QUALITY CONTROL OF

NEUTRACEUTICALS

MONIKA SUHAG (251503020)

INTRODUCTION

 Algae are a diverse group of aquatic organisms

that have the ability to conduct photosynthesis.
Certain algae are familiar to most people; for
instance, seaweeds (such as kelp or phytoplankton),
pond scum or the algal blooms in lakes. However,
there exists a vast and varied world of algae that
are not only helpful to us, but are critical to our
existence.
GENERAL CHARACTERISTICS

 HABITAT
• Generally aquatic
• Can survive on land
• Mostly live independently in different forms
• Can also be in symbiotic relation- ciliates, sponges, molluscs ,
fungi as lichens
 NUTRITION
• Generally photosynthetic
• Can be heterptrophic (osmotrophic , phagotropic , auxotropic)
 CLASSIFICATION
• Cyanobacteria (prokaryotic)
• Eukaryotic algae
 IMPORTANCE
• Neutraceuticals
• Biofuels
• Sewage treatment
• Industry
• Fertilisers
• Food
CHLORELLA
 Chlorella is a type of green algae that grows in fresh water.
 It posseses chlorella growth factor responsible for high
reproduction rate.
 Chlorella is a good source of proteins , fats, carbohydrates,
fiber, chlorophyll, vitamins and minerals.
 Can also act as antioxidant.
 Researches have shown its benefits for reducing cholestrol and
blood sugar levels , enhance immune system, help managing
respiratory diseases .
SPIRULINA
 It is a blue green algae used as nutritional supplement .
 it contains many nutrients including B vitamins, iron, calcium,
magnesium, manganese, potassium, zinc and protein.
 Spirulina is 65% protein by its compostion in which they
contain all essential amino acids and some non essential ones.
 It is helpful in treatment of oral leukoplakia(a recancerous
condition), promotes weight loss , alleviate symptoms of
sinusitis and asthama.
 Also used as an energy booster and memory booster as well .
MEDIA PREPAPRATION
 BG11 and ZARROUCKS MEDIA are specific for growth of
chlorella and spirulina respectively .
BG11 (FOR CHLORELLA)
SLNO. QUANTITY PER LITRE
NUTRIENT

1.
NaNO3 1.5 g

2.
K2HPO4 0.04 g

3.
MgSO4·7H2O 0.075 g

4.
CaCl2·2H2O 0.036 g

5.
Citric acid 0.006 g

6.
Ferric ammonium citrate 0.006 g

7.
EDTA (disodium salt) 0.001 g

8.
Na2CO3 0.02 g

9.
Trace metal 1.0 ml

10.
Agar (if needed) 10.0 g
TRACE METALS
SLNO NUTRIENT QUANTITY PER LITRE

1.
H3BO3 2.86 g

2.
MnCl2·4H2O 1.81 g

3.
ZnSO4·7H2O 0.222 g

4.
NaMoO4·2H2O 0.39 g

5.
CuSO4·5H2O 0.079 g

6.
Co(NO3)2·6H2O 49.4 mg

Stocks can be prepared for trace metals .

Stocks prepared by us
 ZARROUCKS(FOR SPIRULINA )
SLNO. NUTRIENT QUANTITY PER LITRE

1. NAHCO3 (Sodium bicarbonate) 16.8g

2. K2HPO4 (Dipotassium hydrogen phosphate) 0.5g

3. NaNO3 (Sodium nitrate) 2.5g

4. K2SO4 (Potassium Sulfate) 1.0g

5. MgSO4 . 7 H2O (Magnesium sulfate) 0.20g

6. CaCl2 (calcium chloride) 0.04g

7. FeSO4 . 7 H2O (Ferrous sulfate) 0.01g

8. EDTA (ethylene diamino tetracetic acid) 0.08

9. Solution A 1 ml
 Solution A
SLNO. Solution A Quantity per liter of water

1. H3BO3 (Boric acid) 2.86g

2. MnCl2 . 4 H2O 1.81g

3. ZnSO4 (Zinc sulfate) 0.22g

4. CuSO4 (Copper Sulfate) 0.08g

5. MoO3 (Molybdenum oxide) 0.01g

SKINNERS TECHNIQUE
 Skinners technique is used to obtain isolated colonies of algae.
 Isolation of algae was carried out from wastewater samples
collected from a facultative pond of wastewater stabilisation
system located at Sanghol Sukhna Lake , Sector-1,
Chandigarh, India.
 Make dilutions and prepare media (BG11+agar), then let it
solidify and incubate .
 Colonies are observed. Then the solidified media is cut and the
colonies are tranferred in liq. BG11 for further growth.
MORPHOLOGICAL IDENTIFICATION

 CHLORELLA
 SPIRULINA
 CLAMYDOMONAS
GROWTH CURVE
 A growth curve is a graphical representation of how a particular
quantity increases over time. Growth curves are used in statistics
to determine the type of growth pattern of the quantity - be it
linear, exponential or cubic.

