Liquid chromatographic determination of coumarin anticoagulants in tablets

A simple and rapid liquid chromatographic method is described for the qualitative and quantitative determination of 5 coumarin anticoagulants in tablet composites and individual tablets. Analyses are carried out on a C18 reverse phase column using tetrahydrofuran-methanol-water-acetic acid (35 + 10 + 65 + 0.1) as mobile phase and photometric detection at 311 nm. The coefficients of variation for 10 consecutive injections of a mixed standards solution ranged from 0.28% for ethyl biscoumacetate to 0.78% for acenocoumarol. Standard recoveries were as follows: acenocoumarol, 99.3%; dicumarol, 99.6%; phenprocoumon, 101.6%; and warfarin sodium, 99.0%. The method was linear between 2 and 8 micrograms of drug injected. Assay results agreed favorably with those of the USP XX methods for dicumarol, phenprocoumon, and warfarin, and the NF XIV method for acenocoumarol. In addition, close correspondence was found with the results previously reported for the same drugs by a semiautomated spectrophotometric method. The content uniformity testing of individual 50 mg dicumarol tablets and 5 mg warfarin sodium tablets by the proposed method gave average (SD) values of 100.32% (0.64) and 101.00% (0.14), respectively, whereas these values were 101.60% (1.81) and 101.80% (0.18), respectively, by the method of USP XX.     

Collaborative study of a semiautomated method for the analysis of acenocoumarol, phenprocoumon, and potassium warfarin tablets.

This collaborative study was undertaken to determine if the anticoagulants acenocoumarol, phenprocoumon, and potassium warfarin could be analyzed by the automated analysis system described in the collaborative study for the analysis of sodium warfarin and dicumarol. Collaborators were supplied with a composited tablet sample of each anticoagulant. Results agreed well with the National Formulary methods for phenprocoumon and potassium warfarin, and an unpublished method for acenocoumarol. For acenocoumarol, coefficients of variation on individual sets of data ranged from 0.30 to 1.94% For phenprocoumon, coefficients of variation ranged from 0.52 to 1.20%. For potassium warfarin, coefficients of variation ranged from 0.54 to 1.79%. The results of this study show that acenocoumarol, phenprocoumon, and potassium warfarin can be analyzed by the official AOAC method for the analysis of sodium warfarin and dicumarol tablets.     

Determination of fluoride in fluoride tablets and solutions by ion-selective electrode: collaborative study

A proposed method using the fluoride (F) ion-selective electrode has been developed for determining the fluoride ion concentration in tablets and solutions containing sodium fluoride. The method has been subjected to collaborative study. Eight samples consisting of 2 authentic fluoride solutions, 2 commercial fluoride solutions, and 4 commercial fluoride tablets were sent to each of 11 collaborators together with a copy of the method. Single assays on the authentic fluoride solutions known to contain 1 mg F/5 mL were performed with average recoveries of 99.5 and 99.6% and relative coefficients of variation (CV) of 2.11 and 1.91%, respectively. Single assays of 2 commercial fluoride solutions declared at 1 mg F/5 mL gave mean values of 0.994 and 0.990 mg and relative CV values of 1.88 and 2.36%, respectively. Single assays of 2 commercial fluoride tablet preparations declared at 0.5 mg F gave mean values of 0.485 and 0.478 mg and relative CV values of 3.12 and 3.71%, respectively. Single assays of 2 commercial fluoride tablet preparations declared at 1 mg F gave mean values of 0.991 and 0.981 mg and relative CV values of 2.99 and 2.85%, respectively. The method has been adopted official first action.     

UV spectrophotometric determination of hydralazine hydrochloride in tablets: collaborative study

A UV spectrophotometric method for the determination of hydralazine hydrochloride in tablets was collaboratively studied by 5 laboratories. The method is based on conversion of hydralazine to a tetrazolo [5,1-alpha]phthalazine derivative which shows an absorption maximum at about 274 nm. Each collaborator received blind duplicate samples of 2 commercial powdered composites from 10 and 100 mg tablets, and 1 synthetic tablet formulation. Each collaborator also received a set of 10 tablets for determination of content uniformity. The pooled mean recovery of hydralazine hydrochloride from the synthetic formulation was 101.2 +/- 0.94%. The mean assay values for 10 and 100 mg tablets were 95.6 +/- 0.98 and 101.0 +/- 0.73% of the declared amounts, respectively, with corresponding CV values of 1.02 and 0.73%. The pooled mean for individual tablet assay was 99.8 +/- 3.26% of the declared value, with a CV of 3.29%. The method has been adopted official first action.     

