Standard Solution:

Any chemical solution with high precise concentration is known as standard solution

Types of Standard Solution:

1. Primary standard solution

2. Secondary standard solution

1. Primary Standard Solution:

The solution which is made with high purity (99.9%) substances that dissolved in known amount of solvent is
called primary standard solution. These primary standard solutions are used for standardize other solution such
as oxalic acid.

1. Secondary Standard Solution:

It is made for any chemical analysis. Its concentration varies with the passage of time. So, before use, it is
necessary to standardize these solutions against primary standard solutions. Examples are hydrochloric acid
and sodium hydroxide (NaOH).

In analytical chemistry, it is an important technique for preparation and standardize of solution that is used in
chemical analysis.

Standard Curves:

Standard curve, which is also known as calibration curve, is a most common technique for predicting the unknown
concentration of a substance in providing unknown sample by comparing it with a set of standard samples of
known concentration.

Significance:

Standard solutions and curves are helpful in laboratories and industrial level such as production of chemicals,
pharmaceutical companies where standardize solution and compounds are used during quality testing and
manufacturing process.

Preparation of standard solution is not only helpful in analytical laboratories and chemical analysis; these techniques
are required in quality testing and forensic laboratories for different analysis.

It is need to diagnose accurately diseases in urine or blood samples and provide better patient care. It maintains
accurate research, and creates accurate public health data that can compared.

It also decreases percentage of error in the analysis. If any solution is contaminated or impurity, this technique is very
helpful for determine those solution. Through this technique, all the laboratory that are used chemicals are
standardize and measure their accurate concentrations.
CHEMICAL OXYGEN DEMAND:

EXPERIMENTAL PROCEDURE
APPARATUS:
 COD flask
 Pipette
 Burette
 Titration Flask
 Reflux
 Glass Beads
 Measuring Cylinder
CHEMICAL REAGENTS:
 Silver/Mercuric Salt
 25mL Oxidizing Agent (K2Cr2O7)
 75mL Concentrated Sulphuric Acid (H2 SO4)
 Distilled Water
 Ferrion Indicator
 0.25N Ferrous Ammonium Sulphate (FAS)
PROCEDURE:
STEP1: SAMPLE DIGESTION
Took a COD flask, added 50mL sample, and 6-8 glass beads for increasing surface area.
Then added a pinch of silver/mercuric salt to stop the interferences of chlorides. These chlorides are
precipitated out in form of silver or mercuric chlorides.
Then took two separate beakers, one beaker filled with 25mL oxidizing agent that may be K 2CrO4, K2Cr2O7 or
KMnO4 and other beaker filled with 75mLConcentrated Sulphuric Acid (H2 SO4).
Then proximately added 10-15mL Concentrated Sulphuric Acid (H2 SO4) in COD flask, this will be
exothermic reaction, and flask heated because of violent reaction.
PRECAUTION: Addition of Sulphuric Acid in COD flask should be slowly.
Then added 25mL of K2Cr2O7 in COD flask and remaining Concentrated Sulphuric Acid. Now the total
volume of flask became 150mL.
Transferred the COD flask on Reflux for 2 hours. This process will provide nascent oxygen for the oxidation
of organic matter.
But if some nascent oxygen will left in the flask which measured by titration process.
If all the organic matter react with nascent oxygen, there will be no left oxygen in flask and green colour
appeared. This is due to the use of concentrated sample so, repeat the process again but sample concentration
is used less than 50mL that may be 10mL or 1 mL.
STEP 2: SAMPLE PREPARATION
If green colour is not appeared in COD flask, then added 200mL distilled water. Now total volume became
350 mL. Now sample is ready for titration.
STEP 3: TITRATION PROCESS:
Took 100mL sample from COD flask in titration beaker.
Added Ferrion indicator in titration flask, sample colour changed.
Titrated it against 0.25N Ferrous Ammonium Sulphate (FAS) till reddish or reddish brown colour appeared.
Noted the total volume that is used for determination of unreactive oxygen in sample. Repeated the titration
process for remaining 250mL sample.
STEP 4: PREPARATION OF BLANK
The whole process is repeated for blank preparation, only sample is not added.
FORMULA:

(B− A) × Normality of titrant × Eq . wt . of Oxygen×1000

COD =
Volume of sample (mL)

BIOLOGICAL OXYGEN DEMAND:

BIOCHEMICAL OXYGEN DEMAND (BOD) MEASUREMENT:
It is measured in two steps: One is measured for DO while collecting sample that is zero day DO level, and
second one is measured after 5 or 7 days which is called BOD5 or BOD7. The difference represents oxygen
utilized by microbes to break down of organics in sample.

