This document discusses nutraceuticals and proximate analysis. It defines nutraceuticals as foods or food components that provide health or medical benefits, including nutrients like dietary fiber, probiotics, antioxidants, and spices. The document then describes various methods for proximate analysis, including determining the moisture, ash, fat, protein, carbohydrate, and crude fiber content. These standard methods involve processes like distillation to measure moisture, ignition in a furnace to measure ash, Soxhlet extraction for fat, and the Kjeldahl method for determining protein content.

1. NUTRACEUTICALS AND PROXIMATE
ANALYSIS IN BRIEF
SHAHNAZ MOL A

2. NUTRACEUTICALS
“ Let food be thy medicine and medicine be
thy food.”
­ Hippocrates
Hybrid of Nutrition and Pharmaceuticals.
Coined by Dr. Stephen DeFelice.
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Dr.Stephen
DeFelice

3. “Any substance that may be considered
a food or part of a food which
provides medical or health benefits
including the prevention and treatment
of diseases.”

4. CLASSIFICATION OF NUTRACEUTICALS
Based on food source :
Dietary Fibre
Probiotics
Prebiotics
Polyunsaturated Fatty Acid
Antioxidant Vitamins
Polyphenols
Spices

5. Birketvedt et al., 2005 found that dietary fibre
lowers serum LDL cholesterol and improves
glucose tolerance.
Leray et al., 2001 states that Omega ­­ 3 ­fatty
acids have three major effects in cardio ­
vascular disease :
Anti­arrhythmic.
Hypolipidemic
Antithrombotic

6. PROXIMATE ANALYSIS
“ Quantitative analysis which
determines nutritional and
biochemical point of view of foods.”
The methods recommended by the
Association of Official Analytical
Chemists (AOAC), 2000 were used for
the determination of Proximate
Analysis.

7. Moisture Ash
Crude
Fat
Crude
Fibre
Crude
Protein
Carbohydrate

8. DETERMINATION OF MOISTURE
Immiscible solvent distillation method.
Reflux distillation.
High boiling point solvent, Toluene (110.6°C).
Apparatus used is Bidwell sterling moisture
trap.

9. Bidwell sterling
moisture tube.
Round bottom flask
Condensor

11. PROCEDURE
10 gm of ground sample was taken in a
round bottom flask.
Toluene was added enough
to cover the sample.
Connect the round bottom flask to the
side arm of Bidwell­ sterling tube.
Bidwell sterling tube was filled with
solvent.
Heating mantle adjusted to 100°C for
1hr.
Note the volume of water distilled.

12. Moisture(
0
0
)=
volumeof water(ml)
weightof sample(g)
×100
CALCULATION

13. DETERMINATION OF ASH
Indicator of minerals
Ash
Ash refers to the inorganic residue remaining
after either ignition or complete oxidation of
organic matter in a foodstuff.
Dry ashing refers to the use of a muffle furnace
capable of maintaining temperatures of 500–
650 C.◦

14. PROCEDURE
1g of dried sample was accurately
weighed into pre­weighed,clean crucible.
The crucible was heated to the point of
charring of the sample on a hot plate.
The crucible with the carbon residue obtained
as a result of ignition, was placed in muffle
furnace at temperature of 650°C until the carbon
residue disappears.
The sample is allowed to cool in a dessicator and
then weighed.

15. Muffle furnace Crucible Dessicator

16. CALCULATION
TOTAL ASH CONTENT (%) =
W2­W X 100 X 100
W1­W 100 ­ M
Where; W = weight of empty crucible
W1 = (Weight of crucible+weight of sample)
W2 = (Weight of crucible + weight of ash)
M = % Moisture content (Dry basis)

17. CRUDE FAT ESTIMATION
●
Determined using Soxhlet extraction method.
●
Semi continuous solvent extraction, the
solvent builds up in the extraction chamber
for 5–10 min.
●
completely surrounds the sample and then
siphons back to the boiling flask.

