CHAPTER 11

LABORATORY PROCEDURES

by

Jim Sequeira
498 Water Treatment

TABLE OF CONTENTS
Chapter 11. LABORATORY PBOCEDURES

Page

OBJECTIVES 500
woBDS..............._.. .. ... ................ 501

LESSON 1

1.I.0 BASIC WATEFI LABOHATOHY PFIOCEDUBES 507

11.00 lmportance of Laboratory Procedures....... 507

11.01 Metric Syslem ........ 507

1 1.02 Chemical Names and Formulas......................... 508

I 1.03 Helplul Flelerences.. 508
1I.1 LABORATOHY EOUIPI\,IENT AND TECHNIOUES ..... 509
1 1.10 Water Laboratory Equipment........................ 509

1 1.1 1 Use of Laboraiory Glassware...,.................... 518

11.12 Chemical Solutions ..... 520
1 1.13 Titralions 520
1 1.14 Data Recording and Flecordkeeping .................. 521

1 1.15 Laboralory Ouality Contro1................................. 521

11.16 Laboralory Salety.... 522

11.160 Laboratory Hazards 522
1 1. 1 61 Personal Safety and Hygiene........... 524

11 .162 Prevention of Laboratory Accidents . 524

1 1.1620 Chemical Storage... 524
11.1621 Movement of Chemicals ...........
'n.1622 Proper Laboratory Techniques.. 525
11.1623 Accident Prevention ....525
'11.163 Acknowledgments.. 527
1'1.164 Additional Fleading. 527
DISCUSSION AND FIEVIEW OUESTIONS .,..,,,. 527
LESSON 2

11.2 SAMPLING 528

11 .20 lmportance of Sampling.. s28

11 .21 Representalive Sampling 524
 Lab Procedures 499

'l
0
1.2'l lmportance of Representative Sampling... ........528
1 1.21 1 Source Water Sampting ...........
'l'1.212 ln-Plant Sampting......................
'11.213 Distribution System Sampting
... ........529
11.22 Types of Samples ...
........530
1 1.220 Grab Samptes........................... ........530
1 1.221 Composite Samptes.................. ........530
11.23 SamplingDevices...
.......530
1 1.24 Sampling Techniques ..............................
.......531
'1
1,240 Surface Samp|ing.....................
53'l
1 1.24't Depth Samptin9........................
531
1 1.242 Water Tap Samptang ...................
533
i j.243 First_Draw Samples....................
533
11.25 Sampting Containers and preservation of Samples ..
533
1 1.26 Reporting................
533
11.27 Additional Reading .
533
DISCUSSION AND REVIEW QUESTIONS
s33
LESSON 3

11.3 WATER LABORATORY TESTS..

535
1. Alkalinity.............
2. Chlorine Residuat.............
537
3. Chlorine Demand
541
DISCUSSION AND FIEVIEW QUESTIONS
543
LESSON 4

4. Colilorm Bacteria
544
DISCUSSION AND REVIEW OUESTIONS
565
LESSON 5

5. Hardness................._........
566
6. Jar Test.......-.....................
567
7. pH ...-,....:..,.......................
570
8. Temperature............_......_.
57'l
9. Turbidity.............................
572
ARITHMETIC ASSIGNMENT
574
AODITIONAL READING........
574

574

574

575
500 Water Treatment

OBJECTIVES
Chapter 11. TABOBATORY PFOCEDURES

Following completion ol Chapter 1 1, you should be able to:

1. Work safely in a laboratory.
2. Operate laboratory equipment.
3. Collect representative samples and also preserve and transport
the samples.
4. Prepare samples for analysis.
5. Describe lhe limilations of lab tests.
6. Becognize precautions to be taken for lab tests.
7. Record laboratory test results.
8. Perform the following lield or laboratory tests-alkalinity, chlo-
rine residual, chlorin€ demand, coliform, hardness, jar test, pH,
temperature, and turbidity.

-{
_r _Ar
6/)'
 Lab Procedures 501

WORDS
Chapter 11. LABOHATORY PROCEDURES

ACIDIC (uh-SlD-ick) ACtDtC

The condition ol water or soil which contains a sutficient amount of acid substances to lower the pH below 7.0,

ALIQUOT (Al-li-kwo0 ALTOUOT

Bepresentative portion ot a sample. Olten an equally divided porlion ot a sample.

ALKALI (Al-ka-lie) ALKALT

Any oI certain soluble salts, principally of sodium, potassium, magnesium, and calcium, that have the property
of combining with
acids to lorm neutral salts and may be used in chemical waier treatment processes.

ALKALINE (AL-Ka-LINE) ALKALINE

The condition of water or soil which contains a sufficient amounl of alkali substances to raise
ths pH above 7,0.

AMBIENT (AM.bee-ent) TEMPERATUFIE 4MBIENT TEMPERATUBE

Temperature of lhe surrounding air (or other medium). For example, temperaiure of the room where gas
a chlorinator is installed.

AMPEHOMETRIC (am-PURR-o-MEI-rick) AMpEFOMETR|C

A melhod of measurement lhat records electric current flowing or generated, ralher lhan recording voltage. Amperometric tiiralion is
a means of measuring concentrations of certain substances in waler.

AMPEROMETRIC (am-PURFt,o-MET-rick) T|TRAT|ON AMPEHOMETRTC T|TRATION

A means of measuring concentrations of certain substances in water (such as strong oxidizers) based on the electric
current that
flows during a chemical reaction. Also see TITRATE.

ASEPTIC (a-SEP-tick) nseprtC

Free lrom the Iiving germs of disease, fermenlation, or putrefaction. Sterile.

BACTEFIIA (back-TEAH-e-ah) BACTERTA

Bacteria are living organisms, microscopic in sjze, which usually consist of a single cell. Most bacteria use organic matter
for their
food and produce waste products as a result ol their Iife process;s.

ELANK BLANK
A.bottle containin-q only dilutaon water or dislilled water; the sample being tested is nol added. Tests are frequenfly
run on a SAM-
PLE and a BLANK and lhe ditferences are compared. The procedure h6lps to eliminata or reduce test resjlt errors that
could be
caused when the dilulion waler or disttlled waler used is contaminaled.

BUFFER
BUFFEF
A solution or liquid whose chemical makeup neutralizes acids or bases without a great change in pH

BUFFEB CAPACITY
BUFFER CAPACITY
A measure of the capacity of a solution or I iquid to neutralize acids or bases. This is a measure of the capacity of water lor oflering a
resislance to changes in pH

CALCIUM CARBONATE (CaCO3) EQUIVALENT cALcIUIiI CAHBoNATE (caco3) EQUIVALENT

An expression of the concentration ol specified constituents in water in terms of their equivalent value to calcium carbonate. For
example, the hardness in water which is caused by calcium, magnesium and other ions is usually describod as calcium carbonate
equivalent. Alkalinity test results are usually reported as mg/L caCO3 equivalents. To converl chloride trc Cibo, equivatents, multi"
ply the concentration of chtoride ions in mg/L by 1.41, and for suttate, huitipty by t.04. "
 502 Water Treatment

CABCINOGEN (CAFI-sin-o-JEN) CABCTNOGEN

Any substance which tends to produce cancer in an organism.

CHLOHORGANIC (kloror-GAN-ick) CHLORORGANTC

Organic compounds combined with chlorine. These compounds generally originate from, or ar€ associated with, life processes
such as those ol algae in water.

COLOBIMETBIC MEASUBEMENT COLOBIMETRIC MEASUEEMENT

A means of measuring unknown chemical concentrations in water by measuring a sample's color intsnsity. The specific color of the
sample, developed by addition of chemical reagents, is measured with a photoelectric colorimeter or is compared with "color stand-
ards" using, or corresponding with, known concentrations of the chemical.

coMPOSlrE (come-PAH-zit) (PFioPOFiT|ONAL) SAMPLE COMPOS|TE (PROPORT|ONAL) SAMPLE

A composite sample is a collection of.individual samples obtained at regular intervals, usually every ons or two hours during a 24-
hour time span. Each individual sample is combined with the others in pioportion to the rate oi flow when the sample was coiiected.
The resulting mi)dure (composite sample) lorms a representative sample and is analyzed to determine thB avera6e conditions dur-
ing the sampling period.

COMPOUND COMPOUND
A pure substance composed of two or more elements whose composition is constant. For example, table salt (sodium chloride,
NaCI) is a compound,

DPD (pronounce as separate letters) DpD

A method of measuring the chlorine residualin water. The residual may be determined by either titrating or comparing a developed
color with color slandards. DPD stands for N,N-diethy!p-phenylene-diamine.

DESICCATOH (DESS-uh-KAY-tor) DESTCCATOR

A closed container into which heated weighing or drying dishes are placed to cool in a dry €nvironment in preparation for weighing.
The dishes may be empty or they may contain a sample.besiccators contain a substance. iuch as anhydroui ciltium cntorioe, wnich
absorbs moislure and keeps lhe relalive humidity near zero so lhai the dish or sample will not gain weight from absorbed moisture.

DISINFECTION (dis-in-FEcT-shun) DTSTNFECTION

The process designed to killor inactivate mosl microorganisms in water, including essentially all pathogenic (disease-causing) bac-
teria' There are several ways to disinfect, with chlorinalion b€ing the most freque;fly used inwat;r trea'iment. Compare with STEH-
ILIZATION.

ELEMENT ELEMENT
A substance which cannot be separated inlo its constituent parts and still retain its chemical identity. For example, sodium (Na) is an
element.

END POINT END POINT

Samples of water or wastewater are titraled to the end point. This means that a chemical is added, drop by drop. to a sample untit a
certain color change (blue lo clear, for example) occurs. This is called the END POtNf of the titration. in addit6n to a cotor cnange,
an end point may be reached by the formation of a precipitate or th8 reaching of a specifisd pH. An end point may bo d€tsctod-by
the use ol an electronic device such as a pH meter, The completion of a desirdd chemical reaifion.

FACULTATIVE (FACK.ul-TAY-tive) FACULTATTVE

Facultative bacteria can use either dissolved molecular oxygen or orygen obtained from food materials such as sullate or nitrate
ions. ln other words, facultative bacteria can live under aeroi:ic or anaer6bic conditions.

FLAME POLISHED FLAME POLISHED

Melled by. a llame to smooth out irregularilies. Sharp or broken edges of glass (such as lhe end of a glass tube) are rotated in
a
flame until lhe edge melts slighly and becomes smooth.

GHAB SAMPLE GHAB SAMPLE

A single sample of water collected at a particular time and place which re presents the composition of lh€ water only at that timg and
place

GHAVIMETRIC GRAVIMETRIC
A means of measuring unknown concentrations of water quality indicators in a sampla by WEIGHING a precipitats or residue of tho
Sample,
 Lab procedures 503
INDICATOB (CHEMICAL)
A substance that gives a visible change,
usually of color; at a desired point in a chemical
reaction, n"""r",,f:'::;:; ::ltfln:l
INOFGANIC
INORGANIC
ffi:T[!il:i"i:""""11; i3,iii,:l;ii,fll[ffhil,',fl:i;Tl"1%;,a5llii:. rnorsanic substances are o, mrnerarorisin,
whereas

INORGANIC WASTE
,i wasre mareriar such as sand, sarr, iron,
carcium, and orher minerar materiars
**'es are chemicar su6"t""""" or.i"".ii
wiici.:re^.Tt s]islrry gtfg*, ,ffijff]:""ffiJn.t
;:I;:?,Ufl""" ";;:il;;;$rg""ic wasres are chemicar subsrances or an animar
or

,It or MOLAR
A molar solulion consists ol one oram molecular it or MOLAFT
werght ol a compound dissolved in enough
water Io make one liter o, solution. A
ej[ I"f:,1?::iJflil' ;,'!p;i;;;f
- -- - '- --'r lf,lr,t "r " "#i,","i, iJii"l Il, ,," ,i"b*[i*iig-h.r ;liruric acio (H.so.)
sorurron. would consist ",",p,", in enouin JitiiiJo'*aieiio
or 98 grams ol H2so. dissorveo
mare one rite, or

MPN (pronounce as separate letters)

MPN
orsanisms per unir vorume or sampre water.
IHY::H,T#:':li3i"Jil]tj",fl[#:i;:*"up Expressed as a densjty or popura-

MENISCUS (meh-NIS-cuss)
The curved surface ot a column of liquid (warer, MENISCUS
oil, mercury) in a small tube,, when the liquid
with water)' rhe curve rorms a varrev wets the sides ol ths container (as
1i'v6jn rn" .ij6. #'l"J*o ,"'r"rrvi, ir,," ;#;;" ,
""riiri.gi-he ro-p o;'in" iiqiJii reuert"".rlr,,
a meniscus forms in a'measurrng oevice, "i. hi, or upward
l'Jfl:"Jj* or rn" sampte is determined by the bonom
of rhe
WATER
MEFCUFY

(READ (READ
BOTTOM) TOP)

:i. : i

MILLIGHAMS PER LITER, mq/L

Ameasure ol the concentration bv wcight MILLIGHAMS PER LITEH, mgr'L
or a sub-stance per un( volume. For practical purposes.
waler is equar to one part per milrion oirts (ppml. one mg/L of a substance in fresh
rnus i iit-er-oi'*"i"i'*iin. .p""irr"
rf
g[rit ;iiii mirigrams.
f8ff[T:]8T#311?iflflllil;li,""iJi::ltf.;*,mEi;i,i'##,ru'g;:,'"Yi'o]fiffiIo"rriter(r-omgiij,
"r1-.b "il,irinion

MOLE

lle molecular weight of a substance, usually

expressed in grams. MoLE

MOLECULAR WEIGHT

is rhe sum or the aromic weishrs orrhe

*:dlti:iln[?3,T,?liS:ilffiyl*ji$".:s
Element
",","n,.,n,n".o,lrooti:'##:|fl]
Atomic Weight Number of Atoms
H 1
Molecular Weight
S 2 2
32 1
32
16 I 64
98
MOLECULE (NTOLL-uh-KULE)
The sma'est division of a compound thar sti, retains MOLECULE
or exhibits a, of the properties o, the substance
fror NORI/AL
A normal solulion contains one orari. cquivalent /V or NORMAL
weight o, reactant (compound) per ljter of
E lhal weight which conlains on; orem solution. The equivalent weight o, an acid
sulluric acid (Hrsoa) is ag (g8 oiiiotd
atom ot ioniziute nffii""'"iii."Jn""rr*r rii"r".prI,
ti?iq'urvarent weigirt ot
ov z uecauie-inei;liJiiri' ,ipll"E"or" hydrogen
"qrir"r""rions). a oni m soiuiil-n or su[uric acid
w.urd consist of 49 grams or H,Soo
dissoiu"o ,
;;r;;io'mIr.e"iiJTit",. or sorution.
"norgh
 504 Water Treatment

NEPHELOMETBIC (NEFF-el-o-MET-rick) NEPHELOMETHTC

A means of measuring turbidity in a sample by using an instrument called a nephelomstsr. A nsphglometor passes tight through a
sample and the amount of light deflected (usually at a 90-degree angle) is then measured.

NONVOLATILE MATTER NONVOLATILE MATTER

l\raterial such as sand, salt, iron, calcium, and other mineral materials which are only slightly atfected by the actions ol organisms
gld are 191_lo_s!9!.ignition of the dry solids at 550"C. Volatile materials are chemical su6stances usually of animal or pla;t origin.
Also see INOBGANIC WASTE and VOLATILE |V|ATTER or VOLATTLE SOLIDS.

OSHA (O-shuh) OSHA

The Williams-Sleiger Occupational Salety and Health Act of 1970 (OSHA) is a tederat law designed to protect the heatth and safety
of industrial workers and also the operalors of water supply systems and treatmenl plants. The Act regulates the design, construc-
tion, operation and maintenance of water supply systems and water treatment plants. OSHA also reJers to the fedeial and state
agencies which adminisler the OSHA regulations.

OHGANIC OHGANTC
Substances that comelrom animal or plant sources. Organic substances always contain carbon, (lnorganic mat€rials are chemical
substances of mineral origin.) Also see INORGANIC.

ORGANISM ORGANISM
Any form ol animal or plant life. Also see BACTEBIA.

OXIDATION (ox-uh-DAY-shun) OXIOAT|ON

Oxidation islhe addition of orygen, removal ot hydrogen, or the removal of electrons from an element or compound. ln the environ-
ment, organic matter is oxidized to more stable substances. The opposite of REDUCTION.

OXIDATION-REDUCTION POTENTIAL (OFP) OXIDATION-REDUCTION POTENTIAL (ORP)

The electrical potential required to transfer electrons from one compound or element (lhe oxidant) to another compound or element
(the reductant); used as a qualitative measure ol the state of oxidation in water treatment systems. OBP is measured in miltivolls,
with negative values indicating a tendenry to reduce compounds or elements and positive values indicating a tendency to oxidize
compounds or elemenls.

PABTS PER MILLION (PPM) PABTS PEH MILLION (PPM)

Parts per million parts, a measuremenl of concentration on a weight or volume basis. This term is equivalenl to milligrams per Iiter
(mg/L) which is lhe prefe.red term.

PATHOGENIC (PATH-o-JEN-ick) OBGANISMS PATHOGENTC OBGANTSMS

Organisms, including bacteria, viruses or cysts, capable of causing diseases (giardiasis, cryptosporidiosis, typhoid, chotera, dysen.
tery) in a host (such as a person). There are many types ol organisms which do NOfcause diseas€. These organisms are aalled
non-pathogenic.

PATHOGENS (PATH-o-jens) PATHOGENS

Pathogenic or disease-causing organisms.

PERCENT SATURATION PEBCENT SATUHATION

The amount ol a subslance that is dissolved in a solution compared with the amount dissolved in the solution at saturation,
expressed as a percent.
ol Substance That ls Dissolved x 100%
Percent saluralion'
'' = -^'nount
eror* o,"ro,*o ,n sol,rtionlr saturation
pH (pronounce as separate letters) pH
pH is an expression of the intensity of the basic or acidic condition of a liquid. Math€matically, pH is the logarithm (base 10) ot the
reciprocal of the lrydrogen ion activity.
I
oH=Loo-
' " tH-]
The pH may range lrom 0 to 14, where 0 is most acidic, 14 most basic, and 7 neulral. Naturalwaters usually have a pH betwsen 6.5
and 8.5.
 Lab procedures EO5

POTABLE (POE-ruh-bu WATER

)
oo"" no' POTABLEWATER
objectionable pollulion, contamination,
H,111;'t "ontain minerars, or infective agents and is considered
satisractory ror

PRECI PITATE (pre-Stp-uh-TATE)

(1) An insorubre, finery divided substance PRECIPITATE
whrch is a product of a chemicar reaction
(2) within a riquid.
The separation from solution of an insoluble
substance.

REAGENT (re-A-gent)
A pureschemical substance that is used to make new producls REAGENT
or is used in chemical tests to measure, detect,
or examine other sub_

REDUCTTON (re-DUCK-shun)
Beduction is the addilion of hvdroocn BEDUCTION
removal ot oxygen, or the addition of
electrons to an element or compound. under
fi:IHTJJ:ri:'."','J""13fl$"^?iJifr'
*rr"
."#p""-,"i. ,'""iJiJ.ioio ooo,-p,ooucins hydrosen ana'robic
surride (H,s) and orher com-

BEPRESENTATIVE SAMPLE

water that is as nearrv idenricar in conrenr

ff,il5l:ffi:t[l# ffi:'.flor and consistency * **-:::ffifi1;:',:',rt":*irT
SOLUTION
A liquid mixture of dissolved substances. ln a solution SOLUTION
it is imposslble to see allthe separate parts.

STANDARD METHODS
STANDARD MErHoDs FoR rHE EXAMtNAfloN oF wArER
AND wASrF!!.111,
can Public Hearth Association A.",i"an wate, wo.is ,i..'".,r'ili ie*wol, zt.";t Eoition. A joint ffi:"T::,Tr:'ff"":
{APHA),
which outlines the acceoled
laboratorir.proceo*". ,."0 io iiiri", eiii,ii,ilii neo"ration gerl
"roii,,"
"rurvi"fi;iil,jl ii1,l_*1,er and wastewaler. Avairabre ,rom American
[1;:H3']:,i:,..?6ii;;3?i;i";:j?iffiffiru*#1,ffi;xJ,","1't'oaozgs oi;";;ii":'ilid;;;['t"ofr.emoers,$rgaso:
STANDABD SOLUTION
A solurion in which the exact concenrration STANDAHD soLUTloN
of a chemicar or compound is known.
STANDAHDIZE
Tocompare with a standard. STANDABDIZE
I
f(l) ln wet chemislrv, to find out the exacl
slrength ol a solution by comparing it with
a slandard of known strength. This information
[ ,^.
rs used to adjust rhe strength by addirg ,"i" ;tJ#.J,""J r"ti'Jrlostrnc" oi.soru"a.

i
' iffi?"'?,iLJ:1fi'fiL?tli:ff#J:fl0,f"','""15,"",g1.,n,s arrows vou ro ad,usr the instrument so thar it
reads accurarery, or
I
sTERtLtzATtoN (STARE_uh_tuh_zAy_shun)
I
TEFllLlzATloN
[]he removal o. destruclion of all microord:nismc in^r,,.ri^^ ^^.!^^^-:^ -,- ,.
' "'-'oorganisms' including pathogenic
Imr, olsrrrrercrior.'r and oiher bacteria, vegetative forms and
spores. compare
I
i SUPERNATANT (sue.per-NAy-tenl)
SUPEBNATANT
luquid removed kom setfled sludoe. Suocrnar:.r commonlv .^,^-^
re,ers .^ .L - ,:, !..
Im,it rr". .rrr""" ^^--^hr, to the liquid between the studge on rhe bottom and th6 scum on
I
"ir"or.in"o";:;;i;j;:f"'n"t"t
isuBFAcrANT (sir-FAc-renl)
for surrace-acrive agent. The active suRFAcTANT
lAbbreviation agent in detergents that possesses a high
creaning ability.

InrRArE
(TlE-trate)
:To flrFArE a sample' ..
a chemical solution oJ known strength
is added drop.by drop untir a certain
TTTBATE
coror change, precrpitate, or pH
#iill"}'lli,i;ffii: iiXffi]:"'"i:H:f:lJX;fl";til*fJ"ffi liul5in! *"'"nu.i;"i;;#,;t iffi;ji i,,:;ements (0 , -i o
506 Water Treatment

TURBIDITY UNITS (TU) TURBIDITY UNITS (TU)

Turbidity units are a measure of the cloudiness of water. l, measured by a nephelometric (deflected ligho instrumental procedure,
turbidity units are expressed in nephelometric turbidity units (NTU) or simply TU. Those turbidity units-obtained by visuil methods
are expressed in Jackson Turbidity Units (JTU) which are a measure of the cloudiness of water; they are used to indicate the clarity
of water. There is no real connection between NTUS and JTUs. The Jackson lurbidimeter is a visual method and lhe nephelometer
is an instrumental method based on dellected light.

VOLATILE (VoL-uh-tull) VOLATILE

(1) A volatile substance is one that is capable of being evaporated or changed to a vapor at relatively low temperatures. Volatile
substanc€s also can be partially removed by air stripping.
(2) ln terms ot solids analysis, volalile refers to materials lost (including most organic matter) upon ignition in a mufllo furnace for
60 minules at 550'C. Natural volatile materials are chemical substances usurlly ol animai ol planlorigin. Manuractured or syn-
thetic volatile materials such as ether, acetone, and carbon teirachloride are highly volatile and not;f plant or animal origin.
Also see NONVOLATILE MATTEB.

VOLATILEACIDS VOLATILE ACIDS

Fatty acids produced during digestion which are soluble in water and can be sleam-distilled at atmospheric pressure. Also called
organic acids. Volatile acids are commonly reported as equivalent to acetic acid.

VOLATILE LIOUIDS VOLATTLE LIQUIDS

Liquids which easily vaporize or evaporate at room temperature.

VOLATILE MATTEH VOLATILE MATTER

Matter in waler, wastewater, or other liquids that is lost on ignition of the dry solids at ESO.C.

VOLATILESOLIDS VOLATILE SOLIDS

Those solids in water or other liquids that are lost on ignition of the dry solids at sSO"C.

VOLUMETRIC VOLUMETRIC
A measurement based on the volume ot some factor. Volumehic titration is a means of measuring unknown concentrations of wat6r
quality indicators in a sample BY DETERMINING THE VOLUME ot litrant or liquid reagent neede-d to complete particular reactions.
i
Lab Procedures 507

CHAPTERll. LABORATORYPROCEDURES
(Lessonlof5Lessons)

I1.O BASIC WATER LABORATOBY PROCEOUBES l\rany limes in the water laboratory we use smaller amounts
than a meler, a lite( or a gram. To express these smaller
11.00 lmportance of Laboraiory procedures amounts, prelixes are added lo the names of the base metric
Water trealment processes cannot be controlled etfeclively unit. There are many pretixes in use; however, we commonly
unless the operatorhas some means to check and evaluate th; use two or three prelixes more than any others in the labora_
quality ol water being treated and produced. Laboratory qualily tory
control tests provide the necessary information to monitor th6
lreatment processes and to ensure a sale and pleasant-tasting Pretix Abbreviation Meonlng
drinking waterfor allwho use it. By relating laboratory resultst;
lreatment operations, the water lreatment or supply syslem c6nti- c 1l'100 oli ot 0.01 times
operalor can first select the most etfective operational proce-
dures, then delermine lhe efficiency of the treatmenl proc- milli- m 1/1,000 oI; or 0.001 times
esses, and idenlify potential problems before they atfect fin- micro- p 1/1,000,000 of; or 0.000001 tim€s
ished walerquality. Forthese reasons, a clear understanding ol
laboratory procedures is a must lor every watenarorks operaior.
Onecentimeter (cm)is 1/100 (on€ hundredth) oIa meter, one
milliliter (mL) is 'll1,000 (ons thousandth) of a titer, and tikewis€,
one microgram (pgm) is 1/1,000,000 (one mi ionth) of a gram.

NOTICE EXAMPLES:
fhe aollection ol a bad san?le or a baa bborat ory (1) Convert 3 grams into milligrams.
rcault i5 abou't aE uselul as no .esullo. To prefinl 1 milligram = 1 mg = 1/1,000 grams
bad results requirest (1) aongtant maintenancc anl
therefore, 1 gram = 1,000 milligrams
calibration ol labontory cquipment, and (2) use ot
correat lab procedures, A!'c, reoultg of lab te1t5 arc (3 gramsx1,000 mq/gram) = 3,OOO mg
of no value to anyone unlesa thcy arc used, (2) Convert 750 milliliters (mL) to lit€rs.

mL = l/1,000 tirer
1

theretore, 1 liler = 1,000 mL

11.01 Metric System
(750 mLXl liter/1,000 mL) = 0.750 liters
The metric syslem is used in the laboralory to express units
ol length, volume, weight (mass), concentration, and tempera- (3) Convert 50 micrograms (pgm) to grams.
lure. The mekic system is based on the decimal system. All
unils of length, volume, and weight use factors ot lO to express 1 pgm
= 1/1,000.000 gram
larger or smaller quantities of these units. Below is a summary thereicre, 1 gram = '1.000,000 pgm
ol metric and English unit names and their abbrevialions.
50 pgm x I gran/'l,000,000 pgm = O.OOOO5 grams

Type ot English Metric Metric Larger amounts than a mete( liter, or gram can be ex-
Measuremenl System Name Abbreviation pressed using such prefixes as kilo- meaning '|,OOO. A kilo-
gram is 1 ,000 grams-
Length inch meter m The Celsius (or centigrade) temperature scale is used in the
foot
yatd
water laboratory rather than the more familiar Fahrenheit
scale.
Temperalure Fahrenheit Celsius
Volume quart
Fahrenhelr (.F) Celsius ("C)
liter L
gallon Freezing point of water 32 O

Weight ounce gram gm Boiling point of watet 212 ,lOO

pound
Concentration lbs/gal milligrams mg/L _ To converl Fahr€nheit to C€lsius, you can use Ihe lollowing
formula:
strength, % per liter
Temperature, "C = 5/9fF - Sz"F)
 508 Water Treatment

EXAMPLE: Converl 68"F to 'C TABLE11.1 NAMES AND FOBMULAS OF CHEMICALS

Temperature, COMMONLY USED IN WATEB ANALYSES
"C = 5/9fF - 32"F)
Chemical Name Chemical Formuls
= 5/9(68.F _ 32"F)
= 5/9(36) Acetic Acid cH3cooH
Aluminum Sulfate (alum) Al(SOo).. 14.3
= 20"c 1'1,9 "

To convert Celsius to Fahrenheit, the following tormula can Amrnonium Hydroxide NHlOH
be used: Calcium Carbonate CaC03
Temperalure,'F = 9/5("C) + 32.F Chlorotorm cHct3
Copper Sulfate CUSOa
EXAMPLE: Convert 35'C to'F
Ferric Chloride FeC13
Temperature, "F = 9/5("C) i 32'F
Nitric Acid HNO3
= 9/5(35'C) + 32'F Phenylarsine Oxide C6HrAsO
= 63+32 Potassium lodide KI
= 95"F Sodium Bicarbonate NaHC03
'11.02 Chemical Names and Formulas Sodium Hydroxide NaOH
Sulfuric Acid HrSOl
ln the laboratory, chemical symbols are used as shorthand
for the names of the elements. The names and symbols for
some of these elements are listed below. ' 14.3 HrO. Alum in the dryform based on 17./. Alroi.

