An eco-friendly liquid chromatographic analysis of the triple therapy protocol of amoxicillin, metronidazole and vonoprazan for H. Pylori eradication: application to combined dosage forms and simulated gastric fluid.

Helicobacter pylori has a big sway when peptic ulcers are concerned. For its eradication, different protocols have been approved. Among which, the tripartite therapy protocol which embraces vonoprazan as potassium competitive acid blocker in combination with amoxicillin and metronidazole as antibiotics. An environmentally benign HPLC method is addressed in order to simultaneously determine amoxicillin (AMX), metronidazole (MET) and vonoprazan (VPZ) in bulk powder and combined tablet mixture. Full separation of AMX, MET and VPZ is accomplished using C8 column, and a gradient mobile phase system, composed of methanol and phosphate buffer of a pH value of 5. Fine linearity in the concentration ranges 50-600 µg mL-1 amoxicillin, 50-400 µg mL-1 metronidazole and 10-100 µg mL-1 vonoprazan was denoted by the high correlation coefficient (0.9999). The method accuracy and precision are confirmed upon analyzing AMX, MET and VPZ triple therapy not only in their synthetic mixtures and combined tablet mixtures but also in their combined tablet mixtures in simulated gastric fluid. AMX, MET and VPZ triple therapy could be routinely analyzed in QC labs, in case of being co-formulated, using the presented method. Three different assessment tools were adopted revealing the benign environmental impact of presented method.

An eco-friendly multi-analyte high-performance thin layer chromatographic densitometric determination of amoxicillin, metronidazole, and famotidine in their ternary mixtures and simulated gastric juice: A promising protocol for eradicating Helicobacter pylori.

Gastrointestinal tract disorders constitute a heavy burden to healthcare providers. To eradicate Helicobacter pylori infection, different triple therapy protocols have been proposed. Among which are combinations of proton pump inhibitors (e.g., omeprazole), histamine-2 receptor antagonists (e.g., famotidine), along with antibiotics (e.g., amoxicillin). In this work, a sensitive and accurate high-performance thin-layer chromatographic method was developed for the simultaneous determination of amoxicillin, metronidazole, and famotidine in bulk powder and laboratory-prepared combined-tablet mixtures. Complete separation of the cited compounds was achieved using pre-coated silica gel plates with a mixture of methanol:chloroform:toluene:water:glacial acetic acid (5:2:1.5:0.5:0.1 v/v/v/v/v) as the mobile phase. The method was fully validated as per the international conference of harmonization guidelines. Good linearity, a correlation coefficient of 0.9991, was obtained in the concentration ranges 0.1-1.6 µg/band (amoxicillin), 0.1-0.9 µg/band (metronidazole), and 0.1-0.9 µg/band (famotidine). Since the method allowed the determination of the three compounds in combined tablets with a high degree of selectivity, accuracy, precision, with cost-effectiveness, it could be used for regular quality control. Moreover, the applicability of the proposed method was extended to the determination of the ternary mixture in simulated gastric juice. Method greenness was assessed using different green metrics.

Chemometrics-assisted spectrophotometric green method for correcting interferences in biowaiver studies: Application to assay and dissolution profiling study of donepezil hydrochloride tablets.

A green, simple and cost effective chemometric UV-Vis spectrophotometric method has been developed and validated for correcting interferences that arise during conducting biowaiver studies. Chemometric manipulation has been done for enhancing the results of direct absorbance, resulting from very low concentrations (high incidence of background noise interference) of earlier points in the dissolution timing in case of dissolution profile using first and second derivative (D1 & D2) methods and their corresponding Fourier function convoluted methods (D1/FF& D2/FF). The method applied for biowaiver study of Donepezil Hydrochloride (DH) as a representative model was done by comparing two different dosage forms containing 5mg DH per tablet as an application of a developed chemometric method for correcting interferences as well as for the assay and dissolution testing in its tablet dosage form. The results showed that first derivative technique can be used for enhancement of the data in case of low concentration range of DH (1-8µgmL-1) in the three different pH dissolution media which were used to estimate the low drug concentrations dissolved at the early points in the biowaiver study. Furthermore, the results showed similarity in phosphate buffer pH6.8 and dissimilarity in the other 2pH media. The method was validated according to ICH guidelines and USP monograph for both assays (HCl of pH1.2) and dissolution study in 3pH media (HCl of pH1.2, acetate buffer of pH4.5 and phosphate buffer of pH6.8). Finally, the assessment of the method greenness was done using two different assessment techniques: National Environmental Method Index label and Eco scale methods. Both techniques ascertained the greenness of the proposed method.

