ENVTRONMENT tNST|TUTE OF MAtAyStA (E\MAS),

DEqARTMENT OF ENVTRoNMENT (DOE), MAAYS'A,

LAB MANUAL FOR |ETS (BP)

Kalibrasi LDÖ Meter

* lkon ? Akan kelihatan sekiranya kali,brasi telah

tarnat tempoh ataupun jika check standard
telah gagal.

Pengukuran Dissolved Oxygen

.þ Letakkan conductivity probe dalam

sample air.
{. Tekan kekunci GREEN/RIGI{T di bawah Read

t Skrin menunjukkan,,Stabilizing....' Dan

progess bar akan diisi dari O tó 100% sehingga
probe stabil

¡'. lkon lock akan kelihatan bila stabil.

'$ Ulangi langkah ini untuk sampel lain.

U
CePIETSO(BP)-201I
65
Orygen Ðemand, Blochemical DOC316.53.01200
Dilution Methodi Method 8043 ( t

Scope and Application: For water and wastewater.

1 Adapted fiom Sandard Methods for the Examination of Water and Wastewaterand Íiom Klein, R.L.; Glbbs,
C. Joumal of Water pollutíon
Control Fedention, 1 97 9, 51 lS), 2257.

'Before starting the test:

The BOD test is a Sday test. Follow all steps carefqlly to make sure that the test does not have lo be repeated.

The dilution wâter for this test must nst have an oxygen demand or any toxins.'When incubated for S.days at 20 "C, lhe
dissolved oxygen concæntration in the dilution water must not change by more than 0.2 mg/L.

Carbonaceous BOD (CBOD) can be determined by the addition of nit¡ification inhibitor. A tèst for GBOD is recommended for
biologically treated effluenE, samples seeded with biologícally treated efiuents and riverwaler.
'lhe Troubleshaoting-Graphical calculation method provides an altemate system for calculating
results and is a convenient
tool for troubleshooting problems in BOD measuremenb. The graphical calculation rnethod is not approved for regulalory
reporting.

Gollect the following items:

Description Quantity
BOD bottles, 30GmL, glass, with glass stoppers and'plastic caps 6
Dilution water containing nutrient buffer and seed (see Dilution water prepanlion) varies

Nitrification inhibitor (for GBOD only) 1 bottle

Pipet, serological 1

lncubator 1

See Consurnables and replacement ítems Íor reorder information.

Oxygen Demand, Biochemical

66
 Oxygen Demand, Biochemical

Dih¡tion method
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1. Prepare the dilutlon 2. Select the sample 3. Stir the sample gently 4. Fillan additional BOD
water using a BOD volumes. See Samp/e size with the plpet. Use the bottle with dilution water
Nutrient Buffer Pïllow. See selectíon. pipet to add the minlmum only. This will be the
Dilt ttion water prcparation. Note: If the minimum sample sample volume to the first dilution water blank.
volune is 3m4 ormotq BOD bottle.
+ determlne the dîssolved
Add the remalning four
ø<wen in the undituted volumes to four
sample; this detelmination sample
can.be onittedvÌ,hen' more BOD-bottles' Mark
analysing sewage and settled the bottles and record the
effiuents known to have a contents of each bottle'
dissolved oxygen content
near 0 mgtL

\Mren analyzing
disinfected sarnples or
industrial effluents, refer to
lnterferences.
:j;.1 ^i'i:!'I:.;:l :ã-'ï:i..i .:.:ì ;jiq". ¡r
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l
ri:i
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5. lf the test is for CBOD, 6, Fill each bottle to just 7. Stopper the bottles 8. Measure the initial
add two portions of below the lip with dilution carefully to prevent air dissolved oxygen
Nitrification lnhibitor water. Allow the dilution bubbles from becoming concentratlon in each
(approximately 0.16 g) to water to flow down the trapped. Tightly twist the bottle. Use a probe and
each bottle. sides of the bottle to stopper into place. Press meter or titraüon. lf a
The oxidation of nitrogen
prevent air bubbles from down on the stopper and titration is used, tr¡rio sets
becoming trapped in the invert the bottles several of BOD bottles must be
compounds will be
bottle. times to mix. prepared.
prevented. Report results
as CBOD. Be sure to measure the
DO of the dilution water
blank.

(_,

Orygen Demand, Biochemical

Page 2 of 14

e7
 Oryglen Demand, Biochemical

Dilution method
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9. Stopper the bottles 10. Place a plastíc cap 1f. Afterfive days, 12. Calculate the BOD
carefully lo prevenl air over the lip of each bottle. measure the re¡naining value (see Calculation
bubbles from becoming Put the bottles in an dissolved oxygen Metñods-Slandard
trapped. Add dilution water incubator at 20 (r1) "C concentration in each Methods).
to the llp of each BOD lncubate for five days. bottle.
bottle to make a water
At least 1.0 mg/L DO
seal.
should be left in each BOD
bottle.

Dilution water preparat¡on

The dilution water must be prepared very carefully to make sure that no source of oxygen demand
or toxins are added. The water that is used to prepare the dilution water must be of vãry high
quality. The water must not have any organic compounds or any toxic compounds such as
chlorine, copper and mercury.
Use the following guidelines to make sure the dilution water is of high qual¡y.
Guidelines
- Use distilled water from an alkaline permanganate distillation for the best resulls.

