High-Performance Thin Layer Chromatographic Methods to Identify the Marker Molecules From EstroG-100^®, a Blend of Root Extracts of <i>Angelica gigas</i>, <i>Cynanchum wilfordii</i>, and <i>Phlomis umbrosa</i>

EstroG-100 , a dietary supplement composed of extracts from the roots of Angelica gigas (AG), Cynanchum wilfordii (CW), and Phlomis umbrosa (PU), has gained global interest for managing menopausal symptoms.In this study, methods were developed using highperformance thin-layer chromatography (HPTLC) to identify the marker compounds of EstroG-100 , thereby confirming that the product is composed of AG, CW, and PU.To this end, decursin (DE) and nodakenin (ND), 4-hydroxyacetophenone (4-HAP) and 2,4-dihydroxyacetophenone (2,4-DHAP), and shanzhiside methyl ester (SM) were selected as marker compounds for AG, CW, and PU, respectively.These compounds were detected from the extracts of the individual plants and EstroG-100 using silica gel 60 RP-18 F 254 as a stationary phase, with different mobile phases depending on the plant material.First, with a methanol/water (5:1, v/v) mixture as the mobile phase, DE and ND were clearly observed from the extracts of AG and EstroG-100 as bands with retention factors (Rfs) of 0.40 and 0.85, respectively.Second, using a mixture of chloroform/ethyl acetate/methanol/acetic acid (5:2:0.05:0.1,v/v/v/v) as a mobile phase, 4-HAP and 2,4-DHAP were detected from the extracts of CW and EstroG-100 as bands with Rfs of 0.7 and 0.8, respectively.Third, a mobile phase of chloroform/methanol/water/acetic acid (70:30:4:4, v/v/v/v) enabled the detection of SM from the extracts of PU and EstroG-100 as a band with an Rf of 0.6.Finally, with a mobile phase of chloroform/ethyl acetate/methanol/acetic acid (7:1:1:1, v/v/v/v), ND, 4-HAP, and SM were simultaneously detected as bands with Rfs of 0.20, 0.87, and 0.06, respectively.Thus, the HPTLC methods developed in this study are expected to serve as efficient tools for verifying the composition of EstroG-100 and for controlling the quality of the product.Particularly, the last condition offers an advantage by being simple and easy to apply, and well-suited to our intended purpose.