Green gas chromatographic stability-indicating method for the determination of Lacosamide in tablets. Application to in-vivo human urine profiling.

A direct, eco-friendly, stability-indicating GC method was developed for the determination of Lacosamide (LCM) in tablet dosage forms in presence of its degradation products as well as in human urine in presence of the co-administered drug Zonisamide (ZON). The assay method in tablets was validated according to the ICH guidelines, while the method for determination of LCM in urine was validated according to FDA; Bioanalytical Method Validation guidance. Linear response (r = 0.9998) was observed over the range of 20-280 µg/mL of LCM, with detection and quantitation limits of 5.871 and 19.57 µg/mL, respectively for the tablet assay method. While (r = 0.9999) was observed over the range of 0.5-20 µg/mL of LCM, with detection and quantitation limits of 67 and 233 ng mL-1, respectively for the urine analysis method. Under various stress conditions, the investigation of LCM forced degradation behaviour was carried out. Furthermore, monitoring of the drug in urine followed by construction of its urine profile was done after the administration of 50 mg tablet of LCM to three healthy volunteers so as to prove the ability of the method to be applied in assaying LCM in human urine. The method showed also successful separation of LCM and the co-administered drug ZON in urine. Finally, the greenness of the method was assessed using National Environmental Methods Index label and Eco scale methods.

A Validated HPLC Method for the Determination of Linagliptin in Rat Plasma. Application to a Pharmacokinetic Study.

A sensitive and reproducible HPLC method for the determination of linagliptin (LNG) in rat plasma was developed and validated using pindolol (PIN) as the internal standard. Both LNG and PIN were separated on a Zorbax Eclipse XDB C18 column kept at ambient temperature using as mobile phase a combination of 75% methanol: 25% formic acid 0.1% pH 4.1 at a flow rate of 1.0 mL min-1. UV detection was performed at 254 nm. The method was validated in compliance with ICH guidelines and found to be linear in the range of 5-1,000 ng mL-1. The limit of quantification (LOQ) was found to be 5 ng mL-1 based on 100 µL of plasma. The variations for intra- and inter-assay precision were <10%, and the accuracy values were ranged between 93.3 and 102.5%. The extraction recovery (R%) was >83%. The assay was successfully applied to an in vivo pharmacokinetic study of LNG in rats that were administered a single oral dose of 10 mg kg-1 LNG. The maximum concentration (Cmax) and the area under the plasma concentration-time curve (AUC0-72) were 927.5 ± 23.9 and 18,285.02 ± 605.76 ng mL-1, respectively.

Development and validation of spectrophotometric and HPTLC methods for simultaneous determination of rosiglitazone maleate and metformin hydrochloride in the presence of interfering matrix excipients.

Two simple methods have been developed and validated for the simultaneous determination of rosiglitazone maleate (ROS) and metformin hydrochloride (MET) in synthetic mixtures and coated tablets in a ratio of 1:250 (ROS:MET). The first method was a spectrophotometric one. The minor component, ROS was determined by measuring the values of absorbance at λmax 312 nm and the D1 amplitudes at 331 nm where MET shows no absorption contribution. However, absorbance interferences from tablet excipients were successfully corrected by D1 at 331 nm zero-crossing technique. Study of spectral interference from tablet excipients was included in the text. Standard curves for Amax and D1 methods were in the concentration range 20.0-80.0 µg mL(-1). The major component, MET was determined both in binary mixtures and tablets by measuring its Amax at 236 nm. Extensive dilution eliminated any absorption contribution from the coexisting ROS or tablet matrix. Standard curves showed linearity in the concentration range 4.0-12.8 µg mL(-1). The second method was based on high performance thin layer chromatography (HPTLC) separation of the two drugs followed by densitometric measurements of their spots at 230 nm. The separation was carried out on Merck HPTLC aluminium sheets of silica gel 60 F254 using methanol:water:NH4Cl 1% w/v (5:4:1 v/v/v) as the mobile phase. Linear calibration graphs of peak area values were obtained versus concentrations in the range of 0.4-2.0 µg band(-1) and 20.0-100.0 µg band(-1) for ROS and MET, respectively. According to International Conference on Harmonisation (ICH) guidelines, different validation parameters were verified for the two methods and presented.

