This slides explains about inulin, and different extraction, qualitative analysis of inulin. This i prepared for my research purpose. Hope you will find it useful.

1. Inulin

2. Inulins are a group of naturally occurring water-soluble prebiotic fiber
or polysaccharides produced by many types of plants like onions, leeks,
garlic, asparagus, Jerusalem artichoke and many other foods.
These are made up of chains of fructose molecules that are linked
together in a way that cannot be digested by our small intestine.

3. Chemical structure of Inulin
Inulin is a linear polymer of D-fructose joined
by β (2→ 1) bond and terminated generally
with a D-glucose molecule linked to fructose
by an α (1 → 2) bond, as In sucrose.
Formula : C6nH10n+2O5n+1

4. How it works
Inulin is not digested or absorbed in the stomach. It travels to the lower gut,
where it functions as a prebiotic, or food source for the beneficial bacteria
that live there.
These beneficial bacteria in turn assist with digestion and absorption of our
food and play a significant role in our immune function.
In our gut, inulin is converted into short-chain fatty acids (SCFAs) that are
then converted to healthy ketones that feed our tissues. SCFAs may also
nourish colon cells and produce more appetite-controlling hormones in our
body.

5. Advantages
• It keeps us full (of fiber).
• slows digestion
• increases fullness
• removes cholesterol as it passes through the digestive tract
• It promotes digestive health.
• fend off unwanted pathogens (bad bacteria)
• prevent infection
• stimulate your immune system

6. Advantages
• It controls blood sugar.
• Inulin slows digestion, including the digestion of carbohydrates. This allows
sugar to be released slowly without spiking, which promotes healthy blood
sugar levels.
• It could potentially lower our cancer risk.
• As an immune system booster, it may also be a good preventative supplement
against cancers of the digestive system.

7. Advantages
• May Help Treat Inflammatory Bowel Disease
• Inulin may also have benefits against inflammatory bowel diseases, including
ulcerative colitis and Crohn’s disease.
• May Improve Mineral Absorption and Bone Health
• A few studies show that inulin may increase absorption of calcium and
magnesium, and improve bone mineralization in children.
• May Support Heart Health
• Several studies indicate that inulin may improve certain risk factors for heart
disease, but the evidence is mixed.

8. Advantages
• Helps Control Diabetes
• High-performance (HP) inulin has been shown to reduce blood sugar levels in
people with diabetes,
• Promotes Weight Loss
• Inulin and oligofructose supplements may help regulate appetite in adults,
leading to weight loss.
• Relieves Constipation
• For many people, inulin can help relieve constipation by causing more
frequent bowel movements and better stool consistency.

9. Manufactured Sources of Inulin
Inulin is found naturally in several foods. It’s also modified for
commercial use, and there are several varieties.
• Native : Extracted from Natural sources.
• Oligofructose: Made by removing the longer molecules from inulin.
• HP: High-performance (HP) inulin is created by removing the shorter
molecules from it.
• FOS: Fructooligosaccharides (FOS) consist of short inulin molecules
synthesized from table sugar.

10. Natural Sources of Inulin
• Although many plants contain only small amounts of inulin, others are excellent
sources. Here’s how much inulin is in 3.5 oz, or 100 grams, of the following foods:
• Asparagus: 2-3 grams.
• Chicory root: 36-48 grams.
• Garlic: 9–16 grams.
• Jerusalem artichoke: 16–20 grams.
• Jicama: 10–13 grams.
• Onions: 1–8 grams.
• Yacon root: 7–8 grams.

11. Extraction method
• Hot water extraction (Conventional extraction)
• Alcoholic extraction
• Ultrasonic extraction
• Accelerated solvent extraction
• Microwave Assisted Extraction
• Supercritical Carbon Dioxide extraction

12. Hot water extraction
• Fresh Sample > Blanching (60 ◦c 15 min)
• Drying in Oven Cabinet (50 ◦c, overnight )
• Grinding and sieving (0.125 mm)
• Extraction using water in hot water bath (1:1 ratio, 90-120 min, ± 80 ◦c )
• Filtration (Paper filter)
• Centrifugation (4500 rpm, 10 min) or Evaporated (to get ±30 brix)
• Freezing for further use

13. Fresh Sample
Blanching
(60c, 15 min)
Drying
(50 ◦c, overnight )
GrindingGrindingExtraction
Filtration Centrifugation Freezing
Hot water extraction Flow chart

14. Alcoholic extraction
• Fresh Sample > Blanching (60 ◦c 15 min)
• Drying in Oven Cabinet (50 ◦c, overnight )
• Grinding and sieving (0.125 mm)
• Extraction using ethanol in hot water bath (ratio varied, 90-120 min, ± 80 ◦c )
• Filtration (Paper filter)
• Centrifugation (4500 rpm, 10 min) or Evaporated (to get ±30 brix)
• Freezing for further use

