ABSTRACT
The method for the determination of primaquine (PQ) and 5,6-orthoquinone primaquine (5,6-PQ), the representative marker for PQ active metabolites, via CYP2D6 in human plasma and urine has been validated. All samples were extracted using acetonitrile for protein precipitation and analyzed using the ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) system. Chromatography separation was carried out using a Hypersil GOLDTM aQ C18 column (100 × 2.1 mm, particle size 1.9 µm) with a C18 guard column (4 × 3 mm) flowed with an isocratic mode of methanol, water, and acetonitrile in an optimal ratio at 0.4 mL/min. The retention times of 5,6-PQ and PQ in plasma and urine were 0.8 and 1.6 min, respectively. The method was validated according to the guideline. The linearity of the analytes was in the range of 25-1500 ng/mL. The matrix effect of PQ and 5,6-PQ ranged from 100% to 116% and from 87% to 104% for plasma, and from 87% to 89% and from 86% to 87% for urine, respectively. The recovery of PQ and 5,6-PQ ranged from 78% to 95% and form 80% to 98% for plasma, and from 102% to from 112% to 97% to 109% for urine, respectively. The accuracy and precision of PQ and 5,6-PQ in plasma and urine were within the acceptance criteria. The samples should be kept in the freezer (-80 °C) and analyzed within 7 days due to the metabolite stability. This validated UHPLC-MS/MS method was beneficial for a pharmacokinetic study in subjects receiving PQ.
Subject(s)
Primaquine/analysis , Primaquine/chemistry , Primaquine/pharmacokinetics , Chromatography, High Pressure Liquid , HumansABSTRACT
In this short communication, we report the use of a second-generation macrolide antibiotic, gamithromycin (Gam), as a novel chiral selector for enantioseparation in capillary electrophoresis (CE). A preliminary analysis of the experiment results shows that Gam is especially suitable for the separation of chiral primary amines. Factors influencing enantioseparations were systematically investigated including the composition of the background electrolyte (BGE), concentration of Gam, the type and proportion of organic solvents, applied voltage, etc. In particular, N-Methylformamide (NMF) was successfully used as a non-aqueous solvent for Gam, and shown to be extremely effective for the separation of primaquine (PMQ) and 1-aminoindan (AMI) when used alone or mixed with other commonly used non-aqueous solvents (e.g. methanol). To our knowledge this was also the first application of NMF as a non-aqueous solvent for antibiotic chiral selectors in CE. The best separations were obtained with 100 mM Tris, 125 mM H3BO3 and 80 mM Gam in methanol/NMF (25:75) solvent for PMQ and AMI, or 80-100 mM Gam in methanol for the other model analytes. Among the analytes, the resolution (Rs) of amlodipine (AML) reached up to 15.65, which is to our knowledge the highest value ever reported in CE studies for this compound (except for using molecularly imprinted polymers technique).
Subject(s)
Anti-Bacterial Agents/chemistry , Electrophoresis, Capillary , Macrolides/chemistry , Amlodipine/analysis , Electrolytes , Formamides/chemistry , Indans/analysis , Methanol/chemistry , Primaquine/analysis , Solvents/chemistry , StereoisomerismABSTRACT
Primaquine (PMQ), an important anti-malarial drug, has been recommended by the World Health Organization (WHO) for the treatment of life-threatening infections caused by P. vivax and ovale. However, PMQ has unwanted adverse effects that lead to acute hemolysis in patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency. There is a need to develop simple and reliable methods for PMQ determination with the purpose of dosage monitoring. In early 2019, we have reported an UV-Vis and naked-eye based approach for PMQ colorimetric quantification. The detection was based on a Griess-like reaction between PMQ and anilines, which can generate colored azo products. The detection limit for direct measurement of PMQ in synthetic urine is in the nanomolar range. Moreover, this method has shown great potential for PMQ quantification from human serum samples at clinically relevant concentrations. In this protocol, we will describe the technical details regarding the syntheses and characterization of colored azo products, the reagent preparation, and the procedures for PMQ determination.