' Do not use deionized water from ion exchange cotumns. The resins in the cartridges
(especially new cartridges) will occasionally release organic materials that have an oryg"n
demand. ln addition, bacteria can grow on the columns and contaminate the dilution water.

' Store the distilled waler in clean jugs in an incubat or al2O "C. F¡ll containers till about
% full
and shake the jugs to saturate the water wilh air, or cap the jugs toosely and store for 24 hours
or more, to allow dissolution of oxygen.

' A small aquarium pump or air compressor can be used lo saturate the water with air. Make
sure that the air is filtered and that the filter does not grow bacteria. Clean the apparatus
before and after use.

' Add the nutrients and seed (if necessary) to the distilled water immedialely before the test.

' The dissolved oxygen concentration in the dilution water must not change by more than
0.2 mglL when incubated for 5 days at 20 "C.
Procedure

1. Prepare and store the dístiiled water at 20 "c (see Guidetines\.

2- Selecl a BOD nutrient buffer pillow from the BOD nutríent butrer pi¡ows lable.
3. Tap the pillow on a hard surface then shake the pillow to mix the contenls.

Qxygen Demand, Biochemical

Page 3 of 14

68
 Orygen Demand, Bioehemical

after incubation, although this ís not ac{ually measured. ln this case, it was equalto 9.ò mg/L and
the DO of the domestic sewage sample was assumed to be zero. lf another type of sample is
used, the DO of an undiluted sample should be measured either by the \M¡nkler titration or with a
luminescent or electrochemical probe.
The Calculation MethodvStandard Methods formula for calculating BOD also can be written as
follows (not approved for reporting purposes):

mLoflargersamplevolume-mLofsmaller x300-Doo+5 = mg/LBoD

Using this informatlon in the example:

rng/L DO remaining with smaller sample volume = 7.50
mg/L ÞO remainlng with larger sample volume = 2.25
mL of larger sample volume = 9,0
mL of smaller sample vofume = 2.0
' 300 = volume (rnL) of tsOD bottle
DOp = mg/L DO of dilution water = 9.0
S = mg/L DO of sample = assumed in this caseto be zero
Therefore:
7'.59 -Z?sx300-9
LO-2.O ' --- 0 = mg/L BOD =
+ 21G mg/L BOD

Using the equation below:

(slope x 300) - Y-lntercept + sarnple DO = mg/L BOD

To determine slope, arbitrarily select point A in Figure 1. At this point the mg/L DO remaining is
I
equal to 3.0 mg/L. The mL of sample at thls point is mL.The difference between the y-
I
intercept of 9.0 mg/L and 3.0 mg/L equals 6 mg/L; 6 mg/L divided by mL = 0.75 mg/L per mL.
slope = 0.75 mg/L per mL
Y intercept = 9.0 mg/L
sample DO = 0 (Because lhe sample is domestic sewage, this is assumed to be zero.)
Therefore:

(0.75 x 300) - 9.0 + 0 = mg/L BOD = 216 mg/L BOD

\
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Orygen Demand, Biochemical
Page l0 of 14

a6
 Oxygen Demand, Biochemical

{r
y lntercept
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7

Et
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o 5
ú.
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3

0 2 4681012
mL of Sample

Figure 1 Dissolved Oxygen per mL of Sample

Orygen Demand, Biochemlcat

Page 11 ot 14

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Orygen Demand, E¡bchernical

Summary of method
J
Biochemical Orygen Demand (BOD) is a measurement of the orygen requirements of munlcipal
and industrial wastewaters and sewage. The test results are used to calculate the effec,t of waste
discharges on the oxygen resources of the receiving waters. The BOD test is of limited value.in
measuring the actual oxygen demand because temperature change, biological population, water
movement, sunlight, oxygen concentration, and other environmental faclors cannot be reproduced

specific effluent and receiving water have been established.

The BOD test is performed by incubating a sealed wastewater sample (or a prepared dilutíon) for
the standard five'day peÉod and then determining the change in dissolved oxygen content. The
BOD value is then calculated from lhe resulfs of the dissolved oxygen tests.

Consumables and replacement items

Requircd reagenb
Description ..,_-. QuanfityfTest
' Unit Catalog number
BOD Nutrient Buffer Pillows, for 3 liters of dilution w¿ter 1píllow 50/pkg 1486166

Required apparatus
Description QuantityfTest Unlt Gatalog number
BOD Bottle, glass-stoppered, 300-mL, unlabelled 6 6/pkg 62106
BOD Bottle Cap 6 6/pks 241906
Bottle, wash, 50GmL I each 62011
Clippers, large 1 each 96800
Fipet, serological:
Pipet, serological, 1-mL 1 each 919002
Pipe[ serological, S-mL 1 each 53237
Pipel serological, 1 GmL 1 each 53238
Pipet Filler 1 eactr 1218900
Dissolved Orygen measurement apparatus

Recommended shndards
Description Unit Gatalog numbe¡
BOD Standard Solutîon, Voluette@ Ampule, 30G'mg/L, 1O-mL 16/pks 1486510
ezGGA BOD Standard Ampules, 450 mg/L, 2 mL 2Olpkg

Orygen Demand, Biochemical

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