Recent research on Analytical methods of Analysis of Artesunate: A Review

Saravanan. R, Bharani Pandilla*, Vijayageetha. R, Kavitha. M, Ashok. P

Department of Pharmaceutical Analysis, C.L Baid Mehta College of Pharmacy, Thoraipakkam,

Chennai - 600097.

*Corresponding Author E-mail: bharaniashok@gmail.com

ABSTRACT:

Artesunate (active metabolite dihydroartemisinin) and artesunate-based combination therapy (ACT) is recommended by the World Health Organization (WHO) for the treatment of severe and multidrug resistant malaria. In the pharmaceutical industry, analytical method development gives important information of drugs potency, bioavailability and its stability. Analytical method development is required for assuring the quality of product. So far, around thirty three analytical methods have been reported for various studies on analysis of artesunate in bulk, pharmaceutical formulations and biological fluids. This review highlights different analytical methods such as chromatography, spectroscopy and hyphenated techniques of artesunate. These techniques are either explored for the quantification, detection of metabolite and also for stability-studies of the artesunate. The present studies revealed that HPLC techniques along with spectroscopic have been most widely explored for the analysis. The brief review may provide information to the researchers who are working in the area of analytical research of artesunate.

KEYWORDS: Artesunate, Analytical methods, HPLC, LCMS, TLC, UV-Spectrophotometry.

INTRODUCTION:

Malaria has been reported to be caused by Plasmodium parasites. The parasite spreads through the bite of infected anopheles Mosquito also called as “malaria vector” and causing an estimated death of two million people annually [1]. Chronically affect over 240 million people and over 120 million new cases every year [2,3]. Antimalarials with natural and synthetic origins have been reported to kill the parasites with mode of action different from that of quinoline-based drugs [4].

Artemisinin and its derivatives are the fast acting antimalarial drugs. Recently, there was an increase in the use of Artemisinin and its derivatives, as they have the potential to delay the drug resistance against multidrug-resistant falciparum malaria when used in combinations with other, long-acting antimalarial drugs like sulfamethoxypyrazine /pyrimethamine or amodiaquine. The artemisinin derivatives are well-tolerated in clinical trials and when used in general [5]

Artesunate (ART) is the sodium salt of the hemisuccinate ester of artemisinin with molecular weight 384.4g (Figure 1). The chemical name is (3R,5aS,6R,8aS9R,10S,12R,12aR)- Decahydro-3,6,9-trimethyl-3,12-epoxy-12H-pyrano[4,3-j]-1,2-benzodioxepin-10-ol, hydrogen succinate [6]. General methods reported that are used in the analysis of artesunate analysis are HPLC, Colorimetric, UV Spectroscopy, TLC, UPLC, Infra-red spectroscopy which are summarized.

Figure.1. Structure of Artesunate

1. High performance of Liquid-Chromatography (HPLC)

HPLC is the advanced analytical technique in pharmaceutical analysis, which is predominantly used in the pharmaceutical industry [7-9] for a large variety of samples. It is the method of choice for determining the purity of new drug candidates, monitoring changes or scale-ups of synthetic procedures, evaluating new formulations, and scrutinizing quality control of final drug products.

