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

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 γ-CD 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-γ-CD

The analysis of Propolis-γ-CD - 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. x150mm (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-γ-CD 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-γ-CD ③ - The determination of total polyphenol -

Principle
Total polyphenol contents were expressed as gallic acid equivalents.

method

1. 
2. mixing
3. add 10% Na2CO3 0.5ml
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-γ-CD 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 γ-CD, oil was adhered to the spatula and the container.
  → Because of dispersibility and improvement of pungency, γ-CD was the best of 3.
• Sensory test of Propolis-γ-CD
  → Propolis-γ-CD showed less pungent significantly than Propolis itself.
• Analysis of Propolis-γ-CD
  → The contents of Propolis-γ-CD was almost same of the theoritical values.
• The problem of the β-CD's safety for human consumption
  In USA and Europe, The Acceptable Daily Intake (ADI) of β-CD was limited to less than 5mg/kg/day in Joint Expert Committee on Food Additives (JECFA).
  But α-CD and γ-CD were not limited.

In this study, γ-CD showed the best performance for the powderization of Propolis extraction.
The industrial handling, dispersibility and pungency of Propolis extraction were improved by the γ-CD.

Further, we study the stability and bioavailability of Propolis-γ-CD, and smell.