STANDARD CUREVE FOR ALGAE

INDOOR

SL NO. DAY OD AVG. OD

S1 S2 S3

1. 0 (11/06/18) 0.01 0.004 0.001 0.005

2. 2 (13/06/18) 0.012 0.004 0.007 0.007

3. 4 (15/06/18) 0.013 0.003 0.013 0.009

4. 6 (17/06/18) 0.016 0.008 0.013 0.012

5. 8 (19/06/18) 0.017 0.018 0.022 0.019

6. 10 (21/06/18) 0.032 0.029 0.035 0.032

7. 12 (23/06/18) 0.034 0.33 0.037 0.137

8. 14 (25/06/18) 0.051 0.062 0.045 0.052

9. 16 (27/06/18) 0.066 0.066 0.089 0.073

10. 18 (29/06/18) 0.065 0.073 0.063 0.067

GRAPH
0.16

0.14

0.12

0.1
OPTICAL DENSITY

0.08

0.06

0.04

0.02

0
0 2 4 6 8 10 12 14 16 18 20
DAY
OUTDOOR

SL NO. DAY OD AVG. OD

S1 S2 S3
1. 0 (11/06/18) 0.015 0.01 0.02 0.015
2. 2 (13/06/18) 0.018 0.011 0.007 0.012
3. 4 (15/06/18) 0.020 0.014 0.011 0.015
4. 6 (17/06/18) 0.027 0.025 0.018 0.023
5. 8 (19/06/18) 0.080 0.057 0.041 0.059
6. 10 (21/06/18) 0.073 0.066 0.057 0.065
7. 12 (23/06/18) 0.099 0.083 0.050 0.077
8. 14 (25/06/18) 0.065 0.091 0.054 0.07
9. 16 (27/06/18) 0.155 0.153 0.090 0.132
10. 18 (29/06/18) 0.180 0.157 0.051 0.129
GRAPH
0.16

0.14

0.12

0.1
OPTICAL DENSITY

0.08

0.06

0.04

0.02

0
0 2 4 6 8 10 12 14 16 18 20
DAY
MASS CULTURE PREPARATION
 Algal cells divide and double in 4hrs. For mass production of
algae mass culture can be prepared in 10L of media .
 PROCEDURE:
• Prepare 10L media in a container
• Inoculate 100ml algal strain in media.
• Place under desirable conditions.
• Maintain ph between 7-7.5 for optimum growth.
MASS CULTURE
DEWATERING ALGAE
 Microalgae are being developed as a source of fuels and/or
chemicals. A processing challenge is dewatering the algae.
Electrical approaches to dewatering include exploiting
electrophoresis or electroflocculation. The reported experiments
show that electrophoresis does occur but is complicated by the
effects of the fluid motion. It appears that the coupling of the
algal cell and the fluid can be sufficiently strong such that fluid
motion effects can influence or dominate behavior.
Electroflocculation appears to be a robust process. It does,
however, inherently leave electrically induced trace metal
flocculants in the dewatered algae.
 Microalgae do not naturally flocculate or coagulate due to their
net negative charge. Algal growth medial also contains only
trace quantities of the divalent and polyvalent cations required.
The accepted mechanism by which flocculation occurs is that
electrochemical reactions at the electrodes create positive,
polyvalent ions. These positive ions attract the negatively
charged algal cells. The algal cells and polyvalent ions create a
growing network of a charged neutral system. The networks can
extend to other neutral systems and continue to grow into larger
assemblies of flocculated algae.
ELECTRICALLY DEWATERING ALGAE
 Quality Control
 Quality control (QC) is a procedure or set of procedures intended to
ensure that a manufactured product or performed services adheres to
a defined set of quality criteria or meets the requirements of the
clients or customers. QC is similar but not identical to QA
(Quality Assurance).
 Quality Assurance is the total process whereby the quality of
laboratory reports can be guaranteed. The term quality control covers
a part of quality assurance, which primarily concerns the control of
errors in the performance of test and verification of test results. All
materials, equipments and procedures must be adequately controlled.
Culture media must be tested for sterily and performance. Each
laboratory must have standard operating protocols.
CAUSES TO USE QUALITY CONTROL
TESTS
 Quality control is an essential operation in the pharmaceutical
industry. Drugs must be marked as safe and therapeutically
active formulations whose performance is consistent and
predictable. New and better medicinal agents are being produced
at an accelerated rate. At the same time, more exacting and
sophisticated analytical methods are being developed for their
evolution. Contamination is also the reason for quality control.
Drugs must be contamination free. If there is contamination of
pathogenic microorganism, it can cause a disease instead of
curing it.
WEIGHT VARIATION TEST

 Weight variation test is carried out to find the deviation in

weight of a given tablet
 PROCEDURE:
• Weigh 10 tablets
• Note down the weight of each tablet.
• Find the average weight.
• Calculate the deviation of each tablet.
• Find the average deviation of tablets.
 OBSERVATIONS
SL.NO TABLET WEIGHT (in g)
1. T1 0.71
2. T2 0.74
3. T3 0.69
4. T4 0.72
5. T5 0.71
6. T6 0.71
7. T7 0.72
8. T8 0.74
9. T9 0.72
10. T10 0.68
Total weight = 7.14