Subject(s)
Antimalarials/analysis , Chemistry Techniques, Analytical/methods , Ethylenediamines/analysis , Primaquine/analysis , Sulfanilamides/analysis , Antimalarials/blood , Antimalarials/urine , Body Fluids/chemistry , Body Fluids/metabolism , Chemistry Techniques, Analytical/instrumentation , Colorimetry/instrumentation , Colorimetry/methods , Humans , Limit of Detection , Microscopy, Ultraviolet/instrumentation , Microscopy, Ultraviolet/methods , Primaquine/blood , Primaquine/urineABSTRACT
BACKGROUND: Pharmaceutical compounding of orphan active ingredients can offer cost-effective treatment to patients when no other drug product is available for a rare disease or during periods of drug product shortages. Additionally, it allows customized therapy for patients with rare diseases. However, standardized compounding formulas and procedures, and monographs are required to ensure the patients' safety. RESULTS: Standardized formulas and compounding procedures were developed for seven orphan active ingredients (L-arginine, sodium benzoate, sodium phenylbutyrate, L-carnitine, chenodesoxycholic acid, primaquine phosphate, pyridoxal phosphate) and one non-orphan molecule (sodium perchlorate) regularly compounded by hospital pharmacists for extemporaneous use. The stability of these formulations was evaluated over 3 months at refrigerated (5 °C) and standard storage conditions (25 °C/60%RH) using HPLC-based assays and a suitable shelf life was assigned to the formulations. Additionally, suitable analytical methods for quality control of formulations of pyridoxal phosphate and sodium perchlorate were developed as monographs for these components were not available in the European Pharmacopeia or United States Pharmacopeia. CONCLUSIONS: Availability of compounding formulas and protocols, as well as stability information, for orphan active ingredients can improve patients' access to treatment for rare diseases. Such data were collected for seven orphan active ingredients to treat patients with rare diseases when no other treatment is available. More efforts are needed to develop standardized formulas and compounding procedures for additional orphan active ingredients whose clinical efficacy is well-known but which are not available as products with a marketing authorization. Additionally, a legal framework at EU level is required to enable the full potential of pharmaceutical compounding for orphan active ingredients.
Subject(s)
Drug Compounding/methods , Rare Diseases , Arginine/analysis , Belgium , Carnitine/analysis , Excipients/analysis , Humans , Pharmacists , Phenylbutyrates/analysis , Primaquine/analysis , Pyridoxal Phosphate/analysis , Sodium Benzoate/analysisABSTRACT
Primaquine (PMQ), a well-known anti-malarial drug, is of increasing importance as people moving toward global malaria eradication. PMQ has serious side effects that it often causes acute hemolytic toxicity in people with glucose-6-phosphate dehydrogenase (G6PD) deficiency. The development of simple and reliable approaches for quantitative dose monitoring is thus becoming important during malarial treatment with PMQ. Herein, an unexpected Griess reaction on PMQ was systematically studied. The reaction happened between substituted aniline and a primaquine molecule in the presence of nitrite. Both experimental measurements and theoretic calculation showed that UV-vis absorption of the azo products varied because of different electron contributing effects of substituents. Based on the optimized conditions, a novel colorimetric method has been developed for PMQ determination with excellent sensitivity and selectivity. The detection limits for PMQ in water and synthetic urine samples were down to nanomolar range. More importantly, this method has been successfully used to quantify PMQ from human serum samples within clinically relevant concentration ranges.
Subject(s)
Antimalarials/analysis , Drug Monitoring , Models, Chemical , Primaquine/analysis , Spectrophotometry, Ultraviolet/standards , Aniline Compounds/chemistry , Antimalarials/blood , Antimalarials/urine , Azo Compounds/analysis , Chemistry, Pharmaceutical , Primaquine/blood , Primaquine/urineABSTRACT
A gold nanourchins modified glassy carbon electrode (AuNu/GCE) was developed for the determination of antimalarial drug, primaquine (PQ). The surface of AuNu/GCE was characterized by electrochemical impedance spectroscopy (EIS), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and cyclic voltammetry (CV). EIS results indicated that the electron transfer process at AuNu/GCE was faster as compared to the bare electrode. The SEM and TEM image confirmed the presence and uniform dispersion of gold nanourchins on the GCE surface. Upon investigating the electrochemical behavior of PQ at AuNu/GCE, the developed sensor was found to exhibit high electrocatalytic activity towards the oxidation of PQ. Under optimal experimental conditions, the sensor showed fast and sensitive current response to PQ over a linear concentration range of 0.01-1µM and 0.001-1µM with a detection limit of 3.5nM and 0.9nM using differential pulse voltammetry (DPV) and square wave voltammetry (SWV), respectively. The AuNu/GCE showed good selectivity, reproducibility and stability. Further, the developed sensor was successfully applied to determine the drug in human urine samples and pharmaceutical formulations demonstrating its analytical applicability in clinical analysis as well as quality control. The proposed method thus provides a promising alternative in routine sensing of PQ as well as promotes the application of gold nanourchins in electrochemical sensors.