S.No.	Stationery phase	Mobile phase	Flow rate and Method of Detection	Results	References
1.	Symmetry C18, 250 x 4.6 mm, 5μ particle size	Phosphate Buffer: methanol 30:70(v/v) (pH 3.0 with 0.5% ortho-phosphoric acid)	1.0ml/min. UV at 225nm	Rt-3.99 min. Accuracy-99.40% LOD-3.24µg/ml LOQ-15.71µg/ml	Odedara M.H et al [10]
2.	C18	Phosphate Buffer: Acetonitrile 30:70(v/v) (pH 3.0 with ortho-phosphoric acid)	UV	Rt- 6.2 min. Accuracy- 98%	Uzondu Akueyinwa et al. [11]
3.	X-Bridge C18 (250 x 4.6 mm i.d., 5 μm particle size)	0.05 M monobasic potassium phosphate (pH adjusted to 3.0 with phosphoric acid): acetonitrile (50:50 v/v)	1.0ml/min. DAD at 210nm	Rt - 9.88 min. Accuracy – 100.09%	Fernando Henrique Andrade Nogueira et al. [12]
4.	Qualisil C8 (250 mm x 4.6 mm i.d.) with particle size 5 μm	Acetonitrile: phosphate buffer (70:30, v/v)	0.8ml/min. At 221nm	Rt – 5.6min. Accuracy – 98.55% LOD – 0.044mg/ml LOQ – 0.133mg/ml	P. S. Jain et al [13]
5.	HiQ_SiL C8 (250 mm × 4.6 mm i.d.)	acetonitrile: 1 M sodium ac_ etate buffer (pH 3 adjusted with o_phosphoric acid); (70: 30, v/v)	1.0ml/min. UV at 220nm	Rt – 4.883min. Accuracy – 99.97% LOD – LOQ – 52.35µg/ml 158.63µg/ml	S. S. Ranher et al [14]
6.	Zorbax Extend C18, 50 × 4.6 mm (dp: 3.5 μm)	methanol: 10 mM ammonium formate buffer pH 2.8 (90:10, v/v)	0.7ml/min. PDA at 210nm	Rt–1.144min. Accuracy – 95% LOD – LLOQ - 124.1µg/ml 3820µg/ml	Védaste Habyalimana et al. [15]
7.	Accucore RP-MS 2.6 μm, 50 x 2.1 mm	60:40 (v/v) water + 0.1% formic acid: acetonitrile + 0.1% formic acid	0.6ml/min. UV at 210nm	Rt – <2min. % RSD Rt – 0.44	Joanne Gartland et al [16]
8.	Syncronis™ C18, 5 μm, 100 x 2.1 mm	40:60 (v/v) water + 0.1% formic acid: acetonitrile + 0.1% formic acid	0.2ml/min. UV at 210nm	Rt - <5min. %RSD – 0.10	Joanne Gartland et al [17]
9.	C-18, 10 cm x 4.6-mm, 3 µ	monobasic potassium phosphate (pH 3.0 with OrthophosphoricAcid): Acetonitrile in the ratio 500:400	0.8ml/min. UV at 210nm	% ASSAY – 99.20%	Manisha Phadke et al. [18]
10.	Hypersil Gold C18 (250x 4.6 mm 5μ)	Acetonitrile: 25 mM potassium dihydrogen phosphate buffer (70:30, v/v)	1.0ml/min. UV at 220nm	Rt – 5.20min. Accuracy – 100.67% LOD – LOQ 87.97µg/m l- 266.57µg/ml	Padmanabh Deshpande et al [19]
11.	Inertsil ODS C18 (250 x 4.6mm, 5μ)	Potassium dihydrogen phosphate buffer (pH 5.8 with ortho-phosphoric acid) acetonitrile: methanol in (50:30:20, v/v/v)	1.0ml/min. UV at 208nm	Rt – 5.09 min. Accuracy – 99.81% LOD –LOQ 2.07 and 6.27	P RajaRao et al. [20]
12.	XBridge BEH C18 Column (50 x 2.1 mm, 2.5μm particle size)	Water (5mM Ammonium Acetate) : Acetonitrile with gradient flow	0.6ml/min. ELSD	Rt – 0.83 min. Accuracy – 100.31% Linearity (r^2) – 0.9993	Patidar Khushwant et al [21]
13.	Inertsil ODS (250mm x 4.6mm x 5μ)	Phosphate buffer (pH 4.2) : Methanol 40:60(v/v)	1.0ml/min. PDA at 291nm	Rt – 3.63 min. Linearity (r^2) – 0.999 LOD – LOQ 2.71µg/ml – 9.03µg/ml	P. Jyothi et al. [22]
14.	Symmetry C18 (250 × 4.6 mm and 5 μm)	Methanol:phosphate buffer (70:30, v/v) (pH 3.2 using phosphoric acid)	1.0ml/min. UV at 220nm	Rt – 7.30min. Linearity (r^2) – 0.9992 LOD – LOQ 1.39µg/ml – 2.27µg/ml	S. M. Sandhya et al [23]

*UV – Ultra-Violet, DAD – Diode-Array Detector, PDA – Photo-Diode Array, ELSD – Evaporative Light Scattering Detector, R_t – Retention time, LOD – Limit of Detection, LOQ – Limit of Quantitation, RSD – Relative Standard Deviation.

2. Ultra-Violet spectroscopy:

S. No.	Detection wavelength	Solvent	Linearity range	LOD	References
1.	220 nm	Ethanol + 0.1M Sodium hydroxide	10-50µg/ml	0.245µg/ml	Emmanuel Etim et al [24]
2.	240 nm	Methanol	10-60µg/ml	0.54µg/ml	T. M. Kalyankar et al. [25]
3.	342.49 nm	Methanol	5-30µg/ml	0.160µg/ml	Kushwaha Ragini et al. [26]
4.	287 nm	Simulated Intestinal Fluid (SIF)	10–200 mg/ml	0.471 mg/ml	C.O. Esimone et al [27]
5.	289 nm	Water + 1M Sodium hydroixide	0.02-0.1mg/ml	0.01mg/ml	Fateh AL Rahman Magbool [28]
6.	242 nm	Ethanol + 0.1N Sodium hydroxide + 0.1M Acetic acid in 20% ethanol	10-50µg/ml	-	Chinyere Okwelogu et al [29]
7.	240 nm	0.1M Acetic acid in 20% methanol	10-60µg/ml	0.54µg/ml	Sarvesh Sharma et al. [30]
8.	300 nm	Ethanol + 1M Soduim hydroxide	0.5-50µgm/ml	0.375µg/ml	Attih E. E. et al [31]
9.	242 nm	Methanol	20-40µg/ml	0.13µg/ml	Akshay D. Sapakal et al [32]

*LOD – Limit of Detection.

