Validated Reversed-Phase Ion-Interaction High-Performance Liquid Chromatography for Quantitation of Nitrate Content ofClausena anisata(Willd.) Hook. f. ex Benth. Leaves

Abstract

This work sought to validate the reversed-phase ion-interaction high-performance liquid chromatography for quantifying the nitrate content in the extract and raw material of Clausena anisata (Willd.) Hook. f. ex Benth. leaves. Three extraction methods (i.e., decoction, infusion, and ultrasound-assisted extraction) were investigated and compared. Furthermore, the effect of the solid-to-solvent ratio and defatting was also evaluated. The validation result showed that the high-performance liquid chromatographic method had a linear response (R2 = 0.9999) in the range of 1–50 μg/mL. The limit of detection and limit of quantitation were 0.25 μg/mL and 0.75 μg/mL, respectively. In addition, the method was specific, precise, and accurate. So the validated method was suitable for determination of the nitrate content in C. anisata leaves. Infusion of a nondefatted sample using a solid-to-solvent ratio of 1 : 10 gave the highest nitrate content in the raw material, 0.251 ± 0.003%. In case of a defatted sample, decoction provided the highest nitrate content, 0.309 ± 0.001%. Increasing the solid-to-solvent ratio and defatting had a huge effect on the nitrate content of C. anisata leaves extracted from decoction. To the best of our knowledge, this is the first report that used the reversed-phase ion-interaction high-performance liquid chromatography for quantifying the nitrate content in C. anisata leaves. Furthermore, the authors suggested that nitrate could be used as a standard marker for quality control of C. anisata leaves’ extract and raw material.

Optimization of dynamic maceration of Clausena anisata (Willd.) Hook. f. ex Benth. leaves to maximize trans-anethole content

“…Infusion of a non-defatted sample using a solvent-to-solid ratio of 10:1 provided the highest nitrate content of the raw material, while decoction of a defatted sample gave the highest nitrate content. Moreover, increasing the solvent-to-solid ratio and defatting of the decoction sample had a huge effect on the nitrate content [17]. Our other previous works optimized the dynamic maceration of C. asiatica.…”

“…is a herbal medicinal plant belonging to the Rutaceae or Citrus family. It has been investigated for several activities including analgesic [1], antidiabetic [2,3], anti-inflammatory [4,5], antimalarial [1], antimicrobial [3,[6][7][8][9], antioxidant [3,10,11], antipyretic [4], cytotoxic [6], larvicidal [12][13][14][15][16], smoking cessation aid [17], and wound healing [11] effects. The five major compounds containing in essential oils of C. anisata leaves growing in North-Central Nigeria are anethole (31.3%), trans-β-ocimene (20.0%), caryophyllene (10.5%), estragole (6.9%), and α-pinene (6.7%) [18].…”