Subject(s)
Antimalarials/analysis , Electrochemical Techniques , Gold/chemistry , Metal Nanoparticles/chemistry , Primaquine/analysis , Antimalarials/urine , Dielectric Spectroscopy , Electrodes , Humans , Limit of Detection , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Primaquine/urine , Reproducibility of ResultsABSTRACT
BACKGROUND: Chloroquine and primaquine are the first-line treatment recommended by World Health Organization for malaria caused by Plasmodium vivax. Since the problem of counterfeit or substandard anti-malarials is well established all over the world, the development of rapid and reliable methods for quality control analysis of these drugs is essential. Thus, the aim of this study was to develop and validate a novel UPLC-DAD method for simultaneously quantifying chloroquine and primaquine in tablet formulations. METHODS: The UPLC separation was carried out using a Hypersil C18 column (50 × 2.1 mm id; 1.9 µm particle size) and a mobile phase composed of acetonitrile (A) and 0.1% aqueous triethylamine, pH 3.0 adjusted with phosphoric acid (B), at a flow rate 0.6 mL/min. Gradient elution was employed. UV detection was performed at 260 nm. UPLC method was fully validated and the results were compared to a conventional HPLC-DAD method for the analysis of chloroquine and primaquine in tablet formulations. RESULTS: UPLC method was shown to be linear (r2 > 0.99), precise (CV < 2.0%), accurate (recovery rates from 98.11 to 99.83%), specific, and robust. No significant differences were observed between the chloroquine and primaquine contents obtained by UPLC and HPLC methods. However, UPLC method promoted faster analyses, better chromatographic performance and lower solvent consumption. CONCLUSIONS: The developed UPLC method was shown to be a rapid and suitable technique to quantify chloroquine and primaquine in pharmaceutical preparations and may be successfully employed for quality control analysis.
Subject(s)
Antimalarials/analysis , Chemistry Techniques, Analytical/methods , Chloroquine/analysis , Chromatography, Liquid/methods , Primaquine/analysis , Tablets/chemistry , Quality Control , Time FactorsABSTRACT
BACKGROUND: Poor-quality life-saving medicines are a major public health threat, particularly in settings with a weak regulatory environment. Insufficient amounts of active pharmaceutical ingredients (API) endanger patient safety and may contribute to the development of drug resistance. In the case of malaria, concerns relate to implications for the efficacy of artemisinin-based combination therapies (ACT). In Papua New Guinea (PNG), Plasmodium falciparum and P. vivax are both endemic and health facilities are the main source of treatment. ACT has been introduced as first-line treatment but other drugs, such as primaquine for the treatment of P. vivax hypnozoites, are widely available. This study investigated the quality of antimalarial drugs and selected antibiotics at all levels of the health facility supply chain in PNG. METHODS AND FINDINGS: Medicines were obtained from randomly sampled health facilities and selected warehouses and hospitals across PNG and analysed for API content using validated high performance liquid chromatography (HPLC). Of 360 tablet/capsule samples from 60 providers, 9.7% (95% CI 6.9, 13.3) contained less, and 0.6% more, API than pharmacopoeial reference ranges, including 29/37 (78.4%) primaquine, 3/70 (4.3%) amodiaquine, and one sample each of quinine, artemether, sulphadoxine-pyrimethamine and amoxicillin. According to the package label, 86.5% of poor-quality samples originated from India. Poor-quality medicines were found in 48.3% of providers at all levels of the supply chain. Drug quality was unrelated to storage conditions. CONCLUSIONS: This study documents the presence of poor-quality medicines, particularly primaquine, throughout PNG. Primaquine is the only available transmission-blocking antimalarial, likely to become important to prevent the spread of artemisinin-resistant P. falciparum and eliminating P. vivax hypnozoites. The availability of poor-quality medicines reflects the lack of adequate quality control and regulatory mechanisms. Measures to stop the availability of poor-quality medicines should include limiting procurement to WHO prequalified products and implementing routine quality testing.
