Quantitative Determination of Chemical Constituents from Seeds of Nigella sativa L. by using HPLC-UV and Identification by LC-ESI-TOF

Abstract

Nigella sativa Linn. belongs to the Ranunculaceae family and is an indigenous herbaceous plant that is more commonly known as the fennel flower plant. The plant is also known as black cumin (English) and black-caraway (USA). The spicy seeds from this plant have medicinal usage dating back to the ancient Egyptians, Greeks and Romans. In Egypt and the Middle East the black seed oil is popularly used for certain cases of chronic cough and bronchial asthma [1,2]. An HPLC method was developed for the simultaneous determination of nine compounds of Nigella sativa L. The separation was achieved within 23 minutes by using C-18 column material, a water/acetonitrile mobile phase, both containing 0.1% acetic acid gradient system and a temperature of 35°C. The method was validated for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ). The LOD and LOQ of nine compounds were found to be in the range from 0.09–10 µg/mL and 0.3–25 µg/mL, respectively. The wavelength used for quantification with the diode array detector was 205 and 260 nm. The seeds of N. sativa and commercial products showed the presence of all nine compounds. LC-mass spectrometry coupled with electrospray ionization (ESI) interface method is described for the identification of compounds in Nigella sativa L. samples. This method involved the use of [M+H]+ and [M+Na]+ ions in the positive ion mode with extractive ion monitoring (EIM). Acknowledgements: This research is funded in part by “Science Based Authentication of Dietary Supplements” Funded by the Food and Drug Administration grant number 2 U01 FD 002071-07.