Simple and rapid micro‐scale quantification of artemisinin in living Artemisia annua L. by improved gas chromatography with electron‐capture detection

Abstract

Malaria threatens 300-500 million people and kills more than one million people annually. Artemisinin has been widely used as part of the artemisinin-based combination therapies against malaria. However, its supply is seriously short due to very small amounts of production of artemisinin in Artemisia annua. Molecular biologic researches aimed at increasing the artemisinin yield in plant have received more and more attention and therefore corresponding quantification methods for artemisinin analysis are urgently needed. A variety of methods for determination of artemisinin have been developed but they cannot be applied when only very little plant material is available or the material should be kept live, which often occurs in molecular biologic researches. The present work developed a simple, fast and low toxic micro-scale analysis procedure for determination of artemisinin in a single leaf or flower of living Artemisia annua using improved gas chromatography with electron-capture detection. The recovery of > > > >95% was achieved by vortex of a piece of fresh leaf in 1 mL ethyl acetate for 2 min at room temperature. This method provides a powerful tool for biosynthesis study of artemisnin, high-throughput screening high-yield clone in an early stage, or real-time quality control of Artemisia annua crop.

Metabolic analysis of the increased adventitious rooting mutant of Artemisia annua reveals a role for the plant monoterpene borneol in adventitious root formation

“…The artemisinin in A . annua is partly degraded into three major compounds under the high temperature produced by GC, and four peaks pertaining to artemisinin appeared on the chromatogram, in agreement with previous reports (Liu et al ). The second most abundant compound in wild‐type was camphor (0.72 mg g −1 dry weight (DW)), whereas borneol, a reduced form of camphor, was the second most prevalent compound in the iar mutant.…”

“…Gas chromatography‐mass spectrometry (GC‐MS) analysis of terpenoids of A . annua leaves was performed according to the method previously reported (Liu et al ) using an Agilent 7890N GC system (Santa Clara, CA) with a flame ionization detector and an electron ionization source.…”

Simultaneous isolation of artemisinin and its precursors from Artemisia annua L. by preparative RP‐HPLC

“…For compound 1 , four peaks were formed (as shown in Fig. ) owing to the thermal degradation of artemisinin, which was consistent with previous reports (Peng et al , ; Liu et al , ). The results showed that the purity of compounds 1–3 was 99.1, 99.6 and 99.3% respectively.…”

“…GC analysis of artemisinin was performed according to the previously reported method (Liu et al , ) using an Agilent 6890N GC system (Santa Clara, CA, USA) with a flame ionization detector. Briefly, 1.0 μL of sample was injected to a HP‐5 capillary column (Agilent).…”

LC–UV/MS quality analytics of paediatric artemether formulations

“…Current analytical techniques describe derivatisation-based methods [22] , gas chromatography (GC) [23] , thin layer chromatography (TLC) [24] , supercritical fluid chromatography (SCFC) [25] , spectroscopic [26] and immunological techniques [27] , [28] , but it is clear that the main-stream methods are mainly based on high performance liquid chromatography (HPLC), coupled to ultra violet (UV), evaporative light scattering detector (ELSD), electron capture detection (ECD) or electrospray ionisation (ESI)–mass spectrometry (MS) detection [29] , [30] , [31] , [32] . Up till now, the focus was directed to biological matrices like plasma for pharmacokinetic information [33] , [34] , to plant derived samples for production reasons [23] , [35] , [36] or for environmental eco-toxicity studies [37] , [38] . However, with the advent of a plethora of newly developed 1,2,4-trioxane derivatives and the urgent demand to develop suitable paediatric formulations, there is a clear need for analytical methods which are suitable for quality purposes for finished drug products (FDP).…”