A novel liquid-liquid-solid microextraction strategy for bio-sample preparation by in situ self-assembly of zeolitic imidazolate framework 8 on hollow fiber membrane.

A novel liquid-liquid-solid membrane microextraction (LLSMME) method which integrates hollow fiber liquid phase microextraction (HF-LPME) and solid phase microextraction (SPME) was developed for bio-sample preparation. The homogeneous zeolitic imidazolate framework 8 mixed matrix membrane (ZIF-8-MMM) was prepared by in situ self-assembly of ZIF-8 on the inner surface of hollow fiber membrane and employed as a flexible LLSMME device. Incorporating the advantages of both HF-LPME and SPME, the as-fabricated ZIF-8-MMM exhibited excellent performance on the extraction and preconcentration of small molecule drugs of different polarity from complex biological matrices. As a case study, ZIF-8-MMM-based LLSMME coupled with UPLC-MS/MS were developed and validated for determination of ibuprofen, simvastatin and ranitidine at trace levels in rat plasma. The method showed good linearity (r2 > 0.99) and repeatability (RSD < 15%), low limits of detection (2-3 ng mL-1) and high relative recoveries (97.42-103.8%). The enrichment factors were between 87.3 and 112.6. Our study provided a promising strategy for developing more efficient, cost-effective and environmentally friendly technique for bio-sample pretreatment.

A new strategy to eliminate sample mixing during in-tube solid phase microextraction.

During in-tube solid phase microextraction, sample mixing with mobile phase contained in the autosampler tubing during extraction may result in some amount of sample becoming entrained in the mobile phase rather than returning to the sample vial or being directed to waste after extraction. In cases where target analytes have relatively low affinity for the sorbent on the wall of the capillary, mixing can impact data quality. Where the sample contains components that may interfere with either the separation (e.g. proteins) or detection (e.g. ions with MS detection), additional difficulties can arise. In the current research, the magnitude of the sample mixing effect was illustrated by analyzing ranitidine and a series of polycyclic aromatic hydrocarbons (PAH). The sample volume equivalent of mixing was calculated as 37 µL for ranitidine and 20 µL for PAHs using the same inner diameter of capillary. To address this issue, a novel approach involving adding a switching valve located between the metering pump and the capillary was developed. Capillary flush conditions, draw/eject speed and extraction time were optimized for ranitidine with the result that in the final method, no mixing of sample with mobile phase was apparent in the detected amounts. To provide information on a compound class with intermediate polarity, two -blockers were also extracted using the optimized washing conditions respectively. The results indicated that the issue of sample mixing had been resolved for these as well. Finally, in-tube SPME calibration of these three analyte classes was shown to be highly linear, providing further indication that sample mixing was not impacting data quality. Available literature on the subject was surveyed, and a discussion on the rational selection of conditions to guide method development was also provided.

Ozonation of the pharmaceutical compound ranitidine: reactivity and kinetic aspects.

Ranitidine has been ozonated under different operating conditions of pH, applied ozone dose, initial ranitidine concentration and presence or absence of free radical inhibitors. Results of ranitidine evolution with time indicate a high reactivity of this compound with molecular ozone. Mineralization levels achieved in the order of 20-25% suggest that the (CH3)2-N-CH2- moiety bonded to the furan ring could be separated from the rest of the ranitidine structure and further mineralized. Only alkaline conditions (pH=11) are capable of increasing TOC conversion up to values close to 70%. Determination of the direct ozonation rate constant for ranitidine by means of competitive kinetics reveals an unacceptable dependence of the aforementioned constant with the reference compound reactivity. It is hypothesised that only reference compounds with reactivity similar to the target species should be used.

Effect of small levels of impurities on the water vapor sorption behavior of ranitidine HCl.