18. Soxhlet extractor
Thimble
Condensor Round
bottom flask

19. Take 10g sample in a thimble and plug the top of
the thimble with a wad of fat ­free cotton.
Drop the thimble into the fat extraction tube of a
Soxhlet apparatus.
Attach the bottom of the extraction tube to a
Soxhlet flask.
Pour approximately 200ml or more of anhydrous
petroleum ether (boiling pointof 60°­ 80°C)
through the sample in the tube into the flask.
Attach the top of fat extraction tube to the
condenser. Extract the sample for 8hr or longer on
a heating mantle.
At the end of the extraction period, removed the
thimble and ether was evaporated from the
collecting round bottom flask by using a rotary
evaporator, which was maintained in a vacuum
of 200Pa at 49 C.

20. Soxhlet extraction mechanism
CALCULATION
Crude fat (%) =
weight of ether soluble material x 100
Weight of sample

21. DETERMINATION OF PROTEIN
Johan Kjeldahl
Nitrogen content was estimated by the Kjeldahl method
which was based on the determination of the amount
of reduced nitrogen present in the sample.
The various nitrogenous compounds were converted
into ammonium sulphate by boiling with concentrated
H2SO4.
The ammonium sulphate formed was decomposed with
an alkali (NaOH), and ammonia liberated was absorbed
in excess of neutral boric acid solution and then titrated
against standard Hcl acid.

22. Nitrogenous organic compound + H2SO4 (NH4)2SO4
(NH4)2SO4 + 2NaOH Na2SO4 + 2NH3 + 2H20
NH3 + H3BO3 NH4 + H2BO3
H2BO3 + H H3BO3

23. Weigh 0.1 to1g of sample was kept in KjelTRON
digestion tube, and to the tube.
PROCEDURE
Tubes were loaded into the digestion block along
with manifolds. Temperature was increased to 100 to
420°C gradually to avoid frothing.
DIGESTION
Tube was left in the digestion blocks for the complete
digestion.
10 ml of con. Sulphuric acid
3g of catalystic mixture (1:5,CuSO4+K2SO4)
was also added
Becomes bluish green then tubes were removed.

24. The liberated gas was collected to 25 ml of 4 % Boric
acid with mixed indicator of methyl red and bromocresol
green.
Distillation
It was then distilled with 40% NaOH which
convert the ammonium salt to ammonia.
The ammonia (the amount of nitrogen
present in the sample) get liberated.
TITRATION
After distillation ammonia collected was taken out
and treated with 0.1 N HCl
End point is Bluish green to pink

25. Kjeldahl equipment Digestion apparatus
Distillation apparatus

26. %of Nitrogen=
14xNormalityof HClxvolumeof Hcl∗100
Wtof Samplex1000
%Protein=%of Nitrogenx6.24
CALCULATION
Where 6.24 is power factor.

27. DETERMINATION OF CARBOHYDRATE
% of carbohydrate = 100 ­ [ Moisture + Ash + Fat
+ Protein ]

28. DETERMINATION OF CRUDE FIBRE
● Organic residue remains after the food sample
treated with petroleum ether, boiling dil. H2SO4
and dil. NaOH and alcohol.
● Crude fibre consists largely of cellulose.

29. PROCEDURE
12 g of fat free sample was taken in a round bottom
flask and add 200ml dil. H2SO4.
Connect with digestion apparatus and reflux for 30
min and filter through ashless filter paper.
Transfer the residue and add 200 ml of dil.NaoH
Connect with digestion apparatus and reflux for 30
min and filter through ashless filter paper.
Filtered residue were washed with 15ml ethanol,
15ml petroleum ether and hot water.

30. Transfer the residue into a pre­ weighed clean
crucible with ash less filter paper.
Incinerate the contents in the electric muffle
furnace at 600+20 °C for overnight.
After ashing, allowed to cool to room temperature
in a desiccator and weighed.
The difference between the weight of empty
crucible and with ash indicates the weight of the
crude fibre.

31. Calculation
Where ; W = Weight of Sample
W1 = Weight of fibre + Weight of Crucible
W2 = Weight of Crucible + Ash
%CrudeFibre=
(W1−W2)
W
x100

32. ESTIMATION OF DIETARY FIBRE