Subject(s)
Antimalarials/analysis , Biosimilar Pharmaceuticals/analysis , Malaria, Falciparum/drug therapy , Malaria, Vivax/drug therapy , Amodiaquine/analysis , Amodiaquine/standards , Amodiaquine/therapeutic use , Amoxicillin/analysis , Amoxicillin/standards , Amoxicillin/therapeutic use , Antimalarials/standards , Antimalarials/therapeutic use , Artemether , Artemisinins/analysis , Artemisinins/standards , Artemisinins/therapeutic use , Biosimilar Pharmaceuticals/standards , Biosimilar Pharmaceuticals/therapeutic use , Chloroquine/analysis , Chloroquine/standards , Chloroquine/therapeutic use , Drug Resistance , Health Facilities , Humans , Papua New Guinea , Primaquine/analysis , Primaquine/standards , Primaquine/therapeutic use , Quality Control , Quinine/analysis , Quinine/standards , Quinine/therapeutic useABSTRACT
Therapeutic efficiency and hemolytic toxicity of primaquine (PQ), the only drug available for radical cure of relapsing vivax malaria are believed to be mediated by its metabolites. However, identification of these metabolites has remained a major challenge apparently due to low quantities and their reactive nature. Drug candidates labeled with stable isotopes afford convenient tools for tracking drug-derived metabolites in complex matrices by liquid chromatography-tandem mass spectrometry (LC-MS-MS) and filtering for masses with twin peaks attributable to the label. This study was undertaken to identify metabolites of PQ from an in vitro incubation of a 1:1 w/w mixture of (13)C(6)-PQ/PQ with primary human hepatocytes. Acquity ultra-performance LC (UHPLC) was integrated with QTOF-MS to combine the efficiency of separation with high sensitivity, selectivity of detection and accurate mass determination. UHPLC retention time, twin mass peaks with difference of 6 (originating from (13)C(6)-PQ/PQ), and MS-MS fragmentation pattern were used for phenotyping. Besides carboxy-PQ (cPQ), formed by oxidative deamination of PQ to an aldehyde and subsequent oxidation, several other metabolites were identified: including PQ alcohol, predictably generated by oxidative deamination of PQ to an aldehyde and subsequent reduction, its acetate and the alcohol's glucuronide conjugate. Trace amounts of quinone-imine metabolites of PQ and cPQ were also detected which may be generated by hydroxylation of the PQ/cPQ quinoline ring at the 5-position and subsequent oxidation. These findings shed additional light on the human hepatic metabolism of PQ, and the method can be applied for identification of reactive PQ metabolites generated in vivo in preclinical and clinical studies.
Subject(s)
Chromatography, High Pressure Liquid/methods , Hepatocytes/chemistry , Hepatocytes/metabolism , Mass Spectrometry/methods , Primaquine/analysis , Primaquine/metabolism , Carbon Isotopes/analysis , Carbon Isotopes/chemistry , Carbon Isotopes/metabolism , Cells, Cultured , Humans , Isotope Labeling/methods , Primaquine/chemistryABSTRACT
Capillary electrophoresis (CE) methods for chiral resolution of five antimalarial drugs (primaquine, tafenoquine, mefloquine, chloroquine and quinacrine) were developed by using a wide selection of neutral and anionic cyclodextrin (CD) derivatives. The use of sulfobutyl-ß-CD and carboxymethyl-ß-CD (CMBCD) resulted in good resolution of quinacrine and tafenoquine, respectively. New results are presented for resolutions of chloroquine and mefloquine. Application of carboxyalkyl- and sulfobutyl-CD derivatives provided improved resolution for primaquine. The impurity in primaquine sample detected by CE was identified as quinocide by MS and NMR. CMBCD provided not only the best separation of primaquine from quinocide but also the simultaneous complete resolution of both compounds.
