ABSTRACT
Controlling the manganese (Mn2+) concentration is important to product quality in the electrolytic manganese industry. Conventional methods for detecting Mn2+ are complex and reagent-consuming, which makes them slow and polluting. It is of great significance to develop a new fast and environmentally friendly method to quantify Mn2+ in electrolyte. The characteristic ultraviolet-visible (UV-vis) absorbance at 401 nm of Mn2+ will vary linearly with the Mn2+ concentration after data correction. Adjusting the pH, calibrating the spectral bandwidth (SBW) and optical path length (OPL), and subtracting the interference from coexisting substances by linear interpolation improve the measuring accuracy. Mn2+ concentration can be determined by direct fast UV-vis absorbance measurement at characteristic peaks without using harmful reagents which facilitates such measurement to be applicated as on-line detection method for electrolytic manganese industry. The method developed in this study will help achieve the goal of improving the detection speed while cutting back on pollutant discharge from concentration analysis process in electrolytic manganese industry.
Subject(s)
Electrolytes/chemistry , Manganese/analysis , Spectrophotometry, Ultraviolet , Calibration , Environmental Pollutants/analysis , Hydrogen-Ion Concentration , Limit of Detection , Manganese/standards , Spectrophotometry, Ultraviolet/standardsABSTRACT
The compound "marennine" is a blue-green pigment produced by the benthic microalgae Haslea ostrearia, with pathogenicity reduction activities against some bacteria and promising potential as a natural pigment in seafood industries. After decades of research, the chemical family of this compound still remains unclear, mainly because structural studies were impaired by the presence of co-extracted compounds in marennine isolates. To improve the purity of marennine extract, we developed a novel extraction method using a graphitic stationary phase, which provides various advantages over the previous procedure using tandem ultrafiltration. Our method is faster, more versatile, provides a better crude yield (66%, compared to 57% for ultrafiltration) and is amenable to upscaling with continuous photobioreactor cultivation. Our goal was to take advantage of the modulable surface properties of the graphitic matrix by optimizing its interactions with marennine. As such, the effects of organic modifiers, pH and reducing agents were studied. With this improvement on marennine purification, we achieved altogether the isolation of a fucoidan-related, sulfated polysaccharide from blue water. Characterization of the polysaccharides fraction suggests that roughly half of UV-absorbing compounds could be isolated from the marennine crude extracts. The identification of sulfated polysaccharides could be a major breakthrough for marennine purification, providing targeted isolation techniques. Likewise, the added value of Haslea ostrearia and the role of polysaccharides in previous marennine chemical characterization and bioactivity studies remain to be determined.
Subject(s)
Diatoms/chemistry , Graphite/chemistry , Phenols/analysis , Solid Phase Microextraction/methods , Magnetic Resonance Spectroscopy/methods , Magnetic Resonance Spectroscopy/standards , Microalgae/chemistry , Osmolar Concentration , Pigmentation/physiology , Pigments, Biological/analysis , Solid Phase Microextraction/standards , Spectrophotometry, Ultraviolet/methods , Spectrophotometry, Ultraviolet/standards , Ultrafiltration/methods , Ultrafiltration/standardsABSTRACT
The azo dyes, Yellow 5 (Y5), Red 2 (R2) and Blue 1 (B1), quantified in solutions and in mixtures of binary dyes, were studied by means of UV-Vis spectroscopy. In this work was used a CIE algorithm developed in Visual Basic for Applications (VBA). The CIE algorithm is based on the tristimulus chromaticity diagram, as an alternative to the shielding effect that arises in dye mixtures, and it can also be applied to complex quantification methods such as HPLC (High Performance Liquid Chromatography). The results obtained through of the algorithm, showed a higher accuracy from 97 to 99% in relation with similar UV-Vis quantification methods. In contrast, linear methods only managed to reach an accuracy from 78 to 98%. Additionally, the algorithm yielded significant similar values to the UHPLC reference method. The results showed that the method CIE algorithm was accessible and reliable to quantify binary mixtures of the dyes used which suggests the possibility to apply this method on other dyes, within the limits of quantification obtained in this study (0.076-24.56 mg/L) and the pH values from 2 to 10.
