Development and validation of a RP-HPLC method for quality control of oxantel pamoate, pyrantel pamoat and praziquantel in tablets
In the present study, simple, rapid and precise HPLC methods
were developed which would be useful for quality control of
pharmaceutical dosage forms containing praziquantel (PRZ),
pyrantel pamoate (PPA) and oxantel pamoate (OPA). The first
method (M1) was developed for the analysis of PRZ; separation
was achieved using a reversed–phase column (4.6 x 150 mm,
5 μm) C18, a mobile phase comprising ACN:MeOH:20 mM
phosphate buffer (0.2 % TEA, pH 4.5) (50:10:40, v/v/v) and
UV detection at 210 nm. PPA and OPA were analysed
simultaneously using a separate method (M2) by employing
the same column and flow rate. In accordance with the second
method (M2), detection wavelength was set at 295 nm and a
mobile phase of ACN:MeOH:20 mM phosphate buffer (0.2 %
TEA, pH 4.5) (12:3:85, v/v/v) was used. Benazepril
hydrochloride (BZP) and paracetamol (PAR) were used as
internal standards (IS) of the methods M1 and M2, respectively.
Both methods were validated based on the parameters such as
specifity, linearity, precision, accuracy, limit of detection
(LOD) and limit of quantitation (LOQ) besides system
suitability tests. Forced degradation studies were performed to
indicate specifity of the proposed methods. The methods were
found linear over the concentration ranges of 0.5–7.5 μg/mL,
1–15 μg/mL and 2–40 μg/mL for PRZ, PPA and OPA,
respectively. Correlation coefficients (r) of the regression
equations were greater than 0.999 in all cases. The precision of
the methods was demonstrated using intra- and inter-day assay
RSD values which were less than 1% in all instances. Accuracy
of the proposed methods was tested on placebo tablets spiked
with known amounts of actives. Resulting recoveries of assays
were in the range of 99.9–101.1 % whereas, those from
commercial tablets were 99.4–100.8 %.
Keywords: Oxantel pamoate, Pyrantel pamoate, Praziquantel,
HPLC, analytical method validation.
WHO technical report series, no. 972. Research Priorities for Helminth Infections. World Health Organization, 2012, [http://apps.who.int/iris/bitstream/10665/75922/1/WHO_ TRS_972_eng.pdf].
Khan AR, Akhtar MJ, Mahmood R, Ahmed SM, Malook S, Iqbal M. LC assay method for oxfendazole and oxyclozanide in pharmaceutical preparation. J Pharm Biomed Anal 2000; 22: 111-4.
Morovján G, Csokan P, Makranszki L, Abdellah-Nagy EA, Toth K. Determination of fenbendazole, praziquantel and pyrantel pamoate in dog plasma by high-performance liquid chromatography. J Chromatogr A 1998; 797: 237-44.
Grandemange E, Claerebout E, Genchi C, Franc M. Field evaluation of the efficacy and the safety of a combination of oxantel/pyrantel/praziquantel in the treatment of naturally acquired gastrointestinal nematode and/or cestode infestations in dogs in Europe. Vet Parasitol 2007;145: 94-9.
Gonzalez-Esquivel DF, Okuno CM, Sanchez Rodriguez M, Solelo Morales J, Cook HJ. Sensitive high-performance liquid chromatographic assay for praziquantel in plasma, urine and liver homogenates. J Chromatogr 1993; 613: 174-8.
Liu J, Stewart JT. High-performance liquid chromatography determination of praziquantel enantiomers in human serum using a reversed-phase cellulose-based chiral stationary phase and disc solid-phase extraction. J Chromatogr B Biomed Sci Appl 1997; 692: 141-7.
Schepmann D, Blaschke G. Isolation and identification of 8-hydroxypraziquantel as a metabolite of the antischistosomal drug praziquantel. J Pharm Biomed Anal 2001; 26: 791-9.
Meier H, Blaschke G. Investigation of praziquantel metabolism in isolated rat hepatocytes. J Pharm Biomed Anal 2001; 26: 409-15.
Ridtitid W, Wongnawa M, Mahatthanatrakul W, Punyo J, Sunbhanich M. LC determination of praziquantel in human plasma. J Pharm Biomed Anal 2002; 28: 181-6.
Goras JT. High performance liquid chromatographic method of pyrantel tartrate in swine feeds and supplements. J Assoc Off Anal Chem 1981; 64: 1291-6.
Allender WJ. High-performance liquid chromatographic determination of oxantel and pyrantel pamoate. J Chromatogr Sci 1988; 26: 470-2.
Piantavini MS, Pontes FL, Uber CP, Stremel DP, Sena MM, Pontarolo R. Chemometric quality inspection control of