Normal phase liquid chromatographic determination of sulfamethoxazole in tablets: collaborative study

A stability-indicating liquid chromatographic method is presented for determining sulfamethoxazole in tablets. The method uses a 10 micron silica column, an isooctane-methylene chloride-2-propanol-acetonitrile-glacial acetic acid (70 + 25 + 5 + 5 + 0.5) mobile phase, and photometric detection at 254 nm. Seven laboratories collaboratively studied this method on powdered composite samples prepared from commercial 500 and 1000 mg tablets and on an authentic tablet mixture containing 83.32% added sulfamethoxazole. Mean assay results for the 500 and 1000 mg tablets were 102.2 and 97.9% of declared, respectively (n = 4). The mean recovery value for the synthetic sample was 99.4% (n = 4). The pooled reproducibility standard deviation (SD) (coefficient of variation (CV)) and pooled repeatability SD (CV) were +/- 1.01 (1.01%) and +/- 0.96 (0.96%), respectively. These results were in good agreement with those obtained by the Associate Referee for the titration method of USP XX. The proposed method can also be used for monitoring the presence of sulfanilamide in sulfamethoxazole by increasing the proportions of both acetonitrile and 2-propanol in the mobile phase. The method has been adopted official first action.     

Liquid chromatographic determination of acetaminophen in tablets: collaborative study

A reverse phase liquid chromatographic method previously reported for the determination of acetaminophen in tablets was collaboratively studied by 5 laboratories. Each collaborator received duplicate samples of a synthetic tablet formulation and 3 powdered commercial tablet composites. The composites represented single-component and multi-component proprietary products and a single-component generic product. The pooled repeatability (CVDo) and reproducibility (CVDx) values for the proprietary tablets were 0.89 and 1.34%, respectively. For the generic tablets, these values were 0.66 and 0.74%, respectively. The pooled recovery value for acetaminophen added to the synthetic formulation was 98.9 +/- 0.7% (n = 10) with a CV of 0.75%, CVDo of 0.37%, and CVDx of 0.78%. The overall repeatability of the method was 0.64%, and the overall reproducibility was 0.95%. The method has been adopted official first action.     

Liquid chromatographic determination of levodopa and levodopa-carbidopa in solid dosage forms: collaborative study

A liquid chromatographic method for the determination of levodopa in tablets and capsules and levodopa-carbidopa in tablets was collaboratively studied by 6 laboratories. Collaborators were supplied with duplicate powdered composites of levodopa (1 synthetic formulation, 1 commercial tablet, and 1 commercial capsule) and levodopa-carbidopa (1 synthetic formulation and 2 commercial tablets), along with individual levodopa-carbidopa tablets for content uniformity determinations. The repeatability coefficient of variation (CVo) and reproducibility coefficient of variation (CVx) for levodopa single component were 0.48 and 0.87%; for levodopa in combination, 0.50 and 0.90%; and for carbidopa, 0.77 and 1.20%, respectively. Overall, the recovery values for levodopa and carbidopa from synthetic formulations simulating tablets were 100.4 and 99.5%, respectively. The pooled CVDo and CVDx values for the individual tablet assays were 2.07 and 2.30% for levodopa, and 1.80 and 2.24% for carbidopa, respectively. The method has been adopted official first action for determination of the active ingredients in levodopa tablets and capsules and in levodopa-carbidopa tablets and for content uniformity testing in the combination dosage form.     

Liquid chromatographic determination of penicillin V potassium in tablets: collaborative study

A liquid chromatographic method for the determination of penicillin V potassium in tablets was collaboratively studied by 7 laboratories. The method uses an octadecyl silane reverse-phase column, a mobile phase of acetonitrile-methanol-0.01 M potassium phosphate monobasic (21 + 4 + 75 v/v/v), photometric detection at 225 nm, and sulfadimethoxine as an internal standard. Each collaborator received 6 samples: powdered composites of 2 commercial tablet preparations and 1 synthetic tablet powder mixture, each with blind duplicates. The mean repeatability and reproducibility relative standard deviations for commercial samples were, respectively, 1.10 and 1.46% (250 mg dosage), and 0.84 and 2.82% (500 mg dosage). The average standard recovery from the synthetic formulation was 99.1%, with repeatability and reproducibility relative standard deviations of 1.30 and 3.66%, respectively. The method has been adopted official first action.     

Liquid chromatographic determination of methyldopa and methyldopa-thiazide combinations in tablets: collaborative study

A reverse phase liquid chromatographic method for the determination of methyldopa, methyldopa-hydrochlorothiazide, and methyldopachlorothiazide in tablets was collaboratively studied by 8 laboratories. Each collaborator received 20 samples that included drug substance, synthetic and commercial tablet compositions. The overall repeatability and reproducibility standard deviations for commercial tablets were 1.11 and 1.75% for methyldopa, 0.96 and 1.62% for chlorothiazide, and 1.21 and 2.15% for hydrochlorothiazide, respectively. The overall recoveries of methyldopa, chlorothiazide, and hydrochlorothiazide added to synthetic tablets were 100.78, 100.70, and 101.34%, respectively. The method has been adopted official first action.     

