Determination of the composition and monosaccharide content of atractylodes polysaccharides using pre-column derivatization and a quantitative analysis of multicomponents by a single marker method

Abstract

Atractylodes chinensis (DC.) Koidz. (ACK), a prominent herb in traditional Chinese medicine, is widely used to treat conditions such as dampness obstruction in the middle Jiao, epigastric distension, diarrhea, and edema. The primary bioactive components of ACK include sesquiterpenes, alkynes, triterpenes, and polysaccharides. In this study, a high-performance liquid chromatography (HPLC) method with pre-column derivatization using 1-phenyl-3-methyl-5-pyrazolone (PMP) was developed. Additionally, a quantitative analysis of multicomponents by a single marker (QAMS) method was established to quantify polysaccharide components in ACK with high precision. Atractylodes chinensis polysaccharides (ACP) underwent acid hydrolysis with hydrochloric acid, yielding stable monosaccharide-PMP derivatives through pre-column derivatization. These derivatives were analyzed by HPLC, with anhydrous glucose as the internal standard. The relative correction factors for glucuronic acid, galactose, and arabinose were calculated using the QAMS method, yielding values of 1.08, 0.97, and 0.75, respectively. The monosaccharide contents determined by QAMS showed no significant difference from those obtained by the external standard method (ESM), with all relative standard deviations (RSD%) <2 %. This method exhibited good linearity—linear ranges: glucuronic acid (0.0250–0.1250 mg/mL, r = 0.9996), anhydrous glucose (0.0094–0.0472 mg/mL, r = 0.9996), galactose (0.0100–0.0500 mg/mL, r = 0.9997), arabinose (0.0250–0.1250 mg/mL, r = 0.9987)—and the RSD% values for the stability of these four components within 48 hours were all <3 %. In terms of reproducibility, multiple injections (n = 6) showed low relative standard deviations (RSD%): glucuronic acid (2.04 %), anhydrous glucose (0.32 %), galactose (0.53 %), arabinose (0.53 %). Regarding stability, within [specific time range], the RSD% values were: glucuronic acid (1.71 %), anhydrous glucose (2.49 %), galactose (2.11 %), arabinose (1.76 %). The accuracy was also satisfactory, with an RSD% of 3.45 %. These findings indicate that the established approach is robust, reliable, and applicable for determining monosaccharide content in hydrolyzed ACP. This methodology provides a promising tool for quality control and further pharmacological studies of ACP.