Selective high-performance liquid chromatographic determination of artesunate and α- and β-dihydroartemisinin in patients with falciparum malaria

doi:10.1016/0378-4347(95)00428-9

Journal of Chromatography B: Biomedical Sciences and Applications

Volume 677, Issue 2, 3 March 1996, Pages 345-350

https://doi.org/10.1016/0378-4347(95)00428-9 Get rights and content

Abstract

A novel solid-phase extraction and a robust high-performance liquid chromatographic (HPLC) separation procedure for artesunate and α- and β-dihydroartemisinin, using post-column alkali decomposition and UV detection, is described. Extraction was performed with Bond-Elut Phenyl solid-phase extraction cartridges and analysis by HPLC was carried out using a Waters Symmetry C₈ 5-μm 150 × 3.9 mm I.D. column. The mobile phase was 50% acetonitrile in 0.1 M acetate buffer (pH 4.8) delivered at a flow-rate of 0.7 ml/min. The column eluate was mixed with 1.2 M potassium hydroxide in 90% methanol delivered at 0.3 ml/min, in a 1-ml reaction coil at 69°C, to form UV-absorbing chromophores which were detected at 290 mm. The recovery of all analytes was greater than 80%. There was no significant difference in the peak-area ratio of α- and β-dihydroartemisinin in plasma. Preliminary pharmacokinetic data from six adult Vietnamese patients who received 120 mg of artesunate by intravenous injection for the treatment of acute falciparum malaria are presented. Despite limited data, the mean half-life of artesunate was approximately 3.5 min while that for dihydroartemisinin was 34 min. These data confirm the relatively rapid clearance of both artesunate and its principal active metabolite, dihydroartemisinin.

References (17)

T.T. Hien et al.
Lancet
(1993)
I. Lee et al.
Pharmacol. Ther.
(1990)
N.J. White
Trans. R. Soc. Trop. Med. Hyg.
(1994)
G. Edwards
Trans. R. Soc. Trop. Med. Hyg.
(1994)
O.R. Idowu et al.
J. Chromatogr.
(1989)
C.G. Thomas et al.
J. Chromatogr.
(1992)
D.K. Muhia et al.
J. Chromatogr. B
(1994)
Z.M. Zhou et al.
J. Chromatogr.
(1987)

There are more references available in the full text version of this article.

Cited by (119)

