeni cE.coligroupshav ebeenassoci atedwitht hef oll
owingf oodty
pes,
•Enter
otoxigenicE.coli(ETEC) :softcheeses, Mexican-st
ylef oods, r
awv eget
abl
esand
water.
•Enter
opathogeni cE.coli(EPEC) :meatpr oductsandcont ami nateddrinkingwater
.
•Enter
ohemor rhagicE.col i(EHEC):under cookedgr oundbeef , r
awmi lk,cold
sandwiches,water,unpast euri
zedappl ejuice,spr
out sandv eget abl
es.
•Enter
oinvasiveE.coli(EIEC) :
hambur germeatandmi lk.
Theenumer
ati
onoft
heECol
icanbedet
ect
edbyusi
ngf
oll
owi
ngAgar
s,
 Eosi
nmet
hyl
enebl
ueAgar
.
 Tr
ypt
oneBi
l
eAgar
.
 E.
Col
ibr
oth.
 EndoAgarBase.
 Lact
oseBr
oth.
 Deoxy
chol
ateLact
oseAgar
.
 MacConkeyBr
oth.
 Tr
ypt
ophanBr
oth.

3.Tot
alCol
if
orm
"Coli
form"wascoi nedt odescribethisgroupofent er i
cbact eria.Coli
formi snota
taxonomi cclassifi
cationbutr atherawor ki
ngdef i
niti
onusedt odescribeagr oupof
Gram- negati
ve, f
acultativeanaer obicrod-shapedbact eri
athatf ermentslactoset o
produceaci dandgaswi t
hin48hat35° C.In1914, theU. S.Publ icHealthServ i
ce
adoptedt heenumer ationofcol if
ormsasamor econv enientstandardofsani tary
signi
ficance.Theconceptofusi ngE.col iasani ndi
recti ndicatorofheal t
hr i
skwas
sound, i
twascompl icat edinpractice,duet othepresenceofot herentericbacteri
al i
ke
Citr
obact er
,Klebsiell
aandEnt erobact erthatcanalsof er
mentl act
oseandar esimilarto
E.coliinphenot ypi
cchar acter
isti
cs, sothattheyarenoteasi lydisti
nguished.Asar esul
t,
theterm" coli
form"f ound.Col iformsar eeasyt odet ectast heirassociati
onwi thfecal
contami nat
ionwasquest ionablebecausesomecol iformsar ef oundnat ural
lyin
envir
onment al sampl es.
Fort
heenumer
ati
onofTot
alCol
i
for
m asol
i
dmedi
um pl
ati
ngmet
hodi
susedi
nwhi
ch
 Bot
tl
edDr
inki
ngWat
erPr
ocess
VioletRedBil
eAgarisused,whichcontai
nsneutr
alredpHindi
cator
,sothatlact
ose
fermentati
onresul
tsi
nformati
onofpinkcoloni
es.Theenumerat
ionofTotalcoli
for
m
canbedet ectbyusi
ngfoll
owi
ngAgar sorCult
ureMedias,
 Vi
oletRedBi
l
eAgar
.
 Br
il
li
antGr
eenBi
l
eAgar
.
 Chr
omogeni
cCol
i
for
m Agar
.
 ChapmanTTCAgar
.
 Tr
ypt
oneBi
l
eGl
cucr
oni
cAgar
.
 Tr
ypt
oseLaur
ylSuphat
eBr
oth.
 Tr
ypt
oneWat
erPH7.
5
 Chr
omogeni
cCol
i
nst
antAgar

St
eri
li
zat
ion.

St
eri
l
izeal
lequi
pment
’sandmat
eri
als:

 aut
ocl
avef
orst
eam st
eri
li
zat
ion:

 1200Cforcul
tur
ebr
oths/AgarMedi
aanddi
l
uti
onwat
erf
or15
minut
es

At15-
20Psipr
essur
e.

 ovenf
ordr
yst
eri
li
zat
ion:

 1700Cf
orpi
pet
tesi
nmet
alcont
ainer
s,20-
30mi
nut
es

Nocr
oss-contaminat
ionofsampl
esandcul
tur
emedi
awi
thmi
cro-
organi
smsdur
ing
t
heautocl
aving.

Mi
crobi
ologyLabSaf
etyPol
icy
:
Microbiol
ogyhasspecif
icsafetyi
ssuesrel
ati
ngt
orisksinvolv
edfrom wor
kingwithinf
ect
ious
agents.Properl
abor
atoryprocedur
es,equi
pmentandfacil
i
tiesneedtobeinplace.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Pur
pose:

Hav
ingt
heproperpr
ocedur
es,
equipmentandfacil
i
tiesinplacewi
l
lel
i
minat
eoratl
eastr
educe
t
heri
ski
nvol
vedwithworki
ngwit
hinfect
iousagent
s/ di
seases.

Saf
etyPr
ecaut
ions;

 Beawaret hatbacteri
ausedinmicrobiol
ogylabcanmakey ouorotherpeopl
esickwho
workwithy ou.I
tispossibl
eforbacter
iatobebr oughti
ntot
heotherworki
ngareas
thr
oughcont aminatedlabcoat
s,pens,notebooksandotheri
temsthatareusedint
he
microbi
ologylaborator
y.

 Personswor
kingwit
hinfect
iousagents,i
ncludi
ngE.Col
i&Salmonellabact
eri
a,mustbe
awareofpot
enti
alhazar
ds,andmustbet r
ainedinbi
osaf
etypract
icesandtechni
ques
requi
redf
orhandl
ingsuchagentssafely
.