Liquid chromatographic determination of diazepam in tablets: collaborative study

A stability indicating liquid chromatographic method for the determination of diazepam in tablets was collaboratively studied by 6 laboratories. The method uses a C18 reverse phase column, a methanol-water mobile phase, p-tolualdehyde as the internal standard, and photometric detection at 254 nm. The collaborators were supplied with a synthetic tablet powder and 3 commercial tablet samples. The mean recovery of diazepam from the synthetic tablet powder was 100.2%. For all samples analyzed, the coefficient of variation was less than 1.5%. The method has been adopted official first action.     

Liquid chromatographic determination of flucytosine in capsules: collaborative study

A liquid chromatographic method for the determination of flucytosine in capsules was collaboratively studied by 7 laboratories. The method uses a C18 reverse phase column, water-methanol-acetic acid mobile phase containing 1-octanesulfonic acid sodium salt, p-aminobenzoic acid as internal standard, and photometric detection at 285 nm. The mean recovery value (+/- SD) of flucytosine from a synthetic formulation representing capsules was 99.2 +/- 1.72% (CV = 1.73%). Composited samples of 250 and 500 mg commercial capsules gave assay values of (mean +/- SD) 103.17 +/- 2.21 and 99.29 +/- 1.29% of declared, respectively. CV values were 2.15 and 1.30%. Reproducibility and repeatability CVs were 2.19 and 1.50%, respectively, for the 250 mg capsules, and 1.34 and 0.63%, respectively, for the 500 mg capsules. The method has been adopted official first action.     

High pressure liquid chromatographic determination of antihistamine-adrenergic combination products: collaborative study

A reverse phase high pressure liquid chromatographic method in which ion-pairing is used for the determination of combinations of pseudoephedrine hydrochloride with triprolidine hydrochloride or chlorpheniramine maleate in syrups and tablets was collaboratively studied by 8 laboratories. Collaborators were supplied with 12 samples including synthetic and commercial syrup formulations and commercial tablet composites. Mean recoveries of pseudoephedrine hydrochloride and triprolidine hydrochloride from synthetic syrup formulations were 100.5 and 99.6%, respectively. Mean recoveries of pseudoephedrine hydrochloride and chlorpheniramine maleate from synthetic syrups were 98.8 and 100.5%, respectively. Mean coefficients of variation for syrups and tablets ranged from 1.68 to 3.07% for pseudoephedrine hydrochloride, from 2.92 to 3.85% for triprolidine hydrochloride, and from 1.34 to 2.15% for chlorpheniramine maleate. The method has been adopted official first action.     

Proton magnetic resonance spectroscopic determination of bethanechol chloride in tablets

A simple and reliable proton magnetic resonance spectroscopic method was developed for the assay of bethanechol chloride in tablets. The proposed method entails a minimum of procedural steps and reagents. The recovery of bethanechol chloride from synthetic formulations was 99.9 +/- 0.4% (n = 15), CV = 0.4%. Mean assay values for 25 and 10 mg commercial tablets were 100.5% (n = 8) and 99.7% (n = 7) of declared, respectively. A detailed interpretation of the proton magnetic resonance spectrum is also presented.     

High pressure liquid chromatographic determination of oxazepam dosage forms: collaborative study

A reverse phase high pressure liquid chromatographic method for the determination of oxazepam in tablets and capsules was collaboratively studied by 9 laboratories. Collaborators were supplied with 6 samples that included synthetic and commercial formulations. Tablet and capsule composites are diluted with methanol and filtered. Oxazepam is determined at 254 nm by using a C18 column. Mean recoveries of oxazepam from synthetic tablet and capsule formulations were 97.2 and 99.0%, respectively. Mean coefficients of variation for tablets and capsules ranged from 1.85 to 2.86%. The method has been adopted official first action.     

Atomic absorption spectrophotometric determination of mercury in mercury-containing drugs: collaborative study

An atomic absorption spectrophotometric (AAS) method applicable to a wide variety of mercury-containing drugs has been developed and subjected to a collaborative study. Samples were digested with a water-HCl-HNO3 (4 + 3 + 1) mixture, and the mercury was determined in solution by AAS. High levels of mercury were measured with a conventional air-acetylene flame, whereas low levels were measured by the flameless technique. Each of 7 collaborators received duplicate synthetic samples of a tincture, an ophthalmic solution, and an antiseptic solution, and duplicate commercial samples of an ointment and an injectable. The overall mean value found by collaborators for mercury in these samples was 100.13%. The corresponding overall repeatability SD (CV, %) and reproducibility SD (CV, %) values were 2.18 (2.18) and 3.38 (3.38), respectively. The proposed AAS method has been adopted official first action.     