Chemical Name Symbol

Calcium Ca Poor results and safety hazards are otten caused by usjng a
chemical from the shelf thal is not exactly the same chemical
Carbon c called for in a particular procedure. The mistake usually
Chlorine ct occurs when the chemicals are not properly labeled or have
similar names or formulas. This problem can be eliminated il
Copper Cu you use both the chemical name and formula as a double
Fluorine F check. The spellings of many chemical names are qujte simi-
lar. These slighl ditferences are critical because the chemicals
Hydrogen H do not behave alike. For example, the chemicals potassium
\( nitrate (KN03) and potassium nitrite (KNOr) are just as ditfer-
lron Fe
ent in meaning chemically as the words fat and flt are to your
Lead Pb )r doctor.

Magnesium Mg 11.03 Helpful Bererences

I\ranganese t\.4n .2. 1. "Meihods for Chemical Analysis ot Water and Wastesl' US
Nitrogen N Environmental Prolectjon Agency. EPA No. 600-4-79-020,
Obtain from National Technical lnformation Service (NTIS),
Oxygen o 5285 Port Boyal Road, Springfield, VA 22161. Order No.
PB84-128677. price, $1'11.00, ptus 96.66 shipptng and
Sodium Na handling per order.
Sulfur S 2. SIMPLIFIED PROCEDURES FOR WATER EXAMINATION
(M12). Obtain from American Water Works Association
A compound is a substance composed of two or more differ- (AWWA), Bookstore, 6666 West Quincy Avenue, Denver,
ent elemenls and whose composition (proportion of ele- CO 80235. Order No. 30012. ISBN '1-58321 -'182-9. Price to
ments) is constant. Generally, all chemical compounds can be members, $79.50; nonmembers, $1'16.50; price includes
divided into two main groups, organic and inorganic. Organic cost of shipping and handling.
compounds are lhose that contain the element carbon (C). 3. STANDARD METHODS FOB THE EXAMINATION OF
There are, however, a few simple substances containing WATER AND WASTEWATEB, 21st Edition. Obtain from
carbon thal are considered to belong to the realm ol inorganic American Water Works Association (AWWA), Bookstore,
chemislry. These include carbon dioxide (OOr), carbon mon- 6666 West Ouincy Avenue, Denver. CO 80235. Order No.
oxide (CO), bicarbonate (HCO3-), and carbonate {CO.r-) as in 10084. Price to members, $198.50; nonmembers, 9266.00;
calcium carbonale (CaCO3). price includes cost of shipping and handling.
Many ditferent compounds can be made from the same two 4. HANDBOOK FOR ANALMCAL OUALIW CONTBOL IN
or three elements. Therefore, you must carelully read the for- WATEB AND WASTEWATER LABORATORIES, US EnvI.
mula and name to prevenl errors and accidenls. A chemical tor- ronmental Protection Agency. EPA No. 600-4-79-019,
mula is a shorthand or abbreviated way lo write the name of a Obtain from National Technical lnformation Service (NTIS),
chemical compound. For example, the name sodium chloride 5285 Port Royal Boad, Springfield, VA 22161. Order No.
(common table salt) can be written "NaCl." Table 1 1.1 lists com- PB-297451. Price, $60.00, plus $6,00 shipping and han-
monly used chemical compounds found in the water laboratory. dling per order.
 Lab Procedures 509
5. ''Microbiological Methods for Monitoring the Envaronment_
GBADUATED CYLTNDERS. Graduated cytinders atso are
Water and Wastes,, US Environmental protection Agency.
basic to any laboratory and come in sizes from 5 mL
EPA No. 600-8-78-017. Obtain from Nationat Techniiat tri-
to 4,000 ml. They are used to measure volumes
(NTt S). 5285 porr Royat Road. Sprin g_
J9r,n'qti.91 _Sejvic_e more accurately than beakers.
lield. VA 22't 61. Order No. pB-290329/2. price. $86.0d. ptG
$6.00 shipping and handling per order
6. WATEB QUALTT)a Obtain from American Water Works As-
I
sociation (AWWA), Bookstore, 6666 West euincy Avenue,
Denver, CO 80235. Order No. i958. |SBN 1-SBd21_2g2_g.
Price lo members, $78.50; nonmembers, $.t 16.50; price in-
cludes cost of shipping and handling.
7. WATER ANALYSTS HANDBOOK, Fitrh Edition. Obtain from Cylinder,
HACH Company, pO Box 399, Loveland, CO 80539-0389. Graduated
Order No. 29547OO. ptice, $95.00, ptus g15.9S shipping
and handling.

I
t
5
/_
P/PEIS. Pipets are used to deliver accurate volumes and
range in size from 0..1 mL to 1OO mL.
I !
I
t
_----.'*_.],_Fri=

Pipet
OUESTIONS (pie-PET),
Write your answers in a notebook and then compare your Volumetric
answers with those on page S7S.
11.0A Why are laboratory quatity control tests important?
11.08 What does the prefix milti- mean? ,'.."'.'5,. 0

11.0C What is-the proper name of the chemical compound,

CaCO3? pipet, Serotogical

11.-I LABORATORY EOUIPMENT AND TECHNIQUES

For accurale lest results, accuracy in pipenino is verv imoor-
11.10 Water Laboratory Equipment Iant. This depends not only on the instrumen'is, me6lariicat
lunclion but also proper pipetting technlque. A regular calibra_
The itemsol equipment in a water laboratory are the opera_ rron Drogram can minimtze the risk of using maltuncfioning
lor's tools of the trade. ln any laboratory, there are certain pipets in critical.lests by quickty identitying -pip€ts operatin;
basrc pieces of equipment that are used routinely lo perform oulside specified tolerances. yet, pipetting lechnique can als6
water analysis tests. The following is a baef descri'plion of sev_ aflect delivered volume. and operator training is necessarv.
eral ol lhe more common items ol gtassware and pieces of Uontact your pipet supplier lor calibratron procedures.
equipment used in the analvsis of waler.
EUFErS. Burets are also used to deliver accurate volumes.
Volumelric. glassware lgraOuateO cyttoders and pipets) is They are especially useful in a procedure called li_
calibraled either "to contain" (TC) or "io detivef tration." Burets come in sizes from 1O to 1.000 mL_
1fd1. Gtass-
ware designed to deliver will do so accurately o;ly when the
inner surlace is so scrupulously clean thal waier
wits lhe sur_
lace immediatety and torms a uniform lilm on the
surlace upon
emptying.

EEAKEFS. Beakers are the most common pieces ol laboratory

equipment. They come in sizes from 1 mL to 4,OOO
mL. They are used mainly for mixing chemicals and to
measure approximale Volumes.
Support, Buret,
and Buret Clamp

1:j
o o
Beaker
 -l-
7
510 Water Treatment

Burel
(byur-RET)
Kjeldahl Flask Flask.
(KELL-doll) Distilling

BOTTLES. Bottles are used to store chemicals, to collect

samples for testing purposes, and to dispense
liquids.
oli

r0r
iri
Automatic
Buret

Bottle, Bottle,
Reag6nt BOO

FL,ASKS. Flasks are used for containing and mixing chemi-

cals. There are many ditferent sizes and shapes. FUNNELS. A funnel is used lor pouring solutions or transfer.
ring solid chemicals. This tunnel can also b6 used
with filter paper to remove solids from a solution.
Flask,
Erlenmeyer
(ERlen-MY-e0
Wde Mouth

Funnel

Flask,
Boaling
. Flat Bottom
A Buchner lunnel is used to separate solids from a mrxture
It is used with a filter tlask and a yacuum.

,R
()
Funnel.
Flask,
Boiling
Round Bottom
p F:\
':! Buchner
with
Short Neck Perforated Plate

Flask,
Filtsring
 Lab Procedures 511

Separaiory funnels are used to separate one chemical OTHER LABWARE AND EQUIPMENT.
mixture. from another. The separated chemical usually is dis-
solved in one or two layers of liquid.

l-)

Separatory Funnel

Condenser

TUBES. Test tubes are used for mixing small quantities of

chemicals. They are also used as containers for
bacterial testing (culture tubes).

K
r(l::--h,
1,.)->:=!::==r'{
''--ru-rrl!-Y
Dish, Petri

rilmt
Test Tube Culture Tube
without Lip

Desiccator
(DESS-uh-KAY-to0
\l

Color Comparison
Tubes, Nessler

Thermometer, Dial I
512 Waler Treatment

'1
I k*.q
yt*,aa {
i
,l

Y,4 Clamp, Beaker,

Salety Tongs
I

s:{61
Oven, Mechanical Convection -.-N

Clamp, Dish,
Safety Tongs

*!-. ,
Hot Plate

Clamp, Flask,
Safety Tongs

.+
1
Clamp, Test Tube
,g
..!

a.
I
o
Muffle Furnace. Electric Clamp Holder
qr
Lab Procedures 5.13

7..
!
I

-
I

.t
Clamp, Utitity

Fume Hood

Clamp

Tripod, Concenlric
Ring

Porlable Dissolved Oxvqen Meter

(with computer dockinj
stafiony
(C.une3y o, SACH Compaiy)

Burner. Bunsen
at lD
-tD

v
Triangle,
Fused Portable pH Meter
(Cou.t€sy ol HACH Coinpany)

-l
5 t4 Water Treatment

Crucible (CREW-suh-bull),
Porcelain

Pump, Air Pressure and Vacuum

Crucible,
Gooch
Porcelain

Pipet Bulb

Dish,
Evaporating

4{ l/
+-s
H fdrion
P" pafl B

5 1
f
E Laboratory Turbidimeter
(ratio mode--€N/OFF-is keypad sel€ctable)
Test Paper, pH 1-11 (Coirriosy ot HACH C{mp.rry)
 Lab Procedures 515

1:-_ Magnetic Stirrer

(Pemis!6n o|n16mlyn6)

i5
1t

( t

Autoclave

lncubator
lPemisio. ot Alue M Elettu)

\
-

ir-
I
DireclReading Colorimeter
(free chlorine residual)
(coudesy ol HAcr Compar'y)

Portable Spectrophotometer
(Coutusy or HACH Comoaiy)
516 Water Treatment

Chlorine Besidual Test Kit

(lree and total chlorine)
(C&nesy ol HACH Conpanll

aaa
t
I
a o I
I
I

:\- .!'.-

>
-

0 0 6.

w.ash! =
E
95.5560 am.

Balance, Analytical
(Pemisslon ol M6nl6D
 Lab Procedures 517

a E

I
I
I

c
I
t
o

..=i:--
,.8t---

Anperomettic Titrator

I {Coudesy ol BACH Company)

518 Water Treatment

11.1'l Use of Laboratory Glassware
BURETS
A buret is used lo give accurate measurements of liquid vol-
umes. The stopcock conlrols the amount of liquid that willflow
from the buret. A glass stopcock must be lubricated (slopcock
grease) and should not be used with alkaline solutions. A
When a buret is lilled with liquid, lhe surtace ol the liquid is
curved. This curve ol the surface is called the meniscus (melF
NIS-cuss). Dependang on the liquid, the curve lorms a valley,
as with wate( ortorms a hill, as with mercury Sjnce most soiu-
tions used in the laboratory are water based, always read h6
bottom of the meniscus with your eye at the same Isv6l (FtgurB
1 1 .1 ). lf you have the meniscus at eye level, the closest marks

Teflon stopcock never needs to be lubricated.
that go all lhe way around the buret will appear as strsjgrt
lines, not circles.
GRADUATED CYLINOERS
The graduated cylinderor "graduate'is one ol th6 most olten
used pieces ot laboratory equipment, This cylind€r is mad€
either of glass or of plastic and ranges in sizes from 1O mL to4
liters. The graduate is used to measure volumes of liquid with
an accuracy less than burets but greater than beakers or flasks.
Graduated cylinders should never be heated in an open flame
because they will break.

Stopcock

Buret

Burets come in several sizes, with lhose holding 10 to 25

milliliters used most frequently.

7- b t_
Lt
'-ot!
t-:
?b?Eg LINE
oF vtrroN 4 : aeu!6cu,h
<9ge9' I
6
Fig. 1 1.1 How to read meniscus

FLASKS AND BEAKERS
Beakers and flasks are used for mixing, heating, and wejgh-
ing chemicals. Most beakers and flasks are not cllibrated with
exacl volume lines; however, they are somelimes marked with
approximale volumes and can be used to estimate volumes.
1gi
:l
Flask Beaker
VOLUMETRIC FLASKS
Volumetric tlasks are used lo prepare solutjons; they come
in sizes from 10 to 2,000 mL. Volumetric flasks should never
be healed. Flalher than slore tiquid chemicats in volumetric
flasks. the chemicals should be transferred to a storage botfle.
Volumelric flasks are more accurate than gradualed crlinders.
Lab Procedures 519
PIPETS
Pipets are used tor accurate volum€ measurements and
transfer. There are three types of pipets commonly used in the
laboratory-volumetric pipets, graduated (measuring) or Mohr
pipets, and serological pipets.
Volumetric Pipet
(=JErl,_B l
Graduated or Measuring Pipet
Serological Pipet
Volumetric pipets are available in sizes such as 1, jO, 25,
50, and 100 mL. They are used to deliver a single volume.
l\reasuring and serological pipets, however, will deliver frac-
tions of the total volume indicated on the pipet.
Volumetric pipets should be held in a vertical position when
emptying and th€ outflow should b€ unrestricted. The tip
should be touched lo the wet surlace of the receivino vessel
and kept in contact with it until the emptying is c;mpt€le.
Under no circumstance should the small amount remaining in
the tip be blown out.
Measuring and serological pipets should be held in the ver-
tical position. After outflow has stopped, the tip should be
touched to the wet surface of the receiying vessel. No drain-
age period is allowed. When the small amount remaining in
the tip is to be blown out and added. this will be indicated by a
frosted band near the top of the pipet.
Use of a pipet filler or pipet bulb (page 514) is recommend-
ed to draw the sample into a pipet. Never pipet chemical re-
agent solutions or unknown water samples by mouth. Use the
following techniques for best results.
1. Draw liquid up into lhe pipet past the calibration mark.
2. Ouickly remove th6 bulb and place dry fingertip over the top
end of the pipet.
3. Wipe excess liquid from the tip of the pipet using laboratory
tissue paper.
4. Litt finger and allow desired amount, or alt, of tiquid to drain_
Pipets can be drained without remoying the pipet bulb.
NOTE: fhere aG pipet bulbs with valves that can control the
llow ol liquid from the pipet without removing the
bulb.
 520 Water Treatment

ACKNOWLEDGMENTS When preparing standard solutions or reagents, the direc-

tions may say to weigh out 7.6992 grams o, a chemical and
Pictures of laboratory glassware and equipment in
this
dilute to one liter with distilled water. To weigh out 7.6992
manual are reproduced with the permission ol VWFI Scientific,
grams ol a chemical, determine the weight of a r,veighing dish
San Francisco, California, with exceptions noted.
and add this weight to the weight ol the chemical. Plac€ lhs
weighing dish on the woighing platform ot an analylical bal-
OUESTIONS ance. (See page 516. Some balances have the weighing plat.
form on top for the weighing dish.) Gently add the chemical to
Write your answers in a notebook and then compare your
lhe weighing dish until you are slightly below the desired
answers with those on page 575.
weight. The weighing mechanism should be ofl while the
1
'1.1A For each type of glassware listed below, describe the chemical is being added and then turned on to determine the
item and its use or purpose. exact lveight. When you get close io the exact weighl, plac€
some of the chemical on a spatula. Gently tap the spatuta to
1. Beakers add very small amounts of chemical to the weighing dish.
2. Graduated cylinders Continue this procedure until you have reached the exact
3. Pipets weight. lf you add too much chemical, remove some ol the
4. Burets chemical wilh the spatula and again repeat the procedure until
't '1
.18 What is a meniscus? you reach the exact weight.

11.1C Why should graduated cylinders never be heated in Anolher procedure is to place approximately the desired
an open flame? weight in the weighing djsh. Weigh this amount exac y, Then
add a proportionate amount of distilled water.
'11.12 Chemical Solutions
EXAMPLE 1
Many laboratory procedures do nol give the concentrations
of standard solutions in grams/liter or milligrams/liter. lnstead, The directions for preparing a standard reagent indicate that
lhe concentrations are usually given as NORMALITY (N),1 you should weigh out 7.6992 grams and dilute to ona liter. You
which is lhe standard designation lor solution skengths in weigh out 7.5371 grams. How much waler should be added to
chemistry. produce the desired concentration or normality ol ihe standard
reagent?
EXAMPLES:
0.025 N H,SO. means a 0.025 normal solution of sulfuric
acid
2 A/NaOH means that the normality of a sodium hy-
droxide solution is 2
The larger the number in front of the M the more concen-
kated the solution. For example, 1 A/ NaOH solution is more
concenlraled than a 0.2 N NaOH solution.
Another method of specitying the concenlration of solutions
uses lhe 'a + b system." This means that "a'volumes of con-
centrated reagent are diluted with "b" volumes of distilled Known Unknown
water to form lhe required solution.
= 7.6992 gm
Desired Weight, gm Water, mL
EXAMPLES: AclualWeight, gm = 7.5371 sm
1+1 HCI means 1 volume of concentrated HCI is diluted The chemical should be dilut€d to how many milliliters?
with 1 volume of distilled water
(Actual Weight, gm)(l,000 mL)
'l + 5 H"SO. means 1 volume of concentraled sulfuric acid Ditule to mL _
Desired Weight, gm
is diluted with 5 volumes of distilled water
(7.5371 gm)(1,000 mL)
When the exact concentration of a prepared chemical solu-
tion is known, it is referrdd to as a "standard solution." Many 7.6992 gm
times, standard solutions can be ordered already prepared
from chemical supply companies. Once a standard has been = 979 mL
prepared, it can then be used lo standardize other laboratory
The 7.5371 grams oI chemical should bB diluted to 979 mL.
solutions. To standardize a solution means to determine and
adiust its concentration accurately, thereby making it a stand-
ard solution. "Standardization" is the process oI using one so-
1l.13 Titrations
lution of known concentration to determine the concentration A titration involves the measured addition of a standardized
of another solution. This action often involves a procedure solution, which is usually in a buret, to another solution in a
called a "litralionl llask or beaker. The solution in ihe buret is rsferred to as the

' N or Nomal. A normal solution conlarns one gram equivalent weight ol reactant (compound) per liler ol solution. The equivalont weight ol an
acid is that weight which conlains one gram atom of ionizable hydrogen or ils chemlcal equivalent. For sxample, th€ equivalent wgight ol sullurlc
acid (H,SO4) is 49 (98 divided by 2 because there are two replaceabl€ hydrogen ions). A one N solution ol sulluric acid would consist of 49 grams
ol HrSOa dissolved in enough water lo make one liler of solulion.
T Lab Procedures 521

"tilrant'and is added to the other solution until there is

a
measurable change in lhe test solulion in the flask or beaker.
Ttis change is lrequen y a color change as a resull ol the
ad_
drtron 0t anolher chemical called an "indicato/'to lhe solution
in lhe flask belore the titralion begrns. The sotution in the
burel
is added slowly to the llask until the change. which is called .+- 2. Flecord level
the'end point, is reached. The entire Drocess is lhe .,titration." w. in buret.
Figve 1,1.2 illustrates the four general steps used during a 14il
chemical titration. lff
1'1.14 Data Recording and Recordkeeping
xg
is
The use of a laboratory notebook and worksheets are a
musl for laboratory analysts and water supply system and
keatmenl plant operators. Notebooks and wo;ksheets help tr
you record data in an orderly manner. Too oflen, hours a-
of wor'k
are wasled when lest results and other data (such as a
sample volume) are written down on a scrap of paper only to
be mrsplaced or thrown away by mistake. NoteOoot<s inO
worksheets help prevent errors and provide a record of your
work. The routine use ol laboratory worksheets and nbte-
books.is the onty way an operalor or i lab person can be sure
lhat all imporlant informalion is properly recorded.

1. Flecord volume
!
ol sample.

I
,/E

is no standard taboratory
lorm. Most operators usu-
.,There
ally develop their own data sheets tor recordtng test results
and olher important data. These data sheets shiould be pre-
<_
pared in a manner that makes it easy for you 3. Add titrant
to record results, until
revrew lhem. and recover these results when jl is necessary. end point is
Each trealment planl wi have diflerent needs tor coffeitinlj reached.
and recording dala and may require several different dala
o-r
worksheets. Figures I 'l .3 and 1 1.4 illustrale two typical labo-
mlory worksheets.

11.15 Laboratory euatity Control

Having good equipment and using the correct methods are
not enough lo ensure correct analytical results. Each operator
must be constantly alert to lactors in the water trealmenl proc-
ess lhat can lead to poor quatity of data. Such tactors in;lude
sloppy laboratory lechnique. deterjorated reagents
and stand-
ards, poorly operating instruments, and catc;tation mistakes.
one ol lhe best ways to ensure quality control in your labora_
10ryis.io analyze re,erence.type samptes to proviie indepen-
denl checks on your analysis. These relerence_type samDles
are available from lhe US Environmental proteition Agency
and hom commercial sources, From time to time, it is
gmd idea lo split a sample wilh one ol your lellow
itso i 4. Record level of titrant
operators or
a4olher laboratory and compare anaiytical results. ln at end point.
addi_
lrequent self-appraisat and evaluaiion-from sampling lo
lion.
pporling resutts---can hetp you gain lull
confidence in y-our
resulls.
Fig 11.2 Titration Steps

I
 522 Water Treatment

WATER ANALYSES REPORT SHEET

(All resulls in mg/L except as noted)

SAMPLE SOUFICE

DATE SAI,4PLED: BY

ANALYST] DATE CON4PLETED:

A. PHYSICAL PAFIAMETERS ADDITIONAL INFORMATION

color (units) Water Tr., Vol. ll, pg. 514

odor (units) Water Tr., Vol. ll, pg. 538

iurbidily (NTU) This N.4anual, p9.572

B. GENEFAL iillNERAL AD D ITI O NA L I N FOB I\,4ATI O N

pH (units) This Manual, p9.570

total alkalinity (CaCO3) This Manual, pg.535

specific conductance, pmhos/cm Water Tr., Vol. ll, pg. 535

total dissolved solids Water Tr., Vol. ll, pg. 543

total hardness (CaC03) This Manual, pg. 566

calcium Water Tr., Vol. ll, pg. 51 I

fluoride Water Tr., Vol. ll, pg. 519

iron Water Tr., Vol. ll, pg. 523

manganese Water Tr., Vol. ll, pg. 526

chloride Water Tr., Vol. ll, pg. 512

sulfale Water Tr., Vol. ll, pg. 535

Fig. 1 1.3 Typical laboratory wo*sheet

OUESTIONS Personnel working in the water lndustry must reallze that a

number of hazardous materials and conditions can exist.
Write your answers in a notebook and then compare your Always be alert and carelul. Be awars of potential dangers at
answers with those on page 575. all times. Safe practice in the laboratory and any time while
'1
1 .1D What is a standard solution? working around chemjcals requires hardly any more etforl
than unsafe practices, with the important benelits from pre-
11.1E What is the primary purpose ol laboratory note- vention of iniury to you and your lellow operators.
books and worksheets?
On specific questions of safety, consult your state's Generai
11. 1F List three sources or causes of poor quality of ana- lndustrial Salety Orders or OSHA regulations.
lyticaldata.
1 1.160 Laboratory Hazards
'11.16 LaboratorySarety Working with chemicals and other materials in the water
treatment plant laboralory can be dangerous. Laboratory haz-
Safety is just as important in the laboratory as it is outside ards include:
the lab. State laws and lhe Occupational Safety and Health
Acl (OSHA) demand lhat proper salety procedures be exer- 1. Hazardous materials 4. Electric shock
cised in the laboratory at all times. OSHA specifically deats 2. Explosions 5. Fire
with "safety at the place of work." The act requires that "each
employer has the general duty lo furnish all employees wilh
3. Cuts and bruises 6. Burns (heat and chBmical)
employment free from recognized hazards causing, or likely to Hazardous materials include: (1) corrosive, (2) toxic, and (3)
cause, death or serious physical harm." explosive or llammable materjals.
 Lab Procedures 523 I

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 524 Water Treatment

1. Corrosive lvlaterials the eyeball and irritate the eye, the National lnstitute of 0c-
cupational Safety and Health (NIOSH) recommends an eye
ACIDS injury hazard evaluation be conducted in accordance with
the safety guidelines listed in Current lntelligence Bulletin
a. Examples: Hydrochloric or muriatic (HCl), hydrofluoric (ClB) 59 2005-1 39 (www.cdc.gov/niosh/docs/2005-1
(HF), glacial acetic (CH3COOH), nitric (NHO3), and sut- 394.
furic (H,SOJ.

b. Acids can be extremely corrosive and hazardous to

human tissue, metals, clothing, cement, stone, and con-
crete.
c. Commercially available spill cleanup kits should be kept Safety Glasses
on hand to neulralize the acid in the event of an acciden-
tal spill. Baking soda (bicarbonate, not laundry soda)
effeclively neutralizes acids. Baking soda can be used on
lab and human surfaces withoul worrying about toxicity.

EASES (Causlics)
a. Examples: Sodium hydroxide (caustic soda or lye,
NaOH), quicklime (CaO). hydrated time (Ca(OH)r), and
alkaline iodine-azide solution (us6d in dissolved oxygen
test).
b. Bases are exlremely corrosive to skin, clolhing, and
leather. Caustics can quickly and permanenty cloud
vision if not immediately flushed out of eyes. Determine
location of safety showers and eye wash stalion before
starting to work wilh dangerous chemicals.
c. Commercially available spill cleanup materials should
be kept on hand for use in the event of an accidental
spill. A jug of ordinary vinegar can be kept on hand to
neutralize bases and it will nol harm your skin.

MISCELLANEOUS CHEMICALS
a. Examples; Alum, chlorine, fenic salts (ferric chloride),
and other strong oxidants.
2. Toxic Materials
Examples:
a. Solids: Cyanide compounds, chromium, orthotolidine,
cadmium, mercury, and other heavy metals. DO NOT PIPET HAZARDOUS
b. Liquids: Chloroform and other organic solvenls. LIOUIDS BY MOUTH.
c. Gases: Chlorine, ammonia, sulfur dioxide, and chlorine
dioxide.
3. Explosive or Flammable Materials 3. Never pipet hazardous materials by your mouth.

Examples: 4. Always wear a lab coat or apron in the laboratory to protect

a. Liquids: Acetone, ethers, and gasoline.
b. Gases: Propane and hydrogen.
11.161 Personal Satety and Hygiene
Laboratory work can be quite dangerous il proper precau-
tions are not taken. Always lollow these basic rules:
1. Never work alone in the laboratory. Someone should
always be available to help in case you should have an ac-
cident that blinds you, leaves you unconscious, or slarts a
fire you cannot handle. lf necessary have someone check
on you regularly to be sure you are OK.
2. Wear protective goggles or eyeglasses at alltimes in th€ lab-
oralory Contact lenses may be worn under safety goggles,
but contact lenses are not eye protective devices, and wear-
ing them does not reduce the requirement for eye and face
protection. Because fumes can seep between the lens and
your skin and clothes.
5. Wear insulated gloves when handling hot objects. It there is
a danger of hot liquid erupting from a container, wear a lace
shield, too.
6. Do not keep lood in a relrigerator that is used for chemical
or sample storage.
7. Good housekeeping is an etfective way to prevent acci-
dents.
11.162 Prevention ol Laborulory Accidents
1 I.I620 CHEMICAL STORAGE
An adequate chemical storeroom is essential for safety in
the water laboratory. Th6 storeroom should be properly venti-
lated and lighted and lard out to segregate incompatible chem-
icals. Order and cleanliness must be maintained. Clearly label
and date all chemicals and bottles of reagents,
 Lab Procedures 525

Store heavy items on or as near to the floor as possible. 1 1.1 622 PBOPER LABOFATORY TECHNIOUES
y1UTlLE LIQIJIOS' that may escape
as a gas. such as
€lher, musl be kepl away from heat sources, sunlight, and . Faulty technique is on€ of th6 chief caus€s of accidents and,
eleckic switches. Store acids and bases in separate-slorage because it involves the human element, is one of lhe most dif_
cabinets designed for acid and base storage. ficult to correct.