 Washhandsfr
equentl
ywhil
eworki
ngi
nandi mmedi
atel
yaf
terl
eav
ingt
hemi
crobi
ology
l
abandfol
lowproperhandwashi
ngpr
act
ices.

 Wearal
abcoatorotherpr
otect
iveuni
for
m ov
erper
sonal
clot
hingwhenwor
kingi
na
mi
crobi
ologyl
abor
atory
.

 Ify
ouwor
kwithE.
Col
i&Sal
monel l
abacter
iai
nami cr
obiol
ogyl
aborat
ory,
watchfor
sy omsofSal
mpt monel
lai
nfect
ion,suchasdi
arr
hea,f
everandabdominal
cramps.

 DonotuseMi crolabapparatusoutsi
det hemicrolabforanyotherpurposeast heyare
pot
enti
all
ycont aminated&considerallqual
it
ycontrolmateri
alsaspotenti
all
yinfect
ious.
 Per
form proceduresthathav
et hepotenti
altogenerateaer
osolsordroplet
sina
bi
ologi
calsafetycabinet
.Keepbi ol
ogicalsaf
etycabinet
sclearofclut
ter.

 Al
way
schangegl
ovesi
ftheyhav
ebecomegr
ossl
ycont
ami
nat
edaf
terev
erysampl
i
ng.

 Proceduresforcl
eaning,di
sinf
ect
ionandst
eri
l
izat
ionofl
abor
ator
yequi
pment
,suppl
i
es,
andenv i
ronmentalsurf
acesshoul
dbeav ai
l
abl
e.

 Cleananddi
sinf
ecttheal
lequipmentofmicr
olabatdai
l
ybasi
saf
tersampl
i
ng
/contami
nat
ionwit
hanybiological
mater
ial
.

Bact
eri
alCommonGr
oups
 Bot
tl
edDr
inki
ngWat
erPr
ocess
•Col
if
orms

 Totalcol
if
orms
 Fecalcol
if
orms
 Escheri
chi
acol
i
•St
rept
ococci
f ecalstr
eptococci
 enterococci
•Spor
eFor mer s
Clostr
idi
um
 Perf
ri
ngens

Bact
eri
aFoundI
nSur
face/
GroundWat
er

Bact
eri
a Di
sease/i
nfect
ion Sy
mpt
oms
Aer
omonas Ent
eri
ti
s Verythi
n,bl
ood-andmucus-
contai
ningdi
arr
hea

Campy
lobact
erj
ejune Campi
l
obact
eri
ose Fl
ue,diar
rhea,headand
stomachaches,fev
er,cr
ampsand
nausea

Escher
ichi
acol
i Uri
narytr
actinfecti
ons, Water
ydiar
rhea,headaches,
fev
er,
neonatal
meni ngit
is, homil
eti
curemia,ki
dneydamage
i
ntesti
naldi
sease
Pl
esi
omonasshi
gel
l
osi
s’
s Pl
esi
omonas-
inf
ect
ion Nausea,stomachachesandwat
ery
di
arrhea,somet
imesf ev
ers,
headachesandvomi t
ing

Ty
phus Ty
phoi
dfev
er Fev
ers

  Sal
monel
l
osi
s Sickness,i
ntest
inal
cramps,
Sal
monel
l
a vomi t
ing,
diarr
heaandsometimes
l
ightfevers
 Bot
tl
edDr
inki
ngWat
erPr
ocess
St
rept
ococcus (
Gast
ro)i
ntest
inal
disease Stomachaches,
diar
rheaandf
ever
s,
someti
mesv omit
ing

Vi
bri
oElTor(
freshwat
er) (
Lightf
orm of
)Chol
era Heav
ydi
arr
hea

Env
ironmentMoni
tor
ing/SwabTestLi
mit
s
Sr
.# Mi
crobi
ologi
calExami
nat
ion Mi
crobi
ologi
calLi
mit
s

