Franz Cell Diffusion Study
HINNAO High-Stability Liposomal Glutathione
An independent absorption study conducted by CC Research (P) Ltd. using the Franz Cell Diffusion method with HPLC analysis, measuring the percentage of glutathione diffused through biological mucosal membrane under physiological conditions.
Methodology
The study used a vertical Franz-type diffusion cell with a biological sheep mucosal membrane, replicating the conditions of sublingual absorption in the human buccal cavity. Samples were collected at 10-second intervals from 0 to 90 seconds and analysed by High-Performance Liquid Chromatography (HPLC).
Materials
- PBS buffer solution (pH 7.4)
- Franz diffusion apparatus
- Sheep mucosal membrane
- HPLC instrument
- Syringe
HPLC Conditions
| Column | INERTSIL ODS C18 150×4.6mm |
| Wavelength | 215 nm |
| Flow rate | 0.8 ml/min |
| Injection vol. | 10 µl |
| Mobile phase | Acetonitrile:water (50:50) |
Cell Parameters
| Temperature | 37°C ± 1°C |
| Stirring | 50 rpm |
| Receptor vol. | 60 ml |
| Diffusion area | 10.18 cm² |
| Sample dose | 5 ml at 1 mg/ml |
Absorption Over Time
Percentage of glutathione diffused through biological mucosal membrane at each time point.
Flux and Permeation Data
Flux (J) measures the rate of glutathione permeation through the mucosal membrane per unit area. Higher flux values indicate faster and more efficient delivery into systemic circulation.
| Time (sec) | Dose (mg/ml) | Concentration | % Drug Release | Flux |
|---|---|---|---|---|
| 10 | 200 | 5038.06 | 25.95% | 66.99 |
| 20 | 200 | 5131.76 | 28.29% | 136.48 |
| 30 | 200 | 5225.46 | 30.64% | 208.46 |
| 40 | 200 | 6162.45 | 54.06% | 327.79 |
| 50 | 200 | 6264.40 | 56.61% | 416.52 |
| 60 | 200 | 7193.04 | 79.83% | 573.91 |
| 70 | 200 | 7202.40 | 80.06% | 670.44 |
| 80 | 200 | 7211.67 | 80.29% | 767.20 |
| 90 | 200 | 7305.28 | 82.63% | 874.30 |
HPLC Linearity Verification
Five concentration levels of L-glutathione were tested to confirm a linear relationship between concentration and HPLC peak area. A correlation coefficient at or above 0.99 is required for analytical validity.
What the data confirms
HINNAO High-Stability Liposomal Glutathione demonstrated rapid and high-efficiency permeation through biological mucosal membrane under physiological conditions. Absorption reached 79.82% at 60 seconds and 82.63% at 90 seconds, confirming that the liposomal delivery system enables near-immediate systemic availability of glutathione via the buccal mucosa.
This is clinically significant for glutathione specifically, as the molecule is a fragile tripeptide that is rapidly degraded by digestive enzymes in standard oral formats. The Franz Cell data confirms that HINNAO's sublingual liposomal technology protects the molecule and delivers it intact, bypassing hepatic first-pass metabolism entirely.
The HPLC linearity study returned a correlation coefficient of 0.998, confirming full analytical validity of the measurement method across all tested concentration levels.
Analysis conducted by: CC Research (P) Ltd. | 20 White Church Lane, London, E1 7QR
Commissioned by: HINNAO Technology
Method: Franz Cell Diffusion Study with HPLC Analysis | Membrane: Sheep mucosal membrane | Temperature: 37°C ± 1°C
Get in Touch
Have a question or need assistance? We'd love to hear from you.