Cap and_secure cytinders of gas
orlipping._They shoutd atso be placed
sources of heat or open llames.
iil
in slorage to prevent rolling Because ol their nature and prevalence in the laboralory
awat from any possibl; acids and other corrosive materials conslitute a series of hai_
ards ranging from poisoning. burning, and gassing through ex_
plosion. Always llush the oulsides ot acid'Oottle-s with "water
belore opening them. Do noi lay the stopper down on the
tl counlertop where a person mjght lay a hand or rest an arm on
iii it. Keep all acids tighfly stoppered when not in use and make
lrl sure no spilled acid remains on the floor, table, or bot e atter
lli use. To avoid splashing of acid, do not pour water into acid;
Always pour acid into water.

. Mercury requires special care. Keep all mercury containers

tightly stoppered. Even a small amount of spilled m€rcury can
llltrll- poison th6 almosphere in a room. To clean up a small meicury
spill (the amount in a thermometer), put on rubber, nitrile, oi
latex glovss..Use a squeegee or, c"idboaro to gaiher the ;iei-
cury beads. Use slow sweeping molions to ke6p the mercury
,rom becoming uncontrollabte. Use a disposabie eyedroppei
lo coltect or draw up the mercury beaOs. btowty ani caret[fiy
squeeze the mercury onto a damp paper towel. place th6
paper towel in a zip-lock bag and secure. Make
sure to label
b as directed by your local health or tire department.
l
l?9 9aS
Never pour mercury down a drain. lt may lodge in the plumb_
llllrl rng ard.cause future problems during ptumbi;g repairs.
lf dis-
charged, it can cause poltution of lhe seplic-lank or lyaste_
waler.keatment pta . Contacl your local health Oepartmeni,
munrcipat wasle. authority, or your local fire department
rr',rilrlil,i for
rh proper drsposat in accordance with local,
state, and federal
illllllilli Iaws.. Femember lo keep the area well ventilatsd
liii iiril iiirii to-fnelri_
sroe (wtndows open and fans in exterior windows runnino) for
:lsprlls,
l:-r:1,?4
hours after your successful iof rilsJr.
CLAMPS, RAISED SHELF EDGES, call your local or slate health or "r""nup.
environmontal agenc!.

AND PROPER ARRANGEMENT 1 1.1623 ACCIDENT PREVENTION

ELECTRIC SHOCK. Whercvet there are etectrical
-
PREVENT ST()CKROOM FALLOUT. wiring. connections. there is a d;S;; ;
outlets.
?]rS: Td
snock. the usual "do's" and ,don,ts. ot prot;ction
;ili;;
against
Follow usual common-sense shock in the home are equalty appticabte in fi,e laOorato-ry.
l
.keeping rules of storage. Good house- do
not use wornor lrayed wires. Replace connections
is a.mosl signiticanl contribution t6*"iO i"ti* when ihere
rs_any sigr- ol,lhinnrng insulation. Ground all
safety campaign. "n apparatus usint
rnree-prong ptugs. Do not continue lo run a motor
atter lioui;
11.1621 MOVEMENT OF CHEMICALS has spitted on it. Turn it ofl immediately anO cfean
analryi-frJ
inside thoroughty before attempting to use it again.
The nexl area ol concern is the transler of
.
l cnemlcats, aDparalus, gases, or other haz- .. Electrical units that are operalgd in an area exoosad to
i ardous malerials from lhe storeroom lo the rammabte vapors should be explosion-proof. All permanent
.19r,9 b9.in:ta ed by an etedrician
laboralory tor use. Use ciadles or tilters to Ia_ l]I9
or tsx caDte to eliminale any danger
w h proper conduil
cililate handling carboys or other larqe chem- o, circuit overloading.
icalvessels_ CUfS. Some of the pieces of glass used in the laboratorv.
Drum Tilter
ln transporting sucn as gtass tubing. thermometers. and funnels, must
,trussed..hand cylinders of compressed gases, use a sened. through rubber stoppers. lf the glass is lorced
be i;_
Iruck. Never roll a cylinder by ils i"fre. imreOi lhrouoh
arery aner they are posrtioned for use, cylrnders :191"9"J!
tl" .tLqper by apptying a tor of pressure. lh€ sta;s
shoutd be usua y breaks. This js one of the most common sourcEs
cramped securety inlo ptace to prevent shifting or of
toppling. cuts in the laboratory.
Carry llammable lquids in safety cans or, in the
case ol re- Use care in making rubber-to-glass connections. Lenoths
agenr-grade chemicals, protect lhe bot e with of
a carrrer.
Arways wear protective gloves. sal€ty shoes, and rubber
.
grass ruDrng shoutd be supported white thev
are bei-no in-
aprons rn case ol accidenlal spilljng o, chemical
containers.
le]red rnto rubber. The ends of the gtass sh;utd Oe ftlUr
POLISHED3 and either wefled or c&ered witfr a
tuOricatinl

U_olatile Liquids. Liqutds which easrly vapoize or evaporate at room lempEralure.

"sil''iiliio,i,ili,ldses
fifl,'JlTfl"g.ryfl::,1?l;li:lUl*l:*gf;Iiii". or srass (such as rho 6nd or a
srass rube) ar€ rorared in a
 526 Water Treatment
jelly for ease in ioining connections. Never use oil or grease.
Wear gloves when making such connections, and hold the
tubing as close lo the end being inserted as possible to pre-
vent bending or breaking. Also, never try to torce rubber tubing
or stoppers from glassware, Cut off the rubber or materials.
A first-aid kit must be available in the laboratory
BURNS. All glassware and porcelain look cold after the red
from heating has dasappeared. The red is gone in seconds but
the glass is hot enouoh to burn for several minutes. Afler heat-
ing a piece ol glass, put it out of the way until cold.
Spattering from acids, caustic materials, and strong oxidiz-
ing solutions should be washed otl immediately with large
quantities of water. Every worker in the water laboratory
should have access to a sink and an emergency deluge show-
er. Keep vinegar and soda handy to neutralize acids and
bases (caustic materials).
Many safeguards against burns are available. Gloves,
safety tongs, aprons, and emergency deluge showers are but
a few examples. Never decide it is too much trouble to pul on a
pair ol gloves or use a pair of tongs to handle a dish or flask
that has been healed-
) iiirl
I'
USE TONGS_DO NOT JUGGLE
HOT CONTAINERS.
Perhaps the most harmlul and painful chemical burn
occurs when small objects, chemicals, or lumes get into your
eyes. lmmediately flood your eyes with water or a special
eyewash solution lrom a sarety kil or from an eye wash sla-
lion or fountain.
TOXIC FUMES. Use a ventilated laboratory fume hood lor
rouline reagenl preparation. Select a hood that has adequate
air displacement and expels harmful vapors and gases at their
source. An annual check should be made ol the entire labora-
tory building. Sometimes, noxious lumes are spread by the
heating and cooling system of the bujlding.
When working with chlorine and other toxic substancsq
always wear a selr-contained breathing apparatus. lt possibls,
lry to clear the almosphere with adequate ven lalon bsfom
entry
WASTE DISPOSAL A good sarety program requires con-
stant care in disposal of laboralory waste. Corrosive materi
als should never be poured down an ordinary sink or drain.
These substances can corrode away the draln pipe or kap.
Corrosive acids should be neutralized and poured down
corrosion-resislant sinks and sew€rs using large quantities ol
water to dilute and flush the acid.
To protect mainlenance personnel, use separato cov€red
containers to dispose of broken glass.
)
o
0
t
}P
I
t\
-l :{ #,1
r: Ir {
n h
: U,I lrll iirtr lr II
DO NOT POUR VOLATILE LIOUIDS
INTO THE SINK.
F/FE The laboratory should be equipped with a fire blanket.
The fire blanket is used to smother clothing fires. Small fires
that occur in an evaporaling dish or beaker may be put out by
covering the container with a glass plate, wet towel, or wet
blanket. For larger fires, or ones that may spread rapidly,
promptly use a tire extinguisher. Do nol use a rire exlinguisher
on small beaker fires because the force ol the spray will knock
over the beaker and spread the rire. Tak6 tim€ to become fa-
miliar with the operation and use ot your fire extinguishers.
The use of the proper type ot extinguisher for each class ol
fire will give the best control of the situation and avoid com-
pounding the problem. For example, water should not be
poured on grease lires, electrical lires, or metal fires because
water could increase the hazards, such as splattering ol the
fire and electric shock. The classes of fires given here are
based on the type of material being consumed.
Fire classifications are important lor determining the type of
fire extinguisher needed to control the fir€. Classifications also
aid in recordkeeping. Fires are classified as A, B, C, or D fires
based on the type of material b€ing consumed: A, ordinary
combustibles; B, flammable liquids and vapors; C, energized
 Lab Procedures 527

electrical equipment; and D, combustible metals. Fire extin-
guishers are also classified as A, B, C, or D lo correspond with
the class of lire each will extinguish.
Class A lires: ordinary combustibles such as wood, paper,
cloth, rubber, many plastics, dried grass, hay, and stubble.
Use foam, water, soda-acid, carbon dioxide gas, or almost
any lype of exlinguisher.
Class B lires: flammable and combuslible liquids such as
gasoline, oil, grease, tar, oil-based paint, lacquer, and sol-
venls, and also flammable gases. Use foam, carbon dioxide,
or dry chemical extinguishers.
Class C fires: energized electrical equipment such as slart-
ers. breakers, and motors. Use carbon dioxide or dry chemical
extinguishers to smother the fire; both types are nonconduc-
lors ol electricity.
Class D fires:
combustible metals such asmagnesium,
sodium, zinc, and potassium. Operators rarely encounler this
type oi fire. Use a Class D extinguisher or use fine dry soda
ash, sand, or graphite to smother the fire. Consult with your
local lire department about the best melhods to use for spe-
cilic hazards that exisl at you. facility.
Calilornia Water Pollution Conlrol Association's OPERATORS'
LABORATORY MANUAL. Some ol the ideas and materlal also
came from lhe FISHEB SAFETY MANUAL.
1 1.164 Actdltlonal Beacllng
'1. FISHER SAFETY CATALOG. Obtain trom Fisher Scientitic
Company, Safety Division,4500 Turnberry Drive, Suite A
(Cuslomer Service), Hanover park, IL 60133 or phone
(800) 7i 2-6733.
2. GENERAL INDUSTRY, OSHA. SAFEW AND HEALTH
STANDAqDS (CFFI, Title 29, Labor pt. 1900-1910 (most
recent edition)). Obtain from the US Gov€rnment Printing
Otfice, PO box 979050, Si. Louis, MO 63't97-9OOO. Order
No. 869-062-001 09-6. Price, $61.00.
3. See SAFEry page 1-38, STANOARD METHODS, ZIst
Edition,2005.
OUESTIONS
Write your ansrwers in a notebook and then compare your
answers with lhose on page 575.

Multipurpose e)dinguishers are also available, such as a 1'1.1G List live laboratory hazards.
Class BC carbon dioxide extinguisher that can be used to 1
'1.1H Why should you not work alone in a laboratoM
smother Class B and Class C fires. A multipurpose ABC
carbon dioxide extinguisher will handle most laboratory fire sit- 11.11 True or False? You may add acid to water, but nevsr
uations. (When using carbon dioxide extinguishers, remem- water to acid.
ber that the carbon dioxide can displace oxygen-take appro-
priate precautions.)
11.'1J How would you dispose of a corosive acid?

There is no single type of fire exlinguisher that is etfective

lor all lires so it is important thal you understand the class of
lire you are lrying to control. You must be trained in the use of fudo( Leaaod of 5la,gqota,6
the ditlerent types of extinguishers, and the proper type should ow
be located near the area where that class of fire may occur.
LAgOgN,eq
11.163 Acknowledgments
Po(ions of this section were taken from material writen by
wocE?uEEq
Greenberg, "Safety and Hygiene;'which appeared in th;
A. E. Please answer the discussion and review questions next,

DISCUSSION AND REVIEW QUESTIONS

Chapler 11. LABORATORY PHOCEOUBES
(Lesson '1
ol 5 Lessons)

At the end of each lesson in this c hapter, you will lind some 3. How are pipets emptied or drained?
dscussion and review questions. The purpose of these ques-
is to indicate to you how well you understand the material 4. How would you litrat€ a test solution?
lhe lesson, Write the answers to these questions in your
5. List as many of the seven basic rules for working in a labo-
mlebook
ratory as you can remember.
, How do the operators of water lreatment plants use th€ re- 6. Why should work with certain chemicals be conducted
sults lrom laboratory tests? under a ventilated laboratory tume hood?
Why must chemicals be properly labeled? 7. Why should waler nol be poured on certain types of firss?
 528 Water Treatment
CHAPTER 11. LABOHATORYPROCEDURES
(Lesson2ol5Lessons)
11.2 SAMPLING
11.20 lmportance ot Sampling
Sampling is a vital part of studying the quality ol water in a
water treatment process, distribution system, or source of
waler supply. The major source of error in the whole process
of obtaining water quality information often occurs during sam-
pling. Proper sampling procedures are essential in order to
obtain an accurate description of the material or water being
sampled and tesled. This lact is not well enough recognized
and cannot be overemphasized.
ln any type of testing program where only small samples (a
liter or two) are withdrawn from perhaps millions of gallons of
water under examination, there is potential uncertainty be-
cause of possible sampling errors. Water treatment decisions
based upon incorrect data may be made if sampling is per-
formed in a careless and thoughtless manner. Obtaining good
resulls will depend to a great eltent upon the lollowing factors:
1. Ensuring that the sample taken is truly representative of
the water under consideration
2. Using proper sampling techniques
3. Protecting and preserving the samples until they are ana-
lyzed
((
e quired. A portable conductivity meter is very useful in select-
The greal,cal, gouraesol eftorg in laboralory tegto
are ifi?ro?er 6afi?ling, ?oot ?rcgerrallon, or laak ol
enouqh rni\ing aurinT tcalinO, The acauracy of your
analysis i9 only as good a9 Lhe care thal wag I.aken
in oblaining a repre^entative aarn?le.'
11.21 BepresentativeSampling
11.210 lmpottance ol Represenletive Sampllng
A representative sample must be collected in order for test
results to have any significant meaninq. Without a representa-
tive sample, the test results will nol reflect actual water condi-
lions.
' Reprcsentative Sampl6. A sample portion ot malerial or waler lhat is as nearly identical in contenl and consistency as possible to that in the
larger body ol male.ialor water being sampled.
The sampling ol a tank or a lake that is completely mixed is a
simple matter. Unfortunalely, most bodies ol waler are not well
mixed and obtaining samples that are truly represenlative of
lhe whole body depends to a great degree upon sampling
technique. A sample that is properly mixed (integrated) by
taking small portions ol the water at poinls dislribuled over the
whole body represents the material better than a sampl6 col-
lected from a single point. The more portions taken, lhe more
nearly the sample represenls the original. The sample error
would reach zero when the size of the sample became equallo
the original volume oI material being sampled but, lor obvious
reasons, this method of decreasing sample error is not practi-
cal. The size of sample depends on which water quality indica-
tors are being tested and how many. Every precaution must be
taken to ensure that the sample collected is as representalive
ol the water source or process being examined as is feasible.
11.211 Source Water Sampling
RIVERS. Io adequately delermine the composition ol a
flowing stream, each sample (or set of samples taken at the
same time) must be representative of the entire flow at the
sampling point at that instant. Furthermore, the sampling
process must be repeated lrequently enough io show signifi-
cant changes of water quality that may occur over time in the
water passing the sampling point.
On small or medium-sized streams, it is usually possible to
find a sampling point at which the composition ot the walsr is
presumably uniform at all depths and across the stream. Ob-
taining representative samples in these streams is relatively
simple. For larger slreams, more lhan one sample may be re-
ing good sample sites.
BESERVOIRS AND LAKES. Water stored in reservoirs and
lakes is usually poorly mixed. Thermal stratification and asso-
ciated depth changes in water composition (such as dissolved
oxygen) are among the most frequently observed effects.
Single samples can therefore be assumed to represent only
the spot of water from which the sample came. Therefore, a
number of samples musl be collected at dilferent depths and
from different areas of the impoundment to accurately sample
reservoirs and lakes.
GROUNDWATER. Most ol the physical tactors that pro-
mot6 mixing in surlace waters are absent or much less effec-
tive in groundwater systems. Wells usually draw water from a
considerable thickness ol salurated rock and often lrom sev-
eral diiferenl slrata. These water componenls are mixed by
the turbulent flow ol water in the well before lhey reach the
surface and become availabls lor sampling. Most techniques
 Lab Procedures 529

for well sampling and exploration are usable only in unlinished

about half a gallon per minute). AIlow the water to llow until the
or nonoperating wells..Usually, the only means of sampling the
water in the service line has besn replaced twice. Since 50
water tapped by a well is the collection ol a pumped sa;ple.
feel ('15 m) of three-quarter inch (18-mm) pipe conlains over
The operator is cautioned to remember that well pumps ;nd
one gallon (3.8 liters), four or five minules will be required to
casings can contribute to sample conlaminalion. lf a oumD
replace the waler in the line twice. Collect the sample.'Be sure
has not run for an extended period ol time prior to sampling,
the sample container does not touch the taucet.
the water collected may not be representative of the normii
water quality. Do not lry to save time by turning the faucet handlo to wide
open lo flush the service ltne. This will disturb sediment and
11.212 ln-Plent Sampling incrustations in the line, which must be flushed out Oeforeihe
sample can be collected.
Colleclion of representative samples wilhin the water treat-
menl plants is rea y no diflerent from sample coltection in
a
slream or river. The operator simply wants Io be sure lhe FORMULAS
water sampled is representative ol the water passing that
poinl many water plants. money is spent t6 pui- To estimate the flow trom a faucet, use a gallon jug and
::lpfesampte ln
cnase pumps and pipjng only to sample from a point
a
watch. ll you w€nt a flow of hatf a gatton per irinute, t"nen ine
lhat rs not represenlative ol lhe passing water. A sample lug should be half lull in one minuto or completely full in two
tap in minutes.
a dead area of a reservorr or on lhe lloor of a process
basin
serves no purpose in hetping the plant operalor with conlrot of Volume, gallons
Ftow. GpM _
111 9!1ltyr
The operator. is urged to find each and every ' 'Time, minut€s
sampte pornt and ensure rt is located to provide a useful and
representative sample. If the sampljng point is nol properly lo_ To calculate the volume of a service line, multiply tho area of
€ted, plan to move the piping to a better location. the pipe in square feet times the tength of the
iiire in feei to
oblain cubic feet. The diameter ol a pipe is given i'n inches, so
11.213 Oistribution System Sampting thrs vatue must be divided by 12 inches p6r foot to obtain a
volum€ in cubic teet. Multiply cubic ,set by 7.4g gallons per
Bepresentatrve
.ndicalion sampling jn the distribution syslem is a frue cubic loot to obtain the volumB in gallons.
of system waler quality. Flesults ol s;mpling should
show il there are quality changes in the entire sysiem;r parts (Pipe Area' sq inxPips Length, ft)
0l the system, and may point to the source ol a problem (such
Pipe Volume, cu tt - 144 sq in/sq tt
as tastes and odors). Sampling polnts should be
selected, in Pipe Volums, gat
parl, to trace the course from the finished water
source (at ihe = (pipe Volume, cu ft)(7.a8 gal/cu ft)
well or plant), through the transmission mains, and' the; To determine the time to allow water to flow lrom a faucet to
through the major and minor arteries of the system. -
llush a service line twice. divide lhe pipe votume in
A sim_ laiioni'oy
pling poinl.on a major artery. or on an active the.JIow in gallons per minute. Then. multipty the ,"jult Uy t16
main direc y con_
p!199 ]9 I would be represenrarive of the warer qualirybeing so the line will be flushed twice.
lurnrshed to a subdivision of thrs network. Generatiy. these pri
are used as otficiat sample points in evaluating qrpg rio]lrn9:-sflXa
l1,y-,prlnt.
prevartlng water qualily. -
Frushino Time. min = Ftow, gaumin
Obtaining a representative sample lrom the distribution
syslem is not as easy as it might seem. One would lhink EXAMPLE 2
almosl any faucel would do, but experience has shown other-
wise. Local conditions at lhe tap and in its connection How long should a three.quarter inch s€rvice line 80 feet
to the
marn can easily make the poinl unrepresentalive Iong be flushed if the flow is 0.S GpM?
ol water
furnished to yoUr consumers.
being
Known Unknoyyn
The truest evaluation of water in adistribution system can be Diameter, in = 3/a in Flushing Tlme, min
oblained.lrom samples drawn directly lrom tne mjin. you migni
lhink lhat samples taken lrom a tire hydrant would prove
sais- = 0.75 in
bclory but this is usually not the case. The problem wilh tire hy- Length, tt = 80 tt
drajlts as.sampting points is that they grve erralic (uneven)
sults due to the way they are constructed and lheirlack
16- Flow, GPM = 0.5 cPM
of use.
Ingeneral, an idealsample station is onethathas ashort,
direct Calculate the pipe volum€ in cubic feet and then in gallons.
connection wilh lhe main and is made of corrosion-resistant
nalerial. Pipe Volume, (Pipe Area, sq inxPipe Length, ft)
_
In.mosl smaller water systems,
cu ft
specialsample laps are not
available. Therelore, customer's laucets must be used to col- (0.785X0.7S in),(80 ft)
lecl samples. The best sample points
are front Vard laucets on
_
homes supplied by short service lines (homes with 144 sq in/sq tt
short ser_
yice lines are located
on lhe same side ol the street as the
mler main). = 0.245 cu tt
ll the customer is home. you should contact the person PipB Volume,
in
h€home and obtain permi ssion to collect the sam ple. Discon- gal = (Pipe Volume, cu ttX7.4B gat/cu ft)
nel fie hose from the faucet il one is a ttached and do not
= (0.245 cu ftX7.48 gat/cu ft)
to reconnect the hose when finished collectin g the
sam-
open the faucet to a convenient Ilow for samp,i ng (usually
= 1.833 gal
 530 Waler Treatment

Calculate the flushing time for lhe service line in minutes Hov!,evei since operators themselves are often the sampler-
analyzer, continuous analysis would leave little lime for any-
Flushing (Pipe Volume, gal)(2)
Time, min - Flow, gal/min
thing but sampling and testing. Except for tests that cannot
wait due to rapid physical, chemical, or biological changss ol
(1.8s3 galx2) the sample (such as tests for dissolved oxygen, pH, and tem-
perature) a lair compromise may be reached by taking sam-
0.5 GPM ples throughoul the day at hourly or two.hour intervals. Each
sample should be r€frigorated immediaiely after it is col.
= 7.3 min Iected. At the end ol 24 hours, a portion of each sample is
mixed with the other samples. The size of the portion is in
direct proportion io the llow when the sample was collecled
or = 7 min + (0.3 min)(60 sec/min) and the total size of sample needed for testing. For example, il
hourly samples were collecled when the llow was 't.2 MGD.
use a 12-mL portion sample, and when the flow was '1.5 i,,lGD,
= 7 min and 18 sec use a 15-mL porlion sample. The resulting mixture of portions
of samples is called a COMPOSITE S/AMPLE tn no case,
QUESTIONS however, should a composite sample be collected ror bacterio-
logical examination.
Write your answers in a notebook and then compare your
answers with those on page 575. When the samples are taken, they can either be s€t aside to
be combined lat6r or combined as they are collected. ln both
1 1.2A What are frequently the grealest sources of errors in cases, they should be stored at a lemperature ol4"C untilthey
laboratory tests? are analyzed.
11.28 Why must a representative sample be collected?
11.2C How are sampling points sel€cted in a distribution
system?

11.22 Types of Samples

There are generally two types ol samples collected by
waleMorks operators, and either type may be obtained manu-
ally or automatically. The two types are grab samples and
composite Samples.

11.220 Grab Samples

A grab sample is a single water sample collected at no spe-
cific lime. Grab samples will show the water characteristics at
the time the sample was taken. A grab sample may be pre-
lerred over a composite sample when;
1. The water to be sampled does not flow continuously.
2. The water's characteristics are relatively constant.
3. The water is to be analyzed for water quality indicators that
may change with time, such as dissolved gases, coliform
bacteria, residual chlorine, temperalure, and pH.
11.221 Comp@ite Samples
ln many processes, lhe water quality is changing from
moment to momenl or hour to hour. A continuous sampler-
analyzer would give the most accurate results in these cases.
11.23 Sampllng Device3
Automatic sampling devices a16 wonderful time-savers but
are expensive. As with anything automatic, problems do arise
and the operator should be on the lookout lor potential ditficul-
ties.
Manual sampling equipment includes dippers, weighted
bottles, hand-operated pumps, and cross-section samplers.
Dippers consist of wide-mouth, corrosion-resistant containers
(such as cans orjars) on long handles that collect a sample for
testing. A weighted bottle is a collection container that is low-
ered to a desired depth. At this depth, a cord or wire removes
the bottle stopper so the bottle can be lilled (see Figure 11.5).
Some wal6r treatment facilities use sample pumps to collect
the sample and transport it to a central location. The pump
and its associated piping should be corrosion-resistant and
sized to deliver the sample at a high enough velocity to pre-
vent sedimentation in the sample line.
N,lany wat6r agencies have designed and installod special
sampling stations throughoul their dislribution systoms (see
Figure 1 1.6). These stations provide an excellent location to
sample the actual quality ol water in your distribution system.
 Lab Procedures 531

coRo
. XNOT
RING r*
sroPPER
-/?

BING -------}.

GLASS SAMPLE
BOITLE

WIRE CAGE

5-POUND
r--z?777_ LEAD WEIGHT

Fig. 1 1.5 Sectional view ol homemade depth sampler Fig. 11.6 Distribution system sampling station

11.24 Sampling Techniques
11.240 Surtace Sampling
A surface sample is obtained by grasping the sample con-
lainer at the base with one hand and plunging the boftle mouth
down into the water to avoid introducing ant materiat floating
on the surlace. Position the mouth of lhe bot e into lhe current
and away from the hand of lhe collector (see Figure I i.7). lf
lhe waler is not flowing, then an artificial current aan be creat-
ed by moving the bottle horizontally in the drrection il is point-
ed and away lrom the sampler. Tip the botUe stighlty upward to
allow arr lo exit so the bottte can fi . Tigh y sto;per and tabel
lhe botlle.
Place the bot e in a weighted frame that holds the botfle se-
curely when sampling from a walkway or other structure above
a body ol water. Flemove the stopper or lid and lower the
device lo the water surlace. A nylon rope, which does not
absorb water and wtll not rot. is recommended. Face the bot e
moulh upstream by swinging lhe sampling device downstream
and then allow it to drop into the water, without slack in the
rcp€, Pull lhe sample device rapidly upstream and out of the
water simulating (imitating) the scooping motion of the hand.
sampling described previously. Take care not to dislodge dirt
0I olher debris that might fall into the open sample container
from above. Be sure to label the container when sampling is
completed.
11.241 Depth Sampling
Several additional pieces of equipment are needed for col-
lection of depth samples from basins, tanks, lakes, and r6s€r-
voirs. These depth samplers require lowering the sample
device and conlainer to the desired water depth, then opening,
filling, and closing the container, and returning the device t,o
the surface. Although deplh measurements are best made
with a pre-marked steel cable, the sample depths can be de-
termined by pre-measuring and marking a nylon rope al inter-
vals with a non-smearing ink, paint, or fingernail polish. One of
the most common commercial devices is called a Kemmerer
sampler (see Figure 11.8). This type ot depth sampler con-
sists ol a cylindrical tube that contains a rubber slopper or
valv€ al each end. The device is lowered into the water in the
open position and the waler sample is trapped in the cylinder
when the valves are closed by the dropped messenger.
Figures 1 1 .5 and 1 1.9 show typicat depth samplers. These
samplers are low€red to the desired depth. A jerk on the cord
will remove the stopper and allow the bot e in the depth sam-
pler to fill. Good samples can be co ectod in depths ol wat6r
up to 40 feet (12 m).
 532 Water Treatment
I
{
I
i
I

c
()
c
o () I

() ()
o
oe

Fig. 11.7 Demonstration ol technique used in grab sampling of suiace waters

.Microbiologicat
{Sourc6: US EPA Meihods For Monitging the Ediloim€,n.. D.cemb€r ,979)

,l

Fig. 11.8 Kemmerer depth samplet. (A) nyton line, (B) nes-
senger, (C) catch set so that the sampler is open, (D) top
rubbet valve, (E) connecting rod between the valves, (F)
lube body, (G) boftom rubber valve, (H) knot at the boftom ot
the suspension line, and (l) rubber tubing attached to the
sp ri ngloaded ch eck valve.
(Source: US EPA 'MicdDlogical Method. For Monhoiin! Fig. 11.9 Depth sanpler
the ENircn@.ni O.cember 1 978) (Pe.frlsson ol HACH Company)
 Lab Procedures 533

11.242 Water Tap Sampling regulatory agency as required by the Safe Drinking Water
Acl.
To collect samples from waler main conneclions, lirst ,lush
lhe service line lor a brief period of time. Samples should not 11.27 Additional Reading
be laken from drinking lounlains, resirooms, or taps that have
aerators. Aerators can change water quality indicators such as 1. See page 1-27, COLLECTION AND PRESERVATION OF
pH and dissolved oxygen, and can harbor bacteda under SAMPLES, STAN DARD M ETHODS, 21st Edition.
some conditions. Do not sample from taps surrounded by ex-
cessive foliage (leaves, flowers) or from taps that are dirty, cor- 2. WATER OUAuW SAMPLING GUIDELINES, Second Edi-
roded, or are leaking. Never collect a sample from a hose or tion, January 2005. Obtaln from American Water Works
any other attachment fastened to a faucet. Care must be taken Association (AWWA), CA-NV Section, 10574 Acacia
to be sure that the sample collector does not come in contact Street, Suite D6, Bancho Cucamonga, CA 91730. Price to
with the laucet members, $32.00; nonmembers, $37.00; price includes
cost of shipping and handling.
11.243 First-Draw Samples
The Lead and Copper Flule calls for lirst-draw or first-flush
QUESTIONS
samples. These are waler samples taken at the custome/s Write your answers in a notsbook and then compare your
tap after the water stands motionless in the plumbing pipes for answers with those on pages 575 and 576.
at leasl six hours. This usually means laking a sample early in
ihe day belore water is used in the kitchen or bathroom. 11.2D What are the two general types of samples collected
by water lreatment personnel?
11.25 Sampling Containers and Preservalion ol Samples 11.2e Lisl three water quality indicators that are usually
The shorter the time that elapses between the actual collec- measured with a grab sample.
lion of the sample and the analysis, the more reliable your re- 11.2F How would you collect a depth sample trom a lake?
sults will be. Samples should be preserved il they are not
going to be analyzed immediately due to remoteness of the 11.2G Samples should not b€ collected ,rom water taps
laboratory or workload. Preservation of some types of sam- under whal conditions?
ples is essential to prevent deterioration of lhe sample. Some
water quality indicators, such as residual chlorine and temper-
11.2H What information should be recorded when a sample
is collected?
alure, require immediate analysis, while others can be pre-
served and lransporled to the laboratory. A summary of ac-
ceplable sample containers, preservative, and maximum time
between sampling and analysis is shown on Table '1 1.2.
Whatever type ol container you use, clearly identify the
fud of L e+e ottZ of 5 l.omow
oyL
sample location, dale and time of collection, name of collector,
and any other pertinent inlormation. L gOgNOeV
'11.26 Reporting wocE?uQE4
The waler system owner (waler utility agency) is respon-
sible for reporting lab resulls at regular frequencies to the Please answer the discussion and review questions next.