Sucralose.

Sucralose is a nonnutritive, zero-calorie artificial sweetener. It is a chlorinated sugar substitute that is about 600 times as sweet as sucrose. It is produced from sucrose when three chlorine atoms replace three hydroxyl groups. It is consumed as tablets (Blendy) by diabetic and obese patients. It is also used as an excipient in drug manufacturing. Unlike other artificial sweeteners, it is stable when heated and can, therefore, be used in baked and fried foods. The FDA approved sucralose in 1998. This review presents a comprehensive profile for sucralose including physical, analytical, and ADME profiles and methods of its synthesis. Spectral data for X-ray powder diffraction and DSC of sucralose are recorded and presented. The authors also recorded FT-IR, (1)H- and (13)C NMR, and ESI-MS spectra. Interpretation with detailed spectral assignments is provided. The analytical profile of sucralose covered the compendial methods, spectroscopic, and different chromatographic analytical techniques. The ADME profile covered all absorption, distribution, metabolism, and elimination data in addition to pharmacokinetics and pharmacological effects of sucralose. Some nutritional aspects for sucralose in obesity and diabetes are also presented. Both chemical and microbiological synthesis schemes for sucralose are reviewed and included.

Kinetic spectrophotometric methods for the determination of artificial sweetener (sucralose) in tablets.

Two simple and sensitive kinetic spectrophotometric methods for the determination of sucralose are described. The first method is based upon a kinetic investigation of the oxidation reaction of the drug with alkaline potassium permanganate at room temperature for a fixed time of 30 min. The absorbance of the green coloured manganate ions produced was measured at 610 nm. The second method is based on the reaction of sucralose with cerium (IV) ammonium sulfate in the presence of perchloric acid with the subsequent measurement of the excess unreacted cerium (IV) ammonium sulfate at 320 nm at a fixed time of 30 min in a thermostated water bath at 60 ± 1 °C. This principle is adopted to develop a kinetic method for sucralose determination. The absorbance concentration plots in both methods were rectilinear over the range 4-16 and 10-30 µg ml(-1) , for the first and second methods, respectively. The different experimental parameters affecting the development and stability of the colours were carefully studied and optimized. The determination of sucralose by rate constant method, fixed concentration method, and fixed-time method was also feasible with calibration equations obtained but the latter method was found to be more applicable. The two methods have been applied successfully to commercial tablets.

Differential pulse, square wave and adsorptive stripping voltammetric quantification of tianeptine in tablets.

Differential pulse polarographic (DPP) and square wave polarographic (SWP) techniques were applied at hanging mercury drop electrode (HMDE) for quantitative determination of tianeptine (TIA) in tablets. The adsorptive stripping voltammetric (ASV) behavior of TIA was also studied. TIA gave a sensitive reduction peaks at -1256, -1244 and -1072 mV for DPP, SWP and ASV, respectively (versus Ag/AgCl) in Britton-Robinson buffer (B-R buffer) at pH 11. The solution conditions and instrumental parameters were optimized for the determination of TIA in tablets. Calibration plots and regression data validation, accuracy, precision, limit of detection, limit of quantitation and other aspects of analytical merit are presented.

Non-parametric linear regression of discrete Fourier transform convoluted chromatographic peak responses in non-ideal conditions.

This manuscript discusses the application of chemometrics to the handling of HPLC response data using a model mixture containing ascorbic acid, paracetamol and guaiphenesin. Derivative treatment of chromatographic response data followed by convolution of the resulting derivative curves using 8-points sinx(i) polynomials (discrete Fourier functions) was found beneficial in eliminating different types of interferences. This was successfully applied to handle some of the most common chromatographic problems and non-ideal conditions, namely: very low analyte concentrations, overlapping chromatographic peaks and baseline drift. For example, a significant change in the correlation coefficient of guaiphenesin, in case of baseline drift, went from 0.9978 to 0.9998 on applying normal conventional peak area and first derivative under Fourier functions methods, respectively. It also compares the application of Theil's method, a non-parametric regression method, in handling the response data, with the least squares parametric regression method, which is considered the de facto standard method used for regression. Theil's method was found to be superior to the method of least squares as it assumes that errors could occur in both x- and y-directions and they might not be normally distributed. In addition, it could effectively circumvent any outlier data points.

Spectrophotometric and titrimetric determination of nizatidine in capsules.

