1. 1 G&H Meeting 4-5 th October, 2001 G&H Meeting 4-5 th October, 2001 Wellesbourne, UK Wellesbourne, UK WP 7 Pharmaceutics WP 7 Pharmaceutics Objectives / Deliverables Results Outlook 1

2. 2 Objectives / Deliverables Objectives / Deliverables Analytical method / dissolution method Analysis of European garlic market products Publication/Report Development of a novel dosage form 2

3. 3 Results Development Analytical Method Results Development Analytical Method HPLC method for all purposes ColumnHypurity Elite ® C18 (150 x 3 mm; 3µm) Pre-ColumnSupelguard ® LC-18, 20 mm Flow rate0,4 ml/min Temperature25 °C UV-Detektion208 nm Eluents A20 mM phosphate buffer pH 2.1 + 10 mM heptane sulfonic acid Eluents B acetonitrile/20 mM phosphate buffer pH 2.1 (50/50) (v/v) + 10 mM heptane sulfonic acid GradientTime [min]Eluent A [%]Eluent B [%] 0 100 0 5 70 30 25 46 54 26-28 0 100 3

4. 4 Results European Market Products Results European Market Products Content of Garlic Ingredients (% m/m) 4

5. 5 Results European Market Products Results European Market Products Content of Garlic Products II (% m/m) 5

6. 6 Results European Market Products Results European Market Products Stability – 18 months storage at ambient conditions 6

7. 7 Results Development Dissolution Method Results Development Dissolution Method Dissolution testing on garlic powder – profile of released compounds (accord. USP: apparatus 2 “paddle method”) Temperature:37 °C Rotation speed: 100 rpm Volume: 500 ml Amount Garlic powder:10 g Dissolution media:66 mM phosphate buffer (for pH 5-9 experiments) 66 mM citrate buffer (for pH 1-5 experiments) Media additives:starch, microcristalline cellulose, cysteine, argenine,tyrosine, glutamic acid, ferric sulfate 7

8. 8 Results HPLC Method “All Purpose” Application

9. 9 Results HPLC Method Results HPLC Method Water/MeOH extraction (enyzme inhibiting conditions) 0.1 0.3 0.5 0.7 0.9 024681012141618202224 Retention time [min] Absorption (AU) 1 Alliin 2 unknown (Allylcysteine) 3  -Glutamyl-S-allylcysteine 4  -Glutamyl-S-trans-1-propenylcysteine 5  -Glutamyl phenylalanine 1 2 3 4 5

10. 10 HPLC Method HPLC Method Water extraction (no enyzme inhibition)

11. 11 Release Profiles Release Profiles Alliin – as a function of pH [min]

12. 12 Release Profiles Release Profiles Allicin as a function of pH [min]

13. 13 Release Profiles Release Profiles Different compounds – without additives [min]