Subject(s)
Aminoquinolines/analysis , Antimalarials/analysis , Chloroquine/analysis , Mefloquine/analysis , Primaquine/analysis , beta-Cyclodextrins/chemistry , Anions , Cyclodextrins/chemistry , Electrophoresis, Capillary , Humans , Magnetic Resonance Spectroscopy , Mass Spectrometry , StereoisomerismABSTRACT
A sensitive electroanalytical methodology for the determination of chloroquine and primaquine using differential pulse voltammetry (DPV) at a Cu(OH)2 nano-wire-modified carbon paste electrode is presented. The cyclic voltammetric and DPV pulse voltammetric techniques are compared. The effects of scan rate and pH on current were investigated and an optimal scan rate of 50 mV s(-1) and a pH 5.5, 0.1 mol L(-1) phosphate buffer solution (PBS), were used. Additions of chloroquine and primaquine using DPV show linear ranges from 0.068 to 6.88 microg mL(-1) with a detection limit of 0.01 microg mL(-1) for chloroquine and 0.58-5.89 microg mL(-1) with a detection limit of 0.25 microg mL(-1) for primaquine. The method was then successfully utilized for the determination of chloroquine and primaquine in a real sample of their tablets and a recovery of 95% was obtained without interference from tablet matrix.
Subject(s)
Antimalarials/analysis , Electrochemical Techniques/instrumentation , Carbon , Chloroquine/analysis , Copper , Electrochemical Techniques/methods , Electrodes , Hydroxides , Nanowires , Primaquine/analysis , TabletsABSTRACT
Malaria is one of the most widespread and deadly diseases on the planet. Every year, about 500 million new cases are diagnosed, and the annual death toll is about 3 million. Primaquine has strong antiparasitic effects against gametocytes and can therefore prevent the spread of the parasite from treated patients to mosquitoes. It is also used in radical cures and prevents relapse. Consequently, primaquine is an often-used drug. In this study the separation of unprocessed primaquine from the contaminant quinocide based on gas chromatography-mass spectrometry with supersonic molecular beam (SMB) is presented and 7.5 mg primaquine diphosphate tablets were analyzed. We present a novel method for fast determination of quinocide which is an isomer of primaquine as the main contaminant in unprocessed primaquine and in its medical form as tablets by gas chromatography-mass spectrometry with SMB (also named supersonic GC-MS). Supersonic GC-MS provides enhanced molecular ion without any ion source related peak tailing plus extended range of compounds amenable for GC-MS analysis. In addition, major isomer mass spectral effects were revealed in the mass spectra of primaquine and quinocide which facilitated the unambiguous identification of quinocide in primaquine tablets. Fast GC-MS analysis is demonstrated with less then 2 min elution time of the drug and its main contaminants.
Subject(s)
Aminoquinolines/analysis , Antimalarials/analysis , Gas Chromatography-Mass Spectrometry/methods , Primaquine/analysis , Drug Contamination , Tablets/chemistryABSTRACT
With the objective of reducing analysis time and maintaining good efficiency, there has been substantial focus on high-speed chromatographic separations. Recently, commercially available ultra-performance liquid chromatography (UPLC) has proven to be one of the most promising developments in the area of fast chromatographic separations. In this work, a new isocratic reverse phase chromatographic method was developed using UPLC for primaquine phosphate bulk drug. The newly developed method is applicable for assay and related substance determination of the active pharmaceutical ingredient. The chromatographic separation of primaquine and impurities was achieved on a Waters Acquity BEH C18, 50 x 2.1mm, 1.7 microm column within a short runtime of 5 min. The method was validated according to the regulatory guidelines with respect to specificity, precision, accuracy, linearity and robustness. Forced degradation studies were also performed for primaquine phosphate bulk drug samples to demonstrate the stability indicating power of the UPLC method. Comparison of system performance with conventional HPLC was made with respect to analysis time, efficiency and sensitivity.