3. Colorimetric:

S. No.	Chromogenic reagent	Solvent	Detection wavelength	Linearity range	LOD	References
1.	i) Safranin O ii) Variamine blue	Water	i) 521nm ii) 556nm	i) 20-140µg/ml ii) 20-140µg/ml	i) 0.0043µg/ml ii) 0.4496µg/ml	Thekke Veettil Sreevidya [33]
2.	p-dimethylamino benzaldehyde	Methanol	540nm	15.4-77µg/ml	5.93µg/ml	Olajire A. [34]
3.	Aniline	Buffer solution at pH 4	420nm	-	-	R. A. Mahgoub et al. [35]
4.	i) Methylene blue ii) Soluble Starch	Ethanol	i) 665.6nm ii) 456.5nm	20-140µg/ml	i) 0.101µg/ml ii) 1.5µg/ml	Lawal, A et al [36]

*LOD – Limit of Detection.

4. Infra-red spectroscopy:

S. No.	Wavenumber	Group	Reference
1.	1870-1550 cm^-1 1225-950 cm^-1 1650-1370 cm^-1 1380-1370 cm^-1 700 cm^-1	C=O C-H (inplane deforming vibrations) C-C (aromatic) C-O CH2 (rocking vibration)	Lawal, A et al [36]

5. Thin-Layer Chromatography (TLC):

S. No.	Derivatization Agent	Stationery phase Mobile phase Detection method	Retention factor (Rf) and LOD	Reference
1.	Iodine	Glass silica gel plates (20 × 20 cm), paraffin : n-hexane (2:3 v/v), Iodine tank	0.04 ± 0.03 0.001 mg/ml	G. O. Adewuyi et al. [37]
2.	Anisaldehyde-sulphuric acid	Pre-coated plate of silica gel 60 F254, (10.0 × 10.0 cm, 250 mm thickness), Toluene:ethyl acetate-acetone (2.5:1.0:0.5, v/v/v) Deuterium-tungsten lamp at 525nm	0.59 ± 0.01 5.67ng/band	S. M. Sandhya et al [23]

*LOD – Limit of Detection.

6. Liquid Chromatography-Mass Spectroscopy (LC-MS):

S. No.	Internal Standard	Sample preparation, Stationery phase Mobile phase,	Flow rate, Detection (m/z) LLOQ	Reference
1.	Artemisin	Liquid-Liquid extraction, XTerra RP-C18 (2.1 x 100mm, 3.5µm)0.15% aqueous formic acid and 0.15% formic acid in acetonitrile (gradient flow)	0.2ml/min SIM – 221,249 and 267 25ng/ml	Stijin A.A. Van Quekelberghe et al [38]
2.	Indometacin	Used Methanol as solvent, XBridge C18 column (50 × 2.1 mm, 5 μm) water/acetonitrile/methanol (30:35:35, v/v/v) with 0.1% formic acid	350µl/min MRM, 407.0 and 261.1 -	Breno M. Marson et al [39]
3.	[^2H4]ARS	Protein precipitaion RP-C18 (4.6 by 50 mm) 63% acetonitrile/ 37% water, both containing 0.1% formic acid	MRM,406.2  267.21ng/ml	Yun Liu et al. [40]
4.	SIL-ARS	Solid phase Extraction Hypersil Gold C18 (150 × 2.1 mm, 5 μ) Acetonitrile: 10mM ammonium acetate (40:60 v/v) (pH 3.5 with 1% Acetic acid) Gradient flow	500µg/ml MRM, 402→267 (ARS), 406→163 (SIL-ARS), 1.38ng/ml	Niklas Lindegardh et al. [41]
5.	-	Phenomenex Kinetex 2.6 μm EVO C18 100 Å LC Column (150x3.0 mm) Acetonitrile with 1% Formic acid (Gradient flow)	10-25ml/min.	David A. Godin. [42]
6	Artemisin	Protein precipitation C18 column, 0.5mM Ammonium acetate buffer (pH 4.5) : Acetonitrile : Water 8% : 40% : 52%	0.4ml/min Single Ion recording, 402, 302, 300, 267 and 284 1.23ng/ml	Paktiya Teja-Isavadharm et al [43]

*LLOQ – Lower Limit of Quantitation, SIM – Selected Ion Monitoring, MRM – Multiple Ion Monitoring, ARS – Artesunate, SIL-ARS – Stable-Isotope Labeled-Artesunate.

CONCLUSION:

The present review discussed about different analytical approach employed for the assessment of ART. Profuse examination have been accomplished including HPLC, UPLC, UV/Vis-Spectroscopy, Colorimetry, LC-MS for evaluation of ART in bulk and in its combination with other drugs from pharmaceutical formulations and also biological fluids.

Liquid chromatography with UV detection has been found to be most studied for estimation of ART in bulk as well as pharmaceutical dosage forms, while hyphenated LC-MS methods reported for determination of ART and its metabolite in plasma and other biological fluids. Few chromatography approaches like stability-indicating HPLC, UPLC, and TLC are also reported. Few simple UV spectrophotometric methods may be used for routine analysis of ART alone and in combination with other drugs. These compiled data may of use for research for further studies in analysis of ART.

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Received on 09.07.2020 Revised on 13.08.2020

Accepted on 08.09.2020     ©Asian Pharma Press All Right Reserved

Asian J. Pharm. Ana. 2021; 11(1): 49-53

DOI: 10.5958/2231-5675.2021.00010.7