Subject(s)
Azo Compounds/analysis , Colorimetry/methods , Colorimetry/standards , Spectrophotometry, Ultraviolet/methods , Spectrophotometry, Ultraviolet/standards , Spectrophotometry/methods , Spectrophotometry/standards , Algorithms , Calibration , Chromatography, High Pressure Liquid , Limit of Detection , Programming LanguagesABSTRACT
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
Teicoplanin is a glycopeptide antibiotic prepared by fermentation from cultures of Actinoplanes teichomyceticus, used as drug of last resort for the treatment of bacterial infections in humans. This study, which is the first in a series of two parts, describes the development of a LC method for the separation of Teicoplanin drug substance and its related impurities compatible with MS detection. The separation conditions for Teicoplanin were set on a LiChrospher 100 RP-18 column under gradient elution with a mobile phase composed of ammonium formate 25 mM at pH 6.00 and ACN. The new method was shown equivalent in terms of selectivity to the one reported in the European Pharmacopoeia Teicoplanin monograph, and was validated according to ICH Q2 R1 guidelines for the drug substance assay. The new method offers similar performance to the compendial one but has the advantage of being fully compatible with MS and it can be proposed as a useful tool also for controlling the quality of Teicoplanin fermentation batches and the occurrence of potential impurities.
Subject(s)
Anti-Bacterial Agents/analysis , Chromatography, High Pressure Liquid , Chromatography, Reverse-Phase , Drug Contamination , Mass Spectrometry , Spectrophotometry, Ultraviolet , Teicoplanin/analysis , Chromatography, High Pressure Liquid/standards , Chromatography, Reverse-Phase/standards , Fermentation , Mass Spectrometry/standards , Quality Control , Reproducibility of Results , Spectrophotometry, Ultraviolet/standardsABSTRACT
While the HPLC/UV (high performance liquid chromatography coupled with ultra-violet spectrometry)-based DPRA (Direct Peptide Reactivity Assay) identifies dermal sensitizers with approximately 80% accuracy, the low selectivity and sensitivity of the HPLC/UV-based DPRA poses challenges to accurately identify the sensitization potential of certain chemicals. In this study, a high performance liquid chromatography coupled with tandem mass spectrometry (HPLC/MS-MS)-based DPRA was developed and validated according to the test guideline (OECD TG 442C). The final results were compared with the results from the traditional HPLC/UV-based guideline DPRA. This HPLC/MS-MS-based DPRA demonstrated similar performance compared to HPLC/UV-based DPRA using known dermal sensitizers and non-sensitizers according to the test guideline (OECD TG 442C). Following the validation, a challenge set of chemicals with either overlapping retention time with peptides, or higher hydrophobicity or chemicals potentially forming non-covalent interactions with peptides were assessed for dermal sensitization potential using both methods and the results were compared to existing in vivo data. The HPLC/MS-MS-based DPRA correctly predicted these chemicals as sensitizers or non-sensitizers; however, the HPLC/UV-based DPRA resulted in false-positive predictions for hydrophobic substances, chemicals with UV peaks overlapping with those of the peptide(s), and compounds that non-covalently interact with the peptides. These findings demonstrate the broader applicability and better sensitivity and selectivity of the LC/MS-MS-based DPRA over the traditional HPLC/UV-based guideline DPRA.
Subject(s)
Chromatography, High Pressure Liquid/methods , Peptides/chemistry , Spectrophotometry, Ultraviolet/methods , Tandem Mass Spectrometry/methods , Calibration , Chromatography, High Pressure Liquid/standards , Cysteine/chemistry , Lysine/chemistry , Peptides/metabolism , Spectrophotometry, Ultraviolet/standards , Tandem Mass Spectrometry/standardsABSTRACT
While color is arguably the most important optical property of evidential fibers, the actual dyestuffs responsible for its expression in them are, in forensic trace evidence examinations, rarely analyzed and still less often identified. This is due, primarily, to the exceedingly small quantities of dye present in a single fiber as well as to the fact that dye identification is a challenging analytical problem, even when large quantities are available for analysis. Among the practical reasons for this are the wide range of dyestuffs available (and the even larger number of trade names), the low total concentration of dyes in the finished product, the limited amount of sample typically available for analysis in forensic cases, and the complexity of the dye mixtures that may exist within a single fiber. Literature on the topic of dye analysis is often limited to a specific method, subset of dyestuffs, or an approach that is not applicable given the constraints of a forensic analysis. Here, we present a generalized approach to dye identification that (1) combines several robust analytical methods, (2) is broadly applicable to a wide range of dye chemistries, application classes, and fiber types, and (3) can be scaled down to forensic casework-sized samples. The approach is based on the development of a reference collection of 300 commercially relevant textile dyes that have been characterized by a variety of microanalytical methods (HPTLC, Raman microspectroscopy, infrared microspectroscopy, UV-Vis spectroscopy, and visible microspectrophotometry). Although there is no single approach that is applicable to all dyes on every type of fiber, a combination of these analytical methods has been applied using a reproducible approach that permits the use of reference libraries to constrain the identity of and, in many cases, identify the dye (or dyes) present in a textile fiber sample.