1 Tot
alPl
ateCountascf
u cm2
100/

2
2 Tot
alCol
i
for
ms o/c
Zer m

2
3 St
aphy
lococcusAur
eous 20/cm

2
4 Yeast&Mol
d 20/cm

Mi
crobi
alLi
mitf
orSol
idSampl
es
Mi
crobi
ologi
calExami
nat
ion

Sr
.# Mi
crobi
ologi
calLi
mit
s

1 5×105
Tot
alPl
ateCountascf
u/g

2 Zer
o/25g
E-
Col
i
 Bot
tl
edDr
inki
ngWat
erPr
ocess
3 Zer
o/25g
Sal
monel
l
a

4 St
aphy
lococcusAur
eous Zer
o/25g

Mi
crobi
ologi
calLi
mit
sforDr
inki
ngWat
er

Sr
.# Mi
crobi
ologi
calExami
nat
ion Mi
crobi
ologi
calLi
mit
s

1 Het
erot
rophi
cPl
ateCountcf
u/ml <500/
100ml

2 Tot
alCol
ifor
ms Zer
o/100ml

3 Feacal
Col
ifor
ms Zer
o/100ml

4 E-
Col
i Zer
o/100ml

5 Pseudomonas Zer
o/100ml

Wat
erGl
ossar
y
Absol
ute
Themicr
onrangeofaf
il
ter
,iti
ndi
cat
est
hatanypar
ti
clel
argert
hanaspeci
fi
csi
zewi
l
l
betr
appedwit
hint
hefi
l
ter.
Absor
pti
on
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Whenasol
i
dtakesupmol
ecul
esi
ntoi
tsst
ruct
ure.
Aci
dicwat
er
pHbel
ow7showst
hatsubst
ancesamountofaci
dcont
ains.
Aci
dit
y
Thequantit
at i
vecapaci
tyofwatert
oneutral
i
zeabase, expr
essedinppm orgm/ L
Calci
um carbonateequi
v al
ent
.Thenumberofhydrogenatom thatar
epresent
determi
nest hi
s.Iti
sactual
lymeasuredbyti
tr
ati
onwithastandardsolut
ionofsodium
hydroxi
de.
Act
ivat
edCar
bon
Thisisthemostcommonl yusedadsor pt
ionmedi um,producedbyheat i
ng
carbonaceoussubstancesorcellul
osebasei ntheabsenceofair.Ithasav er
yporous
str
uctureandiscommonl yusedt oremov eorganicmatteranddissolvedgassesf
rom
water.It
sappearanceissimilart
ocoal orpeat.Avai
labl
eingranular,powderorbl
ock
for
m; inpowderformithast hehighestadsorpti
oncapacity.
Adsor
pti
on
Separ
ati
onofl
iqui
ds,
gasses,
coll
oidsorsuspendedmat
terf
rom amedi
um by
adher
encet
othesurf
aceorporesofasoli
d.
Aer
ati
on
Techniquethatisusedwi t
hwatertreatmentthatdemandsoxy gensupply,commonly
knownasaer obicbiologicalwaterpuri
fi
cati
on.Eit
herwat eri
sbroughtintocont
actwit
h
waterdropl
etsbyspr ayingorairisbroughti
ntocont actwit
hwat erbymeansof
Aerat
ionfacil
it
ies.Airispressedthroughabodyofwat erbybubblingandthewateri
s
suppl
iedwithoxy genwhi lethet
ankt hati
susedt oinjectai
rint
owat eri
sknownas
Aerat
iontank.
Al
kal
ini
ty
Alkal
init
ymeanst hebuff
eringcapacit
yofwat er
;thecapacit
yofwatertoneut r
ali
ze
i
tself
.Itprevent
st hewaterpHl evel
sfrom becomingtoobasicoracid.I
talsoadds
carbontowat er.Alkal
i
nit
yst abi
li
zewat eratpHlevel
saround07.Howev er
, whenthe
acidi
tyishighinwat er
,Alkali
nit
ydecreases,whi
chcancausehar mfulconditi
onsfor
aquaticli
fe.
Inwaterchemist
ryAlkal
init
yisexpressedinppm ormg/Lfr
om equi
val
entt
ocalci
um
carbonat
e.Total
Alkali
nit
yofwat eristhesum ofal
lthr
eesor
tsofAl
kal
ini
ty;
car
bonates,
bicar
bonatesandhydroxideAlkal
ini
ty.
Aquat
ic
Gr
owi
ngi
nwat
er,
li
vi
ngi
nwat
erorf
requent
ingwat
er.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Aqueous
Somet
hingmadeofwat
er.
Aqui
fer
Al
ayeri
nthesoi
lthati
scapabl
eoft
ranspor
ti
ngasi
gni
fi
cantv
olumeofgr
oundwat
er.
Bact
eri
a
Mi
croscopi
cal
l
ysmal
lsi
ngl
e-cel
lor
gani
sm,
thatr
epr
oducebyf
issi
onofspor
es.
Bi
ochemi
calOxy
genDemand(
BOD)
Theamountofoxygen(measur
edinmg/L)t
hatisrequi
redfordecomposi
ti
onof
organi
cmatt
erbysingl
ecell
organi
sms,
undertestcondi
ti
ons.Iti
susedtomeasur
ethe
amountofor
ganicpoll
uti
oninwast
ewat
er.

Bi
oci
de
Achemical
thatistoxi
ct omicr
oorgani
sm.Bi
oci
desar
eof
tenusedt
oel
i
minat
ebact
eri
a
andot
hersingl
e-cel
lorgani
sm fr
om water
.
Bi
ofi
lm
Populat
ionofv
ari
ousmicr
oor
gani
sm,
trappedi
nal
ayerofsl
i
meandexcr
eti
onpr
oduct
s,
at
tachedtoasurf
ace.
Bi
ologi
calCont
ami
nant
s
Li
vingorgani
sm suchasvir
uses,
Bact
eri
a,Fungi
,andmammal
andbi
rdant
igenst
hat
cancauseharmfulheal
theff
ectt
ohumans.
Boi
li
ngpoi
nt
Thetemper at
ureatthevaporpressureofali
qui
dequalthepressur
eofit
ssurface.The
l
iqui
dwi l
lthanvapori
zedifthepressureoft
heli
qui
dv ar
ies,
theactual
boil
ingpoint
vari
es.Forwatertheboi
li
ngpointis100degreeCel
sius.
Br
ine
Highlysal
tyandheav
il
ymi
ner
ali
zedwat
er,
cont
aini
ngheav
ymet
alandor
gani
c
contaminant
s.
Buf
fer
Asubstancehatr
eact
swi
thhy
drogenorhy
droxy
lionsi
nasol
uti
on,
inor
dert
opr
eventa
changeinpH.
Cal
cium hy
pochl
ori
te
Achemi
cal
thati
swi
del
yusedf
orwat
erdi
sinf
ect
ion,
fori
nst
ancei
nswi
mmi
ngpool
sor
 Bot
tl
edDr
inki
ngWat
erPr
ocess
waterpur
if
icat
ionpl
ant
s.Iti
sespeci
all
yusef
ulbecausei
tisast
abl
edr
ypowderand
canbemadei ntot
abl
ets.