Four simple and accurate methods are described for the determination of nizatidine (NIZ) in pharmaceutical preparations. The first method is based on the formation of an ion-pair complex between the drug and either of bromocresol purple or picric acid with subsequent measurement of the developed colors at 411 and 400 nm, respectively. The second method depends on the condensation of mixed anhydrides of citric acid/acetic anhydride, with the tertiary amino group of the drug, where the developed color is measured spectrophotometrically at 545 nm. The oxidation of nizatidine by N-bromosuccinimide was utilized as a basis for the titrimetric method for its assay in capsules. The last method depends on the oxidation of nizatidine by ammonium cerium IV sulfate in the presence of perchloric acid with subsequent measurement of the absorbance at 314 nm; this principle is adopted to develop a kinetic method for the determination of NIZ in capsules. All the reaction conditions have been studied. The detection limits were varied from 0.44 to 0.78 microg ml(-1). The proposed methods were successfully applied to the assay of nizatidine in capsules.

Spectrophotometric and fluorimetric determination of aztreonam in bulk and dosage forms.

Spectrophotometric and fluorimetric determination of aztreonam were achieved through its reaction with cerium (IV) in acidic medium. The spectrophotometric method involves the quantitation of the amount of ceric equivalent to aztreonam by measuring the absorbance at 317 nm and the corresponding first-derivative value at 284 nm for the blank solution against the reaction solution. Beer's law is obeyed over the concentration ranges of 1.5-4 and 1-4 microg ml(-1), respectively. Meanwhile, in the fluorimetric method, higher sensitivity was achieved by measuring the fluorescence intensity of the formed cerium (III) at lambda(em)=357 nm (lambda(ex)=257 nm) within a concentration range 150-350 ng ml(-1). Study of the reaction conditions and reaction stoichiometry were presented. Interference from L-arginine, which is frequently co-formulated with aztreonam, was tested. The proposed procedures were applied successfully to the determination of aztreonam in pure form and in presence of arginine both in laboratory mixtures and commercial vials. The proposed methods are sensitive, accurate and precise as compared with the official USP 24 HPLC method.

Spectrophotometric determination of binary mixtures of pseudoephedrine with some histamine H1-receptor antagonists using derivative ratio spectrum method.

A derivative spectrophotometric method is developed for the assay of three binary mixtures of pseudoephedrine with fexofenadine (mix I), cetirizine (mix II) and loratadine (mix III). The method is based on the use of the first derivative of the ratio spectrum. The ratio spectrum was obtained by dividing the absorption spectrum of the mixture by that of one of the components. The concentration of the other component was determined from its respective calibration graph treated similarly. Moreover, the influence of Deltalambda for obtaining the first derivative of the ratio spectra and the effect of the divisor concentration on the calibration graphs were studied. The described method was applied for the determination of these combinations in synthetic mixtures and dosage forms. The results obtained were accurate and precise.

Spectrophotometric determination of omeprazole, lansoprazole and pantoprazole in pharmaceutical formulations.

The compensation method and other chemometric methods (derivative, orthogonal function and difference spectrophotometry) have been applied to the direct determination of omeprazole, lansoprazole and pantoprazole in their pharmaceutical preparations. The methods have been validated; the limits of detection were found to be 3.3x10(-2), 3.0x10(-2) and 3.5x10(-2) microgram ml(-1) for the three drugs, respectively. The repeatabililty of the methods were found to be 0.3-0.5%. The linearity ranges were found to be 0.5-3.5 microgram ml(-1). The proposed methods have been applied to the determination of the three drugs in their grastro-resistant formulations. The difference spectrophotometric (DeltaA) method is unaffected by the presence of acid induced degradation products; hence can be used as a stability indicating assay.

Determination of some histamine H1-receptor antagonists in dosage forms.

Three simple and accurate methods are presented for determination of Cetirizine, Fexofenadine, Loratadine and Acrivastine in pure form and commercial dosage forms. The first method is based on the reaction of the above cited drugs with bromocresol purple dye to form ion-pair complex extractable with chloroform and subsequently measured spectrophotometrically. Secondly, eosin gives with these drugs ion-pair complex, measurable directly without extraction both spectrophotometrically and spectrofluorimetrically. The last method involves the base-catalysed condensation of mixed anhydrides of organic acids (citric acid/acetic anhydride) where as the tertiary amino group in the above-cited drugs acts as the basic catalyst. The product of condensation is measured spectrophotometrically. All the reaction conditions for the proposed methods have been studied.