Nat Prod Sci.  2016 Jun;22(2):77-81. 10.20307/nps.2016.22.2.77.

Simultaneous HPLC Analysis of Three Flavonoids in the Extracts of Artocarpus heterophyllus Heartwoods

• Septama AW
• Panichayupakaranant P

• KMID: 2328870
• DOI: http://doi.org/10.20307/nps.2016.22.2.77

Abstract

A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of three antibacterial flavonoids, artocarpanone, artocarpin, and cycloartocarpin in ethyl acetate extracts from Artocarpus heterophyllus heartwoods. Separation was achieved using a TSK-gel ODS-80Tm column (5 µm, 4.6 × 150 mm) at 25℃ with a gradient elution system of methanol and water as follows: 0-8 min, 60:40; 8-27 min, 80:20; 27-35 min, 60:40, v/v, at a flow rate of 1 mL/min, and a quantitative UV detection at 285 nm. The method was validated by measuring the key parameters, including specificity, linearity, sensitivity, accuracy, repeatability and reproducibility. A high degree of specificity and sensitivity was achieved. The calibration curves for all three flavonoids showed good linearity with a coefficient of determinations (R²) of ≥ 0.9995. The recoveries of the method were from 98-104%, with good reproducibility and repeatability (RSD values of less than 2%) were also achieved. Ethyl acetate was the best solvent for extraction of these three flavonoids using the heat reflux conditions for 1 h. This optimized sample preparation and HPLC method can be practically used for a routine standardization process of the extracts from the A. heterophyllus heartwoods.

Keyword

Artocarpus heterophyllus; Artocarpanone; Artocarpin; Cycloartocarpin; HPLC

MeSH Terms

Artocarpus*
Calibration
Chromatography, High Pressure Liquid*
Flavonoids*
Hot Temperature
Methanol
Methods
Sensitivity and Specificity
Water
Flavonoids
Methanol
Water

Figure

• 

  Fig. 1. Chemical structures of artocarpanone (1), artocarpin (2), and cycloartocarpin (3).

• 

  Fig. 2. HPLC chromatograms of (A) authentic compounds an (B) heartwood extract of A. heterophyllus. (1: Artocarpanone, 2 Artocarpin, 3: Cycloartocarpin).

• 

  Fig. 3. HPLC chromatograms of hexane, chloroform, ethyl acetate, and methanol extracts from of A. heterophyllus heartwoods.

Reference

(1). Salguero C.P.; A Thai herbal traditional recipes for health and harmony; Findhorn press; Scotland. 2003; 119.

(2). Arung E. T., Yoshikawa K., Shimizu K., Kondo R.Fitoterapia. 2010; 81:120–123.

(3). Sato M., Fujiwara S., Tsuchiya H., Fujii T., Iinuma M., Tosa H., Ohkawa Y. J.Ethnopharmacol. 1996; 54:171–176.

(4). Ko F. N., Cheng Z. J., Lin C. N., Teng C. M.Free Radic. Biol. Med. 1998; 25:160–168.

(5). Zheng Z. P., Chen S., Wang S., Cheng K. W., Wu J. J. J.Agric. Food Chem. 2009; 57:6649–6655.

(6). Septama A. W., Panichayupakaranant P.Pharm. Biol. 2015; 53:1608–1613.

(7). Wei B. L., Weng J. R., Chiu P. H., Hung C. F., Wang J. P., Lin C. N. J.Agric. Food Chem. 2005; 53:3867–3871.

(8). Dej-Adisai S., Meechai I., Puripattanavong J., Kummee S.Arch. Pharm. Res. 2014; 37:473–483.

(9). Arung E. T., Wicaksono B. D., Handoko Y. A., Kusuma I. W., Shimizu K., Yulia D., Sandra F. J.Nat. Med. 2010; 62:423–429.

(10). Kirchmair J., Rollinger J. M., Liedl K. R., Seidel N., Krumbholz A., Schmidtke M.Future Med. Chem. 2011; 3:437–450.

(11). Chung M. I., Weng J. R., Wang J. P., Teng C. M., Lin C. N.Planta Med. 2002; 68:25–29.

(12). Shabir G. A. J.Chromatogr. A. 2003; 987:57–66.

(13). ICH. Guideline Q2(R1)-Validation of Analytical Procedure: Text and methodology. ICH;Geneva: 2005. p. 1–13.

(14). Zheng Z. P., Chen S., Wang S., Wang X. C., Cheng K. W., Wu J. J., Yang D., Wang M. J.Agric. Food Chem. 2009; 57:6649–6655.