CFU
Col
onyfor
minguni
ts.Thi
sisameasur
ethati
ndi
cat
est
henumberofmi
croor
gani
smsi
n
wat
er.
Chl
orami
nes
Achemical
compl exthatconsistofchlorineandammoni a.I
tserv
esasawater
di
sinf
ect
antinpubli
cwat ersuppli
esi nplaceofchlor
inebecausechl
ori
necancombi
ne
wi
thorgani
cstoform dangerousr eacti
onpr oduct
s.
Chl
ori
nat
ion
Awaterpurif
icati
onprocessi
nwhichchl
ori
nei
saddedt
owaterfordi
sinf
ect
ion,
fort
he
cont
rolofpresentmicroor
gani
sm.Iti
sal
sousedi
ntheoxi
dat
ionofcompound
i
mpurit
iesinwat er
.
Chl
ori
necont
actChamber
Thepar
tofawat
ert
reat
mentpl
antwher
eef
fl
uenti
sdi
sinf
ect
edbychl
ori
ne.
Concent
rat
ion
Theamountofmat
eri
aldi
ssol
vedi
nauni
tofsol
uti
on,
expr
essedi
nmg/
L.
Condensat
e
Wat
erobt
ainedbycondensat
ionofwat
erv
apor
s.
Condensat
ion
Thechangeofst
atef
rom agast
oal
i
qui
d.
Conduct
ivi
ty
Theamountofel
ect
ri
cit
ythatwat
ercanconduct
.Iti
sexpr
essedi
nachemi
cal
magni
tude.
Cool
ingTower
Lar
getowerusedtot
ransfert
heheatincooli
ngwaterf
rom apowerorindust
ri
alpl
ant
tot
heatmosphereei
therbydi
rectev
aporati
onorbyconvect
ionandconducti
on.
Cor
rosi
vi
ty
Abi
l
ityofwat
ert
odi
ssol
veorbr
eakdowncer
tai
nsubst
ances,
par
ti
cul
arl
ymet
als.
Cr
ypt
ospor
idi
um
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Ami croor
ganism i
nwatert
hatcausesgastroi
ntest
inal
il
lnessi
nhumans.I
tis
commonl yfoundinuntr
eat
edsurfacewaterandcanber emovedbyf
il
tr
ati
on.i
tis
resi
stantt
odisinf
ectant
ssuchasChlori
ne.
De-car
bonat
ion
Theprocessofr
emov
ingcar
bondi
oxi
def
rom wat
er,
usi
ngcont
actt
ower
sorai
r
scr
ubbers.
Di
ff
user
Acomponentoft
heozonecontact
ingsy
stem i
nanozonegener
atort
hatal
l
ows
di
ff
usi
onofanozonecontai
ninggas.
Di
ff
usi
on
Themov ementofgasmol
ecul
esoraer
osol
sint
oli
qui
ds,
causedbyaconcent
rat
ion
gradi
ent
.
Di
sinf
ect
ion
Thedecont aminat
ionofflui
dsandsurfaces.Todisi
nfectaflui
dorsur
faceavar
iet
yof
techni
quesar eusedsuchasOzonedi sinf
ecti
on.Oftendisi
nfect
ionmeansel
i
minati
ng
thepresentmicroorgani
smswithabiocide.
Di
sti
ll
ati
on
Watertr
eatmentmethodwherewaterisboi
l
edtosteam andcondensedi
nasepar at
e
r
eservoi
r.Cont
aminantswi
thhigherboi
li
ngpoi
ntst
hanwat erdon’
tvapor
izedand
r
emainintheboil
i
ngf l
ask.
Ev
apor
ati
on
Thepr
ocessoft
hepassageofwat
erf
rom l
i
qui
dtov
apor
.
Fi
lt
rat
ion
Separ
ati
onofasol
i
dandal
i
qui
dbyusi
ngapor
ousmedi
um t
hatonl
ylet
sthel
i
qui
dto
passthr
oughi
t.
Fi
ssi
on
Repr
oduct
ionofmi
croor
gani
smsbymeansofcel
ldi
vi
sion.
Fl
oc
Afloccul
entmasst hati
sf or
medi ntheaccumulat
ionofsuspendedparti
cles.I
tcan
occurnatural
l
y, butisusuall
yinducedi
nor dert
obeabletoremov ecer
tai
npar ti
cles
fr
om wast ewater.
Fl
occul
ati
on
Theaccumul
ati
onofdest
abi
l
izedpar
ti
clesandmi
crof
lakes,
andsubsequent
lyt
he
 Bot
tl
edDr
inki
ngWat
erPr
ocess
f
ormat
ionofsizeabl
efl
akes.Onemustaddanotherchemi
cal
cal
l
edf
loccul
enti
nor
der
t
ofaci
l
itat
etheformati
onofflakescal
l
edf
locs.
Fl
ux
Ther
ateatwhi
char
ever
seosmosi
smembr
aneal
l
owwat
ert
opasst
hrought
oit
.

