Alkaloids from the New Species Papaver kuvajevii

Abstract

A new Papaver species, P. kuvajevii Schaulo et Sonnikova, which is a rare species of the flora of western Sayan (relict, endemic), was discovered in 2003 at the Sayano-Shushenskii Natural Biosphere Reserve and described [1]. An investigation of the chemical composition of P. kuvajevii would prompt its industrial production while preserving the genetic resource of this natural endemic species if valuable biologically active constituents were found.The goal of the present work was to study the alkaloid composition of P. kuvajevii. Herein we communicate results from a GC-MS investigation of the alkaloid composition of this plant.Plant material (aerial part, stems, leaves, flowers, pods) was collected in July 2012 at Sayano-Shushenskii Natural Biosphere Reserve. Herbarium specimens are preserved at the Reserve Herbarium. The yield of substances from air-dried and ground aerial parts (285 g) that were extracted by EtOH in a Soxhlet apparatus was 12.6% (36.0 g). The EtOH extract was worked up with dilute H 2 SO 4 and separated by fractional extraction into fractions according to basicity using the classical method [2].Extraction by CHCl 3 produced alkaloid fraction AC (36.9 mg, 0.01% relative to starting air-dried raw material) after basicification by Na 2 CO 3 (pH 8) and alkaloid fraction B (83.8 mg, 0.03%) after basicification by NaOH (pH 11). The fractions were analyzed by GC-MS on an HP 5890 Series II GC with an HP 5971 mass-selective detector (EI, 70 eV) and HP-5 capillary column (5% diphenyl-95% dimethylsiloxane, 30 m u 0.25 mm, 0.25 Pm) using He carrier gas at 1 mL/min. The column temperature regime was 50°C (2 min), 50-300°C (10 °C/min), and 300°C (30 min). The vaporizer temperature was 280°C; ion-source, 170°C. The scan rate was 1.2 scan/s in mass range 29-650 amu. Constituents were identified by comparing total mass spectra with those in the NIST08 and W8N08 databases.The principal alkaloids in both fractions were amurensinine (1) and mecambridine (syn. oreophylline) (2). In addition to these, minor alkaloids amurensine (3) and D-allocryptopine (4) were also identified (Table 1).Fraction AC was separated by preparative TLC on SiO 2 F 254 plates (Merck) using CHCl 3 -EtOH (15:1, v/v) to afford six fractions that were analyzed by GC-MS. Fractions AC-4 and AC-5 were enriched in amurensinine (1). Fraction B was also separated by preparative TLC under the same conditions and afforded seven fractions that were analyzed by GC-MS. Fraction B-3 was enriched in mecambridine (2); fraction B-5, 1. Fractions AC-4, AC-5, and B-5 were combined. PMR spectra of additionally purified 1 (2.3 mg, 0.00080%) and 2 (3.1 mg, 0.00108%) were recorded. The structures of isolated alkaloids 1 and 2 were confirmed by PMR spectral data and comparison with those reported [3,4].