Subject(s)
Antimalarials/analysis , Chromatography, High Pressure Liquid/methods , Primaquine/analysis , Reproducibility of Results , Sensitivity and Specificity , Spectrophotometry, UltravioletABSTRACT
The drug primaquine diphosphate is used for causative treatment of malaria. Using HPLC-MS and GC-MS, this research group was previously able to show that the main contaminant of primaquine is the positional isomer quinocide [I. Brondz, D. Mantzilas, U. Klein, D. Ekeberg, E. Hvattum, M.N. Lebedeva, F.S. Mikhailitsyn, G.D. Soulimanov, J. Roe, J. Chromatogr. B: Anal. Technol. Biomed. Life Sci. 800 (2004) 211-223; I. Brondz, U. Klein, D. Ekeberg, D. Mantzilas, E. Hvattum, H. Schultz, F. S. Mikhailitsyn, Asian J. Chem. 17 (2005) 1678-1688]. Primaquine and quinocide are highly toxic substances which can have a number of side effects upon use in medical treatment. A standard for quinocide is not typically commercially available. In the present work, supercritical fluid chromatography-mass spectrometry (SFC-MS) with two different columns was used to achieve a shorter analysis time for the separation between the positional isomers quinocide and primaquine in primaquine diphosphate and to elucidate additional information about differences in their MS fragmentation. Unlike using HPLC-MS, it was possible to achieve the differential fragmentation of positional isomers at branching points using the SFC-MS technique. The desired short analysis time was achieved using SFC equipped with a Discovery HS F5 column and the differential fragmentation of positional isomers during SFC-MS provides information on the differences in the structure of these substances. Using a Chiralpak AD-H chiral column, it was possible to resolve the enantiomers in primaquine and separate quinocide from those enantiomers.
Subject(s)
Antimalarials/analysis , Primaquine/analysis , Aminoquinolines/analysis , Chromatography, High Pressure Liquid , Chromatography, Supercritical Fluid , Mass Spectrometry , Spectrometry, Mass, Electrospray Ionization , Spectrophotometry, Ultraviolet , StereoisomerismABSTRACT
An enantioselective capillary electrophoresis method for the simultaneous determination of primaquine (PQ) and carboxyprimaquine (CPQ) in rat liver mitochondrial fraction, suitable for in vitro metabolism studies is presented. The drug and metabolite were extracted by liquid-liquid extraction using ethyl ether. The enantiomers were resolved in a fused-silica capillary, 50 microm inside diameter (ID) and 24 cm of effective length, using an electrolyte solution consisting of a 20 mmol/L sodium phosphate solution, pH 3.0, and 10% w/v maltodextrin. Hydrodynamic sample injection was used with a 10 s injection time at 50 mbar pressure. The applied voltage was 22 kV and the capillary temperature was controlled at 20 degrees C. Detection was carried out at 264 nm. Under these conditions, the enantiomeric fractions of the drug and of its metabolite were analyzed within 6 min. The extraction procedure was efficient in removing endogenous interferents and low values (<10%) for the coefficients of variation and deviation from theoretical values were demonstrated for both within-day and between-day assays. The method described allows the determination of PQ and CPQ enantiomers as low as 100 and 40 ng/mL, respectively. After validation, the method was used for an in vitro metabolism study of PQ. The results showed that the enantiomer (-)-PQ was preferentially metabolized to (-)-CPQ.
Subject(s)
Electrophoresis, Capillary/methods , Primaquine/analogs & derivatives , Primaquine/analysis , Primaquine/chemistry , Animals , Buffers , Humans , Hydrogen-Ion Concentration , In Vitro Techniques , Liver/metabolism , Male , Molecular Structure , Polysaccharides , Primaquine/metabolism , Rats , Rats, Wistar , StereoisomerismABSTRACT
The main contaminant of primaquine (CAS 90-34-6) has been tentatively identified, by using two liquid chromatography (LC) methods and liquid chromatography-mass spectrometry (LC-MS), as the positional isomer quinocide (CAS 525-61-1). The first LC system was equipped with a chiral Chirex (S)-VAL and (R)-NEA column and the second system was equipped with an Adsorbosphere Nucleotide-Nucleoside 7 micro column. Comparison of the main contaminant of primaquine with an authentic quinocide standard by using co-chromatography in both LC systems and LC-MS (mass fragmentation) supported the hypothesis. The toxicity of quinocide batch 17172, primaquine batch 16039, and the drug primaquine diphosphate batch 20107 used in pharmaceutical industry, and the effect of the substances on respiratory and electron transport chain were compared in the eucaryotic unicellular fresh water green alga Chlamydomonas reinhardtii as a model system. These studies suggest that minor amount of other related substances can contribute more to the toxicity of the drug primaquine diphosphate than the positional isomer quinocide.
