Simultaneous Determination of the Main Organic Acids in Anatolian Black Pine by HPLC with DAD Detector

Abstract

The versatile characterization of organic materials in Anatolian black pine (Pinus nigra)
especially the significant organic acids is great importance. A rapid, simple, and sensitive
HPLC–DAD method was developed and presented for a simultaneous detection of six
organic acids. This method was thoroughly validated to determine the most abundant
organic acids present in Anatolian black pine and their distribution in branches, leaves
and cones. Separation of six organic acids were developed and achieved on ODS C18
column (250mm×4.6mm, 5µm particle size), using a mixture of (A) water (pH: 2.0,
adjusted by H2SO4) and (B) acetonitrile in the ratio of (95:5, v/v) at wavelength of 210
nm with a flow rate of 0.8 mL min-1 within 14 minutes. The dynamic range was between
5 to 1000 mg L-1 with relative standard deviation less than 0.73%, (n=4). Limits of
detection and the recoveries for oxalic acid, tartaric acid, formic acid, lactic acid, acetic
acid and citric acid were ranged 1.85 mg L-1 (87.75-99.36%), 3.61 mg L-1 (100.89-
119.55%), 4.01 mg L-1 (83.57-119.64%), 1.16 mg L-1 (82.29-112.19%), 1.19 mg L-1
(80.86-110.42%) and 2.87 mg L-1 (96.96-117.74%), respectively.