DISCUSSION AND REVIEW QUESTIONS

Chapter'11. LABOHATOBYPROCEDURES
(Lesson2of5Lessons)

Write the answers to these questions in your notebook. The

queslion numbering continues from Lesson 1.

8. Why are prop6r sampling proceduros important?

9. What is meant by a representative sample?
10. Generally speaking, how would you obtain a representa-
tive sample?
11. Under what conditions and why would you preserve a
sample?
 534 Water Treatment

TABLE 11.2 RECOMMENDATION FOB SAMPLING AND PRESEBVATION OF SAMPLES ACCOFDING TO MEASUHET/IENT'

Vol. Beq. Max. Holding

Measuremenl (mL) Conlainerb Preservative Timec

PHYSICAL PROPEBTIES
Color 500 P,G Cool,4"C 48 hours
Conductance 500 PG Cool, 4'C 28 days
Hardnessd 100 EG HNO3 to pH <2, 6 months
H,SO4 to pH <2
Odor 200 G only Cool,4"C 6 hours
pHd NG Det. on site lmmediately
Flesidue, Filterable 100 qG Cool, 4"C 7 days
Temperature 1,000 EG Det. on site lrnmediately
Turbidity '100 PG Cool,4'C 48 hours
METALS (Fe, Mn)
Dissolved or Suspended 200 EG Filter on site, 6 months
HNO3 to pH <2
Tolal 100 PG Filter on site. 6 months l
HNO3 to pH <2
I
INORGANICS, NONMETALLICS
Acidity 100 EG Cool, 4'C 14 days
Alkalinity 200 EG Cool, 4'C 14 days
Bromide 100 RG None Req. 28 days
Chloride 50 P,G None Fleq. 28 days
Chlorine, Total Residual 500 BG Det. on site lmmediately
Cyanide, Total and Amenable 500 qG Cool, 4'C, 14 days
to Chlorination NaOH to pH >'12,
0.6 gm ascorbic acid€
Fluoride 300 D None Req. 28 days
lodide 100 EG Cool, 4'C 24 hours
Nitrogen
Ammonia 500 P,G Cool, 4"C, 28 days
H2SO. to pH <2
Kjeldahl and Organic 500 P,G Cool, 4"C, 28 days
HrSO. to pH <2
Nilrale-Nitrite 204 qG Cool, 4"C, 28 days
H,SO{ to pH <2
Nitrate 100 RG Cool, 4'C 48 hours
Nitrite 100 P,G Cool, 4'C 48 hours
Dissolved Oxygen
Probe 300 G with top Det. on site lmmediately
Winkler 300 G with top Fix on sile, store 8 hours
Phosphorus in dark
Orthophosphate 50 P,G Filter on site, 48 hours
Cool. 4'C
Elemental 50 Cool, 4"C 48 hours
Total 50 BG Cool,4'C, 28 days
H,SO4 to pH <2
Silica 50 P Cool, 4'C 28 days
Sulfate 100 P,G Cool, 4"C 28 days
Sulfide 100 EG Cool,4'C, 7 days
add zinc acetate plus
HrSO4 to pH >9
Sullite 50 PG Det. on site lmmediately

"Required Containers, Preservation lechniques, and Holding Times.' CODE OF FEDERAL REGUL,4IONS, Protection of the Environment, 40, parts
136'149, 2007. Ihis publicat;on is available Irom the US Government Printing Oflice, Superintendent of Documenls, pO Box 979050, St. Louis, MO
63197-9000. Order No. 869-062-00160-6. Price, $61.00.
Polyethylene (P) or Glass (G). For melals, potyethylene with a polypropylene cap (no liner) is preferred.
Holding limes lisled above are recommended tor properly preserved samples based on currentli available data- lt is recognized lhat for some
sample types, exlension ol lhese limes may be possible while lor other lypes, lhese limes may be loo tong. Where shippind regulalions prevent
the use of lhe proper preservalion lechnique or the holding time is exceeded, such as the caie of a 24-hour composiid, tn-e fiial reportio oata
for these samples should indicate the specific variance.
Hardness and pH are usually considered chemical properties of water rath6r than physical properties.
Use ascorbic acid only i, residual chlorine is present.
SPECIAL NO|E:whenever you collecl a sample lor a bacteriological test (coliforms), be sure to use a sterile plastic or glass bot €. ll the sample
conlains any chlorine residual, sutticieft sodium thiosulfaie should be added to neutralize all ol th€ chlorine residual. Usually, two d.ops (b.l
mL) of ten percenl sodium lhiosullate lor every 100 mL o, sample is sufficient, unless you are disinfecting mains or storage tanks.
 Lab Procedures 53S

CHAPTER 11. LABORATORYPROCEDURES

(Lesson3ofsLessons)

11.3 WATER LABOBATORY TESTS

2. Sulfuric acid (H,SOJ, 0.1 N.
1. Alkalinity 3. Slandard sulfuric acid, 0.02 M Dilute 2OO mL O.lO N
A. standard acid to 1 liter using a votumelric ftask. To
Oiscussion deie;_
mrne lhe volume to be diluted, use the lollowing lormula:
The.alkalinity ol a water sample is a measure
ol lhe water.s (Slandard, 0.02 N) x (1,OOO mL
capacrty to neulralize acids. ln nalural and treated Vo,ume Diluled, mL
waters, al- =
kalinity is the result of bicarbonates. carbonates. (Calcul ated Normality. O.'lO N)
and hvdiox-
roes ot the metals of calcium, 63nn""1rrn,
sodium. (0.02 N) x (1,000 mL)
"nd
Many of the chemicals used in water treatment,
. such as (0.10 N)
alum.,chlorine. or lime. cause changes in af Uf inrty.
ffte affrii
rrnrty determination is needed when calculatng
chemical doi_
ages used in coagutation and water softenjnglaff,afinity = 200 mL
muit
also be known to calculate corrosrvjty and lo 6stimate
the car_ E. Procedure
:g!g]" Atkatiniry is usua y expressed in
tetms ot.nLd."::i_gt-yater.
cALctuM cAREoNATE (CaCq) EOUTUALENTI Total and Phenolphthalein Alkalinity

There are ,five alkalinity condiljons posstble in a water 1. Take a clean beaker and add 1OO ml ol samDle (or olher
.
ple.(l) bicarbonate atone, (2) bicarbonale and carbonale.sam_t3)
sample volume that will give a titration voluma ol less
caroonate atone, (4) carbonate and hydroxide, and (5) than 50 mL of acid titrant)
hvdroxl
oe arone. Ihese five conditions may be distinguisifrei 2. Place of pH meter into beaker
and containing
quantities determined from the resutrs ot -electrodes
aciO fitritons OyiiiJ sample.
method given below.
3. Stir sample slowly (with a magnetic stirrer if possible).
B. What ls Tested
4. Check pH of sampte. ll pH is 9.3 or betow, then there is
Sample Common Range, mg/L no phenotphthalein alkaljnity present and you can go to
Step 6.
Raw and Trealed Surface Water 20-300 5. ll the pH is grealer than 9.3, hlrate very carelullv to a oH
We Water 80-500 of 8.3 wilh 0.02 ,V H,SO4. Hecord th; arorni ot i6iJ
used to this point.
C. Apparatus Required
6. Continue lo titrate to pH 4.5 with O.O2 N H,SO..6 Flecord
pH meter graduated cylinder (tOO mL) rhe totat amount of acid used lrom starting
reference electrode ioinilo finish.
buret (25 mL)
glass electrode buret support
7. Calculate total and phenotphthalein (if presenl) atkatini-
magnetic stirrer ties.
beaker (250 mL)
magnetic slir-bar wash bottle E Example
analytical balance desiccator
Results rrom atkalinity titrations on a ,inished water
D Reagents were as follows:
sample

NOfE Standardized solutions are commercially available Sample size '100

lor most reagents. Relet lo STANDARD UefUbOS yoi = mL
tt
wish to prepare your own reagents. mL titrant used to pH 8.3, A
= 0.5 mL
Sodium carbonate (Na,CO3) solution, approximatety lotal mL of titrant used, B = 6.8 mL
0.Os
Acid Normatity, N = 0.02 NH,SO.

calcium cabonate (caco) Eouivatenl' An expressron

of the concentration ot specilied constiluents in water
in t€rms ol their equivalent value
lo calcrum
iJ";;;;U;i"t;.
carbonate' For example, the hardn".i in *"r", *t,i"r'
."gnesiuh and orher ions is usua y describod as catcium
ii"ii3'd.""1,1,,,",en,s, mu,,ip,y
ffiiiq"i"li'l'.Jj]:idlil-fA;?'.ii;iif::llffo":1ifl*ll;%'rli"."d.:;;i'"i",it"rio
reconmends
"#"",t "iiJill
rikatins ro a pH or 4 5 ,or rout;ne o' auromared anarys€s. However.
ffil:;H#"t"1*"r"":;;l"tJ5:[::"?:?-"",fiil] orher pH

,
 536 Water Treatment

(Alkalinity)

OUTLINE OF PROCEDURE FOF ALKALINITY

"' f-".
pHIETEi pH lilETEn
oao olo

,
IEE
1. Add 100 mL ol 2. Place electrodes' of pH 3. Tikate down to pH 8.3 (if
sample. meler in beaker necessary), with
0.02 N H2SO4.

--,>

9H EA
oao '.E

4. Continue lo titrate
to pH 4.5.

. Some pH meters
have a single .combination,, electrode.
 Lab Procedures 537

(Chlorine Ftesidual)
G. Calculations
2 99! !1gu 310-i.1 METH1DS FOR CHEMTCAL
Phenolphthalein Alkalinily, mg,/L as CaCO3 SIS OF WATER AND rrylASIES, March - ANAL4-
" 1919.-
_AxNx5O,OOO
ml of Sample QUESTIONS
Write your answers in a nolebook and then
(0.5 mL) x (0.02 N x 50,000 compare your
) ansiwers wilh those on page 576.
x 100 mL
1 1.3A What chemicals used in water treatmenl
will cause
changes in atkatinity?
= 5 mg/L
Total Alkalinity, mg/L as CaCO3 1 1.38 Why is tt important to know the alkatinity ol
a water
sampl6?
_ BxNx50,OO0
mL of Sample 2. Chlorine Bestdual
(6.8 ml ) x (0.02 N ) x 50,000 A. Discussion
100 mL Chlorine is not only an excellent disinfectant
, but also serves
l"-
j"::t_ rlh .iron, manganese, protei,
= 68 mg/L ano many taste_ and odor-producing compounds "rOitu""e.-.-JiiO",
fi H. lnterpretation of Besults
the test results and the informalion given
prove the quatity of treated water. t"
conrrot microorganrsms rhat mioht
and flocculalion. keeps lilter me;ja tree
.t"rti"
to helo im_
iooir.'.,, in[rir" n![J'ii
*iir, Jo-"jri5ioi
,.,From
diff€rent types
below, the of sf ir" gro*t"h;, irj
ol alkalinjty contained in a water sample helps bleach out undesirable color
d€lermined.
can be
There
are two general types o, residual chlorine produced
, in
Alkalinity, mg/L as CaCOJ chlorinated water. They ari: (1) free residual
chlorine en.t r2l
combtned residual chlorine. Free resiOuat ctrtorine
Titration r#rs'-tJ
Result Bicarbonate Carbonate :l:XX, l.p, llq*hrorus. acid fsoclt. a,j r,e iiipociiJ,iil
Hydroxide ,. uomDrnect residual chtorine genera[y
:"Jj-,_y_y, rdfers to the
cnronne-ammonia- compounds of .orocnfor"ri.r,ne (NHrcDl
P=0 T 0 drchtoramine (NHctJ. and trichtoramine
P is less
0 tllcr, ol' nitrijeiiii_
chloride). Both types ol residuals act as dGinfectanis.
f but
lhan !,4 r -2P 2P 0
differ in their capacity to produce a gerr+ree
,raiei s;pi[
P = ,/zT 0 during the same contact time.
2P 0
P is greater
]t,o, a the

I
to positive aspects o, chtonnatron,
lhan th T 0 2r -2P 2P -T _]::9
may be some adverse effects. potentially carcinoge;ic
there
P=T 0 0 T organjc compounds such as chtorotorm ;;;r:
;nO ortreiir_ifr,fs
where P = phenolphthalejn alkalinity be formed.during lhe chlorination pro"""". io,ini.i.! m-av
| = total atkatinity "ii
:9y."T9 "fi"gr:. the operator shouta le ramiriar wirii tir-e-co#
centrattons
Example:The example in ,,G, above gave ol free and combined residual cntorine proOuced
the following resulls
phenolphlhalein atkatinity S mO/L ::^11"a191
Iremety
suo.pty.rg owing chtorinarion. eorh resiOuais
rmportanl jn producing potable ii! e-xl
= a water ttrat is n.,t
sale to dnnk but is also free of objectronaOte
tastes ""i,,
tolal alkalinity = 68 mg/L oOor"l
Since. the phenolphthalein atkalinity (5
B. What ls Tested "nO
mg/t) is tess lhan
one half of lhe toral atkatinity (6S
mg/ti fro; r;"i;;, i;;; Common Flanoe
liere is bicarbonate and carbbnate
atiatinlty in fne watei. Residual Chlorine, mo/L
Source Free -
--Tle.bicarbonate alkalinity in this case is egual to T _ 2p or Total
68 mg/L - (2 x 5 mg/L) = SS mg/L asCaCO" Chlorinated Raw Surface
Tte carbon_ate atkatinity is equat to
mg4 as CaC03.
2p or 2 x S mg/L = 10 Water (prechtorination) 0.3 to 3 0.5 to s
Chlorinated Finished
L Precautions Surface Water
l (Post-Chtorination)
l l. The sample should be analyzed
O.2 lo 1 0.3 to 'l.5
as soon as possible, at Well Water 0.2 to 'l
leasl within a few hours afteicollectron. 0.2 lo 1
l
2. Thesampte should nol be agilated.
C. [4ethods
warmed, fjltered, di_
luted, concentrated, or allerei in any Thereare-eightm€thods listedlormeasuring
way. residualchlorine
nlhe 21sl Edrtron ot
J. Reference
STANDARD METHODi.
mos-l_practicat and appropriate procedure
il;;t"; i;;
;i
in any particuiai
1. rn-starce generally depends upon
See page 2-27, STANDARD METHOD,S, 2.lst Editjon. the characteristics;ithe water
oerng examined. The AMpEROMETB\CT titrarion
mertroO iii
I Arnperomettic (am"p Un.o+,lEt-ricr1.
,| A method ot measurement that records
etectric current . generaled, rather than recording volt-
ol measulng concentrar'tions ot cenaln sutslinceJ-* flowino or
Amperomekic ti lralion is a means .
age.
---"'Y
 538 Water Treatment

(Chlorine Residual)
standard of comparison for delerminlng free or combined DPD Colorimstric Msthod (Field Comparator Kit)
chlorine residual. This method is relatively free ol interferences
but does require greater operator skill to obtain good results. ln Use reagents suppliod by kit manufacturer.
addition, Ihe titration instrument is expensive.
DPD Titrimetric Method
The DPD$ methods are simpler to perform than ampero- 1. 1 + 3 H2SO.. Carefully add 10 mL concentrated sulfurjc
metric titration but are subject to interferences due to manga- acid to 30 mL distilled waler. Cool.
nese. Field comparator kits are available from several sup-
pliers such as Orbeco-Hellige, Wallace & Tiernan, and HACH. 2. Phosphate buffer solution.
3. DPD indicator solutaon.
4. Standard ferrous ammonium sulfate (FAS) titrant, O.OO2B2

5. Potassium iodide, Kl, crystals.

F. Procedure
Amperometric Titraiion Method
Follow manufaclurer's instructions.
DPD Colorimetric Method (Field Comparator Kit)
To measure chlorine residuals you should follow the direc-
tions provided by the manufacturer of the equipment or instru-
ment you are using,
6{_* lf you are disinfecling clear wells, dislribution reservoirs, or
mains and very high chlorine residuals must be measursd. a
D. Apparatus Bequired drop-dilution technique can be used to estimate the chlorin€
residual. The procedure is as follora,s:
l. Amperometric Titration Niethod
1. Add 10 ml of distilled water and one powd€r pillow of
See page 4-62, STANDARD METHODS,2lst Edition DPD reagent (or 0.5 mL ol DPD solution) to the sample
and amperomelric titrator's instruction manual. tube of the test kit.
2. DPD Colorimetric Nrethod (Field Comparator Kit) 2. Add a sample of the waler being tested drop by drop to
the sample tubs until a color is produced.
Field Comparator
Sample cells 3. Flecord the number of drops added to the sample lube.
Assume one drop equals 0.05 mL,
3. DPD Titrimetric Method,

Graduated cylinder (100 mL)

4. Determine the chlorine rBsidual in the sample as a result
of the color produced and record the residual in milli.
Pipets (1 and 10 mL) grams per Iiter.
Flask, Erlenmeyer (250 mL)
Buret ('10 mt)
EXAMPLE 3
Magnetic stirrer
Magnetic stir-bar The recorded chlorine residual is 0.3 mg/L. Two drops of
Balance, analytical sample produced a chlorine residual of 0.3 mg/L in 1O mL of
distilled water Assume 0.05 mL per drop.
E. Reagents
NOTE: P'epa?d reagents may be purchased from lab-
Knorrn unknown
oratory chemical supply houses. Chlorine Bssidual, mglL = 0.3 mglL Actual Chlorine Residual,
Sample Volume, drops = 2 drops nglL
Amperometric Titratiod Method
1. Standard phenylarsine oxide (pAO) solution, 0.00564 N
Distilleo Water, mL ='10 mL
CAU T I O N : Toxic-avojd ingestion. Calculat€ the actual residual in milligrams per lit€r.
2. Acetate buflersolution, pH 4. Actual Chtorine (Chlorine Residual, mg/t)(Distilted Water, mL)
Hesrduat, mo/L
3. Phosphate butfersolution, pH 7. (Sampls Volume, dropsxo.os mLldrop)
(0.3 mg/Lxl0 mt)
4. Potassium iodide solution.
(2 dropsxo.os mlldrop)
Siore in brown glass, stoppered botfle, preferably in the
refrigerator. Discard when solution becomes yellow.
= 30 mg/L

0
OPD (pronounce as separate leliers). A method of mea€uring the chlorine residual in waler. Th€ residual may be determined
by either titrating
or comparing a developed color wilh color standards. DpD sta;ds for N,N-diethyl-p-phenytene_diamine.
^s Some regulalory
agencies require the use ol the oPD Tilrimetric Metnoo f cntorine iesioual lesting is used in ptace ol some ot th€ colilorm tosts.
 Lab Procedures 539

(Chlorine Besidual)

[*,,*,* )!,)t
"n,.,,r,
1. Place 5 mL each of buffer reagent and DPD indicator in a
mL ol sample
Reading A, mL
Fieading B, mL
Beading C, mL -,
00 mL
.4 mL
.6 ml
.7 mL
250-mL flask and mix.
Calculalion
2. Add 100 ml of sample and mix.
READING CHLORINERESIDUAL
3. Tilrate rapidly with standard FAS titrant until red cotor is
discharged (disappears).
A = mg/L kee residual chlorine
B-A = mg/l monochloramine
4. Record amount of FAS used (Fleading A). lf combined re- C-B = mg/L dichloramine
sidual chlorine fractions are desired, continue to next C-A = mg/L combined available chlorine
t slep. C = mg/L lotal residual chlorine
FOR COMBINED RESIDUAL CHLORINE Example: The concenlrations ol the ditterent types of re-
sidual chlorine presenl can be calculated trom the
Monochloramine
inlormalion given in (H).
5. Add one very small cryslal ol Kl and mix. ll monochlor- READING CHLOBINE BESIDUAL
amine is present, the red color will reappear.
1.4mL = 1 .4 mg/L free residual chlorine
6. Continue titrating carefully until red color again disap- B-A = 1.6-1.4 = 0.2 mg/L monochloramine
pears.
C-B = 1.1 mg/L dichloramine
7. Becord reading of FAS in lhe buret used to this point (this C-A = 2.7 - 1.4 = 1.3 mg/L combined available chlorine
includes amount used above). This is Fleading B. 2.7 mL = 2.7 mgy'L total residual chlorine
t oichloramine J. lnterpretalion of Besults
8. Add several crystals Kl (about 1 gm) and mix until dis- 1. Any chlorine taste and odor that may result from chlorina-
solved. tion would generally be from the dichloramine or nitrogen
trichlorid6 nuisance residuals.
9. Lel stand 2 minutes.
2. Free residual chlorination produces the best results when
10. Continue titrating until red color again disappears. Record the lree r€sidual mak6s up more than 80 percent ol the
amount of FAS used to this point. This includes amounts total residual. Howeyer, this will not be the case if am-
used in two previous titrations for lree and monochlor- monia is added to the water being treated to torm chlor-
amine. This is Reading C. amines in ordsr to prevent the formation o, THMs.
iIOIE Manufacturers of laboratory equipment are continu- K. Reference
ally developing faster and more accurate ways to
measure chlorine residual. lf you have new equip- See page 4-56, STANDARD METHODS, 21st Edition.
ment, Iollow the manufacturer's procedures.

G. Precaulions

1. For accurate results, careful pH conlrol is essential. The

pH ol lhe sample, buffer, and DPo indicator togelher
should be between 6.2 and 6.5.
2. lf the sample contains oxidized manganese, an inhibitor
f; must be used.
I 3. Samples should be analyzed as soon as possible atter
collection.
4. ll nilrogen trichloride or chlorine dioxide are present, spe-
cial procedures are necessary.
5. Operators using lhe DPD colorimelric method to test
water lor a free chlorine residual need to be aware of a
potential error that may occur. lf the DPD test is run on
water containing a combined chlorine residual, a precipi-
tale may form during the test. The particles of precipitated
material will give the sample a turbid appearance or the
appearance of having color. This turbidity can produce a
positive test result for lree chlorine residual when there is
aclually no chlorine present. Operators call this error a OUESTIONS
"false positive' chlorine residual reading. Write your answers in a notebook and thsn compar6 your
answers with thos€ on page 576.
H. Example
A sample laken after prechlorination at a filtration plant
11.3C List some of the important benefits of chlorinating
water.
was lested for residual chlorine using the DPD Titrimetric
Method. 11.3D What is a polential adverse etfect from chlorination?
540 Water Treatment

(Chlorine Besidual)

OUTLINE OF PFOCEDUBE FOR CHLOBINE RESIDUAL

ML FAS T_

FBEE RESIDUAL
CHLORINE

1. Place 5 mL bufler 2. Add 100 mL 3. Titrate wlth

and OPD indicator of sample. FAS.
into flask and mix.

ML FAS

COMBINED RESIDUAL
CHLORINE
l\,lonochloramine
\ 1
4. Add one crystal Kl
A
5. Titrate with FAS until red
color disappears.

Dichloramine
ML FAS

I 0

6. Add sevoral crystals Kl. 7. Titrate with FAS until red

color disappears.
 Lab Procedures 541

(Chlorine Demand)
3. Chlorine Demand stock solution, add 25 mL gl the doslng solution. Note:
A. Discussion ln measuring the volume of the dosing solution, notice
that '1 mL of the 0.025 N thiosullate titrant is equal to
The chlorine demand of water is the difference between the 0.9 mg chlorine.
amount of chlorine app|ed (or dosed) and lhe amount of tree,
combined. or total residual chlorine remaining at the end of a d. Tilrate with standardized 0.025 N thiosulfate titrant
specifjc contact period. The chlorine demand varies with the until the yellow iodine color almost disappears.
amount of chlorine applied, tength of contacl time. pH, and e. Add 1 to 2 mL starch indicator solution.
lemperature. Also, lhe presence of organics and reducing
agents in water will inf,uence the chlorine demand. The chlo: f. Continue to titrate until blue color djsappears.
rine demand test should be conducted with chlorine gas or
wilh granular hypochlorite, depending upon which tor;t you (mL Thiosulfale used) v Nx 35'45
mg/L cl as cl'/ml
' -
usuallv use lor chlorination mL ot Dosing Solution

The chlorine demand test can be used to determine the where fV = normality ol Thiosulfate Titrant
best chlorine dosage to achieve specific chlorinalion objec- 3. Acetic acid, concentrated (glacial).
tives. The measurement of chlorine demand is pertormed by
treating a series of water samples in question witir tnown but 4. Potassium iodide, Kl, crystals.
varying amounts of chlorine or hypochlorite. After the desired
contact time, calculation of residual chlorine in the samples
5. Slandard sodium thiosulfate 0.025 N.
will demonstrale which dosage satisfied the requiremenG of 6. Chlorine demand-free water. prepare chlorine demand_
the chlorine demand in lerms of the residual desiied. free waler lrom good-quality distilled or deionized water
by adding sutficient chtorine to give 5 mg/L free chtorjne
B. What Is Tested residual. Atter standing 2 days. this soluiion should con_
tain al least 2 mg/L lree residua, chlorjne. If not, discard
Common Range
Chlorine Demand, mg/L and obtain better quality water. Bemove remaining lree
chlorine by placing the sotution jn the direct sunlight:After
SurfaceWater O.S to 5 several hours, measure total chlorine residual, Do not use
Well Water 0.1 to 1.3 until last trace of chlorine has been removed.
C. Apparatus 7. Starch indicator
ln addition to the apparatus describecl under one of the E. Procedure
melhods for chlorine residual, the following jtems are required:

Flasks, Erlenmeyer
1. lvleasure a 500.m1sample into each of five to ten 1,OOO-
(1,OOO mL) mL flasks or botfles.
Pipets (5 and 10 ml) 2. To the first ftask, add an amount ol chlorine that leaves no
Graduated cylinder (5OO mL) residual at the end of the contact time. Mix.
Flask, volumetric ('1,OOO mL) 3. Add increasing amounts of chlorine to successive por_
Flask, Erlenmeyer (250 mL) tions of the sample and mix.
Buret (25 mL) 4. Measure residual chlorine after the specific contact time.
Flecord resulls.
0. Reagents
addition to the reagents described under the method you
5. O-n graph paper, plot the residual chlorine (or the amount
.ln of chlorine consumed) versus chlorine dosage.
will use to determine chlorine residual, the following items;re
also needed: E Precautions
1, Stock chlorine solution. Obtain a suitable solution from a 1. Dose sample porlions at time intervals that will leave you
chlorinator solution line or by purchasing a botfle ol enough time for chlorine residual testing at predei€r_
household bteach (.Clorox, or simitar produit). Store in a mined contact times.
dark, cool place to.malntain chemical strenoth. House_
hold bleach products usua y contain approximately live 2. Conduct test over the desired contact time. lf test objec_
percent available chtorine, which is about 5O,OOO mg7l. tive is to dupiicate your plant contact time, lhen mitch
plant detention time as closely as possible.
2. Chlorine dosing solution. lf using household bleach, care-
tully pipet about 1O mL of the bte;ch into a .t ,oOO-mL votu_ 3. Keep samples in the dark, protected from sunlight.
melric flask. Fill to the mark with chlorine demand-free 4. Keep temperature as constant as possible.
waler and slandardize. lf usinq a stock chlorine solution
obtained from a chlorinator solution line, simply standard- 5. Sterilize all glassware if test has a bacteriologic objec-
ize this solulion direc y. tive.