Foul
ing
Thedecomposi
ti
onofor
gani
cmat
teront
hemembr
anesur
face,
whi
chcauses
i
neff
ici
enci
es.
Gal
lon
Auni
tthati
snowent
ir
elyoutofdat
e.I
tisequi
val
entt
o3.
785l
i
ter
s.
Henr
y’sLaw
Awayofcal
cul
ati
ngt
hesolubi
l
ityofagasi
nli
qui
d,basedont
emper
atur
eandpar
ti
al
pr
essur
ebymeansofconst
ants.
Hy
drogenSul
fi
de(
H2S)
Agasemit
tedduri
ngorgani
cdecomposi
ti
onbyasel
ectgr
oupofbact
eri
a,whi
ch
st
rongl
ysmel
lsl
ikerot
teneggs.
Hy
drol
ysi
s
Thedecomposi
ti
onofor
gani
ccompoundsbyi
nter
act
ionwi
thwat
er.
Hy
drophi
li
c
Hav
inganaf
fi
nit
yforwat
er.
Hy
drophobi
c
Hav
inganav
ersi
onf
orwat
er.
I
mmi
sci
bil
it
y
Thei
nabi
l
ityoft
woormor
esol
i
dsorl
i
qui
dst
oreadi
l
ydi
ssol
vei
ntooneanot
her
.
Per
meabl
e
Easi
l
ypenet
rat
edbywat
er.
I
nfl
uent
Thest
ream ofwat
ert
hatent
ersi
nanysy
stem orwat
ert
reat
mentuni
t.
Ki
net
icEner
gy
Ener
gypossessedbymov
ingwat
er.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Lami
narFl
ow
Af
lowi
nwhi
chr
api
dfl
uct
uat
ionsar
eabsent
.
Osmosi
s
Watermolecul
espassi
ngthr
oughmembranenat
ural
l
y,t
othesi
dewi
tht
hehi
ghest
concent
rat
ionofdi
ssol
vedimpur
it
ies.
Oxi
dat
ion
Achemi
cal
react
ioni
nwhi
chi
onsar
etr
ansf
err
ingel
ect
ronst
oincr
easeposi
ti
vev
alence.
Oxy
genDepl
eti
on
Ther
educt
ionoft
hedi
ssol
vedoxy
genl
evel
inawat
erbody
.
Ozone
Anunstableoxi
dizi
ngagentthatconsi
stofthr
eeoxygenatomsandcanbefoundinthe
ozonelayeri
natmosphere.I
tisproducedbyelect
ri
caldi
schar
gethr
oughoxygenorby
speci
all
ydesignOzonegenerator
.
Par
tsperBi
ll
ion
Expr
essedasppb;
auni
tofconcent
rat
ionequi
val
entt
otheµg/
L.
Par
tsperMi
ll
ion
Expr
essedasppm;ameasur
eofconcent
rat
ion.Oneppm i
soneunitweightofsol
ute
permil
l
ionuni
twei
ght
sofsolut
ion.I
nwat
eranalysi
stheppm i
sequiv
alenttomg/L.
Past
eur
izat
ion
Theeliminati
onofmi cr
oorgani
sm byheatappli
esforacer
tainper
iodofti
me.The
pasteur
izati
onprocessiscarri
edoutfora(cont
inui
ngforalongti
me)prol
ongedti
meat
around75-80CTemper at
ureandissupposedtokil
lonl
yheatsuscept
ibl
eorgani
sm and
thei
rspores.
St
eri
li
zat
ion
Ster
il
izat
ionprocessworksatat
emper
atur
eof121andsupposedt
oki
l
lal
lpat
hogens
andnon-pathogensbact
eria.
Pat
hogens
Di
sease-
produci
ngMi
croor
gani
sms.
Per
meabi
li
ty
Theabi
l
ityofamedi
um t
opassaf
lui
dunderpr
essur
e.
Pr
eci
pit
ate
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Ani
nsol
ubl
ereact
ionpr
oducti
nanaqueouschemi
cal
react
ion.
Reser
voi
r
Anat
ural
orar
ti
fi
cial
hol
dingar
eausedt
ost
orewat
er.
Resi
due
Thedr
ysol
i
dsr
emai
ningaf
tert
heev
apor
ati
onofasampl
eofwat
erorsl
udge.
Sedi
ment
ati
on
Set
tl
ingofsol
i
dpar
ti
clesi
nal
i
qui
dsy
stem duet
ogr
avi
ty.
Subl
imat
ion
Thetr
ansit
ionsofwat
erdi
rect
lyf
rom t
hesol
i
dst
atet
othegaseousst
atewi
thout
passi
ngthrought
heli
qui
dstat
e.
Tur
bidi
ty
Ameasur
eofnon-
tr
anspar
encyofwat
erduet
othepr
esenceofsuspendedmat
ter
s.
Tur
bul
entFl
ow
Af
lowt
hatcont
ainsmayr
api
dfl
uct
uat
ions.
Vapor
Thegaseousst
ateofasubst
ancesuchaswat
er.
Vapor
ize
Conv
ersi
onofal
i
qui
dint
ovapor
s.
Wei
r
Aspi
l
lov
erdev
iceusedt
omeasur
eorcont
rol
thewat
erf
lows.
S
Additi
v esandi ngr
edients
Subst ancesaddedt ofoodstoinf
luencethei
rcondi
ti
onortobr
ingaboutspeci
fic
characterist
icsoreff
ects(hel
pmanuf act
ure,
preser
ve,andi
mprovepal
atabi
li
ty,
eye
appeal, conveni
ence-e.g.
Emul
sif
ier
s,

Fl
avor
s,thi
ckener
s,curi
ngagent
s,humect
ant
s,col
ors,
vit
ami
ns,
miner
als,
mol
ds,
yeast
s,
andbact
eri
alinhi
bit
ors)
.

All
ergy
Ar eacti
onprovokedbyanti
gencont
actwi
thant
ibodi
es.Symptomsincl
udewideningof
capil
lar
ies(
reddeningofski
normucousmembranes)
,incr
easeinper
meabil
i
ty(local
 Bot
tl
edDr
inki
ngWat
erPr
ocess
swel
l
ing)andsecr
eti
on(
tear
s,sput
um,
rhi
nit
is)
,it
chi
ng.

