HPLC‐DAD method for the detection of five annopurpuricins in root samples of Annona purpurea

• Published in: Phytochemical analysis Vol. 31; no. 4; pp. 472 - 479
• Main Authors: Hernández‐Fuentes, Gustavo A., Peraza Campos, Ana L., Ceballos‐Magaña, Silvia G., Muñiz‐Valencia, Roberto, Parra‐Delgado, Hortensia
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01.07.2020
• Subjects: Annona purpurea; annopurpuricins; Data analysis; Dichloromethane; Economic analysis; Fractionation; High performance liquid chromatography; HPLC‐DAD; isocratic method; Kidney diseases; Liquid chromatography; Maceration; marker metabolite; Metabolites; Polarity; Separation; Species; Annona purpurea; isocratic method; annopurpuricins; HPLC-DAD; marker metabolite
• ISSN: 0958-0344; 1099-1565
• DOI: 10.1002/pca.2910

Introduction Annona purpurea is a species known in Mexico as “cabeza de negro”. In folk medicine A. purpurea root is used to treat patients with kidney diseases and cancer. Our recent studies demonstrated that this species contains five acetogenins named annopurpuricins A–E, which are active against tumoural cell lines in a subnanomolar range. Objective To develop an analytical method using a high‐performance liquid chromatography diode array detector (HPLC‐DAD) to quantify annopurpuricins A–E in different A. purpurea root samples. Methodology To quantify the five annopurpuricins A–E a sample treatment was carried out, which consisted of fractionation by means of cold and hot maceration; using solvents of ascending polarity: hexane, dichloromethane, methanol and water. The resulting extracts were subject to HPLC‐DAD analysis. The optimised chromatographic separation on a XBRIDGE C18 column achieved separation of all compounds in around 30 min. Results The developed method was validated according to ICH (International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use) validation guide. The developed analytical method was found fast, economic, robust, sensitive, linear and precise. The dichloromethane extract of A. purpurea contains annopurpuricin A in quantities 2‐ to 25‐fold higher than annopurpuricins B–E. This optimised method identified and quantified five annopurpuricins, highly bioactive molecules, in A. purpurea root. Conclusions The fingerprint of the dichloromethane extracts of A. purpurea was obtained at 210 nm. The results analysis allowed to quantify annopurpuricins A–E that are present in different collection batches of medium polarity extracts. After data analysis, annopurpuricin A could be establish as the metabolite marker of the root of the species. 1. A new analytical method using an HPLC‐DAD allowed the detection and/or quantification of annopurpuricins A‐E in different Annona purpurea root samples. 2. Annopurpuricin A is the metabolite marker of the root of A. purpurea. 3. The analysis of the aqueous matrices allowed determining that the annopurpuricins A and C are extracted by the traditional preparations of the species.