Slandardization: G. Example
a, Place 2 mL acetic acid and 20 mL chlorine demand- A raw water sample was collected trom a river to delermine
free water in a 250-mL flask. chlorine demand.
b. Add about 1 gm Kl crystals. Contact Time = 30 minutes (plant Detention Time)
c. Measure into the tlask a suitable volume of chlorine PH = 76
dosing solulion. lf using household bleach as your Temperature = 15"C
542 Waler Treatment

(Chlorine Demand)
OUTLINE OF PBOCEDURE FOB CHLOBINE DEMAND

.NO. 1. . t{o, 2' . xo. 3. NO. a NO.3

m
1. Measure 500 mL water sample into each container.

IrlO. t' lilo.2 t{o.3 NOi 4 ilo: 3'

2. Add desired dosage of chlorine to

each container.

3. Atter end of desired

contact time, measure
chlorine r€sidual. Plot
results on graph paper

0
 Lab Procedures 543

(Chlorine Demand)
Flesults from the chlorino demand test we.e as follows: Figure 11.1'l is a plot of the chtorine demand curve. Note
ChtorineAdded, Total Residual that the far right end of the curve is flat, which indicates that
Flask No. mglL Chlorine after 30 min, mg/L the chlorine d€mand has b€en satisfied. The curv6 reveals
that lhe chlorine demand will increase as chlorine is added l;
1 0.5 0.36
water until you have gone past the breakpoint.
2 1.O 0.82
3 1.5 1.14
cHLOiTNE DE[|AriD, rc/a
4 2.O 0.60
c 2.s o.75
6 3.0 1.25
H. Calculation
Calculate the chlorine demand by using the formula:
Chlorine Oemand,
_ Chlorine Added, Total Residual Chlorino,
mglL-mg/L-mg/L
Added. Totat Residual Chtorine
Chtorine
Frask No. mglL Chtorine. mg/L = Demand, mo/L

1 0.5 0.36 0.14

2 1.0 0.82 0.18
3 1.5 .14
1 0.36
4 2.0 0.60 1.40
5 2.5 0.75 1.75
6 3.0 1.25 1.75
I
Fiq. 1 l.t 1 plot ot chlorine demand
on gruph paper
,lestV€luable knowtedge can be gained by plotting the data from
results_ Figure 1 1.1O
is a plot
of thsintorin; aOOeO versus
me lree chtorine residual after 30 minutes.
l. Fleference
Bv drawino a
betwgen lhe ptoning poinrs, a rypi;;l 0,""'iliini See page 2-41, STANDARD METHOOS, 2.1st Ed ion.
:l]:og[]rl_e
cnonnalton curve is produced.
OUESTIONS
Write your answers in a notebook and then compare your
t answers with ihose on page 576.
11.3E What conditions can cause variations in the
chlorinB
demand ol water?

a
11.3F How do€s lhe operator use the results Irom lhe
chlo_
rine demand test?
- 11.3G Calcxlate the chlorine demand lor a raw water
sam_
ple if chtorine is added at Z.O mg/t anO
lhe-free
i0r residual chlorine after 30 minutes w;s 0.4 ngL. --

fudrtLoaaothof bl.o*0rc
olL
c}ltoRDtElooEo, nc,r u\goeffioce
Fig. 11.10 Briakpoinl chlorination curve
wocEPucth
Please answer the discussion and review questions
next.

DISCUSSION AND REVIEW OUESTIONS

Chapter 1l. LABORATOBY PROCEDURES
(Lesson 3 ofS Lessons)

Wrile the answers to these questions in your notebook.

-
queslron The 13 What could be th6 causos of tastes and odors
numbering conlinues from Lesson 2. lhat might
result from chlorination?
12. Whal precautions must be taken with the sample when
conducling the alkalinity test? 14 What precautions should be exercised when performing
a
chlorine demand test?
 544 Water Treatment
CHAPTERll. LABORATOHYPROCEDURES
(Lesson4ol5Lessons)
4. Colirorm Bacteria
A. Discussion
An improperly treated or unprolected water supply may con-
tain microorganisms that are pathogenic, that is, capable of
producing disease. Tesling for specific pathogenic micro-
organisms (pathogens lhat cause diseases such as typhoid,
dysenlery cryptosporidiosis, or giardiasis) is very time-
consuming and requires special techniques and equipment.
So, instead ol testing for specific pathogens, water is generally
analyzed lor the presence of an "indicator organism," ihe coli
Iorm group ol bacteria.
The presence or absence of coliform bacteria is a good index
ol the degree ol bacteriologic safety of a water supply. ln gen-
eral, coliform bacleria can be divided into fecal or non-fecal
groups. Fecalcoliform bacteria occur normally in the intestines
of humans and other warm-blooded animals. They are dis-
charged in great numbers in human and animal wastes.
Coliforms are generally more hardy than true pathogenic bac-
teria and theirabsence from water is thus a good indication that
the water is bacteriologjcally safe for human consumption. The
presence of coliforms, however, indicates the potential pres-
ence ol pathogenic organisms that may have entered the water
with them, and suggesls that the water is not safe to drink.
The colilorm qroup ol bacteria includes all the aerobic and
10
FAC U LTATI V E anaerobic gram-negative, nonspore{ormin g,
rod-shaped bacteria lhat ferment lactose (a sugad within 48
hours at 35'C (human body temperature). The fecal colilorm
can grow at a higher temperature (45.C) than the nonjecal
coliform.
The bacteriological quality of water supplies is subject to
control by federal, state, and local agencies, all of which are
governed by the rules and regulations contained in the Sale
Drinking Water Act. This law stipulates the methods to be
used, the number of samples required, and the maximum
levels allowed for colilorm organisms in drinking water sup-
plies. The number ot samples required is generally based on
population served by ihe water system.
The primary standards (t CLs) tor colilorm bacteria have
been established to indicate the likely presence of disease-
causing bacteria. The Total Coliform Flule uses a presence/
absence approach (ralher than an estimation ol how many
coliforms are present) to determine compliance with the
standards. The Maximum Contaminant Level Goal (MCLG)for
colilorm is zero-
Compliance under the Total Colilorm Rule is determined on
a monlhly basis. ln general, coliform must be absent in at least
95 percent of the samples lor those larger systems lhat collect
more lhan 40 samples per month. Smaller systems lhat collect
to Facullative (F
AcK'ul-TAY'tive). Facullalive bacleriacan use eilherdissolved molecularorygen oroxygen obtained lrom lood materials such as
sulrale or nitrale ions. ln olher words, facullative bacteria can Jive under aerobic or anaerobia aonditions.-
fewer than 40 samples cannot have coliform-posilive results in
more than one sample per month.
Whenever a routine coliform sample is coliform-positive, th€
regulation calls for determination ol the presence ol tecal coli-
lorm (E. coli) andlot repeat sampling. Whenever fBcal colilom
(E col, are presenl in the rouline sample and lhe repeat sam.
ples are colitorm-positive, there is a violation of lhe i,4CL, addi-
tional repeat sampling is required. and notification ot bolh the
state and the public is required.
Also see ,yAIEE TREATMENT PLANT OPERATTON,
Volume ll, Sections 22.22,'Microbial Standardsl' and 22.220,
'Total Colilorm Rule.'
B. Test Methods
There are a number of approved water testing procedures
lor total colilorm bacteria. This lesson will provide detailed dis-
cussions for the following procedures: (1) the Multipl6 Tube
Fermentation Method (sometimes called the Most Probable
Number or MPN procedure), (2) the Membrane Filler (MF)
Method, (3) the Membrane Filter Method-Escherichia coli
(EPA Method), (4) the Presence/Absence (P/A) Method, (S)
the ColilertrM Method, and (6) the Cotisure Method. Other
approved testing procedures include the E'ColiterM Test, the
m-ColiBlue24@ Test, the Beadyculto Colilorms 100 Presence/
Absence Test, the ChromoculP Colilorm Agar Pressncd
Absence Membrane Filter Test, and the ColilagrM presence/
Absence (PiA) Test. Additional information on these tests is
provided at the end ol this lesson (See G. "Additional Test
Methods," page 563).
1 . Multiple Tube Fermentation Nrethod (or MPN Procedure)
The muliiple tube coliform test has been a standard method
lor determining the colitorm group ol bacteria since 1936. ln
this procedure, tubes ol lauryl tryptose broth are inoculated
with a water sample. Lauryl tryptose broth contains lactose,
which is the source of carbohydrates (sugar). The coliform den-
sity is then calculated lrom statistical probability lormulas that
predict the most probable number (MPN) of coliforms in a 100-
mL sample necessary to produce certain combinations ol gas-
positive (gas{orming) and gas-negative tubes in the series ol
inoculated tubes.
There are three distinct test states lor colilorm testing using
the mulliple tub€ fermentation method-the presumptive test,
the confirmed tesl, and the compleied test. Each test makes
the coliform test more valid and specific. These tests are de-
scribed in detail in the following paragraphs.
2. Membrane Filter (MF) Method
This method was introduced as a tsntativo m€thod in 1955
and has been an approved test for coliform bacteriasincs 1960.

The basic procedure involves filtering a known volume of water
through a membrane lilter of optimum pore size for full colilorm
bacteria retention. As the water passes through the micro-
scopic pores, bacleria are entrapped on the upper surlace of
the filter. The membrane lilter is lhen placed in conlact wilh
either a paper pad saturated with liquid medium or directly over
an agar (gelatin-like) medium to provide proper nutrients tor
bacterial groMh. Following incubation under prescribed condi-
tions ol time, lemperalure, and humidity, the cultures are exam-
ined lor colilorm colonies, which are counted directly and re-
corded as a density ol coliforms per 100 mL of water sample.
There are certain important limitations to membrane filter
methods. Some types of samples cannot be filtered because
ol excessive turllidity, high non-colilorm bacterial densities, or
heavy metal (bactericidal) compounds. ln addition, coliforms
conlained in chlorinated supplies sometimes do not give char-
acteristic reactions on the media and hence special proce-
dures must then be used.
3. Membrane Filter N,lethod-Escherichia cor(EPA N4ethod)
This method combines information from a 1985 EPA publi-
cation (Test methods tor Escherichia coll and Enterococci in
Water by the Membrane Filter Procedure, EPA 600-4-85-076)
and a subsequent fiIarch 2000 manual (lmproved Enumera-
lion Methods lor the Recreational Water Quality lndicators:
Enterococci and Escherichia cofi, ePN821lR-971004). This
method is a membrane filter (MF) procedure for identifying
and counting E. coli in walet. The method incorporates the
use of a selective and differential medium, mTEC agar, fol-
lowed by incubation with urea substrate medium.
The E coli test is used as a measure ol recrealional water
quality. Epidemiological studies have led to the development
ol criteria that can be used to promulgate recreational waler
slandards based on established relationships between heallh
ellecls and water quality. The signilicance ol finding E coli in
recreational water samples is the direct relationship between
lhe density ol E coll and the risk of gastrointestinal illness as-
sociated with swimming in the water. This tesl can be applied
lo kesh, estuarine, and marine waters. Since a wide range of
sample volumes or dilutions can be analyzed by the MF tech-
nique, a wide range ol E, coll levels in water can be idenlified
and counted.
1. Presence/Absence (P/A) Method
The presence/absence (P/A) method for total coliform is a
simple modificalion of the multiple tube (or MPN) method.
Eolh lactose broth and lauryl tryptose broth are the essential
irgredients of the P/A media. ln addition, bromocresol purple
isadded as a pH indicator
The procedure involves the addition of '100 mL of sample to
50 mL ol sterile P/A broth in a 250-mL milk dilution bottle. This
iroculaled media is then incubated at 35"C and inspecled
allet 24 and 48 hours for an acid reaction. A distinct yellow
mlor forms in the media when acid conditions exist lollowing
klose lermentation. ll gas is also produced, gently shaking
he bottle will result in a toaming reaction. Any amount of gas
oracid (yellow color) constitutes a positive presumptive test re-
qjidng confirmation in the same manner as the IVIPN method.
t ColilertrM Method
Colilert, a registered trademark of IDEXX Laboratories,
flovides simultaneous deteclion, specific identification, and
oflfirmation ol total coliforms and E coli in water. Colilert is a
&r,ter A solution or liquid whose chemical makeup n€ukalizes acids or bases without a great change in pH
Lab Procedures 545
(Coliform)
specifically designed reagsnt formulation that is specific only
to coliform bacteria. lt provides a specific indicator for the
target microbes, lotalcoliforms, and E coli As the medias nu-
trients are metabolized, yellow color and fluorescence are re-
leased, confirming the presence of total colilorms and E coll
respectively.
6. Colisure Method
The Colisure test is a presence/absence test for colilorm
bacteria. A sample of waler is edded to a dehydrated mBdium
and, atter 28 to 48 hours, the medium is examined for the
presence ol colitorms. (Th€ Colisure test is available from
IDEXX Laboratories, '1 IDEXX Driva, Westbrook, ME 04092.)
C. Whal ls Tested
Common Range
Total Colitorms per 100 mL
Surlace Waters 50 to 1,000,000
Treated Water Supplies 0
Well Water 0to50
D. Materials Required
1. Sampling Bottles
Botlles of glass or other mat€rial that are watertight, resist-
ant to the solvent action ol water, and capable of being steri.
lized may be used for bacteriologic sampling. Plastic bottles
made of nontoxic materjals are also satisfactory and eliminate
the possibility of breakage during transport. The bottles
should hold a sutficient volume of sample tor all tests, permit
proper washing, and maintain the samples uncontaminated
until examinations are complete.
Before sterilization by autoclave, add 0.1 mL 10 percent
sodium thiosulfate per 4-ounce bottle (120 ml). This will neu-
tralize a sample containing about 15 mg/L residual chlorine. lf
the residual chlorine is not nautralized, it would continue to be
toxic to the colilorm organisms remaining in the sample and
give false results.
When filling botlles with sample, do not llush out the sodium
thiosulfate or contaminate the bottl€ or sampl6. Fill boilles ap-
proximately three-quarters full and start the test in the labora-
tory within six hours. ll the samples cannol be processed
wilhin one hour, they should be held below 10'C for not longer
lhan six hours.
2. Glassware
All glassware must be thoroughly cleansed using a suitablg
detergent and hot waler (160'F or 71'C), rinsed with hot water
('180"F or 82'C) to remove alllraces of residual delergent, and
finally rinsed with distilled or deionized water.
3. Water
Only distilled water or demineralized water that has been
tested and found free from traces of dissolved metals and bac-
tericidal and inhibitory compounds may be used lor prepara-
tion ot culture media.
4, Butfered" Dilution Water
Prepare a stock solution by dissolving 34 grams ol KHZPOT
in 500ml distilled water, adjusting the pH lo 7.2 with 1 N
NaOH and dilute to one liter. Prepare dilution wat6r by adding
545 Water Treatment

(Coliform)
1.25 mL of the stock solution and 5.0 mL magnesium sulfate media stor€d in the refrigerator should be used within
(50 qrams MgSO". z H,O dissolved in one liter of waler) to 1 96 hours.
liter distilled water. This solution can be dispersed into various
size dilution blanks or used as a sterile rinse lor the membrane
The rosolic acid solution should be stored in the
lilter test. dark and discarded atter two weeks or sooner il its
color changes from dark red to muddy brown.
5. Media Preparation
M E D IA_P R E S E NC qABS E N C E ( P/A) M ETHO D
Careful media preparalion is necessary lor meaningful bac-
teriological lesting. Attention must be given to the quality, mix- a. P/A broth
ing, and sterilization ofthe ingredients. The purpose ot this care Prepare this media lollowing the instructions on the
is to ensure thal il the bacteria being tested for are indeed media bottle label.
present in the sample, every opportunity is presented lor the
development and ultimale identification. Bacteriological identi- b. Brilliant green bile broth
lication is often done by noting changes in the medium; conse- Same as for MPN method.
quently, the composition of lhe medium must be standardized.
Much otthe tedium of media preparation can be avoided by pur- c. EC broth
chase of dehydrated media (Difco, BBL, or equivalent) from
local scientilic supply houses. The operator is advised to make Same as for MPN method.
use of these products, and, if only a limited amount of lesting is M ED I A--COLI LERT TM
M ETHOO
to be done, consider using tubed, prepared media.
a. Available lrom: IDEXX Laboratori€s
MEDIA_MPN (TOTAL COLIFOBM) 'l IDEXX Drive
Westbrook, ME 04092
a. Lauryl tryptose broth
(800) 321-0207
For the presumptive coliform test, dissolve the rec-
ommended amount ol lhe dehydraled media in distilled 6. Media Storage
water. Dispense solution into fermentalion tubes con- Culture media should be prepared in balches of such size
taining an inverted glass vial (see illustration of the tube that lhe entire batch willbe used in less than one we€k.
with vial on page 549). For 1o-ml water portions from
samples, double-strength media is required while ali 7. Autoclaving
other inoculations require single strenglh. Direclions
for preparation are given on the media bottle label. Steam auloclaves are used for the sterilization of the liquid
media and associated apparatus. They sterilize (kill all organ-
b. Brilliant green bile (BGB) broth isms) at a relalively low temperature of 121"C within ',5 min-
utes using moist heat.
For the confirmed colilorm test, dissolve 40 grams of
the dehydrated media in one liter of distilled water. Dis- Components of the media, particularly sugars such as lac-
pense and sterilize as with lauryl tryptose broth. tose, may decompose at higher temperatures or longer heat-
ing times. For this reason, adherenc6 to time and temperatu16
c. EC broth schedules is vital. The maximum elapsed tim6 for exposure ot
Prepare this media following the instructions on the the media to any heat (kom the time lhe autoclave door is
bottle. closed to unloading) is 45 minutes. Preheating the autoclavo
can reduce total heating time.
MEDIA-MEMBBANE FILTER M ETHOD (TOTAL
coLtFoRM) Autoclaves operale in a manner similar to the familiar kitch-
en pressure cooker:
a. M-Endo broth
a. Water is heated in a boiler to produce steam.
Prepare this media by dissolving 48 grams of the de-
hydrated product in one liter oI distilled water that con- b. The steam is vented to drive out air.
tains 20 mL of ethyl alcoholl'? per liter. Heat solution to c. The steam vent is closed when the air is gone.
boiling only--do not autoclave. Promptly remove solu-
tion lrom heat and cool. Prepared media should be d. Continued heat raises the pressure io 15 lbs/sq in
stored in a refrigerator and used within 96 hours. (103.4 kPa or 1.05 kg/sq cm); (at this pressure, pure
steam has a temperature of 12'1'C at sea level only).
b. LES Endo agar
e. The heat and pressure are maintained for 1 5 minutes
Prepare this media, used lor the two-step proce-
dure, following lhe instructions on the boltle. f. The heat is turned off.
c. l\4-FC media g. The steam vent is opened slowly to vent steam until at-
mospheric pressure is reached. (Fast venting will
Flehydrate in dislilled water containing 10 mL ol 1
cause the liquids to boil and over,low lubes.)
percent rosolic acid in 0.2 N NaOH. Heat just to boiling
then cool to below 45"C. Do not autoclave. Prepared h. Slerile material is removed to cool.

''? ln some states, the ethyl alcohol used in bacteriological m6dia pr€paralion cannot be lhe speclally denatured alcohol sold by supply houses to
people withoul an alcohol permit. One way to obtain suitable ethanol undor lhese circumslances is lo buy a brand ol gthanol sold lor human con.
sumption.

ln autoclaving termentation tubes, a vacuum is formed in
the inner iubes. As the tubes cool, the inner lubes are filled
with sterile medium. Capture ol gas in this inner tube from the
culture ol bacleria is the evidence of fermentation and is re_
corded as a positive test.
E. Procedure for Testing Total Coliform Bacteria
irulliple Tube Fermentation Method
1. Genoral Discussion
The tesl for colirorm bacteria is used to measure the suit-
ability of a water for human use. The test is not only uselul in
determining the bacterial quality ol a linished water. but il can
be used by lhe operator in the treatment plant as a guide to
achieving a desired degree ot treatment.
Colilorm bacteria are detected in water by placing portions
of a sample of the water in lauryl tryptose broth. L;urvl trvo_
lose broth is a standard bacteriological media containing lac_
tose (milk) sugar in tryptose brolh. The colitorm bacterii are
those that can grow in this media at 35"C temperature and are
able lo ferment and produce gas from the lactose within 48
hours. Thus, to detect these bacteria, the operator need onlv
inspecl the lermentalion lubes lor gas. A schematic of lhe coli:
Iorm test procedure is shown in Figure .11.12.
Dinkjng water coliform samples must be jOO mL in volume,
so the fivelube multiple tube fermentation method is not ap-
propriate. The lest uses len tubes with a 1O-mL sample in
each tube.
2. Materials Needed
FOP UNTFIEATED WATEB SAMPLES
a. Fitleen sterite tubes containing .lO
mL ol lauryl tryp-
tose broth are needed for each sample. Use fiv; iubes
lor each dilution.
b. Dilution tubes or btanks containing 99
ml of sterile
bulfered distilled water.
c. A quantity of 'l-ml and 1O-mL serotogical pipets. The
1-mL pipets should be gradualed in 0.1_mL incre-
ments.
d. lncubator set at 35" j 0.5.C.
e. Thermometer verified to be accurate by comparison
with a National lnstitute of Standards and Technology
(NIST) certified thermomeler
FOR DRINKING WATER SAMPLES
a. Ten sterile tubes_of 1O-mL, double-strength lauryl tryp_
lose broth are needed if lO mL of sample is adOed io
each lube. Ten mL of tauryl tryptose broth is required
in alltubes containing one mL or less of sample.
b. Sterile 10-mL pipet for each sample.
c. lncubator set at 35" t 0.5"C.
d. Water bath set at 44.5. !O.2"C
e. Thermometer verified to be accurate by comparison
I 3.
with a National lnstjtute of Standards and Technology
(NIST) certified thermomeler, or equivalent. ''
Technique for lnoculation ol Sample (Figures 11.13 and
11 14)
All inoculations and dilutions ol waler samples must be ac-
flilale and made so that no contaminants from the air, equip-
ment, clothes, or fingers reach lhe sample, either direcfly dr 6y
-.
Lab Procedures 547
(Colilorm)
way.ol a coniaminated pipet. Clean, sterile pipets must be
used lor each separate sampls.
FOB UNTREATED WATER SAMPLES
a. Shake the samplo bot e vigorously 20 times before re-
moving sample volumes.
b. Pipet 10 ml of sample direc y into each ot the lirst livs
tubes. Each tube must contain 10 mL lauryl tryptoss
broth (doubl€ skength).
NOIE You must realiz€ that the sample voluma
applied to the first fiv€ tubes will depend upon the typs
of water being tested. The sample volume apptiei to
each tube can vary from 1O mL for high-quality waters
to as low as 0.00001 mL (applied as 1 mL of a dilut€d
sample) for highly polluted raw water samples.
NOIE When delivoring the sample into the cullure
medium, deliver sample portions of i mL or less down
into the cullure tube nearth6 surface ofthe medium. Do
not deliver smallsample volumes at the top of the tube
and allowthem to run down inside the lube:too much ol
the sample willfail to reach the culture medium.
A/OTE; Use 1o-ml pipets lor 1O-ml sample porlions,
and l-ml pipets for portions ol .t ml or iess. Handle
sterile pipet only near the mouthpiece, and protect the
delivery end from enernal contamination.
c. Pipet 1 mL ol water sample into each of the next five
lauryl tryptose broth (single strength) tubes.
d. Pipet 1/10 mL (0.1 mL) ol water sample into each ol
the next.five lauryl tryptose broth (single strength)
tubes. This makes a 0.1 (1 to tO) dilution.
At this point, you have t5 tubes inoculated and can
place these three sels of lubes in the incubator; how-
ever, your sample specimen may show gas production
in all fermentation tubes. This means your simple was
not diluted enough and you have no usable resulls.
e. To make a 1/100 (0.01) ditution, add 1 mL ol well_
mixed water sampl€ to 99 mL of sterile butfered dilu-
tion water. Mix thoroughly by shaking. Add 1 mL lrom
this bottle direc y into each of five more lauryt tryptos€
broth tubes.
f. To make a 1/1.000 (0.001) dilution, add 0.1 mL from
the 1/100 dilution bonle direc y into each of live more
lauryl tryptose broth tub€s.
The first time a sample is analyzed, 25 tubes of tauryl tryp-
.
tose brolh shoutd be prepared. Once you find out wtrit dill.
tions give usable results for determining the MpN index. you
will only _need to prepare t5 tubes to analyze subsequint
samples lrom the same source.
548 Water Trealment

(Colilorm)

OUTLINE OF PFOCEDUFIE FOFI TOTAL COLIFOFIM

1. Presumptive Test

lnoculate in lactose or lauryl lryptose; incubale 24 t 2 ht

at 35" t 0.5'C

No gas Gas produced

incubate 24 hr more continue with confirmed test
(and lecal coliform tesl)

No gas
no colilorm present
discard tubes

2. Conlirmed Test

lnoculate (with loop or applicator stick) brilliant green

bile broth. lncubate 48 hr i 3 hr at 35'r 0.5'C

No gas Gas produced

colilorm absent colilorm group conlirmed
calculate confirmed MPN

FECAL COLIFORM
1. Presumptive Test
(same as for TOTAL COLIFOHM)
2. Fecal Coliform Test

lnoculate (with loop or applicator stick) EC broth.

lncubate 24 hrs t 2 hrs al 44.5" ):0.2"C

No gas Gas produced

lecal coliform absent fecal coliform confirmed
calculate f6cal MPN

Fig. 11.12 Schematic outline ol tesl procedure for totalcolilorm and fecal coliform-qultiple tube lermenlation method
lrqr
Lab Procedures 549

(Coliform)
q!{19! 4 lncubation (Totat Coliform)
I0mLIOEACHTIjBE
a. 24-Hour Lauryl Tryptose (LT) Broth presumptive Test
-l NO DILL]TION
Place all inoculated LT broth tubes in 35" t O.S"C incu-
0 0 0 0 0 bator. After 24 t 2 hours have elapsed, examine each
tube lor gas rormation in inverled vial (inner tube). Mark
plus (+) on report form such as shown in Figure 11..1S for
all tubes that show presence of gas. Mark mlnus (-) for all
tubes showing no gas formation. lmmediately perform
confirmation test on atl positive (+) tubes (See Faiagraph
0 0 0 0 0 c. below.'24-Hour Brilliant green bite (BGB) Confirma-
lion Test"). The negative (-) tubes musl be reincubated
for an additional24 hours.
b. 48-Hour Lauryl Tryptose Broth presumptive Test
0 0 0 0 0 Record both positive and negative iubes at the end of
48 t 3 hours. lmmediately perlorm total coliform contir-
mation test on all new positive tubes. For drinking water
samples, all positive tubes should also be tested for fecal
coliforms fE. colr.
0 0 0 0 0 c. 24-Hour Brilliant green bile (BGB) Confirmation Test
0l mt TO EACH TUB€
,Confirm all presumptive tubes that show gas at 24 or
DILUTION

48 hours. Transfer, with the aid ot a sterile 3-mm platinum

BOTTLE B
wire loop (sterile wood appljcator or disposablo loops may
be used also), one loopJutt of the broth from the tauryil
0 0 0 0 0 tryptose broth tubes showing gas, and inoculale a corre-
sponding tube of BGB broth by mixing lhe loop of brolh in
INCUBAIE AL! TUBES AT 35' } 0.5" C FOB 2' FIiOUFS the BGB broth. Discard all positive lauryl tryptose broth
tubes after lransrening is completed.