Ambientt
emperat
ure
Sur
roundi
ngtemperat
ure;
usual
l
yref
erst
oroom t
emper
atur
e.

Anti
biotics
Secondar ymet aboli
tesofmi
croor
gani
sm whi
ch,
insmal
lquant
it
ies,
cani
nhi
bitor
l
ethall
yhar m another
microorganism.

Bact
eri
a
Adiver
segroupofsi
ngl
e-cel
l
edorgani
smst
hatar
enei
therpl
ant
snorani
mal
s.Some
bact
eri
aar
euseful
;ot
hersarehar
mful.

Botul
ism
Neuromuscul arintoxi
cat
ioncausedbyCl ostri
dium botulinum toxi
n.Whenthe
veget
ati
v ecel
lsgr owinaf oodtheyproduceapot entexotoxinwhichcausesbotul
ism
wheningested.Itisthemostdanger oustypeoff oodpoi soningandisusual
lycausedby
under
-processedcont aminatedcannedorv acuum- packedf oods.

CodexAl iment
ar i
usCommi ssi
on(Codex)
TheCommi ssionwasj oint
lyest
abl
ishedbyFAOandWHOt odevel
opfoodstandar
ds
guideli
nesandr el
atedtextssuchascodeofpractice.It
smainpurposesarethe
protecti
onof
consumerhealth, ensur
ingfai
rtr
adepract
icesinthef oodt
radeandprovi
dingco-
ordinat
ionofallfoodstandardsdevel
opedbyinternati
onal
,gover
nmentalandnon-
governmentalorganizat
ions.

Cri
ti
calCont
rolPoi
nt(CCP)
Astepatwhichcontr
olcanbeappl
iedandisessent
ial
topr
eventorel
i
minat
eaf
ood
saf
etyhazar
dorreduceitt
oanacceptabl
elev
el.(
Codex)

Contaminant
Anybiologi
calorchemi
calagent
,for
eignmat
terorot
hersubstancesnotint
ent
ional
l
y
addedtofoodthatmaycompromisefoodsaf
etyandsui
tabi
li
ty.(Codex)

Cont
aminati
on
Theint
roduct
ionoroccur
renceofacont
ami
nanti
nfoodorf
oodenv
ironment
.(Codex)

CrossContami nation
Acont aminati
onoccur ri
ngduringtheproducti
on,
processi
ngorprepar
ationoff
ood,
ei
therthr
oughdi rectcontactofuncontaminat
edmateri
alswit
hcontaminatedmater
ial
s
orthr
ought r
ansmi ssionbyav ehi
cle.
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Dangerzone
Thetemperatur
erangemostconduci
vet
othemul
ti
pli
cat
ionofbact
eri
a(5oCt
o60oC)
.

Decontaminati
on
Sanit
ation,
disi
nfect
ion.Dr
ast
icr
educt
ionoft
hemi
crobi
alpopul
ati
on.

Deepf
reezi
ng
Amethodtoextendt
heshel
fli
feorkeepi
ngqual
it
yofaf oodpr
oductbyst
ori
ngi
tat
o
t
emperat
uresbel
ow-18Cunti
liti
sdeli
ver
edtotheconsumer.

Dehy
drat
ion
Lossofwater
.

Diar
rhea
Referstopassingaflui
dstool
swi t
hahi
ghf
requency
.Thi
sist
hemostt
ypi
cal
sympt
om
associ
atedwithfoodbornei
nfect
ion.

Disi
nfecti
on
Ther educti
on,bymeansofchemicalagent
sand/orphy
sical
methods,ofthenumberof
microorgani
smsi nt
heenvi
ronment,t
oal ev
elt
hatdoesnotcompromisefoodsafet
yor
suit
abil
ity
.(Codex)

Endemic:
Adiseasethatpr
evai
l
sorr
ecur
sfr
equent
lyi
nal
ocal
i
tyorpopul
ati
on.

Epidemic
Anout br
eakofani
nfect
iousdi
seaset
hataf
fect
smanypeopl
eatonet
imei
nthesame
area.

Extr
insi
c
Factorsext
ernal
toafoodthatmaybeappli
ed(e.
g.bytheprocessor)forext
endingt
he
shel
flif
eorkeepi
ngquali
tyofafood(
e.g.t
emperat
ure,
preservat
ives,stor
age).

Fermentat
ion
Adesi r
abl
epr ocessofbi
ochemical
modi
fi
cat
ionofpr
imar
yfoodpr
oduct
sbr
ought
aboutby
microor
ganismsandt hei
renzy
mes.

Foodhandl er
Anyper sonwhohandl espackagedorunpackagedfood,foodequi
pmentandutensil
s
orsurfacesincontactwi
thfoodandisther
eforeexpectedtocomplywi
thfoodhygiene
requi
rement s.(
Codex)
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Foodhy gi
ene
Al l
condit
ionsandmeasuresnecessar
ytoensur
ethesaf
etyandsui
tabi
l
ityoff
oodatal
l
stagesofthefoodchai
n.(
Codex)

Foodbor nedi sease

Adi sease, usuall
yeitheri
nfect
iousortoxicinnatur
e,causedbyagent
sthatent
ert
he
bodyt hrought heingesti
onoffood.Thet er
m“ food”i
ncludesdr
inki
ng-
water
.Someti
mes
thi
sisi ncorrectl
yreferr
edtoasf oodpoisoni
ng.

Foodpoi
soning
Thisi
sater
mt hati
sof
tenusedt
oref
ert
ofoodbor
nei
l
lness/
diseasebutWHOdoesnot
recommendit.