Pharmacokinetics and antimalarial activities of reduction-responsive releasing dihydroartemisinin prodrug self-assembled nanoparticles in rodents
2021, Journal of Drug Delivery Science and Technology
Artemisinin and its derivates (ARTs) exert antimalarial effect by virtue of unique endoperoxide bridge, which is generally considered to generate oxygen free radicals and then form carbon-centered free radicals to damage parasites. The known high level of glutathione (GSH) in parasites plays a key role in maintaining redox balance to protect parasites from oxidative stress damage, which may be a risk factor for the failure of ARTs-based combination therapy and drug resistance. Nevertheless, the highly reducing environment in cytoplasm of malaria parasites provides an available trigger for intracellular nanoparticle disassembly to responsively release drug. In this study, a reduction-responsive releasing dihydroartemisinin (DHA) prodrug (C₁₄-SS-DHA) was synthesized by conjugating DHA and tetradecylamine (C₁₄–NH₂) with disulfide bond (–SS–) as a linker. As a control, a DHA carbon-carbon bond (–CC–) prodrug (C₁₄-CC-DHA) was synthesized using succinic anhydride as a –CC– linker. To note, DHA prodrugs were prepared as self-assembled nanoparticles (C₁₄-SS-DHA NPs and C₁₄-CC-DHA NPs). The size and zeta potential of C₁₄-SS-DHA NPs were 121 ± 1.582 nm and −29 ± 0.635 mV, and those of C₁₄-CC-DHA NPs were 137.5 ± 0.5132 nm and −41 ± 0.377 mV, respectively. The C₁₄-SS-DHA NPs can release DHA in a reductive media in vitro. The pharmacokinetics study showed that the area under the curve (AUC_(0-t)) of plasma concentration versus time of DHA from C₁₄-SS-DHA NPs was 20737.2 ± 6028.3 h μg L⁻¹, which was much higher than that of C₁₄-CC-DHA NPs (6254.5 ± 2578.8) or DHA solution (857.6 ± 103.7 h μg L⁻¹). The pharmacodynamics study presented that the ED₉₀ of C₁₄-SS-DHA NPs was 3.81 ± 0.35 μmol/kg, which was lower than that of C₁₄-CC-DHA NPs (12.70 ± 1.63 μmol/kg)) or DHA solution (17.67 ± 3.38 μmol/kg). C₁₄-SS-DHA NPs showed significantly higher systemic exposure and better antimalarial activity in comparison with C₁₄-CC-DHA NPs or DHA solution (P < 0.05).
Rapid quantification of artemisinin derivatives in antimalarial drugs with dipstick immunoassays
2020, Journal of Pharmaceutical and Biomedical Analysis
Substandard antimalarial drugs will result in unsatisfied therapeutic efficacy and increase the risk of resistance development. The point-of-care, qualitative, or semi-quantitative dipstick immunoassays cannot differentiate the substandard drugs with confidence. A rapid and quantitative analytical method that can be used under field conditions is needed. Here, three lateral flow immunoassays (LFIAs) based on colloidal gold nanobeads (CGN) as labels were developed for quantification of artemether, dihydroartemisinin and artesunate contents in antimalarial drugs with the aid of a portable optical scanner. Also, time-resolved fluorescent nanobeads (TRFN)-LFIA, coupled with a portable fluorescent lateral flow reader, was developed for quantification of artesunate. Commercial antimalarial drugs were used to validate these LFIAs with comparison to the gold standard high-performance liquid chromatography (HPLC) method. The drug contents estimated with these CGN-LFIAs were in the range of 85.5–109.3% of the contents determined by HPLC with a coefficient of variation (CV) of 4.5–13.0%. The TRFN-LFIA results were in the range of 93.7–108.4% of contents determined by HPLC with a CV of 5.2–8.9%. There were no significant differences between the results of CGN-LFIA and TRFN-LIFA (P = 0.5277, t-test). Both types of LFIAs with portable readers may be used for quantitation of active ingredients in antimalarial drugs and for screening substandard antimalarial drugs in resource-limiting settings.
Old wine in new bottles: Drug repurposing in oncology
2020, European Journal of Pharmacology
Increasing costs, much time consumption and high risk of failure associated with the process of de novo development of new anticancer drugs have prompted the pharmaceutical industry to seek alternative strategies that may facilitate and accelerate the whole process. In particular, the repurposing strategy, known also as repositioning or reprofiling strategy, is a potential source of new treatment options for cancer patients with high unmet medical needs. However, it should be noted that the repurposing strategy, being still a new trend in drug development, should only complement the process of discovering new anticancer drugs, and should not be its alternative. The best repurposable oncological drug candidates are the agents whose original patent protection has already expired, and for which there is a possibility to create a formulation enabling, together with a new therapeutic indication, new patent protection. In this review article we discuss the advantages of the repurposing strategy, and provide an overview of a number of promising candidates, such as artesunate, aspirin, cimetidine, doxycycline, ivermectin, metformin, rapamycin (sirolimus), and thalidomide, that have the potential to be repurposed as anticancer drugs both in cancer prevention and therapy. In addition, we highlight some of the studies regarding the signalling pathways and molecular targets altered by these drugs, and describe the biological mechanisms underlying their anticancer effects.
