Comparison of Main Components in Agrimonia pilosa Ledebour and Agrimonia coreana Nakai. by HPLC
Background : Agrimonia pilosa (A. pilosa) Ledebour has been registered in The Korean Herbal Pharmacopeia (KHP). In the recent study, A. Coreana showed antioxidant and anti-inflammatory effect. However, Studies of components in Agrimonia coreana (A. Coreana) Nakai was not much. So, we compared A. pilosa and A. coreana by High performance liquid chromatography (HPLC). we perfomed Thin layer chlromatography (TLC) including the anatomical characteristics by using microscope. Methods and Results : The anatomical characteristics of A. pilosa were similar to them of A. coreana. But, fascicular fivers of A. Coreana was broader than it of A. pilosa. TLC were performed to identify the Rutin and Apigenin-7-glucuronide compound. In the extract of Agrimoniae Herba, they were identified on the spot of Rf 0.2, 0.4 in Ethyl acetate - Formic acid – Water (8 : 1 : 1). The Rutin and Apigenin-7-glucuronide were analysed by HPLC/UV with Thermo Column (5 μm, 4.6 × 250 mm, C18), the column temperature at 40 ℃ and a diode-array detector (DAD) seted at 255 nm and 338 nm. The mobile phase was composed of water and acetonitrile containing 0.1% formic acid with the flow rate 1 mL/min. All compounds showed good linearity (R2>0.999) within test ranges. Agrimoniae herba was extrated by four kinds of extraction methods: MeOH, 50% MeOH, EtOH and water. The highest extraction rate occurred, when it was treated with 50% Methanol for refluxing extraction (60min). Content of Rutin was found to be 0.07±0.00 mg/g in A. pilosa and 0.02±0.00 mg/g in A. coreana. Content of Apigenin-7-glucuronide was found to be 0.12±0.00 mg/g in A. pilosa and 0.11±0.00 mg/g in A. coreana. Conclusion : The anatomical characteristics of A. pilosa were similar to them of A. coreana. Contents of Rutin and Apigenin-7-glucuronide in A. pilosa was higher than them in A. coreana slightly. But there were observed the similar patterns of Agrimonia pilosa Ledebour and Agrimonia coreana Nakai on the finger print anelysis.