Recently, barley is gaining renewed interest for food due to nutritional and functional characteristics. and many breeder developed new barley varieties containing nutritional and functional components. Anthocyanin have been well characterized for its bioactive properties, anti-oxidant, anti-cancer, anti-proliferative and anti-inflammatory properties. Some colored varieties with purple lemma that is a kind of anthocyanin were developed. We have to find out the harvesting time containing maximum anthocyanin content. In this study, we analyzed anthocyanin content at intervals of 4-6days after heading date. We analyzed Delphinidin-3-glucoside(Del3g), Cyanidin-3-glucoside (Cya3g), Pelargonidin-3-glucoside(Pel3g), Peonidin(Peo3g), Malvidin-3-glucoside(Mal3g) using the HPLC with Xterra C18 column and UV Detector. Anthocyanin was extracted with 80% MeOH solution containing 1% HCl for 24hr. Mobil phase composition of HPLC was 45% Acetonitrile solution with 10% Formic acid. Anthocyanin content was increased rapidly from 30day after heading date to 35day. At 35-37day, anthocyanin content was maximum. Thereafter, it was decreased rapidly to 40day.