PURPOSE: Deliquescence is the process by which a solid undergoes dissolution by sorbing moisture from its surroundings when a characteristic relative humidity, RH0, is reached. For mixtures of two or more deliquescent solids, RH0 will generally be lowered. The goal of this research was to investigate the effect of small amounts of impurities or degradants on RH0 for a model deliquescent pharmaceutical salt. MATERIALS AND METHODS: The model salt chosen for this work was ranitidine HCl, which has two polymorphic forms. Moisture sorption profiles for each polymorph and samples of different purities were obtained using a gravimetric water vapor sorption balance. RESULTS: Polymorphs of a similar purity yielded virtually identical moisture sorption profiles. In contrast, samples containing higher levels of impurities had both enhanced moisture sorption below RH0 and a lowered value of RH0. CONCLUSIONS: It was concluded that small levels of degradants and/or impurities can drastically affect the moisture sorption profile of a deliquescent material, both through affecting the deliquescence relative humidity and by altering the overall interaction of the substance with moisture. Such changes in behavior may have significant effects on both active pharmaceutical ingredient and drug product stability during both processing and storage.

The application of sub-2-microm particle liquid chromatography-operated high mobile linear velocities coupled to orthogonal accelerated time-of-flight mass spectrometry for the analysis of ranitidine and its impurities.

Impurity profiling of pharmaceutical drug substances or dosage formulations require methods involving high sensitivity and resolution from LC and MS alike as well as an acceptable analysis time. While throughput can be increased, it is usually at the expense of chromatographic resolution. The application of sub-2-microm stationary phases and high mobile linear velocities has been combined with orthogonal acceleration (oa)-TOF MS for the impurity structural characterization analysis of small-molecule pharmaceutics. A pharmaceutical drug substance was forcefully degraded and used to test the proof of concept of developing an impurity profile method by ultra performance liquid chromatography (UPLC). Optimum conditions were identified by use of method development simulation software as well as traditional approaches of method scouting with columns and a varied range of pH. Further analysis illustrated the effectiveness of applying oa-TOF MS techniques to assist in achieving exact mass coupled with MS/MS to define the structural characterization of the related substances relative to the pharmaceutical active ingredient and identification of any unknown impurity substances. The barriers with trade-offs between resolution and speed are overcome by the application of UPLC, whereas the increased sensitivity provides for superior exact mass oa-TOF MS.

Biodegradability and toxicity of pharmaceuticals in biological wastewater treatment plants.

In this experimental study both biological treatability of pharmaceuticals and their potential toxic effect in biological processes were evaluated. The pharmaceuticals were selected among those that are present at higher concentration in the Italian wastewater treatment plant effluents and widely used as antiulcer (ranitidine), beta-blocker (atenolol) and antibiotic (lincomycin). The present paper is the continuation of a work already presented,[1] which used a synthetic wastewater fed to laboratory scale SBR (Sequencing Batch Reactor) operated with different sludge ages (8 and 14 days), different biochemical conditions (aerobic or anoxic-aerobic mode) and several influent drug concentrations (2, 3 and 5 mg/L). In this case a real municipal wastewater was used as influent to the SBR. In parallel, batch tests were conducted to determine the removal kinetics of drugs and nitrogen. Toxicity tests using a titrimetric biosensor to verify possible inhibition on microorganisms were also performed. Finally, the possible adsorption of the pharmaceuticals on activated sludge was evaluated. The drugs under investigation showed different behaviours in terms of both biodegradability and toxicity effect on nitrifiers. Ranitidine showed generally low removal efficiencies (17-26%) and a chronic inhibition on nitrification. Atenolol showed generally higher removal efficiencies than ranitidine, even if the fairly good efficiency obtained in the previous experimentation with synthetic wastewater (up to 90%) was not attained with real wastewater (36%). No inhibition on nitrification was observed on both acclimated and non acclimated microorganisms with a high nitrification activity, whilst it was present with activated sludge characterised by a lower nitrification activity. Consistently with his pharmaceutical properties, lincomycin showed significant inhibition on nitrification activity.

Ranitidine hydrochloride: development of an isocratic stability--indicating high-performance liquid chromatographic separation.

The development of a stability-indicating assay for ranitidine hydrochloride using a mobile phase added ion-interaction reagent was achieved. The assay easily separated all known and unknown impurities/degradants. This assay may be used for the determination of purity, identity and strength for the active ingredient and finished dosage forms. Placebo samples were analyzed for all of the dosage forms and did not interfere with the separation. The system was found to be linear over a range of 0.056 to 44.4 microg/g, with a limit of detection of 0.028 microg/g and a limit of quantitation of 0.056 microg/g. The system precision was determined to be 0.7%. The development of the stability-indicating assay and the effect that each chromatographic variable had on the separation will be discussed.