Separation and purification of five alkaloids from Aconitum duclouxii by counter‐current chromatography

Abstract

C19 -diterpenoid alkaloids are the main components of Aconitum duclouxii Levl. The process of separation and purification of these compounds in previous studies was tedious and time consuming, requiring multiple chromatographic steps, thus resulted in low recovery and high cost. In the present work, five C19 -diterpenoid alkaloids, namely, benzoylaconine (1), N-deethylaconitine (2), aconitine (3), deoxyaconitine (4), and ducloudine A (5), were efficiently prepared from A. duclouxii Levl (Aconitum L.) by ethyl acetate extraction followed with counter-current chromatography. In the process of separation, the critical conditions of counter-current chromatography were optimized. The two-phase solvent system composed of n-hexane/ethyl acetate/methanol/water/NH3 ·H2 O (25%) (1:1:1:1:0.1, v/v) was selected and 148.2 mg of 1, 24.1 mg of 2, 250.6 mg of 3, 73.9 mg of 4, and 31.4 mg of 5 were obtained from 1 g total Aconitum alkaloids extract, respectively, in a single run within 4 h. Their purities were found to be 98.4, 97.2, 98.2, 96.8, and 96.6%, respectively, by ultra-high performance liquid chromatography analysis. The presented separation and purification method was simple, fast, and efficient, and the obtained highly pure alkaloids are suitable for biochemical and toxicological investigation.

Isolation and purification of two antioxidant isomers of resveratrol dimer from the wine grape by counter‐current chromatography

“…To obtain an efficient separation of target compounds, the previous articles on CCC demonstrated that the two‐phase solvent system should meet the following requirements: (i) sufficient sample solubility (<30 s); (ii) short settling time; (iii) appropriate partition coefficient ( K ) values (usually between 0.5 and 2); (iv) no decomposition or denaturation of the sample; and (v) satisfactory retention of the stationary phase. According to the previous reports, two‐phase solvent systems composed of n ‐hexane/ethyl acetate/methanol/water and light petroleum/ethyl acetate/methanol/water were widely used and also very efficient for CCC separation of polyphenol. The two‐phase solvent system was selected according to the K values of each target compound.…”

Aconitum Diterpenoid Alkaloid Profiling to Distinguish between the Official Traditional Chinese Medicine (TCM) Fuzi and Adulterant Species Using LC-qToF-MS with Chemometrics

“…The first RTEM pair indicated that its retention time and exact mass were 3.53 min and m/z 470.2540, and the values of the second RTEM pair were 4.93 min and m/z 484.2693. Using our inhouse ADA-database, these two markers were tentatively identified as contorsine (30) and sczukitine (40) (Figure 7A and Table 3), and these markers account for the chemical variance of A. hemsleyanum species from the other Aconitum species. Similarly, this strategy was employed with four other species, A. nagurum var.…”

A new tropane alkaloid from the leaves of Erythroxylum subsessile isolated by pH‐zone‐refining counter‐current chromatography

“…The reverse displacement pH-zone-refining counter-current chromatography technique was chosen due to its ability to rapidly and easily separate weak bases, like alkaloids, with low solvent consumption. Despite the small number of studies on the isolation of tropane alkaloids by CCC [19], many other types of alkaloids have been separated by conventional HSCCC, with or without the use of pH gradients and pH-zone-refining CCC techniques [15][16][17][18][22][23][24][25]. However, the great chemical diversity of alkaloids requires the development of enhanced solvent systems for each sample.…”