Subject(s)
Antimalarials/analysis , Drug Contamination , Primaquine/analysis , Aminoquinolines/analysis , Aminoquinolines/toxicity , Animals , Antimalarials/toxicity , Biological Assay , Chlamydomonas reinhardtii/drug effects , Chlamydomonas reinhardtii/growth & development , Chlamydomonas reinhardtii/metabolism , Chlorophyll/analysis , Chlorophyll/metabolism , Chromatography, High Pressure Liquid , Circular Dichroism , Electrodes , Electron Transport/drug effects , Light , Mass Spectrometry , Oxygen/chemistry , Oxygen/metabolism , Photosynthesis/drug effects , Primaquine/toxicity , Reference Standards , Spectrophotometry, Ultraviolet , StereoisomerismABSTRACT
A combination Kit for antirelapse treatment of P. vivax malaria, consisting of Chloroquine phosphate tablets and Bulaquine capsules has been recently developed, and marketed under the trade name Aablaquine. Bulaquine is prepared from Primaquine. Several methods of analysis are reported for each drug separately as well two drugs in combination but no method for simultaneous estimation of these three drugs is known. Therefore, the present study was undertaken to develop a sensitive and reproducible high performance liquid chromatographic as well as high performance thin layer chromatographic assay method for the simultaneous estimation of Chloroquine, Primaquine and Bulaquine.
Subject(s)
Antimalarials/analysis , Chloroquine/analysis , Primaquine/analogs & derivatives , Primaquine/analysis , Chromatography, High Pressure Liquid/methods , Chromatography, Thin Layer/methodsABSTRACT
A new simple, selective and reproducible high-performance liquid chromatographic method for the determination of quinine in plasma, saliva and urine is described. The ion-pair method was carried out on a reversed-phase C18 column, using perchlorate ion as the counter ion and ultraviolet detection at 254 nm. Quinine was well resolved from its major metabolite, 3-hydroxyquinine, and the internal standard, primaquine. The limit of detection was 10 ng/ml and the recovery was greater than 90% from the three biological fluids.
Subject(s)
Quinine/analysis , Saliva/chemistry , Chromatography, High Pressure Liquid , Humans , Indicators and Reagents , Primaquine/analysis , Primaquine/blood , Primaquine/urine , Quinidine/analogs & derivatives , Quinidine/analysis , Quinidine/blood , Quinidine/urine , Quinine/blood , Quinine/urine , Spectrophotometry, UltravioletABSTRACT
A number of drugs have been studied for their effect on the metabolism of the antimalarial drug primaquine by human liver microsomes (N = 4) in vitro. The only metabolite generated was identified as carboxyprimaquine by co-chromatography with the authentic standard. Ketoconazole, a known inhibitor of cytochrome P450 isozymes, caused marked inhibition of carboxyprimaquine formation with IC50 and K(i) values of 15 and 6.7 microM, respectively. This finding and the dependency of metabolite formation on NADPH indicates that cytochrome P450 isozyme(s) catalysed metabolite production. Of compounds actually or likely to be coadministered with primaquine to malaria patients, only mefloquine produced any inhibition (K(i) = 52.5 microM). Quinine, artemether, artesunate, halofantrine and chloroquine did not significantly inhibit metabolite formation. It seems unlikely that the concurrent administration of mefloquine, or other antimalarials, with primaquine will lead to appreciably altered disposition.
Subject(s)
Antimalarials/pharmacology , Microsomes, Liver/metabolism , Primaquine/analogs & derivatives , Primaquine/metabolism , Cytochrome P-450 Enzyme Inhibitors , Cytochrome P-450 Enzyme System/metabolism , Drug Interactions , Humans , Isoenzymes/antagonists & inhibitors , Isoenzymes/metabolism , Kinetics , Microsomes, Liver/drug effects , NADP/metabolism , Primaquine/analysisABSTRACT
Two methods are described for the determination of amodiaquine in pharmaceuticals. In the first method, amodiaquine was treated with periodate at pH 7.5-8.5 to yield a chromogen, which is believed to be a 1,4-benzoquinone imine derivative, and extracted into chloroform for spectrophotometric assay. The maximum molar absorptivity was found to be 2.17 x 10(3) l mol-1 cm-1 at 442 nm. Primaquine and chloroquine, if present also, do not interfere. The second method, which was used for the simultaneous determination of amodiaquine, primaquine and chloroquine, is based on high-performance liquid chromatography on an octadecylsilane column using a mobile phase of methanol-water (80 + 20) containing 0.5% anhydrous acetic acid and an over-all concentration of sodium dodecyl sulphate of 0.5 mM, with detection at 340 nm.