$
Fig. 11-13 Colitorm bacteria test-raw water
FOR UNTREAIED WAIER

OATE NO 50uRca

28 Rivar
Raw

l0 mrTO EACH TUBE D]LUTION

I'RE5I]MPTiVE CONFIRMEO

88 o
k:l 21
l0 0.1
24
l
---oT-l
ITTr.I,l; TII] rr, rrrl 14
g !ll!
6^\ 0 0 0 toltol i 66 tol
NO DILUTION 4E 48 ITt-F

5 INCUBATE ALt TU8E5 AT ]5' ! 0.5'C FOR 24 HOURS

n( '
24
48 '-
ma OF SAMPLE: I0 Ma, I Mt.
R€SULTS AFTER 24 HOURS OF INCUBATION
RE5ULTS AFTTR 4E HOIJRS OF INCUEATION
and O.t rt

cA5 rN TNNER viAt

15A+Tt5TRESUIT

$
Fig. 11.14 Colilorn bacteria test-4rinking water
.OR ORIt]KING WATER

OATE NO. sOURCE

City
70
Hall
FOR DBINKING WATEB SAMPLES

a. Shake the sample bottle vigorously 20 times before re- PNESUMPTIVI CONFIRMTO MPN/r00 mt
moving a sample volume. t0
b. Pipet 10 mL ol sample direc y into each ol ten tubes 24 24 I T o
containing '10 mL of doubte-strength lauryl tryptose
48 48 I !lll I
broth. Fig. 11.15 Becorded cotilorm test rcsults
550 Water Treatment

(Coliform)
Always sterilize inoculation loops and needles in ,lame
immediately before transter ol culture; do not lay loop
down or touch it to any nonsterile object belore making
the lransfer. After slerilization in a flame, allow sufficient
(7) As soon as results are recorded,
time lor cooling, in the ai( to prevent lhe heat of the loop
from killing lhe bacterial cells being transferred. Sterile
wood applicator sticks also are used to transfer cullures, (8)
especially in the lield where a flame is not available for
sterilization. lf using hardwood applicators, sterilize by
autoclaving belore use and discard after each transrer.
Aiter 24 hours have elapsed, inspect each of the BGB
tubes for gas formation. Those with any amount of gas (9)
are considered positive and are so recorded on the data
sheet. Negative BGB tubes are reincubated for an addi'
lional 24 hours.
production tor each tube. Gas in any quantity is a
positive test.
discard all
tubes. This is a 24-hour tesl for EC broth inocula-
tions and not a 48-hour test.
Transfer any additional 48-hour gas-positive
tubes lrom the presumptive tesl to corrgspond-
ingly labeled tubes ol EC broth. lncubato tor 24 t
2 hours at 44.5" ! O.2"C and record results on
data sheel.
Codily results using the same procedure as lor
total colilorms and determine MPN of fecal coli-
forms per,I00 mLof sample (Figure'11.15).

d. 48-Hour Brilliant Green Bile Confirmation Test OAMPLE:

(1) Examine tubes for gas at the end ot the 48 t 3 mL portion 10 1 0.'l
hour period. Record both positive and negative
tubes.
Readings 521
Bead MPN as 70 per 100 mL lrom Table 11.3
(2) Complete reports by determining MPN index and
recording MPN on worksheets. Report resuhs as MPN
ol lecal colilorms = 70 p€r 100 mL
5. Fecal Colilorm (FC) Test
6. Recording Results
For drinking water samples, all presumptive tubes lhat
show gas at 24 or 48 hours must be tested for fecal coli- Results should be recorded on data sheets prepared es-
form. This tesl more reliably indicates the potential pres- pecially for this test. Examples are shown in Figure 1 1 .15.
ence ol pathogenic organisms than do tests lor tolal coli-
form group of organisms. The procedure described is an 7. Method of Calculating the lvlost Probable Number ([IPN)
elevated temperature test for fecal colilorm bacteria. Select the highest dilution or inoculation with all positive
tubes, belore a negative tube occurs, plus the next two di-
a. l\,,laterials Needed
lulions.
Equipment required lor the lests are the same as
those required for lhe 24-hour lactose brolh presump- EXAMPLE 1 (UNTREATED WATERFSelecl the under-
live test, plus a water bath sel at 44.5 t 0.2'C, EC lined inoculations.
broth media, and a thermometer certified against an
NIST thermometer. mL of sample 10
,1
0.1 0.01 0.001
dilutions 0 0 -1 -2 -3
b. Procedure positive lubes 5 1 U 00
Flead MPN as 30 pe|I00 mL from Tabl6 1 1.3
(1) Run lactose broth or lauryl tryptose broth pre-
Fleporl MPN as 30 per 100 mL
sumptive test.
(2) Atter 24 hours, temporarily retain all gas-positive EXAMPLE 2 (UNTREATED WATEBI'SeIect the under
tubes. lined lnoculations (see Figuro 11.16).
(3) Label a lube of EC broth to correspond with each mL ofsample 10 1 0.1 0.01 0.001
gas-posiiive tube of broth lrom presumptive test. dilutions 0 0 -'l -2 -3
(4) Shake or mix positive presumptive tubes by rotal-
positive tubes 552 0 0

ing them. Transler one loop-full of culture lrom Read MPN as 50 per 100 mL from Table 1't.3
each gas-positive culture in presumptive test to Fleport results as 500/100 mL
the correspondingly labeled tube ol EC brolh. we added one zero to 50 because we started with the 1

(5) lncubate EC broth tubes 24 t 2 hours at 44.5" t

mL sample and Table 1'1.3 begins with one dilution
column to the left.
0.2"C in a water bath with water depth sutlicient
to come up at least as high as the top ol the cul-
ture medium in the tubes. Place in water balh as EXAMPLE 3 (DRINKING WATEFI SAMPLE)_(SEE
soon as possible atter inoculation and always Figure 11.17).
within 30 minutes atter inoculation. mL ol sample 10
(6) After 24 hours, remove the rack of EC cultures positive tubes 1

lrom the water bath, shake genlly, and record gas Flead MPN as 1.1 perl00mLfromTabl6 11.4
 Lab Procedures 55'l

(Coliform)
,I
TABLE 1.3 MPN INDEX FOB VARIOUS COMBINATIONS OF POSITIVE AND NEGATIVE BESULTS
lN A PLANTING SERIES OF F|VE 1O-mL, F|VE .t-ml, AND
FIVE 0.1-mL pORTIONS OF SAMpLE

Number ot tubes giving positive

reaction out ol MPN lndex
Five 10-mL Fivel-ml Five 0.1-mL (organisms)
portions portions portions per 100 mL
0 0 0 <2
0 0 1 2
0 0 2 4
0 1 n 2
0 ,1

1 4
0 1
5
0 0 4
0 2 1 6
0 3 0 6
1 0 0 2
l 0 1 4
1
,l
0 2 o
0 8
1 1 0 4
1 1 1 6
1 1 2 8
2 0 6
2
2
'I
I
'10
2
3 n 8
3 1 t0
1 4 0 11
2 0 0 4
2 0 1 7
2 0 2 o
2 0 3 2
2 1 0 7
1
,l
1 9
2 2 12
2 2 0
2 2 ,,
12
2 2 2 14
2 3 0 12
2 3 1 14
2 4 0 15
3 0 0 8
0 1 11
0 2 13
3 1 0 11
3 1
,l
14
3 1 2
3 1
20
3 2 0 14
3 2 1 17
3 2 20
 552 Waler Treatment

(Coliform)
TABLE 11.3 MPN INDEX FOR VARIOUS COMBINATIONS OF POSITIVE AND NEGATIVE RESULTS
IN A PLANTING SEBIES OF FIVE 1O-mL, FIVE 1.m1, AND
FIVE 0.1-mL PORTIONS OF SAMPLE (conrnued)

Number ot tubes giving positiv€

reaction out ol MPN lndex

Five 10-mL Five l-ml Five 0.1-mL (organisms)

portions portions portions per 100 mL

3 3 0 17
3 3 1
3 4 0
3 4 1 24
3 0 25
4 0 0 '13
4 0
,1
17
4 0 2 21
4 0 3 25
4 1 0 17
,1
4 1 21
4 1 2 26
4 2 0 22
4 2 '1
26
4 2 2
4 3 0 27
4 1

4 3 2 39
4 4 0 34
4 4 1 40
4 5 0 41
,|
4 5 48
5 0 0
,1
5 0 30
5 0 2 40
5 0 3 60
5 0 4 80
5 0 30
,l
5 50
5 60
5 3 80
5 2 o 50
2 1 70
5 2 2 90
5 2 3 130
5 2 4 150
5 2 180
5 3 0 80
'1
3 1 10
5 3 2 140
3 3 170
5 3 4 210
5 3 250
5 4 0 130
4 1 170
5 4 2 220
5 4 280

f-.
 Lab Procedures 553

(Colilorm)
TABLE 11.3 MPN INDEX FOR VARIOUS COMBINATIONS OF POSITIVE AND NEGATIVE HESULTS
IN A PLANTING SERIES OF FIVE 1O.mT, FIVE l.mL, AND
F|VE 0.1-mL pOBT|ONS OF SAMpLE (continued)

Number ol tubes giving positive

reaction out ol i,4PN lndex

Five 1o-ml Five 1-mL Five 0.'l-mL (organisms)

portions porlions portions per 100 mL
5 4 4 350
5 4 430
5 5 0 240
5 5 1 300
5 5 2 500
5 5 3 900
5 5 4 1,600
5 5 >1,600

For Example No. 2 For Example No. 3

FESULTS INOCULANO FESULIS

G^S GIS GAS GAS CrS NO NO NO NO NO NO NO NO NO
GAS GAS GAS GAS GAS GAS GAS GAS CAS GAS
10 mL
0 B 0 B 0 a
crs 6ls cls crs 6rs
U 0 0 .0 0 0 0 0 0 !
'10 mL 1 ouT oF 10
1mL
Fig. 11.17 Results ot colilorm test-drinking water
0 0 0 0 0
GAS XOGrc NOGAS XO CrS GrS
TAELE 11.4 MPN INDEX FOR VARIOUS
COMBINATIONS OF POSITIVE AND NEGATIVE BESULTS
0.1 mL 2OUTOF5
WHEN TEN I0-mL PORTTONS ARE USED
0 BB6E Number of tubes giving posiiive
reaction out of ten '!o-ml portions
MPN lndex
per 100 mL
0 <1.1
0.01 mL

66BB
1 1.1
0 2 2.2
3.6
4 5.1
5 6.9
6
0.00'l mL O OUTOF 5
7 12.O
0 0 0 0 B 16.1
I 23.0
Fig. 11.16 Besults ol colilorm test-untreated water 1 0 >23.0
 554 Water Treatment

(Coliform)
When you wish to summarize with a single MPN value the 3. Determine the mL of sample in all tubes.
resulls from a series of samples, the arithmetic mean, the me-
dian, or the geometric mean may be used (see following sec- Number mL of Sampte
tions for formulas and examples).
Sample Size, ml Tubes
ol in Att Tubes
10 ml 5 (10 mrxs)
1.0 ml 5 (1.0 mL)(5)
FOBMULAS 0.1 mL 5 (0.1 mLXs) = 0.5 ml

The IVIPN for combinations not appearing in the table given

Total = 55.5 ml
or for olher combinations ol tubes or dilutions may be esli- 4. Estimate the MPN/'|00 mL.
mated by the formula:
[rPN/100 mL = (No. of Positive Tubes )(100)
(No. ol Positive Tubes )(1 00 mL Sample in
MPN/I00 mL = I .. imL Sample in)
mL Samole in \ / mL Samole in\ Negative Tubes l ' ( All Tubes .l
Negative Tubes J " I ett trU"" i (4)(1oo)
=
Coliform resulls may be summarized with a single l\.4pN "@s m alrlsrs mD
value by using the aritltmetic mean, median, or geometric
mean. The arithmetic mean is often called the average. 400
s!. ,! lt9.:lll'"!:" 36.87
Mean = Number of ltems or Values

The median is the middle value in a set or group of data. = 10.8 MPN Coliforms/'l00 mL

There are just as many values larger than the median as there
are smaller than the median. To determine the median, the
dala should be written in ascending (increasing) or descend-
ing (decreasing) order and lhe middle value identified.
= 11 MPN Coliforms/'|00 mL

EXAMPLE 5
Median = Middle Value ol a Group of Data
Flesults from IVIPN tests during one week were as follows:
The median and geometric mean are used when a sample
contains a few very large values. This frequen y happens DAySMTWTHFS
when measuring the MPN of raw water. lf X is a measurement i\.4PN/100 mL 2 8 14 6 10 26 4
(MPN) and n is the numberof measurements, then
Estimate the ('1) mean, (2) median, and (3) geometric mean
Geometric Mean .................................. (X")],, of the data in MPN/100 mL.
= [(X,X&XX.)
1. Calculate the mean.
This calculation can be easily performed on many electronic
calculators using the power tunction, y'. sum of All MPNs
t4ean. MPN/'1oo ml -
Number of MPNS
EXAMPLE 4 2+8+ 14+6+ 10+26+4
_
The results from an MPN test using five ,ermentaiion tubes 7
for each dilution on a sample of water were as follows; 70
Sample Size, mL 10 1.0 0.'1 7

Number of Posilive Tubes

Out of Five Tubes 3+ 1+ O+
= 10 MPN/100 mL
2. To determine the median, rearrange the data in ascend-
Determine the information necessary to solve the lormula: ing (increasing) order and select the middle value (three
(No. of Positive Tubes )(100 will be smaller and three will be larger in this €xampte).
IVPN/I00 mL =
mL Sample in r , .nL Sample in Order 1 2 4 56 7
Negative Tubes' Alt Tubes ' MPNi'100 mL 2 4 6 8 '10 14 26
1
1 Number of positive lubes. There were 3 positive tubes
lvedian, MPN/100 mL = Middle value of a group of data
with 10 mL of sample and 1 positive tube with 1 mL of
sample. Therefore, 3 + 1 = 4 positive tubes. = 8 MPN/100 mL
3 Calculate the geometric mean for the given data.
2 Determine the mL of sample in negative lubes
Number ot lnl o, Sampt€ in ""fi[lr1||.Hi"' = (x]xx,xxJ(x1xxsxx")(x?)r,,
Sample Size, lna Negative Tubes Negalive Tubes
= (2X8X14)(6)(10)(26)(4)li'
10 mL 2- (10 max2) 20 ml
1.0 mL (1.0 mr)(4) 4mL = ['t,397 ,76oln
0.t ml (0.1 mL)(5) 0.5 mL
Total ,+5 *L = 7.5 MPN/100 mL

4. Summary
1. Mean = 10 MPN/100 mL
2. l,4edian 8 MPN/100 mL
= enrichment technique).
3. Geometric Mean = 7.5 N4PN/100 mL
As you can see trom the summary the geometric mean
more nearly describes most of the MPNS. This is the
reason why the geometric mean is sometimes used to de-
scribe the results of MPN tests when there are a lew very
large values that would distort the mean.
8. Belerence
See page 9-48, STANDARD \IETHODS, 21st Edition.
Membrane Filter Method
1. General Discussion
ln addition to the fermenlation tube test lor coliform
bacteria, anolher test is used for these same bacteria in
water analysis. This test uses a cellulose ester filler,
called a membrane filter, the pore size ol which can be
manufactured to close tolerances. Not only can the pore
size be made to selectively trap bacteria from water fil-
tered through lhe membrane, but nutrients can be dif-
lused (lrom an enriched pad) through the membrane to
grow these bacteria into colonies. These colonies are rec-
ognizable as coliform because the nutrients include fuch-
sin dye, which peculiarly colors the colony. Knowing the
number of colonies and the volume ot water liltered, the
operator can then compare the water tesled with water
quality standards.
A two-step pre-enrichment technique for chlorinated
samples is included at the end of this section. Chlorinated
bacteria that are still living have had their enzyme sys-
lems damaged and require a 2-hour enrichmenl media
before contact with the selective M-Endo media.
2. Materials Needed
a. One sterile membrane filter having a 0.45p pore size.
b. One sterile 47-mm pelridish wjth lid.
c. One sterile tunnel and support stand.
d. Two sterile pads.
e. One receiving flask (side-arm, 1,000 mL).
f. Vacuum pump, trap, suction or vacuum gauge, con-
nection sections of plastic tubing, glass T-hose clamp
to adjust pressure bypass.
g. Forceps (round-tipped tweezers), alcohol, Bunsen
burner, grease pencil.
h. Sterile, buffered, distilled water tor rinsing
i. M-Endo media.
Slerile pipets-two s-ml graduated pipets and one 1-
mL pipetlor sample or one 10-mL pipet lor larger sam-
ple. Quantity of 1-mL pipets if dilution of sample is nec-
essary. Also, quantity of dilution water blanks if dilu.
lion of sample is necessary.
rr ,4rqoot (AL-li'kwot). Represenlalive portion of a sample. Ollen
J t
Lab Procedures 555
(Coliform)
k. One moist incubator at 35"C; auxiliary incubalor dish
with cover.
l. Enrichment media-lauryl tryptose broth (for pre-
m. A binocular, widejield, dissecling microscope is rec-
ommended for counting. The light source should be a
cool white, f luorescent lamp.
3 Selection of Sample Size
The size of the sample ot ALIQUATI1 will be governed
by the expected bacterial densiry An ideal quantity will
result in the groMh oI 20 to 80 coliform colonies, but not
more than 200 bacterial colonies of all types. The table
below lists suggested sample volumes for MF lotal coli-
form testing.
Ouantities Filtered (mL)
100 '10 1 0.1 0.0'1
Well Water x
Drinking Water x
Lakes x x x
Fiivers x x xx
When less than 20 mL of sample is to be filtered, a
small amount ot sterile dilulion water should be added to
the funnel before filtration. This increase in water volume
aids in unlform dispersion of the sample over the mem-
brane filter.
4. Preparation of Petri Dish for Membrane Filter
a. Sterilize forceps by dipping in alcohol and passing
quickly lhrough Bunsen burner flame to burn off the al-
cohol. An alcohol burner may be used also.
b. Place sterile absorbent pad into sterile petri dish.
c. Add '1.8 to 2.0 mL lvl-Endo medium to absorbent pad
using a sterile pipet. Flemove excess media.
5. Procedure for Filtration of Unchlorinated Samples
All liltrations and dilutions of water samples must be ac-
curate; no contaminants from the air, equipment, clothes,
or fingers should reach the specimen either directly or by
way oI the contaminated pipet.
a. Secure tubing trom pump and bypass to receiving
flask. Place palm of hand on flask opening and start
pump. Adjusl suction to % atmosphere with hose
clamp on pressure bypass. Turn pump switch to OFE
b. Set sterile filter support stand and funnel on receiving
flask. Loosen wrapper. Rotate funnel counterclockwise
to disengage pin. Secure wrapper.
c. Place petri dish on bench vrith lid up. Write identifica-
tion on lid with grease pencil.
d. Unwrap sterile pad container. Light Bunsen burner.
e. Unwrap membrane filter container.
f. Sterilizo forc€ps by dipping in alcohol and passing
quickly through Bunsen burnsr to burn off th6 alcohol,
an equally divided porlion of a sample
 556 Water Treatment

(Coliform)
g. Center membrane filter on lilter stand with forceps
after lifting funnel. l\.4embrane filter with printed grid
should show grid uppermost (Fig. I, next paqe).
h. Replace funnel and lock against pin (Fig. ll).
i. Shake sample or diluted sample. Measure proper ali-
quot with sterile pipet and add to funnel.
j. Add a small amount o, the sterile dilution water to fun-
nel. This will help check for leakage and also aid in dis-
persing smallvolumes (Fig. lll).
k. Now start vacuum pump.
SPECIAL NOTE:
lnexperienced persons often have great difiiculty with
connected colonies, with mirror reflections ol fluorescent
tubes (which are confused with metallic sheen), and with
water condensate and particulate matter, which are occa-
sionally mistaken for colonies. Thus, there is a lendency
for inexperienced persons to make errors on the high sida
in MF counts. Al least live apparent coliform colonies
should be transterred to lauryl tryptose broth tubes for
verification as coliform organisms.

l. Flinse filter wilh three 20- to 3O-mL portions of sterile

dilution water.
m. When membrane filter appears barely moist, switch
pump to OFE
n. Sterilize forceps as before.
o. Fiemove membrane filter with forceps after first remov_
ing funnel as belore (Fig. t).
p. Center membrane filter on pad containing M-Endo
media with a rolling motion to ensure water seal. ln-
specl membrane to ensure no captured air bubbles
are present (Fig. lV).
q. Place jnverted petri dish jn incubator for 22 !.2hours.
lncubate at 35.C.
6 . Procedure for Counting Membrane Filter Colonies
a. Flemove petridish rrom incubator.
b. Remove lid from petridish.
c. Turn so that your back is to window
d. Tilt.membrane filter in base of petri dish so that green
and yellow-green colonies are most apparent. airect
sunlight has too much red to facilitate counting.
e. Count individual colonies using an overhead fluores-
cent light. Use a low-power (10- to '15 magniiications),
binocula( widejield, dissecting microscope or olher
similar optical device. The typicat colony has a pink to
dark red color with a metallic surface sheen. The
sheen area may vary from a small pin-head size to
complele coverage of the colony sudace. Onty those
showing this sheen should be counted.
f. Fleport total number of coliform colonies on work-
sheet. Results are often reported as ,,colony lormation
units" or CFUs. Use the membranes that show from 20
lo 90 colonies and do not have more than 2OO colonies
of all types (including non-sheen or, in other words,
non-coliforms).
7. Procedure for Filtration of Chlorinated Samples Using En,
richment Technique
a. Place a sterile absorbent pad in the upper half of a
sterile petri dish and pipet 1.8 to 2.0 mL sterile lauryl
tryptose broth. Carefully remove any surplus liquid.
b. ASEPTICALLYU place the membrane filter through
which the sample has been passed on the pad.
c. lncubate the filter, without inverting the dish, forl r,, lo
2 hours at 35'C in an atmosphere of 90 percent hu-
midity (damp paper towels added to a plastic contain-
er with a snap-on lid can be used to produce lhe hu-
midity).
d. Remove the enrichment culture from the incubalor,
Place a fresh, slerile, absorbent pad in the bottom half
of the petri dish and saturate with '1.8 to 2.0 rnL M-
Endo broth.
e. Transfer the membrane fjlter to the new pad. The used
pad oI lauryl tryplose may be discarded.
f. lnvert the dish and incubate for 20 to 22 hours at 35. t
0.5"c.
g. Count colonies as in previous method
8. Procedure for Fecal Coliform
For drinking water samples, all positive total coliform
colonies must be tested for fecal coliform, When the
membrane filter method is used, groMh from th€ positive
total coliform colony should be lranslerred to a tube of EC
media to determine the presence of fecal coliform.
a. Transfer, with the aid of a sterile 3-mm platinum wire
loop, groMh from each positive total coliform colony to
a corresponding tube of EC broth.

EXAMPLE: b. Incubate inoculated EC broth tube in a water bath at

44.5" 10.2"C for 24 hours.
A total ol42 colonies grew after filtering a 1O-mL sample.
c. Gas production in an EC broth culture within 24 hours
No. ol Colonies Counted x 100 mL or less is considered a positive fecal colilorm reaction.
Bacterirl00 mL =
Sample Volume Filtered, mL x 1OO mL
d. Calculate total number of fecal colitorm colonies.
_ (42 Colonies)(1oo mL)
(10 mLX100 mL) EXAMPLE:

(4.2X100 mL) For 100 mL of sample, 2 colonies gave a positive total

100 mL coliform result. Only 1 of the 2 gave positive results when
growth was transferrod to EC broth.

= 420 per 100 mL Fecal Coliform/100 mL = 1 Coliform/1OO mL.

'a Aseptic (a-SEP-tick). Free lrom the tiving germs ol disease, lermentation, or putrefaclion. Sterile,
 Lab Procedures 557

(Colilorm)
OUTLINE OF PFIOCEOUFE FOFI INOCULATION OF MEMBHANE FILTER

Fig. I Fig. ll
1. Cenler membrane lilter on 2. Place funnel
filter holder. Handle mem- onto filter
brane only on outer 3/r6 holder
inch with forceps sterilized
betore use in ethyl or methyl
alcohol and passed lightly
through a llame.

Fig. lll Fig. lV Fig. V

I
(r,

5. lncubate in
inverted
4. Flemove membrane filter lrom
position for
filter holder with sterile
22 t 2 houts.
forceps. Place membrane
3. Pour or pipet sample on pad. Cover with petri 6. Counl coliform-
aliquot into funnel. top. appearing colonies
Avoid spattering. Alter on membrane.
suction as applied, rinse
lour times with sterile,
bullered, dislilled water.
558 Water Treatment

(Coliform)
9. Alternate Nlembrane Filter Procedure for FecalColi{orm EXAMPLE:
The following method is a way to test for fecal coliform di- A total of 2 colonies grew after tiltering a 100-mL
reclly without first running the tolal coliform test as a separate sample.
procedure. This membrane filter procedure for recal coliform
uses an enriched lactose medium (M-FC broth) that depends Fecal Coliforms Fecal Colonios Counted x'100 mL
on an incubation temperature of 44.5't 0.2"C for its selectiv- per 100 mL Sample Volume Filtered, mL x 100 mt
ity. Since the temperature is critical, incubation takes place in (2 Colonies)(100 mL)
a water bath using watertight plastic bags. = rco ,rxroo -L)
a. Materials Required (0.02x100)
(1) M-FC media. 100 mL

(2) Culture dishes with tight-fitting lids.

= 2 Per 100 mL
(3) Membrane filters. 10. Reference
(4) Watertight plastic bags. See page 9-56, SIANDARD METHODS,21st Edition.
(5) Water bath set at 44.5" t 0.2'C. The thermometer
must be checked against a National Institute of Membrane Filter Melhod-Esche chla cori (EPA Method)
Standards and Technology (NIST) certified ther- 1. General Discussion
mometer to ensure the accuracy of lhe water bath
temperature. This method describes a membrane filter (MF) procedure
for identifying and counling Escherichia cor'- Because the bac-
(6) Use a sample size of 100 mL or whatever size is terium is a natural inhabitant only of the intestinal tract of
recommended by your regulatory agency. warm-blooded animals, its presence in water samples is an in-
b. Preparation ol Culture Dish dication ol lecal pollution and the possible presence of enleric
(intestinal) pathogens.
Place asterile absorbent pad in each culture dish and
pipet (sterile) approximately 2 mL of N4-FC medium to This MF method provides a direct count of bacteria in water
saturate the pad. Caretully remove any surplus liquid. based on the development ol colonies on the surface of the
membrane filter. A water sample is filtered through the mem-
c. Filtration of Sample brane that retains the bacteria. After filtration, the membrane
containing the bacterial cells is placed on a specially formulat-
Observe the procedure as prescribed for lotal coli ed medium, mTEC, incubated at 35'C for 2 hours to revive in-
form using membrane filters. jured or slressed bacteria, and then incubaled a|44.5'C lor 22
d. lncubation hours. Following incubation, the filter is translerred to a filter
pad saturated with urea substrate. After 15 minutes, yellow,
Place the prepared culture dishes in waterproof yellow-green, or yellow-brown colonies are counted with the
plastic bags and immerse in water bath set at 44.5'i aid of a fluorescent lamp and magnifying lens.
0.2'C for 24 hours. Allculture dishes should be placed
in the water bath within 30 minutes atter filtration. 2. lvlaterials and Reagents Required

e. Counting a. Glass lens with magnificalion of 2 to 5x, or stereo-

scopic microscope.
Colonies produced by fecal coliform bacteria are
blue. The nonjecal coliform colonies are gray to b. Lamp with a cool, white lluorescent tube.
cream colored. Normally, few non]ecal coliform colo- c. Flasks.
nies will be observed due to the selective action of the
elevated temperature and the addition of the rosolic d. Autoclave for sterilization of media and glassware.
acid to the l\.4-FC media.
e. Culture dishes with tight-fitting lids.
Examine the cultures under a low-power magnifica-
tion. Count and calculate fecal coliform density per'100
f. Test tubes, screw-cap.
mL. g. Membrane filtels,
Fecat Cotiforms Fecal Colonies Counled x 100 mL h. lncubator maintained at 35' r 0.5"C.
per 100 mL - Sample Volume Fillered, mL x 100 mL
i. Whirl-Pale bags.

<.\
j. Water bath maintained at 44.5'r 0.2'C
k. Absorbent pads.