15. Accelerated solvent extraction
• Fresh Sample > Blanching (60 ◦c 15 min)
• Drying in Oven Cabinet (50 ◦c, overnight )
• Grinding and sieving (0.125 mm)
• Extraction cell (20 min, 80 ◦c, 1500 psi)
• Filtration (Paper filter)
• Centrifugation (4500 rpm, 10 min) or Evaporated (to get ±30 brix)
• Freezing for further use

16. Determination of inulin content
• Phenolsulphoric acid method of Dubois et al method of Dubois et al.
using inulin (Raftiline GR) as standard)
• Reducing sugar determeniantion (dinitrosalicylic acid method using
D()-Fructose (Mw = 180.16, Fluka) as standard)
• Inulin extraction yield (%) = (total carbohydrate content − reducing
sugars content) × volume of extraction liquid(after extraction)/mass
of sample used) × 100.

17. Physical analysis
• Determination of pH value : pH meter
• Appearance, flavor, taste and sweetness
• Solubility: Inulin solubilities was determined (Leite, 2004) by using 5
gm sample per 100 ml of water at temperature about 25 ºC.
• Determination of color: Colorimeter
• Determination of Viscosity: using the Brookfield rotational
viscometer, using No.4 spindle at 20 rpm.
• Scanning Electron Microscopy (SEM)
• Fourier Transform Infrared Spectrometry (FT-IR)

18. Scanning Electron Microscopy (SEM)
A scanning electron microscope (SEM) is a type of electron microscope
that produces images of a sample by scanning it with a focused beam
of electrons.
In addition to topographical, morphological and compositional
information, a Scanning Electron Microscope can detect and analyze
surface fractures, provide information in microstructures, examine
surface contaminations, reveal spatial variations in chemical
compositions, provide qualitative chemical analyses and identify
crystalline structures.

20. Fourier Transform Infrared Spectrometry (FT-IR)
FT-IR stands for Fourier Transform InfraRed, the preferred method of
infrared spectroscopy. In infrared spectroscopy, IR radiation is passed
through a sample. Some of the infrared radiation is absorbed by the
sample and some of it is passed through (transmitted). The resulting
spectrum represents the molecular absorption and transmission,
creating a molecular fingerprint of the sample.
Like a fingerprint, no two unique molecular structures produce the
same infrared spectrum. This makes infrared spectroscopy useful for
several types of analysis.

21. FT-IR provide:
• It can identify unknown materials
• It can determine the quality or consistency of a sample
• It can determine the amount of components in a mixture

22. Chemical analysis
• Moisture, protein, fat, ash and fiber were determined according to
the methods described in the AOAC
• Total carbohydrates: were determined by the phenolsulphoric acid
method of Dubois et al., (1956).
• Reducing sugars: were estimated by 3, 5- dinitrosalicylic acid (DNS)
method using D-fructose (Mw= 180.16) as standard (Miller, 1959).

23. Total carbohydrate determination
(phenolsulphoric acid method)
• An aqueous solution of carbohydrate(s) is transferred using a pipette
into a small tube.
• A blank of water also is prepared.
• An aqueous solution of phenol is added, and the contents are mixed.
• Concentrated sulfuric acid is added rapidly to the tube so that the
stream produces good mixing. The tube is agitated.
• A yellow-orange color results.
• Absorbance is measured at 490 nm.
• The average absorbance of the blanks is subtracted, and the amount
of sugar is determined by reference to a standard curve.

24. Carbohydrate Solution
Phenolsulphoric acid method Flow chart
Addition of Phenol & Mixing
Addition of sulfuric acid & mixing Results yellow-orange color
Measurement of absorbance at 450 nm
Amount of sugar calculated using Standard curve

25. Total Reducing Sugar
(Somogyi–Nelson Method alternative to 3, 5- dinitrosalicylic acid method)
• A solution of copper(II) sulfate and an alkaline buffer are added by
pipettes to a solution of reducing sugars(s) and a water blank.
• The resulting solution is heated in a boiling water bath.
• A reagent prepared by mixing solutions of acidic ammonium
molybdate and sodium arsenate is added.
• After mixing, dilution, and remixing, absorbance is measured at 520
nm.
• After subtracting the absorbance of the reagent blank, the A250 is
converted into glucose equivalents using a standard plot of
micrograms of glucose vs. absorbance.

26. Carbohydrate Solution
Somogyi–Nelson Method Flow chart
Copper(II) sulfate and an alkaline buffer are added
Solution heated in a boiling water bath
ammonium molybdate and sodium arsenate is added
After mixing, dilution, and remixing, absorbance is measured at 520 nm.
Amount of reducing sugar is calculated using Standard curve

27. Thank you