Foodsafety
Assurancethatf
oodwil
lnotcausehar
mt otheconsumerwheni
tispr
epar
edand/
or
eat
enaccordingt
oit
sint
endeduse.(Codex)

Foodspoil
age
Foodbecomesunf i
ttoeatasar esul
tof:
growthandacti
v i
ti
esofmicroorgani
sms,
i
nsecti
nfest
ation,andacti
onofenz ymes,chemical
react
ions,andphy
sicalchanges(
e.g.
fr
eezi
ng,burni
ng,dryi
ng,pressur
e,andhumi di
ty)
.

Foodtoxin
Compoundsnat ural
lypresenti
nfoodthatar
etoxi
corcar ci
nogeni
corhave
pharmacologi
caleff
ects.Thi
sincl
udesnatur
alcar
cinogensandnutri
ti
onal
inhi
bit
ors.
FungusSeeMol d.

Gastr
o-ent
eri
ti
s
Alsoknownasguti nf
ecti
on.I
nflammatorychangeoftheli
ningofstomachand
i
ntest
ineusual
lycausedbymicroorgani
smsingestedwit
hthef oodorwater
.

Gutmicr
obes
Micr
oorgani
smst
hatl
i
vei
nthegast
roi
ntest
inal
tractofmanorani
mal
s.

HACCP
“Hazar
dAnaly
sisandCri
ti
calCont
rolPoi
nt”
.Asyst
em thati
dent
if
ies,
eval
uat
es,
and
cont
rol
shazar
dsthatar
esigni
fi
cantf
orfoodsaf
ety
.(Codex)

Hazard
Abi ol
ogi
cal
,chemicalorphysi
calagenti
n,orcondi
ti
onof
,foodwi
tht
hepot
ent
ial
to
causeanadverseheal
theffect
.(Codex)
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Heatst
eri
li
zat
ion
Commonlypract
icedmet
hodf
ort
hedest
ruct
ion(
kil
l
ing)ofal
lvi
abl
emi
croor
gani
sms.

Heav
ymet
als

Withaspecif
icgrav
ityof>5,
suchasir
on,
lead,
copper,
zinc,
nickel
,chr
omi
um,
molybdenum,
cadmium,sil
ver
,aswellassemi-
metal
ssuchasarseni
c.

High-ri
skf ood
Thesei ncludefoodsthathavebeenli
nkedepidemiologi
cal
l
yt ofoodbornediseaseor
that,duetot hei
rnat
ure,pr
eparat
ionorstor
age,presentagreat
erriskoffoodborne
diseasethanot herf
oods.
Inter
nationalOrgani
zati
onf orSt
andardi
zati
on(ISO)
Agl obalnetworkofnationalst
andardsinst
it
uti
ons, wor
kinginpart
nershi
p
withinter
nationalor
ganizati
ons,gov
ernment s,
industr
y,busi
nessandconsumer
representat
ivestoestabli
shint
ernat
ionall
yrecognizedquali
tyandgoodpract
ice
standardssuchasI SO9000.

I
mmunit
y
Par
toft
hedef
ensesy
stem ofmanandani
mal
agai
nsti
nfect
ions.

I
ncubati
onper i
od
Theti
mei ntervalbet
weenexposuretoapathogenort
oxi
nandtheappear
anceofthe
f
ir
stcli
nical symptoms(e.
g.7-21day sf
orenter
icf
ever
).I
tdi
ff
ersconsi
der
abl
ybetween
f
oodbornei nfecti
onsandint
oxicat
ion.

I
ndi
cat
ororgani
sm
Amicr
oorgani
sm usedt
ocheckt
heef
fect
ivenessofGHP.

Infecti
on
Ent ryandcol
oni
zat
ionofani
nfect
iousmi
croor
gani
sm inali
vi
ngmi
croor
gani
sm (
host
).
Disease
doesnotalwaysdevel
opbutt
hehostbecomesa“carr
ier
”.

I
rr
adiat
ion
Treat
mentwi
thi
oni
zi
ngr
adi
ati
ont
orenderf
oodsaf
eori
ncr
easei
tsshel
f-
li
fe.

Met
abol
ism
Bi
ochemical
processi
nal
ll
iv
ingcel
l
s.Upt
akeofnut
ri
ent
sandassi
mil
ati
oni
nthecel
l
.

Met
aboli
te:
Anint
ermediat
eorendpr
oductofmi
crobi
almet
abol
i
sm par
ti
all
yexcr
etedi
nfood,
wit
h
 Bot
tl
edDr
inki
ngWat
erPr
ocess
adesir
edor
undesi
redef
fecte.g.or
ganicacids,CarbonDioxi
deandothergases,
ethanol
,ant
ibi
oti
cs,
mycotoxi
ns,
flavorandothersubstancesinf
luenci
ngthet
aste.

Microor
ganisms( microbes)
Simpleli
vingcreat
urescompr i
singv
iruses,
bact
eri
a,al
gae,
prot
ozoaandf
ungi
(fungi
i
ncludeyeastandmol ds).
Moni
tor
ing(
HACCP)
Theactofconduct
ingapl
annedsequenceofobservat
ionsormeasur
ement
sofcont
rol
par
ameter
stoassesswhetheraCCPisundercontr
ol.(
Codex)

Mold
Referstoanyf
ungusthatnormall
yformsamatofbr anchedel
ongat
edcel
l
s.Several
moldsareusef
uli
nthepreparat
ionoffood(e.
g.cheeseandTempe)butmanycause
spoi
lage.Somemoldsproduceharmfulmycot
oxins.