L mTEC agar (Difco 0334)

Preparation: Add 45.3 grams ol dohydrated mTEC
agar to 1 L of reagenlgrade distilled water in a flask
and heat to boiling until the ingredients dissolve. Auto-
clave at 121"C (15 lbs pressure) ror 15 minutes and
cool in a 50'C water bath. Pour the medium into each
 Lab Procedures 559

(Colilorm)
9- x 50-mm culture dish to a 4- to s-mm depth (approx- with urea substrale medium and allow to sit at room
imately 4 to 6 mL) and allow to sotidify. The tinal pH temperature lor 15 to 20 minutes.
should be 7.310.2. Store in a refrigerator
k. After incubation on the urea substrate at room lemper-
m. Urea substrate medium ature, count and record the number ol yellow, yellow-
green, or yellow-brown colonies on the membrane fil-
Preparation: Add dry ingredients to 100 mL of re-
agent-grade distilled water in a flask. Stir to dissolve ters, ideally containing 20 to 80 colonies.
and adjust to pH 3 lo 4 with 1 N HCl. The substrate so- 4. Calculalion of Besults
lution should be a straw-yellow color at this pH.
Select the membrane filter with an acceptable number of
n. Nutrient agar (Dirco 0001, BD431r472) yellow, yellow-green, or yellow-brown colonies (20 to 80) on
Preparation: Add 23 grams ol nutrient agar to 1 L of the urea substrate and calculate the number ol E coli per 1OO
. reagent-grade distilled water and mix well. Heat to
mL according to the lollowing general formula:
boiling to dissolve the agar completely. Dispense in _ No. E coli Colonies Counted x 100 mt
screw-cap tubes and autoclave al 121.C ('15lbs pres- E. coli llOO mL
Volume in mL of Sample Filtered x 100 mL
sure) fo|l5 minutes. Remove the lubes and slant. The
finalpH should be 6.8 ! 0.2. 5. Reporting Besults
3. Procedure Fleporl the results as E coliper'100 mL of sample.
a. Prepare the mTEC agar and urea subsirate medium 6. Example
as directed in (l) and (m), respectively.
A total ol 40 E coll coloni€s were count€d atter filtering a
b. Mark the petri dish and report form wiih the sample 50-mL sample.
identification and voiume.
E. coti llOO mL _ No. E 6oli Colonies Counted x .100 m L
c. Place a sterile membrane filter on the tilter base, grid Sample Volume Filtered, mL x 100 mL
side up, and attach the funnel to the base; the mem-
40 E. coli 100 mL
brane filter is now held between the funnel and the
(50 mL)(100 mL)
base.
0.8 E. colilmL 100 mL
d. Shake the sample bottle vigorously about 25 times to '1
00 m L
distribute lhe bacteria uniformly and measure the de-
sired volume of sample or dilution inlo the funnel.
= 80 E. coli /10O mL
e. For ambient surlace waters and wastewaters, select 7. References
sample volumes based on previous knowledge of pol-
lution level to produce 20 to 80 E coll colonies on the EPA Test Method: Escherichia coli (E. coli) in Water by Mem-
membranes. Sample volumes of 'l io 100 mL are nor- brane Filtration Using l\rembrane-Thermotolerant Escherichia
mally tested at hall-log intervals, lor exampte, 100, 30, colAgar (mTEC), Method 1 '103.1, 2002.
10,3 mL.
See pages 9-48 and 9-56, STANDARD METHODS,2lst
t l. Smaller sample sizes or sample dilutions can be used Edition.
I lo minimize the interference of turbidity or for high bac-
lerial densities. Multiple volumes of the same sample Presence/Absence Method
or sample dilutions may be liltered and the results may 1. General Discussion
be combined.
The presence/absence (P/A) test for the coliform group
g. Filter the sample and rinse the sides of the funnel at in drinking wat6r is a simple modilication of the multiple
least twice with 20 to 30 mL ol sterile buffered rinse tube procedure described above. One large test portion
water. Turn off the vacuum and remove the funnel lrom (100 mL) in a single culture bottle is used to obtain quati-
I the filter base.
tative information on the presence or absence ol coli-
h. Use sterile lorceps to aseptically remove the mem- forms. The media used js a mixture of laclose and lauryl
brane filter from lhe filter base and roll it onto the tryptose broths with bromocresol purple added to indicate
mTEC agar to avoid the formation of bubbles between pH. Following incubalion, gas or acid (yellow color oI
Ihe membrane and lhe agar surface. Fleseat the mem- media) is produced if colilorms are present. The p/A test
brane if bubbles occur. Close the dish, invert, and incu- procedure is outlined in Figure 11.18.
bate ai 35' 10.5" lor 2 hours. 2. Materials Needed
i. Alter a 2-hour incubation at 35. t 0.5"C. transler the
a. P/A broth. Prepare according to the instructions on the
plate to a Whirl-Pakd bag, seal the bag, place the bag
bottle.
with lhe plate inverted in a test-tube rack, and pul the
rack in a 44.5" I 0.2"C water bath for 22 to 24 hours. b. 250-mL screw-cap. milk dilution bottle with 50 mL ster-
ile triple-strength P/A broth.
j. After 22 to 24 hours, removo th6 plate from the water
bath. Place an absorbenl pad in a new petri dish or the c. Autoclave for sterilization of m€dia and glassware.
lid of the same petri dish and salurate the pad with
urea substrate medium. Aseptically hansfer the mem- d. lncubator set at 35" t 0.5"C.
brane lrom mTEC agar to the absorbent pad saturated e. Sterile, '100-mL graduated cylinder.
560 Water Treatmenl

(Coliform)
OUTLINE OF PBOCEDURE FOR TOTAL COLIFORM

1. Presumplive Test

lnoculate 100 mL in a P/A broth and incubate

24 ! 2 ht at 35" I 0.5'C

No gas or yellow color- Gas or yellow color produced-

incubate 24 hr more continue with confirmed total colilorm
and fecal coliform tests

No gas or yellow color-

no coliform present

2. Conlirmed Tesl

lnoculate (with loop or applicator stick) brillianl green bile brolh

lncubate 48 hr 1 3 hr at 35" t 0.5oC

No gas Gas produced

colilorm absent coliform group present

FECAL COLIFORM

lnoculate (with loop or applicator slick) EC broth

lncubate 24 ht t 2 fi al 44.5" t O.2'C

No gas Gas produced

fecal coliform absent fecal coliform present

Fig . 1 1 . 1 8 Schematic outline ol test procedure for total colitorm and lecal colifonn-presence/absence melhod
T Lab Procedures 56,

f. fhermometer verified to be accurate

by comparison
(coliform)
wtth a Nalional lnstitule o, St.
and rechnologv ,o
of a srerire pratinum wire
tocr,o,*,Jprnoioil;#?Xn"J,.Jil,:"ff
rr.rrsrt certirieJ ihJrml'mli"ll"no"to"
i"o"TIJ:ii;
t f,"J'iil'"fl"":irti5"T?$.
t'"ou'" as insrructed under
briltiant green bite (BGB) broth.

Afler 24 hours have elapsed, inspect

the tube for
h. Water bath set at 44.5" sas Jormarion. Any amount'ot gas foi"iiiij.il..iJi"lJl
L 0.2C. ered po_silive and so ,ecoraeo
6n1n" oa-rr".;;";:;,i"J:
i. EC brolh. prepare according
to instructions on bot e. :I/e.
BGB tubes are rerncubareo r",
nours "".iiiilirriZro
FOB DRINKING WATER SAMPLES
f. 48-Hour Brilliant Green Bile
Confirmation Test
a. Shake sample approximalely 25
times. Examine tubes tor gas at the
b. Inoqrlate 1OO mL into p/A culture
botfle and mix thor- period. Record both p-ositive
end o, the 4g r 3 hour
and negative-tu;;.-
,-"
oughty.
g. Becording Results
c. lncubate at 35" t0.5"C and inspect afiet 24 and
hours for acid reactions (yellow 48 production in the BGB
cotor). Record ..Gas brolh culture within 48
posrtlve and negative both hours.conrirms the presence of com*, O""i"rll
cultu;e bonles.
Heport resutt as presence/absenc"
d. A distinct yellow color forms negative for totat cotitorms in I
i;;i Gil;,;;
cond-itions exist gas
in the culture when acid oo miois-am"piJ..,'- "'
Itrs atso O"ing pr.ojr""jlo"iiiJ
snakrng ot rhe bome wil resutr rn 6 (oRtNKtNG wArER sAMPLEHsee
a f6aminJil#. il;
amount ot gas or acid constitutes
rve tesl requiring contirmation ,or
a po" ,i" ii".rrrl i{iy:* Fisure
tolalcoftfolh-, Delermine if tecal and total
uon to turther lesting tor fecalcoljform.
"iJ_ presence/absence '-*' *"-"""
colilorm are present using the
e. 24-Hour Bri iant Green Bile te"i
Conrirmation Test
ml of sample, 1OO mL
presumptive cutture bofltes Presence/Absence Method
^^911f,1,
gas .ull
or acid at 24 or 48 hours. Transrer that show
liqriJ, ;irh ,i; Becord test results.

lnoc u latio n
I00 mL
O No gas P/A pres umptive
but yellow color pos
present ltive

NO CAs

Fecal E NO GAS

Coliform BCB confirmed

Negative
U P/A negative
0

Date No
Total Colif orm Fecal
t Sou rce Presumptive Confirmed Coliform
4/8/o8 29 jty Halt
C
+

Fig . 1I. 1 9 Record presence/absence cotiforn

test resulls

I
I
i

i
562 Water Treatment

(Coliform)
ColilertrM Method
1- General Discussion '10 mL
Colilert provides a media that contains specific indica- lillline
tor nutrients for total colilorm and E. coli. As these nutri-
ents are metabolized, yellow color and fluorescence are
released confirming the presence of total coliform and E.
coli respectively. Non-coliform bacteria are chemically
suppressed and cannot metabolize the indicator nulri-
ents. Consequently, they do not inlerlere wilh the identifi-
cation ot the target microbes. Total colilorms and E. coli \ t)
are specifically and simultaneously detected and identi-
fied in 24 hours or less.
Although the Colilert method can yield presence/
absence results within 24 hours and is less cumbersome
to perlorm than the m€mbrane filtration (NlF) method, op-
erators should be aware of the limitations ol these tests in e. Cap ths tubes tightly.
evalualing samples lor regulatory purposes. The results f. Mix vigorously to dissolve the reagent by repeated in-
lor the Colilert and MF tests are not always comparable, version of carrier.
which may be due to:
. /VOIE; High calcium sall concenlrations in cerlain walers
lnterferences in the sample lhat may suppress or may cause precipilat€. This will not atfect test results.
mask bacterial groMh
. Greater sensitivity of lhe Colilert media
. Added stress to organisms related to filtering
. The fact that ditlerent media may obtain betler groMh
for some bacteria 4
Both test methods are approved by the US Environ-
mental Proteclion Agency for reporting under lhe Safe
Drinking Water Program. When these tests produce con-
flicting bacteriological results, however, the safest course
of action is to increase monitoring and treatmenl efforts
until the results for both tests are negative.
2 l\,4aterials Needed

a. 10 culture tubes each containing Colilert reagent for

10 mL of sample (available from IDEXX Laboratories,
1 IDEXX Drive, Westbrook, ME 04092. Phone (800) g. lncubate inoculated reagent tubes in the carrier at
321-O2O7). 35"C lot 24 hours. lncubalion should begin within 30
b. Sterile '1o-ml pipets. minutes of inoculalion.

c. lncubator at 35'C. NOIE lncubation exceeding 28 hours should be

d. Long wavelength ultraviolet lamp.
e. Color and fluorescence comparator.
3. Test Procedure
a. Remove lhe front panel of the Colilert tube box along
4 Test Results and lnterpretation
the perforation line to allow easy access and removal
ol tube carriers.
b. Flemove a 1o-tube carrier lrom the kit and label each
1o{ube MPN test as indicated on the carrier.
c. Crack the carrier along the front perforation and bend
lhe carrier along the back crease line. Sland the car
rier on the table top.
d. Aseptically fill each Colilert tube with sample wat€r to
the level ot the back of the lube carrier (10 mL).
avoided.
h. Flead tubes in the carrier at 24 hours. lf vellow color is
seen, check for lluorescence. Color should be unilorm
throughout the tube. lf not, mix by inversion belore
reading.
a. Compare each tube against the color comparator. ll
the inoculated reagent has a yellow color greater or
equal to the comparator, the presence ol total coli-
lorms is confirmed.
b. lf any of the tubes are yellow in color, observe each tube
Iorfluorescence by placing carrier2 to 5 inches irom the
long wavelength. ultraviolet lamp. lf the lluorescence ot
the tube(s) is greater or equal to the comparator, th€
presencs ol E coll is specilically confirmed.
 Lab Procedures 563

(Cotiform)
c. Samples are negative for total colitorms if no color is Head MPN as 2.2 per 1OO ml for totat cotirorm and <.1..1 for E
observed at 24 hours. Should a sample be so ligh y colifrom Table 1 1.S.
yellow alter 24 hours' incubalion that vou cannot d;fin-
itively read it relative to the posilive;omparalor tube,
you may incubate it up to an additional 4 hours. lf the
lnoculation Results
sample is coliform positive, the color will intensify. ll it
does not intensify, consider the sample negative. 10 mL
2 ol 10 - yellow
d. To ,ind theconcentration of total coliforms or E. colipet 0 of 10 - fluorescence
100 ml, compare the number of posjtjve reaclion
tubes per sample set (.10 tubes) to the standard MpN
Yellow Yellow
(Mosl Probable Number) chart shown on Table 1 1.5 or no fluorescence no fluorescence
in STANDARD METHODS.

TABLE 11.5 MPN INDEX FOH VARIOUS COMBINATIONS

OF POSITIVE AND NEGATIVE RESULTS WHEN
TEN 10-mL pORT|ONS ARE USED

Number of lubes giv;ng posiiive l\rPN lndex

reaction out of len 1O-mL portjons per 100 ml
0 <1.1
1 1.1
2 2.2
3 3.6
4 5.1
5 AO
6 9.2
7 12.O
8 16.1
9 23.0
10
>23.O
5. Becording Fesults
Hesults should be recorded on data sheets prepared
especially for this tesl. An example is shown in Figure
11.20.
6. Melhod ol Calculating the Most probable Number (MpN)
EXAMPLE 7 (DRINKING WATER SAMpLEI_(see
11.21).
Fig. 11.21 Results ol Colitert test-drinking water
Colisure Method
The Colisure test is a presence/absence test for coliform
bacteria. A sample ot water is added to a dehydrated medium
and, after 28 to 48 hours, the medium is eiamined for the
presence of coliforms. (The Colisure test js available from
IDEXX Laboratories, 1 TDEXX Drive, Westbrook, ME 04092.)
F. Comparison of Coliform Tests
The total coliform bacteria test includes both Escherichia
and Aerobacter coliform bacteria youps. Aetobacer and
some Escheichia can grow in soil. Therelore, nor all coliforms
found in the totar coliform test come from human wastes. Es-
cheichil cg!!.(E: coti) apparcn y arc alt of tecal origin. How-
ever. rt is difticult to determine E co, wilhoul meas-uring soil
coliforms too. The fecal coliform and Escherichia cotitesis arc
Figure used in an attempt to more specjtically determine the extent of
human wastes in water

mLof sample 100 mL G. AdditionalTestMethods

positive tubes 2 For a summary of approved and accepted microbiological
yellow color yes methods for EPA water monitoring regutalions, see E-pA's
,luorescence no websile al www,epa.gov/safewater/methods/rules*micro.html,

DATE
MPN/]00 mL
NO SOURCE 24 HRS TOTAL COLIFOHM E. COU
1701 ----+----+---- Yellow
4t16t0B x29 Main St 2.2 <l.1
Fluorescence

Fig. 11.20 Record colilorm test resutts using Cotilert method

 Lab Procedures 565

(Coliform)
.11,3L
QUESTIONS Estimate the most probable number (MPN) ol coli-
lorm group bacteria for a raw water sample from th€
Write your answers in a notebook and then compare your following test results:
answers with those on page 576.

11.3H Why are drinking waters not tested for specific patho-
ml ol sample 1 0.1 0.01 0.001
gens (such as Cryptosporidium ot Giadia)2 dilutions 0 -1 -2 -g
readings (+ tubes) 5 5 3 't
11.31 To perform coliform lests, how would you decide how
'l
many samples to test? 1.3M How is the number of colilorms estimated by the
membrane filter method?
11.U Why should sodium thiosulfale be added to coljform
1 1.3N What are the incubation conditions for chlorinated
sample bottles?
samples when using the membrane filter method
11.3K Steam autoclaves sterilize (kill atl organisms) at (enrichment technique)?
a
pressure of _psi and temperature ol_"C 11.30 Why is the lotal coliform tesl not an ad€quate meas-
during a time period (at sea level). ure of lhe extent of human wastes in wateaT

-minute

Efid o( Leaa o*4 of 5 Lo*ottd

olL
LAgOe\lce\t
?gocEquQE4
Please answer the discussion and review questions next

DISCUSSION AND REVIEW QUESTIONS

Chapter ll. LABORATORY PROCEDURES
(Lesson4of5Lessons)

Write the answers to these queslions in your notebook. The

queslion numbering continues lrom Lesson 3.

15. What is the purpose oi the coliform group bacteria lest?

16. How would you determine the number o, dilutions for an

MPN test?
17. What factors can cause errors when counting colonies on
membrane filters?
18. How can questionable colonies on membrane filters be
( \\

verified as coliform colonies?

566 Water Treatment

CHAPTERll. LABORATORYPROCEDURES
(Lesson5ol5Lessons)

5. Hardness Funnel
Hot plate
A. Discussion Flask, volumetric (1,000 mL)
Hardness is caused principally by lhe calcium and magne-
sium ions commonly present in water. Hardness may also be
D, Fleagents
caused by iron, manganese, aluminum, strontium, and zinc if NOIe Standardized solutions are available already pre-
present in significant amounts. Because only calcium and pared from laboratory chemical supply compa-
magnesium are present in significant concentrations in most nles,
waters. hardness can be defined as the total concentration of
calcium and magnesium ions expressed as the calcium 1. Bufler solulion
carbonate (CaCO3) equivalent. 2. Standard EDTA or CDTA titrant. EDTA is disodium
There are two types or classifications oI water hardness: ethylene-diaminetetraacetats dihydrate, also called (ethyl-
carbonate and noncarbonate. Carbonate hardness is due to enedinitrilo)-tetraacetic acid disodium salt. CDTA is diso-
calcium/magnesium bicarbonate or carbonale. Hardness that dium-CDTA (1,2 cyclohexanediaminetetraacetic acid).
is due to calcium/magnesium sulfate, chloride, or nitrate is 3. lndicator solulion.
called noncarbonaie hardness.
Hard water can cause incrustations (scale) when the water
evaporales, or when heated in household hot water heaters
and piping. Hardness-producing substances in water also OUTLINE OF PROCEDURE
combine with soap lo lorm insoluble precipitates. The
common method ol minimizing lhese and other problems due mr EOT
to hardness is water supply softening. This procedure is dis-

f 1
cussed in Volume ll, Chapter 14, "Softeningl'
B. What ls Tested
Common Range
6
mg/L as CaCOr

Surface Water 30 to 500'

Well Water 80 to 500'
1 Add 50 mL of 2. Add 2 mL ol buffer 3. Ikat€ wilh EOTA
' Levels of hardness depend on local conditions samPle lo and 2 drops of to bl0e end poinl.
clean beaker. indicator
C. Apparatus
Buret (25 mL)
Buret support E. Procedure
Graduated cylinder (100 mL)
1. Take a clean beaker and add 50 mL ol sample.
2. Add 2 mL of bulfer solution.
u 3. Add 2 drops indicator solution.
t 4. Titrate with standard EDTA solution until the last reddish
tinge disappears lrom th6 solution. The solution is pur6
* blue when lhe end point is reached.
5. Calculate total hardness.
F. Example
Besults from waler hardness testing ol a w6ll wet6r
sample were as follows:
Beaker (250 mL)
lvlagnelic stirrer
SampleSize = 50 mL
Magnetic stir-bar ,.i,?J,:il},A=ro5mL
Flask, Erlenmeyer (500 mL)
 Lab Procedures 567
I

G. (Jar Test)
Calculation
3. Beakers (1,OOO mL)
Hardness, mg/t as CaCO3 _ A x 1,000 4. Pipers (10 mL)
mL ot Sample
5. Flask, volumetric (l,OOO mL)
_ 10.5 mL x 1,OOO
6. Balance, analytical
50 mL o, Sampte
D. Beag€nts
= 2_tO mg/L '1. Stock Coagulant Solulion
H. Precautions
a. Dry alum, Alr(SO,)3 . 14.3 HrO.15 Dissolve
1. Some melal ions interfere with this procedure by arum (r / percent) jn 600 ml distilled water
1O.O om drv
causing con-taine;
lading or indistinct end points. ln tnese in a 1,000-mL votumelric ftask. Filt to miri.
reagent should be used. you mav Iitrate
cases, an inhibitor
wiff, tion contains 1O,OOO mgr'l or iO mg/mt.
ilil;;:
et'
ard CDTA solution or EDTA sotution. "iit "i"ni-
2. The titration should be completed withjn
o
!E:,0, :!f, .Ar,(SoJ"
.
4e.6 H,o.,5 rhe operator
tve minules lo :11r,,",_y:r]ry,rh"
strength of rhe atum with a hydrom-
minimize CaC03 precipitation. erer. Lrqutd atum is usually shipped as 8.0
to 8.5 oer_
3. A sample volume should be selected that requires cent Al2Oj and conlains a6out b.g6 pound";;;li;-
less minum surate (48.5 percent orvl pii
than 15 mL of EDTA titrant to be used.
gravity 1.325). This converts ro leb,OO6l"rri,i flplJiii"
4. For titralions ol samples containing fore, add,'l5_6 mL tiquid atum to a f
mg/t:fi;;;:
low
hardness concen_ .OOO-mf"vof umeirt
(anons (tess Ihan 150 mg/L,
as CaCO.) a larger sampte lasx and ,i to mark. This solution contains IO,OOO
votume should be used- mglLot t0 mg/mL.
L Reference c. Table I 1.6 indicates the slrengths of stock solulions lor
varlOUS ClOSageS.
See page 2-37, STANDARD METHOD.S, 2jst Edition.
TABLE 11.5 DRY CHEMICAL CONCENTRATIONS
USED
QUESTIONS FOR JAR TESTING
'
Write your answers in a notebook
- ers with and then compare your 1 ml
Added to
ans those on page 576. Approx. Do3age GramVLlter 1 Litor Stock Sotution
Required, mg/Lb lo Preparec Sample Eq!.ts
11.3P What^are the principal hardness_causing Conc., mg/t (%)
ions in
l-10 mg/t
water? l gmtL 1 mllL 1,000 mg/a (0.17c)
10-50 mg/t 10
11.3Q What problems are caused by hardness EnlL 10 mgtL 10,000 mg/a (1.0?,
50 500 mg/L 1o0 g.t/L 1o0 mg/L
in waler? 100,000 mgla fi0.0%)

6. e Ftom JAF| fESf by E. E. Arasmith. Unn-Benton

Jar Tesl Community Col-
lege, Albanv. OR.
A. Discussion D Use lhis colrmn,
which indicates lhe approxrmale dosage
by raw wal6r. to doterrnine the triat dosages required
,:"
tesrs designed to show rhe effectiveness fo Oe usei in ;-ai til
^,111::t^. of c
memrcat trealment in a water trealment factlity. fr,l""V Tirs_column indicates tho grams of dry chemical
cnemicals.w€ add to waler can be evatuated
tn" that should b6
oi #iril"i"l "i used when preparing th€ srock sotutio;.
rne sroci sotuiioi i#
sists ol the chemical plus enough wal6r lo m"f,"
fP-? :"glg
by the use or a jar tesr. n," ,o"r "lrJoiLiiir o*_fil"i Jr-
mese chemicals are those used for coagulation, such as
atum
"
jar lest. the operaror can approximiie
*!E fy.T:.j:ir,g !he
corred coagulanl dosaoe lor ntant use
wni:i varyinl
amo"unts of turbidity,
color. or-other factors lndicate raw water
E. Proceduro
qualily changes. The jar test is atso a very usetul tool
in evalu_ 1. Collect a two-ga on (B-titer) sampte ol the water to be
Sllng new coagulants or polymers
being considered for use on tested.
r plant scale.
2. lmmediatety measure six l,OOO-mL quantilies
and place
8. What ts Tested into each of six 1.OOO-mt beakers.
Baw water, lor op limum coagulant
dose, which
ing on coagulan t(s) used and water quality.
va ries de- 3. Place all six beakers on stirring apparatus.
4. With a measuring pipet, add increasinq
dosaoes of coa.!_
Apparatus uranl sotutrcn to the beakers asrapidv;s po"liOle. Set"it
A slirring machrne with six paddles a series ol dosages so thal the first beakar will
capabte of variable reoreseni
speeds from 0 to IOO revolutions per miriute -'- an underdose and the last an overdose.
lnefvf).
An illuminator located undernealh the stirring 5. Iiih-lllnq paddtss towerod into the boakers, starl sflr-
mochanism r]ng_appalatus and op€rat6 it for one mlnut6
(optional). at a spgod o,
80 BPM,'6

Values ol 14.A H and 49.6 H2O were obtained lrom

purchased by your water healmenl planl ALIJMINUM SULFAfE published by
alum Staufrer Chsmicals. Aciual values lor commercial
may vary stigh tly
Use slirring speeds and tjmes
thal are si milar to actual conditi ons in your
watertrcatment planl
568 Water Treatmenl

(Jar Test)

6. Beduce the stining speed for the ne)t 30 minules to 20 There are a number of tests lhat can be performed to im-
RPM.16 prove the jar test and the interpretation of lhe results. These
tests include:
OUTLINE OF PROCEDUFE 1. Alkalinity (before and afler)
2. pH (before and atter)
\I 3. Turbidity of SUPEFNATANT" (before and after)
4. Filtered turbidity ol supernatant
See Figure 1 1.22 for a helplul jar test data sheet.
NO 'NO 2 lro. s; NO,1 NO.5 NO6
After estimating the optimum coagulant dosage, run another
jar test with the optimum coagulant dosage constant, bui vary
the pH. These results will give you the optimum pH.

1. Add 1,000 ml lo each of 6 beakers Alkalinity must be monitored very carelully belore and atter
the jar test. Alkalinity must always be at least half of the coagu-
lant dose. For example, if the opiimum coagulant dose is 50
mg/l. the total alkalinity must be at least 25 mg/|. lI the natural
alkalinity is less than 25 mg/l, adjust lhe total alkalinity up to
25 mg/L by adding lime. For additional inlormalion on how to
calculate the amount ol lime needed lo increase the alkaliniry
0 see page 172, Section 5.243, "Arithmetic for Solids-Contact
Clarilication;'and page 173, Example 6.

NO. r NO,2 NO.3 NO.4 5 NO.6 G. Example

A sample of river water was collected for jar test analysis to
determine the optimum alum dosag€ ror etlective coagulation.

2. Add increasing dosages of coagulant The following series ol alum dosages were added:

Beaker No. mL Alum Solution Added Alum Added, mg/L

G (7 ) 1 1.0 IO
2 1.2 12
3 1.4 14
,li )
H
lr,-. 4 1.6

etl a,9
a
5
6
1.8
2.O
18
20

3. Stir for appropriate time period. Evaluate floc quality H. lnterpretation of Besults
Results of the above jar testing were recorded as follows:
7. Observe and evaluate each beaker as to lhat specific
dosage's floc quality. Record results. Beaker No. Alum Dose, mg/L Floc Ouality

8. Stop the stirring apparatus and allow samples in beakers 1 10 poor

to settle lor 30 minutes.'6 Observe the lloc settling charac- -12
2 fair
teristics. A hazy sample indicates poor coagulation. Prop- 3 14 good
erly coagulated water contains floc particles that are well- 4 16 excellent
formed and dense, with the liquid between the particles 5 18 excellent
cleat Describe results as poo( fair, good, or excellent. 6 20 good
F. Precaulions
Without going lo extreme measures, it is very difficult to du- The above results seem to indicate that a dose of 16 or 18
plicate in thejartest exactly what is happening on a plant scale. mg/L would be oplimum. The operator should, however, verify
The jar test, therelore, should be used as an indication of what this result with visual observation of what is actually happen-
is to be expecied on a larger scale in a water treatment plant. ing in the tlocculation basin.

r5
Use stirring speeds and times lhat are similar to aclual condilions in your water trealmenl planl.
t7
Supemalant \sue-perNAY.lent). Liquid removed lrom settled sludge. Supernatant commonly relgrs lo thB liquid bolwesn the sludge on the
bottom and lhe scum on lhe water surlace ol a basin or conlainer.
 Lab Procedures 569

=o
-aE
=<
=
za.
il4sr
z Az2
= =H?
=
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J;Y
=.; iii
> -.'!