Subject(s)
Coloring Agents/analysis , Spectrophotometry/methods , Spectrum Analysis, Raman/methods , Textiles/analysis , Forensic Sciences/methods , Forensic Sciences/standards , Humans , Microspectrophotometry/methods , Microspectrophotometry/standards , Reference Standards , Spectrophotometry/standards , Spectrophotometry, Infrared/methods , Spectrophotometry, Infrared/standards , Spectrophotometry, Ultraviolet/methods , Spectrophotometry, Ultraviolet/standards , Spectrum Analysis, Raman/standardsABSTRACT
The NISTmAb Reference Material (RM) 8671 is intended to be an industry standard monoclonal antibody for pre-competitive harmonization of best practices and designing next generation characterization technologies for identity, quality, and stability testing. It must therefore embody the quality and characteristics of a typical biopharmaceutical product and be available long-term in a stable format with consistent product quality attributes. A stratified sampling and analysis plan using a series of qualified analytical and biophysical methods is described that assures RM 8671 meets these criteria. Results for the first three lots of RM 8671 highlight the consistency of material attributes with respect to size, charge, and identity. RM 8671 was verified to be homogeneous both within and between vialing lots, demonstrating the robustness of the lifecycle management plan. It was analyzed in concert with the in-house primary sample 8670 (PS 8670) to provide a historical link to this seminal material. RM 8671 was verified to be fit for its intended purpose as a technology innovation tool, external system suitability control, and cross-industry harmonization platform. Graphical abstract The NISTmAb Reference Material (RM) 8671 is intended to be an industry standard monoclonal antibody for pre-competitive harmonization of best practices and designing next generation characterization technologies for identity, quality, and stability testing.
Subject(s)
Antibodies, Monoclonal/chemistry , Immunoglobulin G/chemistry , Animals , Biosimilar Pharmaceuticals/chemistry , Chromatography, Gel/methods , Chromatography, Gel/standards , Drug Stability , Dynamic Light Scattering/methods , Dynamic Light Scattering/standards , Electrophoresis, Capillary/methods , Electrophoresis, Capillary/standards , Humans , Microscopy/methods , Microscopy/standards , Models, Molecular , Peptide Mapping/methods , Peptide Mapping/standards , Protein Stability , Quality Control , Reference Standards , Spectrophotometry, Ultraviolet/methods , Spectrophotometry, Ultraviolet/standards , Tandem Mass Spectrometry/methods , Tandem Mass Spectrometry/standardsABSTRACT
Carotenoids and tocopherols were characterised in the meso- and exocarp of wild-growing Costa Rican Acrocomia aculeata fruits. Comprehensive profiling of these lipophilic micronutrients in fruits of three varying maturity stages was conducted for the first time. A method for the simultaneous extraction and quantitation of carotenoids and α-tocopherol was developed and validated. Detailed HPLC-DAD-APCI/ESI-MSn analyses enabled the identification of α-tocopherol and 25 carotenoids. The latter comprised antheraxanthin, ß-carotene, lutein, luteoxanthin, neoxanthin, phytoene, phytofluene, violaxanthin, zeaxanthin, and several (Z)-isomers of the aforementioned compounds. Quantitation by HPLC-DAD/FLD revealed total carotenoid concentrations of 872±178 and 3075±407µg/100g fresh weight in the meso- and exocarp of fully ripe fruits, respectively. In both fruit fractions, progressing maturation resulted in the accumulation of phytoene, phytofluene, (all-E)-zeaxanthin, (all-E)-antheraxanthin, and (all-E)-violaxanthin. Carotenoid profiling was supported by multivariate data analysis. Carotenoid precursors and xanthophyll cycle pigments characterised Macauba fruits of full maturity.
