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Fabrication Of Conjugated Linolenic
acid (CLnA) Nanoemulsion
Colloidal Delivery System : Its
Mitigative Role Against Alloxan
Induced Oxidative Stressed Diabetes
Mellitus RatsDebjyoti Paul
University of Calcutta
About bitter gourd seed oil….• Bitter gourd (Momordica charantia) seed oil
(BGO) is a unique oil which contains 9cis, 11trans, 13trans-conjugated linolenic acid (9c,11t,13t-CLN) commonly known as α-eleostearic acid (ESA) at a high level of >50%.
• Fatty Acid Composition (w/w %) of Bitter Gourd Seed Oil as obtained by Gas-Liquid Chromatography:
C14:0 C16:0 C18:0 C18:1 C18:2 C18:3
BITTER GOURD
SEED OIL- 2.4 % 31.3 % 8.0 % 7.9 % 50.4 %
Conjugated linolenic acidConjugated linolenic acid (CLnA) refers to a group of octadecatrienoic acid isomers that have three double bonds in conjugation.
Attributed with numerous health benefits.
Antioxidative Anti-Cancer
HypoglycemicHypolipidemic
Cytotoxic Apoptosis•Downregulation
bcl-2 gene •Upregulation of bax gene
• Upregulation of Peroxisome proliferatorActivated receptor proteinPPARs
• Down regulation of ApoB100 • Increase in Serum TG • Increase fatty acid mobilisation• Enhanced activity of mitochondrial peroxisomal β-oxidation
• Scavenging action against lipid peroxidation• Genoprotective role against oxidative stress
• Lipid peroxidation of membrane lipids• Alteration in membrane fluidity
• Antiangiogenesis Down regulation of VEGF,IL-4, Up-regulates MIF
Conjugated linolenic acid
OBJECTIVES OF THE STUDY
To prepare CLnA from seeds of Momordica charantia
(bitter gourd)
Develop colloidal delivery system in the form of nanoemulsion of the CLnA rich bitter gourd oil (BGO) to
increase bioavailability in in vivo system
Study the effects of the formulated system on alloxan induced diabetic rats
Here’s why…..• Reports suggest that o/w nanoemulsions (d <
200 nm) confers a distinct advantage over other delivery systems due to:
enhanced long term stability of the particles in the delivery system;
high optical clarity;
increased bioavailability
Extraction & refining of Bitter gourd seed oil
1. C r u s h e d2. S o l v e n t ex t ra c ti o n u s i n g n - h exa n e3. M i s c e l l a r e fi n i n g ( 2 : 1 )
Bitter Gourd Seed
Refined BGO
BGO Nanoemulsion(20-200 nm)
Preparation of bitter gourd seed oilnanoemulsion
High pressure homogenization
BGO nanoemulsion characterization for droplet size after preparation.Result represented as DelsaTM Nano (Beckman Coulter, Inc., USA) software generated data.
BGO nanoemulsion characterization for droplet size after 12 weeks.Result represented as DelsaTM Nano (Beckman Coulter, Inc., USA) software generated
data.
Animal experiment
ALLOXAN INJECTION (i.p.)
TYPE II DIABETES INDUCED
DIABETIC RATS FORCE FED WITH THE FORMULATED BGO-NE & BGO-CE AT
DIFFERENT DOSES (1% & 0.5%)
AMELIORATION IN CONDITION OF DISEASED STATE DETERMINED ON
THE BASIS OF PHYSIOLOGICAL,
BIOCHEMICAL AND HISTOPATHOLOGICAL
PARAMETERSHEALTHY RAT
Effect on blood glucose
0 5 10 15 20 25 30
40
60
80
100
120
140
160
BLO
OD
GLU
CO
SE
LE
VE
L(m
g/d
L)
NUMBER OF DAYS
NORMAL CONTROL ALLOXAN 1 % BGO NE 0.5 % BGO NE 1 % BGO CE 0.5 % BGO CE
Lipid peroxidation studies of Plasma, Erythrocyte membrane & tissue lipids
GROUP FATDIET
PLASMA LIPID PEROXIDATION
(nmole of MDA/mL)
ERYTHROCYTE MEMBRANE LIPID
PEROXIDATION(nmole of MDA/mL)
LIVER TISSUE LIPID PEROXIDATION
(nmole of MDA/mL)
CONTROL(NON-DIABETIC) SO 10.71 ± 0.414# 6.64 ± 0.44# 1.62 ± 0.34#
DIABETICGROUPS
SO27.21 ± 0.35 15.23 ± 0.26 3.4 ± 0.62
SO+0.5 % ESA NE a 5.34 ± 0.25* 3.26 ± 0.17* 1.42 ± 0.81*
SO+0.25 % ESA NE b 4.8 ± 0.33* 2.83 ± 0.91* 1.28 ± 0.33*
SO+0.5% ESA CE c 5.2±0.32* 4.23±0.42* 1.73±0.63*
SO+0.25 % ESA CE d 6.12±0.27* 4.4±0.31* 2.5±0.24*
[MDA] COMPARISONAMONG DIET GROUPS
ALX(SO)>CON(SO)>0.25 % ESA CE>0.5 % ESA NE>0.5 % ESA CE>0.25% ESA NE
THUS,THE FORMULATED BGO NE: A POTENT NUTRACEUTICAL
AGAINST TYPE II DIABETES
THE FORMULATED BGO-NE
DIABETIC PATIENT PATIENT RECOVERS
THE ATTEMPT OF FABRICATING A STABLE BGO-NE COLLOIDAL DELIVERY SYSTEM SUCCESSFULLY
INCREASED THE EFFICACY OF ESA MULTITUDINALLY WHEN ADMINISTERD IN ALLOXAN INDUCED
OXIDATIVELY STRESSED DIABETES MELLITUS CHARLES FOSTER STRAIN RATS (IN VIVO SYSTEM)