9onI
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iio
o
(,

Ntht st

Nlv! 0e

4 Ntht 9z q)
o e
F N ll/'l 0Z
I o
z NIIEt Is
F 9 1
F
o a NIW OI .q
l- G
.9
F o NI S q
IC
F.'
lst
Hd
\
oflsntov
lsoo lr-
orv -cvo3
SSOO
cvo 3
A.tlO t€ a nl
o3all rB
>".3k
r >,.3k
tr
F
>",3a
: >"8e
I ZIS
3td htvs
cc
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'l

lsnfov Hd 1N V',t n CvoS

otv I Nvtn0vo3 :3 !1t1
ls l-'tYNv f .Lvo :ftHnos
570 Water Treatment

(pH)

OUESTIONS B. What ls Tested

Write your answers in a notebook and lhen compare your Common Range
answers with those on page 576. Surface water 6.5 to 8.5
11.3R What is the purpose of the jar test? Finished waier 7.5 to 9.0
Well water 6.5 to 8.0
11.3S What stirring speeds are used during the jar tests lor
optimum alum dosage?
C. Apparatus
1. pH meter
2. Glass electrode
3. Fleferenceelectrode
4. Magnelic stirrer
5. Magnetic stir-bar
D. Reagents
1. Butfer tablets for various pH value solutions (available
from laboratory chemical supply houses).
2. Distalled water.
7. pH
E. Procedure
A. Discussion
Due to the ditference between the various makes and
The pH ol a water indicates the intensity of its acidic or models ol pH meters commercially available, specific in-
basic strength. The pH scale runs irom 0 to 14. Water having structions cannot be provided for the correct operatjon of
a pH of 7 is at the midpoint of the scale and is considered all instruments. ln each case, follow the manufactureds in-
neutral. Such a water is neither acidic nor basic. A pH of structions for preparing the electrodes and operating the
grealer than 7 indicates basic waler The stronger the basic instrument.
intensity, the greater the pH. The opposite is true of lhe acidi-
ly. The stronger the intensity of the acidity, the lower will be 2. Standardize the instrument (pH meteo against a butfer
solution with a pH close to that of the sample.
the pH.
pH SCALE 3. Rinse electrodes thoroughly with distilled water after re-
moval trom butler solution.
O+ INCREASINGACID --_-7 - _- . INCREASING BASE -}14 4. Place electrodes in sample and measure pH.
5. Bemove electrodes lrom sample, rinse thoroughly with
, Neulral. distilled water.
Mathematically, pH is the logarithm (base '10) of the recipro- 6. lmmerse electrode ends in beaker of pH 7 buffer storage
cal ot the hydrogen ion activity, or the negative logarithm of lhe solution,
hydrogen ion activity.
1
7. Turn meter to standby (or OFF).
pH = Log *-
lH'l
= -Los [H*]
F. Precautions
'1,
For example, if a water has a pH of then the hydrogen ion 1. To avoid laulty instrument calibration, prepare fresh buffer
activity [H.] = 10 1 = 0.1. lf the pH is 7, then [H.] = 10i = solutions as needed, and ai least once per week (from
0.0000001. A change in the pH of one unit is caused by commercially available buffertablets).
changing the hydrogen ion level by a lactor ol 10 (10 times).
2. pH meter, buffer solution, and samples should all be near
ln a solution, bolh hydrogen ions [H.] and the hydroxyl ions the same temperature because temperature variations
IOH-I are always present. At a pH of 7. the activity of both hy- will give somewhat erroneous results. Allow a few minutes
drogen and hydroxyl ions equals 10-7 MOLES'8 per liter. for the probes to adjust to the butfers before calibrating a
When the pH is less than 7, the activity of hydrogen ions is pH meter to ensure accurate pH readings.
greater than lhe hydroxyl ions.
3. Watch for erratic results arising from electrodes, faulty
pH plays an important role in the water treaiment processes connections, or fouling of electrodes with aily precipi
such as disinfection, coagulation, soflening, and corrosion tated matter. Films may be removed from electrodes by
control. The pH test also indicates changes in raw and fin- placing isopropanol on a tissue or a Q-tip and cleaning
ished water quality. the probe.

's Mole- The molecular weight of a substance, usually expressed in grams.

 Lab Procedures 571

(Temperature)
4. The temperature compensator on the pH meter adjusts gas concentrations. and water slability with respect lo calcium
the meler lor changes in electrode response with te;per- carbonate, rn addition lo its acceptability by consumors tor
alure. However, the pH ol water also changes with t;m_ drinking.
peralure and the pH meter cannot compensate for this
change. B. What ls Tested
5. We. recommend standardizing lhe pH meter using a pH Common Flange, oC
buller sotution close to the pH ol the sample. nowevei it Baw and Treated Surface Water
you use another butfer solution with a ditferent pH to de- 5to25
termine the calibration of the pH meler for a range ol pH Well Water 10 to 20
values and the pH meter does not give the pH of ihe
second butfer, follow the manufacturer,s directions to C. Apparatus
adiu_st the, pH meter. This procedure is calted adjusting
the "slope' of the pH meter. 1. One National lnstitute of Standards and Technology
(NIST) thermometer lor calibration ot the other thermori-
6. lt.you are measuring pH in colored samples or samples eters.
with high solids content, or if you are taking measure_
ments that need to be reported to the US EpA;you should 2. One Fahrenheit, mercuryJilled, '1" subdivided thermom-
use a pH electrode and meter instead ol a cblorimetric eter.
method or test papers. pH meters are capable of provid- 3. O-ne Celsius (Iormerly called Centigrade), mercuryJilled,
t
ing 0.1 pH accuracy in most applications. ln contrast, 'l' subdivided thermometer.
colorimetric lests provide : 0.1 pH accuracy onty in a lim-
iled range. pH papers provide even less accuracy. 4. One m6talcas€ to fit each lhermometer.
G. lnterpretation ol Flesutts NOIEr There are three types of thermom€ters and two
scales.
The pH ol water has a very imporlant tnfluence on lhe effec-
lr!€ness of chlorine disinfeclion. Chlorination is a chemical re- SCALES
action in which chlorine is an oxidizing agent. Chlorine is a more
efleclive oxidant at lower pH values. Simply slated. a chlorine
1. Fahrenheit, marked "F.
Esidual.ol 02 mg,/t at a pH of 7 is
'10.
iust as etfective as I mg/L at 2. Celsius, marked "C (formerly Centigrade).
a pH ol Therefore. live times as much chlorine is requiied to
do the same disinfecting job at pH 10 as it does at pH 7. THERMOMETER STYLES

The finished water of some treatment laciljties is adjusted 1. Total imm€rsion. This type of thermometer must be totally
or caustic soda to a slighfly basic pH tor the purpose
with.lime immersed when read. Readings with this type of thermom. I
dmrnrmrztng corrosion in the distribution system. eter will chang€ most rapidly when removed from the liquid
to be recorded.
H. Rererence
2. Parlial immersion. This type thermotneter will have a solid
See page 4-90, STANDARD METHODS. 21st Edirion. line (water-level indicator) around the stem below the point
where the scale starts.
OUEST]ONS 3. Dial. This type has a dial that can be easity read while the
i Wrile your answers in a notebook and then compare your thermometer is still immersed. Oial thermometer readjngs
0,$vers with thoso on page 576. should be checked (calibrated) against the NIST thermo;-
eter. Some dialthermometers can be recalibrated (adjusted)
il,3l What does the pH of a water indicate? to read at a set temperature against ihe NIST thermometei
ll.3u What precautions should be exercised when using a D. Beagents
i pH meter?
None required.

(, (l
i
E, Procedure I

1. Collect as large a volume of sample as is practical, The

temperaiure will have less chance to change in a large
/_ volume than in a small conlainer.
2. lmmerse the thermometer to the proper depth. Do not
touch the boflom or sides of the sample container with the
thermometer.
rl
3. Record temperature to the nearest fraction of a degree
lhat can be estimated from the thermometer available:

t, Temperature
4. When measuring th€ temperature of well water samples,
allow the waler lo continuously overllow a small container
Discussion (a polystyrene cotfee cup is ideal). place th8 thermometer
in the cup. After ther€ has b6€n no change in th6 tempsr-
Temperature is one ol the most frequen y taken tests in the ature reading for ono minuts, record th6 tsmp€rature. ih6
industry Accurate water temperature readi ngs are impor- t€mperature ot waler samples collected from a distrlbu.
not only for historical purposes but also b ecauss of its in-
tion system mainly depends on th6 soiltemp6rature at the
on chemical reaction rates, biological growth, dissolved depth of the water main.
 572 Water Treatment

(Turbidity)
F. Precautions suspended matter such as silt, linely divided organic and inor-
ganic matter, and microscopic organisms such as algae.
To avoid breaking or damaging a glass thermometer, store it
in a shielded metal case. Check your thermometer's accuracy The accepted method used to measure turbidity is called
against the NIST-certilied thermometer by measuring lhe tem- the nephelometric method. The nephelometric turbidimeter or
peralure ol a sample \,vith bolh thermometers simultaneously. nephelomeler (Figure 1'1.23) is designed to measure particle-
Some of the poorer quality thermometers are substantially in- reflected light at an angle of 90 degrees to the source beam.
accurate (off as much as 6'F or 3"C). The greater the intensity of scattered light, the higher lhe tur.
bidity.
G. Example
To measure the temperature of treated water, a sample was
obtained in a gallon bottle, the thermometer immediately im- NEPHELOMETEF
mersed, and a temperature ol 15"C recorded after the reading
METER
(-
became constanl.
c
H. Calculation
Normally, we measure and record temperatures using a ther- PHOTOCELL
momelerwith the proper scale. We could, however, measure a TURBIOITY PARTICLES
temperature in "C and convert to'F. The following formulas are
LAMP
I! SCATTER LIGHT
used lo convert temperalures from one scale to lhe other.
LENS
I 7II
1. Il we measure in 'F and wanl 'C,

"C=11"P-.r"t,
9
SAMPLE CELL
2. lf we measure in "C and want "F,
{vEBTTCAL VrEW}

"r=9t"o*sz"r
5
Fig. 11.23 Nephelometer
(Pemission ol I^IACH Compar,y)
3. Sample Calculation
The measured treated water temperature was 15'C. Whal fhe TURBIDITY UNlf (IU) r'g is an empirical quantity lhat is
is the lemperature in "F? based on the amount ol light that is scattered by particles ol a
polymer reference standard called formazin, which produces
"F = 9/s("C) +32"F particles that scafter light in a reproducible manner. Formazin.
= 9/5(15"C) + 32'F the primary turbidity standard, is an aqueous (waler-based)
suspension of an insoluble polymer formed by the condensa-
tion reaction between hydrazine sulfate and hexamethylene-
tetramine.
= 59"F
l. Fleference Secondary turbidity standards are suspensions of various
materials formulated to match the primary formazin solutions.
See page 2-61, STANDARD METHODS, 21st Edition. These secondary standards are generally used because of
their convenience and the instability ol dilute lormazin primary
QUESTIONS slandard solutions. Examples of these secondary standards
include standards that are supplied by the turbidimoier manu-
Write your answers in a notebook and then compare your facturer with the instrumenl. Periodic checks ol these second-
answers with those on page 577. ary standards against the primary lormazin slandard are a
must and will provide assurance of measurement accuracy.
1 1.3V Why are temperature readings important?
Th€ turbidity measurement is one of the most important
11.3W Why should the lhermometer remain immersed in
tests the plant operator performs. The Safe Drinking Water Act
the liquid while'being read?
stipulates specific monitoring requiremenls lor turbidity. Tur
11.3X Why should thermometers be calibrated against an bidity of treated surface water musl be measured every four
accurate NIST-certified thermometer? hours or conlinuously and the results reported to the appropri-
ate authority. Turbidity testing is the most critical tool in recog-
9. Turbidity nizing changes in raw water quality, detecling problems in co-
agulalion and sedimentation, and troubleshooting filtration
A. Discussion problems.
The term turbidity is simply an expression of the physical The maximum contaminant level (MCL) tor turbidity is one
cloudiness of water. Turbidity is caused by the presence ol TU with five TU allowed under certain circumstances. Water

1'1
Turbidity Unils (TU). Turbidity units are a m€asure ol th6 cloudin6s6 oI wat€r. ll measursd by a neph€lomskic (delloctod lighl) lnslrum€ntal pro-
cedure, turbidity units are expressed in nephelomelric turbidity units (NTU) or simply TU. Those turbidity units obtained by visual molhods ar6 6x.
pressed in Jackson Turbidity Unils (JTU) which are a measure of th6 cloudiness ol water; they are used to indicats the clarity ol water. Thers is no
real connection beh/veen NTUS and JTUS. The Jackson lurbadimeler is a visual method and lhe nephelom8ler is an inslrumental method based on
dellected lighl.
 Lab Procedures 523

(Turbidity)
lreatment plant operators should strive to produce
a finjshed the mark and mix. The lurbidity of this suspension is
water with a turbidity of 0.1 TU or less.
considered 400 NTU exacfly.
. For.additional intormation
lurbidity requirements
on monitoring requirements and d. Prepare solutions and suspensions monthly.
of the Surface Wa'ier teatment Buie
ll[Il.^=, rrllel. chaprer zz, ,,ortnitng witei
Erons. Seclion 22.221, and the poster includeO wrtn tiis
hd; 3. Slandard lurbidjty suspensions: Dilute .tO.OO
mL stock
rurototty suspension to 100 mL with turbidityjree water.
manual. Prepare weekly. The turbidity of rnis suspeirsion ts
Oe-
lined as 40 NTU-
B. What ls Tested
4. Oilute_ turbidity standards: Dilute portions of the
standard
Common Range, NTU turbidity suspension wilh turbidity-free *ut"l' u. *lrirrO.
Untreated Surlace Waler '1 Prepare weekly.
to 300
Filtered Water 0.03 to 0.50 E. Procedure
Well Water 0.05 to 1.0+ 1. Turbidimeter calibration: The manulaclurer.s ooeratino in-
structions should be followed. Measure your sianOarO"so-
C. Apparatus lutions on the turbidimeter covering the iang" ot int"i;.
lf the inslrument is already calibral;d in sta;dard turbiditu
1. Turbidimeter: To minimize ditferences in turbidity
meas- units, this procedure will check the accuracy of the
I urements, rigorous specilications lor turbidimeiers cali'-
are bration scales. At least one standard shouid be;u;;n
necessary The turbidimeter should have the lollowing
im- each. instrument range lo be used. Some in"tirr"ni"
portant characteristics: permit adjustments of sensitivity so that
scate vatueJwi
a. The turbidimeter should consist ol a nephelomeler correspond to turbidities. Fleliance on an initrJmeni
with a light source illuminating the sample.'and manufaclurer's scattering standards lor cafiOrafin!
one oi tfie in_
more photoelectric detectors to indicale the intensity not an acceptabte practice unless rhe-y are in
scaflered light at a 90. angle wilh a readout device.
ol :tM91j
acceplabty" close agreement with prepared standaids.
lf a
precalibrat€d scale is not supplied, tiren
b. The light source should be an intense tungsten curves should be prepared for each usable r.ange
fila- ""liOr"tion
oi ih"
ment lamp. rnslrumenl-
c. The tolal drslance lraveled by the lighl through the 2. Turbidities less than 40 units: Shake the sample
to thor-
sampte should be less than about 5 ceitimelers: oughly disperse the suspended sotids. Wait ,ntit ai,
OrU-
d. The instrument should have several measurement ores otsappea( lhen pour the sample into the turbidimeter
ranges. The instrument should be able to measure ruoe. Head the turbidity direc y lrom the instrument scale
lrom 0 to 1OO turbidity units, with sufficient or trom lhe appropriate calibration curve.
sensitivitu
{on the lowest scale) to detect dif,erences ol 0.O2 or 3. Turbidities exceeding 40 uniis: Dilute the sample with
one
tess in. filtered walers having turbidilies ol less or more volumes of turbidity{ree water until ihe lurbiditv
tha;
one unit rals below 40 units. The turbidity of the orioinal samole il
2. Sample lubes. These are usually provided with the rnstru- then compuled from lhe turbidity ot tne dituted sampte
ment. and the dilution tactor. I

F. Example i
0 Reagents
sampte was coltected and the lurbidity was
l. Turbidityjree water: pass dis lled water through a ,^.tj?'?.oi,
round to be greater than 40 units when first checked. 30 mL of
mem-
brane tilteriaving a pore size no grealer than this sampte was then diluted with 60 mL of turbiOity-11g;;fter.
012 microns
llrlElte lroT taboralory suppty houses). Discard the
lirsl200 mL coltected. lf lillration does not reduce
This diluted sampte showed a turbidity of 25 units.
turbiditv
olthe disti ed water. use unfittered disti ed wa;; - - - ' NTU Found in Diluted Sample, A = 25 NTU
2. Slock formazin turbidity suspension:e mL of Dilution Water Used, B = 60 mL
a. Solution I-Dissotve 1.OOO gm hydrazine sullate
in dis-
mL ol Sample Volume Taken lor Dilution. C = 30 mL
tr ed water and dilule to I OO mL in a volumetnc ,lask. G. Calcutation
b. Solution.ll--Drssotve 10.00 gm hexamelhylenetetra-
mrne rn distiled water and dilute to 100 mL in Nephelometric Turbiditv Units (NTU) (A,*?
a volu- =
metric Ilask.
c. ln a 100-mL volumetric llask. add (using _ (2s NTU)X(60 mL + so mL)
S_mL volumet-
rc prpels) 5.0 mL Sotution I and 5.0 m7 of Sotutron 30 mL
ll.

il Mix and allow to stand 24 hours at 25.C. Then,

dilute r;
= 75 NTU

sloc* secondary standard lurbidity suspensions thal require no preparation

are availabre from comm€rcial supptiers and approved ror
use
574 Water Treatment

(Turbidity)
H. lnterpretalion ot Flesults Section A.139, "Laboratory Procedurss." Check all ol the
arithmetic in this section using an electronic pock6t calcula'
Report turbidity results as lollows tor. You should be able to get the sams answers.
Turbidity Reading Record to Nearest
I1.5 ADOITIONALREADING
0.0 lo 1.0 0.05
1. NEw YORK MANIJAL, Chaplet 4,' 'water Chemistry,'
1 to 10 0.1
Chapter 5,' "Microbiology,' and Chapter 21,' "Laboratory
10 to 40 1
Examinations ol Waler."
40 to 100 5
2. TExAs MAN|)AL, Chapler 6,* 'Water Chemistry,' and
100 to '1 ,000 10 Chapter 12,"Laboratory Examinations.'
>1,000 100
' Depends on odition.

Noles and Precautions 11 .6 WATER LABORATORY TESTS IN VOLUME II'

Sample tubes must be kept scrupulously clean, both WATEN TREATMENT PLANr OPERATION
inside and out. Discard them when they become Laboratorv orocedures for the rollowing tests are provided in
scratched or etched. Never handle them where the light vollme II, WATER TBEATMENT PLANT OPERATION, C\AP.
slrikes them. ter 21, 'Advanced Laboratory Procedures.'
2 Fill the tubes with samples and standards lhal have been 1. Algae Counts 10. Metals
agitated thoroughly, and allow sutlicient time for bubbles
to escape. 2. Calcium 11. Nitrate
3. chloride 12. PH
3 Tubidity should be measured in a sample as soon as
oossibla to obtain accurate resulls. The turbidity ol a 4. color 13 SPecilic
conductance
sample can chanoe atter lhe sample is collecled Shaking 5. Dissolved oxygen
the sample will not re-create the original turbidity. 14- sultate
6. Fluoride
Reference 7. Iron 15. Taste and Odor
J
8. Manganese 16 ?ihalomethanes
see page 2-8, STANDARD METHODS, 21st Edition.
17. Total Dissolved Solids
9. t/arb]e Test
OUESTIONS
Write your answeG ln a notebook and then compare your
answers with those on Page 577. hddLaaeouSolslnnsofib
'11.3Y What are the causes of turbidity in water? dv1,

11.32 How is turbidity measured? L goe&,eg

1T.4 ARITHMETICASSIGNMENT
wocEoucEc,
Turn to the Appendix, "How to Solvo Water Treatment Plant
Arithmetic ProOtemsl'at the back of this manual and read Please answer the discussion and review questions next'

OISCUSSION AND REVIEW QUESTIONS

Chapter 11. LABOFATORY PROCEOUBES

(Lesson5of5Lessons)

Write the answers to these questions in your notebook. The 2'1. what precautions should be exercised when taking tem-
question numbering continues from Lesson 4. perature measuremgnls?

19. What precautions should be considered when perform- 22. Why should turbidity be measured in a sampl€ as soon as
ing lh€ hardness determination on a water sample? possibl€?

20. How could you estimate the most etfective dose ol alum
or a polymer in a water treatment process?
.T
Lab Procedures 575

SUGGESTED ANSWERS
Chapter 11. LABORATORY PROCEDURES

ANSWERS TO OUESTIONS IN LESSON 1 Answers to questions on page 527.

Answers to questions on page SO9. '11.1G Six laboratory hazards are:
11.0A Laboratory quality control tests are important because 1. Hazardous materials 5. Fir€
they provide the necessary information to monilor the 2. Explosions 6. Burns (heat and
lreatment processes and ensure a sate and pleasant_ 3. Cuts and bruises chemical)
tasting drinking waier. 4. Electric shock
11.08 The prefix milli means 1/1,000 (O.OO.I) oI the unit fot- 'l
1.1H Never work alone in the laboratory Someone should
lowing. always be available lo hetp in cass you should have
11.0C The proper name of the chemical compound CaCO3 an accident that blinds you, leaves you unconscious,
is calcium carbonate. or siarts a fire you cannol handle. lf necessary hav6
somaone check on you regularly to be sure you are
Answers to questions on page 520. oK.
11.1A Descriptions ot laboratory glassware and their use or 11.'l I True. You may add acid to water, but neverthe reverse.
purpose.
1 1.1J Dispose of small amounts of corrosive acids by pour-
ing the neutralized acid down corrosion-resistanl
Item Description Use or Purpose sinks.and sewers using large quantities of waler to
l. Eeakers Shorl, wide cylinders in sizes Mixing
dilute and llush the acid.
trom 1 mL to 4,000 mL chemicals
ANSWERS TO QUESTIONS IN LESSON 2
2. Graduated Long, narrow cylinders in M€asuring
cylinders sizes lrom 10 mt to 4.000 mL
i
volumes Answers to questions on page 5gO_
3. Pipels Small-diameter, graduated
lubes, with a poinled tip,
Oelivering
accurale
1'l .2A The greatest sources of errors in laboratory tests are: I

an sizes from'1.0 ml to 100 mL

(1) improper sampling, (2) poor sample preserva-
volumes I
tion, and (3) lack of enough mixing during testing. i
4. Burets Long tubes with graduated Delivering and
walls and a stopcock in measuring 11.28 A representative samplo must be collected in order I

sizes lrom 10 mL to accurate for test results to have any significant meaning. With-
1,000 mL volumes used out a representative sample, the test results will not
in titralions reflect actual water conditions.

11.18 A meniscus 1'1.2C Sampling points in a distribution system shoutd be

is the curve of the surface of a liquid in a
smalltube.
11.1C Never heat graduated cylinders in an open flame
because they may break.
Answers to questions on page SZ2.
11.10 A standard solution is a solution in which lhe exact
concentration of a chemical or compound in a solu,
lion is known-
11.1E Laboratory notebooks and worksheets help record
data in an orderly manner.
11.1F Four sources or causes of poor quality of analytical
data are:
1. Sloppy laboratory technique
2. Deteriorated reagents and standards
3. Poorly operating instruments
4. Calculation mistakes
selected in order to trace the course from the finished
waler source (at the well or plant), through the trans-
mission mains, and then through ths major and
minor arteries ol the system.
Answers to questions on page 539.
11.2D The two general types ot samples collected by water
lreatment personnel are: (1) grab samples, and (2)
composile samples.
11.2E Water quality indicators that are usually measured
wilh a grab sample include: (.1) dissolved gases, (2)
coliform bacteria, (3) residual chtorine, (4) tempera-
ture, and (5) pH.
11.2F Depth samples are collected by the use of a
Kemmerer sampler or similar device. The sampllng
device and contain€r ar6 lower€d lo th6 deslrod
depth, then opened, filled, closed, and returned to
the surface.
 I
576 Water Treatment I

11.2G Samples should not be collected from taps sur- 't

1.3K Steam autoclaves sterilize (kill all organisms) at a
rounded by excessive foliage (leaves, flowers) or pressure of ,5 psi and temperature of ,2l"C during a
trom taps that are dirty, corroded, or are leaking. /gminute lime period (at sea level).
11.2H When collecting a sample, record the sample loca- 't1.3L mL o, sample 0.1 0.01 0.001
tion, date and time of collection, name of collector,
and any other pertinent information.
dilutions -1 -2 -3
readings (+ tubes) 5 3 1

ANSWERS TO QUESTIONS IN LESSON 3 MPN = 11,000/100 mL

Answers to questions on page 537.
'l 1.3A Chemicals used in water treatment that will cause
changes in alkalinity include alum, chlorine, and lime.
11.38 Alkalinity determination is needed when calculating
chemical dosages for coagulation and water soften-
ing. Alkalinity must also be known to calculate corro-
sivity and to estimate carbonate hardness ol water.
Answers to questions on page 539.
't
1 .3i,4 The number ol coliforms is determined by counting
the number of colilorm-appearing colonies grown on
the membrane filler.
'1 '1.3N lncubate the filter, without inverting the dish, ror 1v,
lo 2 hours at 35'C in an atmosphere of 90 percent
humidity. Transfer the membrane filter to a new pad
enriched with M-Endo broth. lnvert the dish and incu-
bate for 20 to 22 hours at 35"t 0.5"C.

'1
1.3C The benefats of chlorinating water include:
1. Disinlection
2. lmproved quality of water due to chlorine reacting
with iron, manganese, protein substances, sul-
fide, and many taste- and odor-producing com-
pounds
3. Control of microorganisms thal might interfere
wilh coagulation and llocculalion
4. Keeps filler media free oi slime groMhs
5. Helps bleach out undesirable color
11.3D A potential adverse effect from chlorination is the
possibility of the formation of carcinogenic chloror-
ganic compounds such as chloroform and other
THMs.
Answers lo questions on page 543.
11.30 The total coliform test is not an adequate measure of
the extent o, human wasles in water because it
measures both fecal colilorms (E col, and soil coli-
lorms. Additional tesls such as the MF-mTEC
method are needed to specifically identity and count
lecalcolilorms.
ANSWERS TO OUESTIONS IN LESSON 5
Answers lo queslions on page 567.
1 1.3P The principal hardness-causing ions in water are caL
cium and magnesium.
'11.3O Problems caused by hardness in water include: (1)
incrustations when water evaporates or scale when
healed. and (2) formation ol precipitates when com-
bined with soap.

'1
1.3E Conditions that can cause variations in the chlorine Answers to questions on pag€ 570.
demand of water include: (1) the amount of chlorine 11.38 The jar test is us€d to: (1) dolermine lh€ correct
applied, (2) length of contacl time, (3) pH, (4) lemper- coagulant dosage, and (2) evaluate new coagulants
ature, (5) organics, and (6) reducing agenls. or polymers.
11.3F The operator uses lhe chlorine demand test to deter-
11.3S Speeds used during the lar lest are as follows:
mine the best chlorine dosage to achieve specific
chlorination objectives. 1. 80 RPN4 for the first minute
1
'1.3G chror,ne 2. 20 RPN,i lor the nexl 30 minutes
Demand. = Chlorne Added. mg/L - Free Residual Chlodne, mg/L 3. Stop stirring (0 RPM) for the next 30 minules
lr,gJL
= 20 nglL- O-a .'..glL These speeds and times should be adjusted if nec-
essary to be similar to actual conditions in the wat€r
= 1.6 mgy'L treatment plant.
ANSWERS TO QUESTIONS IN LESSON 4 Answers to questions on page 571.

Answers to questions on page 565
11.3H Drinking waters are not tested for specific pathogens
because the tests are very time-consuming and
require special techniques and equipment.
'1
1.31 The number ol samples required for colilorm tests is
generally based on the population served by the
water system.
11.3J Sodium thiosulfate should be added to sample bot-
tles for coliform tests to neulralize any residual chlo-
rine in the sample. lf residual chlorine is not
neutralized. it will continue to be toxic to the coliform
organisms remaining in lhe sample and give false
(negalive) results.
'l 1.3T The pH o, a h/ater indicales lhe intensity ol its basic
or acidic strength.
11.3U Precautions to be exercised wh€n using a pH meler
include:
1 .
Prepare tresh butfer solution weekly for calibration
purposes.
2. Have pH meter, samples, and butfer solutions all
near the same temperature,
3. Watch lor erratic results arising from olectrodes.
faulty connections, or fouling of electrodos with
interrering matier.
4. The pH of water changes with temperature.
5. lf testing waters with a rangs of pH values, the
slope of th€ pH meter may require adjusting.

t
 Lab Procedures 577
t
Answers to questions on page 572 Answers to questions on page 574.
11.3V Temperature readings are important because tem- 11.3Y Turbidity in waler can be caused by the presence ol
peralure influences chemical reaction rates, biologi-
cal groMh, dissolved gas concentrations, and water
suspended matter such as silt, finely divided organic
and inorganic matter, and microscoptc organisms
I
stability with respecl to calcium carbonate. Also, con- such as algae,
SUmers are sensitive to the temperature of the water
they drink. 11 .32 Turbidity is measured by the nephelometric method.
E
11.3W The thermometer should remain immersed in the
liquid while being read for accurate results. When

11.3X
removed from the liquid, the reading will change.
All thermometers should be calibrated against an
Qt E
accurale National lnstitule of Standards and Technol
ogy (NIST) lhermomeler because some poorer qual-
ity thermomelers are substantially inaccurate (otf as
much as 6"F or 3'C). I
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