Subject(s)
Arecaceae/chemistry , Carotenoids/analysis , Chromatography, High Pressure Liquid , Fruit/chemistry , Spectrometry, Mass, Electrospray Ionization , Spectrophotometry, Ultraviolet , alpha-Tocopherol/analysis , Arecaceae/growth & development , Calibration , Chromatography, High Pressure Liquid/standards , Costa Rica , Fruit/growth & development , Reference Standards , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization/standards , Spectrophotometry, Ultraviolet/standardsABSTRACT
BACKGROUND: The development and introduction of combined therapy represent a challenge for analysis due to severe overlapping of their UV spectra in case of spectroscopy or the requirement of a long tedious and high cost separation technique in case of chromatography. Quality control laboratories have to develop and validate suitable analytical procedures in order to assay such multi component preparations. METHODS: New spectrophotometric methods for the simultaneous determination of simvastatin (SIM) and nicotinic acid (NIA) in binary combinations were developed. These methods are based on chemometric treatment of data, the applied chemometric techniques are multivariate methods including classical least squares (CLS), principal component regression (PCR) and partial least squares (PLS). In these techniques, the concentration data matrix were prepared by using the synthetic mixtures containing SIM and NIA dissolved in ethanol. The absorbance data matrix corresponding to the concentration data matrix was obtained by measuring the absorbance at 12 wavelengths in the range 216 - 240 nm at 2 nm intervals in the zero-order. The spectrophotometric procedures do not require any separation step. The accuracy, precision and the linearity ranges of the methods have been determined and validated by analyzing synthetic mixtures containing the studied drugs. CONCLUSION: Chemometric spectrophotometric methods have been developed in the present study for the simultaneous determination of simvastatin and nicotinic acid in their synthetic binary mixtures and in their mixtures with possible excipients present in tablet dosage form. The validation was performed successfully. The developed methods have been shown to be accurate, linear, precise, and so simple. The developed methods can be used routinely for the determination dosage form.
Subject(s)
Chemistry, Pharmaceutical/standards , Niacin/analysis , Simvastatin/analysis , Chemistry, Pharmaceutical/instrumentation , Chemistry, Pharmaceutical/methods , Drug Combinations , Niacin/chemistry , Reproducibility of Results , Simvastatin/chemistry , Spectrophotometry, Ultraviolet/instrumentation , Spectrophotometry, Ultraviolet/methods , Spectrophotometry, Ultraviolet/standardsABSTRACT
Troxerutin (TRX) is a mixture of semisynthetic hydroxyethylrutosides (Hers) arising from hydroxyethylation of rutin, a natural occurring flavonoid. TRX is commonly used for its anti-oxidant and anti-inflammatory properties in chronic venous insufficiency and other vascular disorders. In recent studies, the protective effects of TRX in Alzheimer's disease, colon carcinogenesis and hepatocellular carcinoma are emerged. However, the chemical stability of TRX has never been studied. Hence, the aims of the work were to study the TRX chemical stability through a forced degradation study and to develop and validate a new stability indicating LC-UV method for determination of TRX. In order to perform the study, TRX stability was tested in various stress conditions analysing the degradation samples by LC-MS. Three degradation products (DPs; D1, D2 and D3, 3',4',7-Tri-O-(ß-hydroxyethyl)quercetin, 3',4',5,7-Tetra-O-(ß-hydroxyethyl)quercetin and 3',4'-Di-O-(ß-hydroxyethyl)quercetin respectively) arising from degradation in acidic conditions were identified and synthesized: among them, D1 resulted the stability indicator for hydrolytic degradation. Furthermore, a stability-indicating LC-UV method for simultaneous determination of triHer (3',4',7-Tri-O-(ß-hydroxyethyl)rutin, the principal component of the mixture) and D1 was developed and validated. The LC-UV method consisted in a gradient elution on a Phenomenex Kinetex EVO C18 (150 × 3â¯mm, 5⯵m) with acetonitrile and ammonium bicarbonate buffer (10â¯mM, pH 9.2). The method was linear for triHer (20-60⯵gâ¯mL-1) and D1 (5.1-35⯵gâ¯mL-1). The intraday and interday precision were determined and expressed as RSDs: all the values wereâ¯≤â¯2% for both triHer and D1. The method demonstrated also to be accurate and robust and the average recoveries were 98.8 and 97.9% for triHer and D1, respectively. Moreover, the method resulted selective and specific for all of the components present in the degradation pattern of TRX (diHer (3',4'-Di-O-(ß-hydroxyethyl)rutin), triHer, tetraHer (3',4',5,7-Tetra-O-(ß-hydroxyethyl)rutin), D3, D1 and D2) and it was successfully applied for the stability studies of both drug substances and drug products.
