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No.26 Powderization of Propolis Extract by the Use of Cyclodextrins

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Summary

Propolis is a natural hive product that exhibits a broad spectrum of biological activities such as antibacterial, antiviral, anti-inflammatory and tumor growth inhibition. However, the viscous characteristic of propolis gives its intractability for a daily intake in foods and beverages. Here we report on the fine powder formation from viscous propolis ethanol extract formulated with γCD for easy handling.

Problems of Propolis Extract and Study Points

●  PUNGENCY ●  SOLUBILITY and/or DISPERSIBILITY in WATER
●  SMELL ●  INDUSTRIAL HANDLING
●  POWDERIZATION ●  BIOAVAILABILITY
●  STABILITY    

Propolis Extract Vendor：Manuka Health

Preparation of Propolis-CD

The dispersibility test of Propolis-CD

The SENSORY TEST OF PROPOLIS with CDs

The SENSORY TEST of Propolis-γCD

The analysis of Propolis-γCD
- Total Propolis contents (p-Coumaric acid equivalents test) -

Definition
The total peak area of the HPLC chromatogram at 330nm from 0 to 30 minutes were expressed as total propolis contents.

HPLC analytical method
Column SunFire C18 5μm 4.6mm I.D. ×150mm (Waters Lot : 0130382611)
Mobile phase : methanol　70 / water 30 / acetic acid 1 (isocratic)
Flow rate : 1.2mL/min
detector : UV (330nm)

Japan Health Food & Nutrition Food Association's method

Analysis of PROPOLIS-γCD ②  - The determination of total flavonoid -

Principle
Total flavonoid contents were expressed as quercetin equivalents.

method
1
2 mixing
3 1h incubation at room temperature
4 The absorbance is measured at 420nm.

S. Kumazawa et al. / Food Chemistry 84 (2004) 329-339

ANALYSIS of PROPOLIS-γCD ③  - The determination of total polyphenol -

principle
Total polyphenol contents were expressed as gallic acid equivalents.

method
1
2 mixing
3
4 mixing
5 1h incubation at room temperature
6 centrifuge (3000rpm, 5min)
7 The absorbance of the supernatant is measured at 760nm.

S. Kumazawa et al. / Food Chemistry 84 (2004) 329-339

Analysis of PE-γCD content  HPLC chromatogram and MS spectrum

Analysis method
column ： CAPCELL PAK C18 ACR (SHISEIDO)
5μm(Median Particle Size)、2mm(Inner diameter)、250mm(Length)
Mobile phase ： phase A : 0.1% formic acid in water
phase B : 0.08% formic acid in acetonitrile
detector ： PDA and MS
interface ： ESI　SIM : negative　m/z 283
temperature ： 40℃
Flow rate ： 0.2 mL / min
program ：

MS spectrum at tR 37.5 min for CAPE

HPLC chromatogram

MS chromatogram of CAPE and quantitative determination.

Discussion

● Which CD was the best for the powderization of Propolis extraction?
→When it was prepared by γCD, oil was adhered to the spatula and the container.
→Because of dispersibility and improvement of pungency, γCD was the best of 3.
● Sensory test of Propolis-γCD
→Propolis-γCD showed less pungent significantly than Propolis itself.
● Analysis of Propolis-γCD
→The contents of Propolis-γCD was almost same of the theoritical values.
● The problem of the βCD's safety for human consumption
In USA and Europe, The Acceptable Daily Intake (ADI) of βCD was limited to less than 5mg/kg/day in Joint Expert Committee on Food Additives (JECFA).
But αCD and γCD were not limited.
In this study, γCD showed the best performance for the powderization of Propolis extraction.
The industrial handling, dispersibility and pungency of Propolis extraction were improved by the γCD.
Further, we study the stability and bioavailability of Propolis-γCD, and smell.
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