Preparation of Henna Leaf Extract
Six hundred grams of Malaysian henna leaves were collected from a single tree in Hospital Universiti Sains Malaysia (USM) in the district of Kubang Kerian. The henna leaves were collected fresh and healthy from the henna treetop without any fungal or bacterial contamination evidence. The leaves were green in colour; neither brown nor yellow were used to avoid contamination.
The henna preparation was conducted at the Pharmacology Department, School of Medical Sciences, USM. The leaves were cleaned in distilled water and dried at 50 degrees Celsius for two days until brittle. The dried leaves were weighed on a scale balance before being pulverized into a fine powder and stored in a tightly sealed glass jar.
Extraction by using a Soxhlet machine was used to prepare the sample. There were two types of solvents used: aqueous and 70% ethanol. After extensive freeze-drying, 12 grams of ethanol henna extract and 15 grams of aqueous henna extract were produced. These powdered extracts were utilized to determine the exact concentration of Henna required for this research, which is 25% aqueous extract and 25% ethanol extract (Figure 1).