Mycotoxi
ns
Toxi
nspr oducedbyfungi
dur
inggr
owth.Themostwidel
ystudi
edmy
cot
oxi
nsar
ethe
af
lat
oxins
pr
oducedbyt hemoldsAsper
gil
l
usfl
avorsandpar
asi
ti
cus.

Nut
ri
ti
on
Thecombinati
onofpr
ocessesbywhi
chali
vingorgani
sm r
eceiv
esandusesthe
mat
eri
alnecessar
ytomaint
aini
tsf
unct
ion,
togrow,andtorenewit
scomponents.

Ochr
atoxi
ns
Agroupofmycotoxi
nsproducedbyAsper
gil
l
aandPeni
cil
l
iagr
owi
ngonf
oodst
uff
s.
Ochr
atoxi
nAisthebestknown.

Packaging
Acont ainerorwrappingdesi
gnedt
oprot
ectfoodcommodi
ti
esfrom mechani
cal
and
cl
imati
ci nfl
uences,andtoactasabar
ri
ertopest
sandmicr
oorgani
sms.

Pasteuri
zation
Heat i
ngaf oodtotemperaturesatleastt
heequivalentof72oCf
or15seconds.
Pasteuri
zati
ondestroy
smostt oxi
nsandv eget
ati
vecel l
sofmicr
oor
gani
smscausing
foodpoisoning.Mostbacteri
alsporessurvi
vepasteuri
zati
on.

Pat
hogenic
Pert
aini
ngtot
heabi
l
ityt
opr
oducedi
sease.

Pat
hogens
Anydisease-
causi
ngmi
croor
gani
sm ormat
eri
al(
bact
eri
a,y
east
,fungi
,andv
iruses)
 Bot
tl
edDr
inki
ngWat
erPr
ocess
t
hatmayaf
fectot
herl
i
vingor
gani
sms,
tot
hedet
ri
mentoft
hei
rheal
th.

Pesti
cides
Allcompoundsusedi
npl
antpr
otect
ionagai
nstpest
s.

pH.
Val
ueMeasur
eoft
he“
aci
dit
y”or“
alkal
i
nit
y”ofawat
er-
cont
aini
ngpr
oduct
.

Preservat
ion
Vari
ousmet hodstoext endtheshelfl
ifeoffood(e.
g.dehydr
ati
on,heatst
eri
l
izat
ion,
fr
eezing,r
adiat
ion,additi
onofpreservati
ves)byi
nhibi
ti
ngthemulti
pli
cati
onand/or
growthofmicroorganismsandbymi nimizi
ngchemicalandsensor
ychanges.

Preser
v ati
ves
Antimicrobialsubst
ancest
hatpr
eventmulti
pli
cat
ionofmicroor
ganismsand
someti
mesal sousedforpr
event
ingothert
ypesofundesi
rableact
ivi
ti
es.

Risk
Af unct
ionofthepr
obabil
i
tyofanadverseheal
thef
fectandt
hesev
eri
tyoft
hatef
fect
,
consequent
ial
to(a)hazar
d(s)i
nfood..

Spore
“Bacteri
alspor
es”
.Extr
emelyr
esist
antsurvi
val
formspr
oducedbybact
eri
a(gener
a
Bacil
l
us,
Clost
ridi
um)undercondi
ti
onsofnutr
ientl
i
mitat
ion.

Steri
lizati
on
I
nt hecont extoff
oodprocessi
ng,
amet hodofpreserv
ati
onbyki
l
li
ngall
microor gani
smsassoci
atedwithafoodst
uffusual
lybyappl
yi
ngheat
,forexampl
ein
bottl
ingandcanning.(
WHO)

Toxic
Poisonous.

Vector
Alsoknownasav ehi
cle.Methodoft
ranspor
tformicr
oor
gani
smst
ohost
sorhabi
tat
s
(e.
g.wind,
wat
er,
insects,r
odents,
pet
s,man,utensi
l
s).

Ver
if
icat
ion
Theappli
cati
onofmethods,
procedur
es,t
est
sandot
herevaluat
ions,
inaddi
ti
ont
o
monit
ori
ngtodeter
minecompli
ancewitht
heHACCPplan.(
Codex)

Vi
ralgast
ro-
ent
eri
ti
s
 Bot
tl
edDr
inki
ngWat
erPr
ocess
Collect
ivetermf orgastro-ent
eriti
scausedbyvir
uses,most l
ytransmitt
edtomanby
water,r
awsal ads,andv egetables.Thefol
lowi
ngaredescribedaspat hogens:
Rot
a
vi
rusesGr oupAandot hers,Norwalkvir
usesandlarge“r
elated”types,(
27-32nm),
Adeno-,Astro-andCalicivi
ruses.

Vir
alhepat
iti
s
Alsoknownasi nfect
iousjaundice.Anacut
einfect
iont
hatiscausedbythetypeA
hepat
it
isvi
rus.TheVi r
usistransmi t
tedf
rom per
sontopersonbythefaecal
-or
alr
out
e,
suchasoncont aminatedf
ood, waterandutensi
ls.

Vir
us
Inter
nal par
asit
esoft hecel
lsofmanyor
gani
sms;
theyar
eunabl
etogr
owout
sideof
theli
vingcell
soft hei
rhost.
Wor l
dHeal t
hOr ganizat
ion(WHO)
TheUnit
edNati
onsspeciali
zedagencyint
endedtopr
ovi
del
eader
shi
pinat
tai
nmentby
al
l
peopl
esoft
hehighestpossi
blel
evelsofheal
th.

Zoonosi
s
Communi
cabl
edi
seasest
hatcanbet
ransmi
tt
edt
omanbyani
mal
s.

Qual
it
yNev
erEnds….