Quantification of Andrographolide in Andrographis paniculata (Burm.f.) Nees, Myricetin in Syzygium cumini (L.) Skeels, and Brazilin in Caesalpinia sappan L. by HPLC Method

Abstract

Introduction: Andrographolide, myricetin, and brazilin are bioactive compounds from Andrographis paniculata, Syzygium cumini, and Caesalpinia sappan plants that have potential as medicinal ingredients. Objectives: To determine the levels of andrographolide in A. paniculata herb extract (APE), myricetin in S. cumini leaf extract (SCE), and brazilin in C. sappan wood extract (CSE) as marker compounds for extract quality control using the HPLC method. Methods: The separation was carried out on a reverse-phase C18 column (150 x 4.6 mm; 5 μm). The isocratic was prepared from methanol - water (50:50 v/v); 0.1% orthophosphoric acid - methanol (60:40 v/v); and 0,3% acetic acid - acetonitrile (85.5: 14.5 v/v) as mobile phase with flow rate 1 mL/min for andrographolide, myricetin, and brazilin determination, respectively and detection using UV detector at a wavelength of 254 nm, 369 nm, and 280 nm, respectively. Results: The linear regression for andrographolide was y = 14113x + 5948.8 (r2= 0.9994); myricetin was y = 87766x – 138895 (r2=0.9996); and brazilin was y = 18520x – 42668 (r2=0.9992). The andrographolide content in APE was found to be 14.4686 %. The myricetin content in SCE was found to be 0.3190 %. The brazilin content in CSE was found to be 2.1280 %. Conclusion: The described HPLC method was successfully used for the analysis of the APE, SCE, and CSE. This method can be used for the identification and quantification of andrographolide, myricetin, and brazilin in herbal raw materials or herbal products containing these three extracts. Key words: Andrographis paniculata, Caesalpinia sappan, HPLC, Marker compounds, Syzygium cumini, Quality control