EXPERIMENTAL PROCEDURE:
APPARATUS:
 BOD bottles
 Pipette
  Burette
 Titration Flasks
 Incubator
 Measuring Cylinder
CHEMICAL REAGENTS:
 Buffer Solution
 Distilled Water
 Nutrients
 Manganese (II) Sulphate (MnSO4)
 Alkali Azide
 Concentrated Sulphuric Acid (H2SO4)
 0.025N Sodium Thiosulfate (Na2S2O4)
 Starch Solution
PROCEDURE:
Took 11 BOD bottles and divided them into three groups
Group 1: A1 A2 A3
Group 2: B1 B2 B3
Group 3: C1 C2 C3
Blank 1 Blank 2
STEP 1: PREPARATION OF DILUTION MEDIA:
Bacteria need nutrients, specific pH, specific temperature and oxygen demand for the decomposition of the
organic nutrients. These nutrients are mixture of 5 to 6 salts that are required for bacterial activity and growth.
Volume of BOD bottle= 300mL
Numbers of Bottles = 11
Total Distilled used for preparation of Dilution Media = 18 Litres
Took 18 litres distilled water and added 18mL buffer solution to keep pH at 8 -9.
Added 18ml nutrients and aerated it for 40-45 minutes.
STEP 2: SAMPLE PREPARATION
Now half- filled of these 11 bottles with dilution media (up to 150ml). Then added
1mL Sample ………. A1 B1 C1
3mL Sample……… A2 B2 C2
5mL Sample………. A3 B3 C3
No Sample Addition ………. Blank 1 Blank 2
Now remaining filled these bottles with dilution media, placed the caps and excess should not be discard in the
BOD determination.
Now transfer B1, B2, B3, C1, C2, C3 and Blank 2 in the incubator at 20 ±2°C for seven days.
STEP 3: DISSOLVED OXYGEN (DO) LEVEL AT ZERO DAY:
Perform DO experiment on these remaining BOD bottles A 1, A2, A3, and Blank 1 and repeat all the
procedures. At the end, DO level is calculated for all of them.
STEP 4: DISSOLVED OXYGEN (DO) LEVEL AT 7th DAY:
Same DO procedure was applied for the remaining BOD bottles that kept in incubator for seven days.
1
DO Level at 7th Day α
Consumption of oxygen by bacterial activity
Difference in DO level at zero and after seven days will indicate bacterial activity for the consumption of
oxygen in water samples.
STANDARDS FOR BOD:
Biochemical oxygen demand should have less than 1mg/L for drinking water. While acceptable treated waste water
should have less than 20mg/L BOD[14]. Allowable concentration for sewer discharge should be around 300mg/L

WHY WE MEASURE BOD?

The inorganic or organic matter decay in water is termed as biochemical or chemical demand of oxygen, while
oxygen demand is the quantity of substances that is oxidized in water and can decline actual DO level of
water.
It is significant parameter of water quality as it provides an indication to investigate the harmful effects of wastewater
discharged on receiving environment. The higher BOD, there is a greatest amount of organic content or food for
bacteria that consume oxygen. If consumption of DO become greater than DO supply, it make unhealthy environment
for sustaining aquatic biota. Further, DO depletion leads toward hypoxia condition and eventually quality of water
becomes poor and unfit for drinking purpose.

Existence of fecal coliform is indicated by High BOD level water can indicate presence of fecal coliform in
wastewater. This type of contamination can cause serious health issues and problematic at industrial level. So,
some authorities controlled water qualities to protect public from health damage and adverse impacts of poor
quality of water.
It is necessary to check water quality before consumption to prevent the risk of possible adverse effects on
heath and other sensitive species such as children.