Purpose: To measure the rosmarinic acid content of eight commercial tinctures derived from fresh (n= 5) and dried (n=3) Melissa officinalis herb.
Methods: Rosmarinic acid and the internal standard (esculin) were purchased from Aldrich Chemical Co. The column used was a Luna C18, 5 um (150 x 4.6 mm I.D., Phenomenex) maintained at ambient room temperature. The HPLC system consisted of a Shimadzu SCL-6B controller, Shimadzu LC-6A pumps, Shimadzu SPD-6A UV single wavelength spectrophotometric detector set to 320 nm and Shimadzu SIL-6B autosampler. Gradient elution of the samples and standard were performed using ammonium formate (0.02 M; pH 6.25 at 27 oC; eluent A) and methanol (eluent B). The gradient elution initial conditions were 2% of eluent B with linear gradient to 60% at 30 min, followed by linear gradient to 90% of eluent B at 31 min, this proportion being maintained for 4 min. The column was then returned to the initial condition at 36 min and maintained until the end of the run at 43 min. The flow rate was 1 mL/min. The assay was validated for sensitivity, accuracy and reproducibility.
Results: The content of rosmarinic acid in commercial tinctures was significantly higher in the tinctures made from dried plant material (2.96 - 22.18 mg/mL) compared to fresh plant tinctures (</= 0.92 mg/mL).
Conclusion: These results have implications both for the manufacturers of commercial tinctures and also for herbal practitioners in the choice of tinctures for treating Herpes simplex infection.