Average weight= 0.714g

SL.NO TABLET VARIATION
1. T1 0.004
2. T2 0.03
3. T3 0.20
4. T4 0.006
5. T5 -0.004
6. T6 -0.004
7. T7 0.006
8. T8 0.03
9. T9 0.006
10. T10 0.034
= -0.168/10
= -0.0168 g

RESULT:
Each tablet on an average show a deviation of 0.02 g approximately.
DISINTEGRATION TIME
 Disintergration of tablets is the time required for a dosage form to
breakup into granules of specified size under carefully specified
conditions. It is a process whereby the oral dosage form falls apart
or disintegrates into smaller aggregates. This test determines whether
the dosage form such as tablet , capsules disintegrate within the
prescribed time when placed in a liquid medium. Disintegration is
defined as the state in which no residue of the unit remains. Time of
disintegration is a measure of quality, this is because if the
disintegration time is too high; it means that the tablets are too
highly compressed and also if the disintegration time is not uniform
in set of samples being analysed, it indicates batch inconsistency and
lack of batch uniformity.
 Start time: 11:03am
Stop time: 12:14 pm
Time required: 1hr 11min 28 sec
 HEAVY METALS BY ICP-MS
 Inductively coupled plasma mass spectrometry (ICP-MS) is a type of mass spectrometry which
is capable of detecting metals and several non-metals at concentrations as low as one part in 1015
(part per quadrillion, ppq) on non-interfered low-background isotopes. This is achieved by
ionizing the sample with inductively coupled plasma and then using a mass spectrometer to
separate and quantify those ions.
 Compared to atomic absorption spectroscopy, ICP-MS has greater speed, precision, and
sensitivity.
 For detection of heavy metals, inductively coupled plasma mass spectrophotometry can also be
used. This being expensive are tested in labs. We went to PUNJAB BIOTECHNOLOGY
INCUBATOR for lead testing in the water sample as the metal concentration can be controlled in
the initial stages of the process.
Testing sample: water
Testing parameter: lead
Microbial tests

 The microbial limit tests are designed to perform the qualitative and
quantitative estimations of specific viable microorganisms present in
samples. It includes testing total viable count (bacteria and fungi).
The tests should be conducted for samples prepared by mixing
multiple portions of randomly chosen from individual ingredients or
products.
 PROCEDURE:
• Take algal biomass and convert it to powdered form
• Mix 1g of the biomass to 100ml of water.
• This mixture is spread on different media for detection of
contamination.
Total bacterial count

 Nutrient agar medium is used to check the bacterial

contamination.

RESULT:
BACTERIAL CONTAMINATION WAS FOUND.
Total fungal count

 Potato dextrose agar is recommended for the isolation and

enumeration of yeasts and moulds.

RESULT:
FUNGAL CONTAMINATION WAS FOUND.
 Detection of pathogenic organisms

 Pseudomonas aeruginosa
Cetrimide agar is a selective medium used for isolation of Pseudomonas aeruginosa.

RESULT:
COLONIES OF Pseudomonas aeruginosa WERE OBSERVED IN THE SAMPLE.
  Salmonella typhi
Xylose lysine deoxycholate (XLD) agar is a selective media recommended for isolation
and enumeration of Salmonella typhi.

RESULT:
VERY FEW OR NEGLIGIBLE COLONIES OF Salmonella typhi
WEREOBSERVED IN THE SAMPLE.
  Staphylococci
Mannitol salt agar is used as a selective media for isolation of pathogenic Staphylococci.
Mannitol is a fermentable carbohydrate, fermentation of which leads to acid production,
detected by phenol red indicator. S.aereus ferment and produce yellow coloured
colonies surrounded by yellow zones.

RESULT:
COLONIES OF Staphylococci WERE OBSERVED
IN THE SAMPLE.
Algal pasteurization
 Pasteurization is a process, named after scientist Louis Pasteur, that applies
heat to destroy pathogens. Once pathogens are detected in the biomass of algae
before the production of tablets, the biomass cannot be unused. Hence, the
biomass is pasteurized to kill the pathogens at 1210C and 15psi.
 PROCEDURE:
• Place the algal biomass in a container.
• Place it in hot air oven.
• After every 1 hour including 0th hour, take 1g of the sample and store it.
• Prepare nutrient agar media and pour it in petri plates.
• Add 1ml water to each 1g of sample and then spread it on plates.
• Observe the growth on the plates.
RESULT:
THE BACTERIAL COLONIES DECREASED IN A SIGNIFICANT AMOUNT IN 5HRS
ONLY.
Labelling of the product
 Product Labeling is a key feature in marketing. It helps to market
the product allowing customers to know about the item and give
necessary messages including ingredients, instructions, and uses.
 Product labeling can be done in a variety of sizes, materials, and
shapes. It plays a key role as a point of sale display in the market
shelves. They can also communicate information about how to handle
a product or how to dispose of it. You can use the labeling for
security reasons so that a product should not be misused. It is for
these purposes the labeling having the logo or the trademark of the
company. All these are different types of uses of the label for a
product in the world of business.
Thankyou..!