Ultraviolet spectrophotometric determination of hydralazine hydrochloride in tablets following derivatization with nitrite

Routine use of the USP XXI spectrophotometric method for the content uniformity determination of hydralazine hydrochloride tablets has shown that tablet excipients can significantly alter the spectral characteristics of the drug and thus cause inaccurate assay values to be obtained. Because of this problem, a simple and reliable alternative spectrophotometric assay method, based on the conversion of hydralazine to tetrazolo [5,1-alpha]phthalazine with nitrite ions under acidic conditions, was developed. The derivative showed an absorption maximum at about 274 nm and obeyed Beer's law over the concentration range 4-40 micrograms/mL. Mean recoveries of hydralazine hydrochloride added to commercial coated and uncoated tablets were 101.0% (n = 10) and 100.8% (n = 8), respectively. The proposed method was found suitable for the assay not only of individual tablets but also of tablet composites.     

Liquid chromatographic determination of carbamazepine in tablets

A simple and rapid liquid chromatographic method is described for the qualitative and quantitative determination of carbamazepine in tablet composites and individual tablets, using the internal standard technique. Analyses were performed on a C-18 reverse-phase column with tetrahydrofuran-methanol-water (8 + 37 + 55) as the mobile phase. A linear relationship was obtained between detector responses at 254 nm and amounts of carbamazepine injected ranging from 0.2 to 1.7 micrograms. The coefficient of variation for 10 consecutive injections of a standard preparation was 0.4%. Recoveries of carbamazepine from 100 and 200 mg tablets averaged 101.4 and 99.7%, respectively. Assay results for commercial tablets analyzed by the proposed method agreed favorably with those obtained by the method of USP XXI. The assay results for individual tablets indicated that deviations from the average value and the range of individual values are much wider with the compendial method than with the proposed method.     

Collaborative study of the analysis of mestranol in combination with norethindrone or norethynodrel

A collaborative study of the analysis of mestranol in combination with norethindrone or norethynodrel by column chromatography and ultraviolet determination is presented. The study consisted of 4 samples which were a commercial norethindrone tablet, a synthetic norethindrone mixture, a commercial norethynodrel tablet, and a synthetic norethynodrel mixture. The samples were analyzed by 13 collaborators. Average mestranol recoveries were 100.4, 99.9, 98.7, and 101.2%, respectively.     

Collaborative study of a spectrofluorometric assay for Rauwolfia serpentina tablets and powdered root.

Reserpine-rescinnamine group alkaloids are extracted from Rauwolfia serpentina preparations into a dimethylsulfoxide (DMSO)-methanol mixture and diluted with 0.5N H2SO4. The chloroform extract of this solution is passed through a 0.1N NaOH-Celite column and then through a silica gel column. The weakly basic alkaloids trapped on the latter column are eluted with a methanol mixture; a portion of the eluate is treated with nitrous acid and the reserpine-rescinnamine content is determined by measuring the intensity of fluorescence of the oxidation product. The following means and standard deviations (11 collaborators) were obtained for the determination of reserpine-rescinnamine group alkaloids in 4 samples of Rauwolfia serpentina (NF reference powder, 100 mg and 50 mg commercial tablets, and a 45 mg synthetic tablet formulation) : 0.174% +/- 0.0112, 0.131% +/- 0.0047, 0.160% +/- 0.0100, and 0.153% +/- 0.0083, respectively.     

Reverse phase liquid chromatographic determination of bisacodyl in dosage forms

A method is described for the determination of bisacodyl in enteric-coated tablets and suppositories by liquid chromatography (LC). The method will also determine the hydrolysis degradation products monoacetylbisacodyl and desacetylbisacodyl. The sample is dissolved in 2-propanol, and the extract is diluted with the mobile phase and injected into a liquid chromatograph fitted with a mu Bondapak C18 column and an ultraviolet detector set at 254 nm. The column is eluted with methanol-acetonitrile-0.01M citric acid (25 + 25 + 50). The pooled mean recovery value for bisacodyl from commercial enteric-coated tablets and suppositories was 99.7% with a pooled coefficient of variation (CV) of 0.72%. For content uniformity assays, the CVs were 0.7 and 1.0% for groups of 10 individual commercial suppositories and tablets, respectively. Differences between assay values by the LC and USP XX methods were 0.2% of declared for enteric-coated tablets (n = 5) and 1.0% of declared for suppositories (n = 2). The LC method can determine as little as 0.015 microgram of the monoacetyl or desacetyl degradation product.