Development of monoclonal antibody-based immunoassays for quantification and rapid assessment of dihydroartemisinin contents in antimalarial drugs
2018, Journal of Pharmaceutical and Biomedical Analysis
Dihydroartemisinin (DHA) is one of the artemisinin derivatives widely used in artemisinin-based combination therapies (ACTs) for malaria treatment. The availability of a point-of-care device for estimation of DHA quantity would allow a quick quality assessment of the DHA-containing drugs. In this study, 9-O-succinylartemisinin was obtained from microbial fermentation of artemisinin, which was hydrogenated to 9-O-succinyldihydroartemisinin as the hapten for DHA. A monoclonal antibody (mAb), designated as 2G₁1G₄, was identified after screening the hybridoma library, which showed 52.3% cross reactivity to artemisinin, but low or no cross reactivity to artesunate, artemether, and several ACTs partner drugs. Based on this mAb, a highly-sensitive, indirect competitive enzyme-linked immunosorbent assay was designed, which showed 50% inhibition concentration of DHA at 1.16 ng/mL, a working range of 0.26–4.87 ng/mL, and limit of detection of 0.18 ng/mL. In addition, a colloidal gold-based lateral flow immunoassay (dipstick) was developed with an indicator range (indicating sensitivity) of 50–100 ng/mL. This dipstick was evaluated for determination of DHA contents in commercial drugs and the results were highly agreeable with those determined by high-performance liquid chromatography.
Application of an LC-MS/MS method for reliable determination of amodiaquine, N-desethylamodiaquine, artesunate and dihydroartemisinin in human plasma for a bioequivalence study in healthy Indian subjects
2016, Journal of Pharmaceutical and Biomedical Analysis
A sensitive and high throughput bioanalytical method has been developed for reliable determination of amodiaquine (AQ), N-desethylamodiaquine (DEAQ), artesunate (AS) and dihydroartemisinin (DHA) in human plasma by LC–MS/MS. The method employs a solid phase extraction procedure without an evaporation step and with optimum use of organic solvents to circumvent degradation of artemisinin derivatives. The analytes and their deuterated internal standards (ISs) were analyzed on Hypersil Gold (100 mm × 4.6 mm, 5 μm) column using acetonitrile and 2.0 mM ammonium formate (pH 2.50) in 80:20 (v/v) ratio as the mobile phase. A triple quadrupole mass spectrometer equipped with an electrospray ionization interface was used to detect and quantify the analytes. The method was established over the concentration range of 0.250–30.0 ng/mL, 1.50–180 ng/mL, 2.00–600 ng/mL and 5.00–1400 ng/mL for AQ, DEAQ, AS and DHA respectively using 250 μL human plasma. The intra-day and inter-day accuracy and precision (% CV) across quality controls varied from 93.3–105.0% and 1.7–8.3 respectively for all the analytes. The stability was assessed in whole blood as well as in plasma samples under different conditions. All four analytes were stable in whole blood up to 2 h on melting ice. The long term stability in plasma was ascertained up to 90 days. IS-normalized matrix factors ranged from 0.988–1.023 for all the analytes. The method was successfully applied to a bioequivalence study using 50 mg artesunate and 135 mg amodiaquine fixed dose formulation in 14 healthy subjects.
Determination and pharmacokinetic studies of artesunate and its metabolite in sheep plasma by liquid chromatography-tandem mass spectrometry
2015, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
A rapid and sensitive high-performance liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated to simultaneous quantify artesunate and its metabolite in sheep plasma. The plasma samples were prepared by liquid–liquid extraction. Chromatographic separation was achieved on a C18 column (250 × 4.6 mm, 5 μm) using methanol: water (60:40, v/v) (the water included 1 mM ammonium acetate, 0.1% formic acid, and 0.02% acetic acid) as the mobile phase. Mass detection was carried out using positive electrospray ionization in multiple reaction monitoring mode. The calibration curve was linear from 1 ng/mL to 400 ng/mL (r² = 0.9992 for artesunate, r² = 0.9993 for its metabolite). The intra- and inter-day accuracy and precision were within the acceptable limits of ±10% at all concentrations for both artesunate and its metabolite. The recoveries ranged from 92% to 98% at the three concentrations for both. In summary, the LC–MS/MS metho described herein was fully successfully applied to pharmacokinetic studies of artesunate nanoemulsion after intramuscular delivery to sheep.

View all citing articles on Scopus

View full text

Regular paperSelective high-performance liquid chromatographic determination of artesunate and α- and β-dihydroartemisinin in patients with falciparum malaria

Abstract

Lancet

Pharmacol. Ther.

Trans. R. Soc. Trop. Med. Hyg.

Trans. R. Soc. Trop. Med. Hyg.

J. Chromatogr.

J. Chromatogr.

J. Chromatogr. B

J. Chromatogr.

Regular paper
Selective high-performance liquid chromatographic determination of artesunate and α- and β-dihydroartemisinin in patients with falciparum malaria