Materials & Methods:
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HPLC conditions:
Brand: Agilent 1100 HPLC Series
Column: 250mm x 4.6mm i.d; stationary phase, LiChrospher RP-18, 5μm
Mobile Phase: flow rate 1.5ml/min, 0.05 M acetic acid-methanol (95/5)
Refractive Index Detector (RID) at 290nm
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Twenty five henna samples were bought at 15 henna salons located at three different cities in the United Arab Emirates. Among the 25 henna samples, there were 11 black and 14 red henna.
Preparation of PPD standard (0.11mg/ml)
Preparation of PPD standard (0.11mg/ml)
Weigh 0.011g of pure PPD standard and dissolve it in 50% aqueous methanol solution.
Preparation of samples
Weigh 1.0g of collected henna samples and dilute it with 50% aqueous methanol solution to the brim in a 50ml volumetric flask. This solution will be filtered after 15 minutes. Transfer 1ml of the filtered solution and dilute it to 5ml with 50% aqueous methanol solution. The PPD solution can now be analysed.
Dilute 1ml of the
standard solution to 5ml with 50% aqueous methanol solution and analyse the
spectrum and retention time. This method is use to confirm the identity of PPD
in the samples.