Subject(s)
Chromatography, High Pressure Liquid , Hydroxyethylrutoside/analogs & derivatives , Spectrophotometry, Ultraviolet , Technology, Pharmaceutical/methods , Calibration , Chromatography, High Pressure Liquid/standards , Drug Contamination , Drug Stability , Hydroxyethylrutoside/chemical synthesis , Hydroxyethylrutoside/chemistry , Limit of Detection , Linear Models , Molecular Structure , Reference Standards , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization , Spectrophotometry, Ultraviolet/standards , Technology, Pharmaceutical/standardsABSTRACT
Pharmaceutical manufacturers have to study the stability of drug products before marketing according to ICH guideline Q1A(R2); data of those investigations aim to set expiry dates. The expiry date on the container of a remedy assures the physician and the patient a stability of the drug in its formulation i.e. within a specification of 95-105%. Only few studies show that shelf-lives of pharmaceutical products are often longer than expiration dates. The objective of the study presented here was determining the content of nine expired ampoules manufactured in the last century and identifying the impurity profile by means of HPLC-UV and HPLC-MS, respectively. The ampoules are part of the "PEAK-collection" of long expired finished pharmaceutical products at IBMP, Nürnberg-Heroldsberg, and consists among others of epinephrine (Suprarenin and Adrenalin in Oil), etilefrine (Effortil®), synephrine (Sympatol®), caffeine and procaine (Impletol), caffeine and sodium salicylate (Caffeinum Salicylicum), dipyridamole (Persantin®), furosemide (Lasix®), and metamizole (Novalgin®). For chromatographic investigations methods of the European Pharmacopoeia for related substances were used; for determining the content, they were validated for linearity, precision, and accuracy. The results were compared to current reference ampoules. Five out of nine ampoules were still within the specified content limits. In Suprarenin and Adrenalin in Oil, both containing epinephrine, Impletol (procaine), and Persantin® (dipyridamole) contents were decreased to 70%, 74%, 79%, and 86%, respectively, and therefore out of specification.
Subject(s)
Chromatography, High Pressure Liquid , Drug Packaging , Pharmaceutical Preparations/chemical synthesis , Spectrophotometry, Ultraviolet , Technology, Pharmaceutical/methods , Calibration , Chromatography, High Pressure Liquid/standards , Dosage Forms , Drug Compounding , Drug Stability , Linear Models , Quality Control , Reference Standards , Reproducibility of Results , Spectrophotometry, Ultraviolet/standards , Technology, Pharmaceutical/standards , Time FactorsABSTRACT
A swift, precise and simple HPLC bioanalytical technique with UV detection was established and validated for quantitative estimation of valsartan in human plasma. The analyte was separated from plasma by protein precipitation with acetonitrile and chromatographically separated on Zorbax SB-C18 (5µm, 4.6mm × 15cm) column. The solvent mixture system consisting of acetonitrile, water and glacial acetic acid (40:59:1 v/v), was pumped using isocratic mode at 1mL/min flow rate. Samples' detection of drug was made spectrophotometrically at a wavelength of 264nm. The analyte response was instituted to be linear from 0.06 to 8µg/mL with a regression value of 0.999. The accuracy of the proposed method was ranged between 97.2-100.3% with 5% RSD. The analytical recovery (>95%) was consistently observed and satisfactory sample stability was also found at different environmental conditions. In conclusion the reported bio-analytical method is easy and robust that was successfully utilized in estimation of valsartan in a pharmacokinetic study.
Subject(s)
Angiotensin II Type 1 Receptor Blockers/blood , Chromatography, High Pressure Liquid , Spectrophotometry, Ultraviolet , Valsartan/blood , Angiotensin II Type 1 Receptor Blockers/pharmacokinetics , Calibration , Chromatography, High Pressure Liquid/standards , Humans , Limit of Detection , Male , Reference Standards , Reproducibility of Results , Spectrophotometry, Ultraviolet/standards , Valsartan/pharmacokineticsABSTRACT
A simple stability indicating UV-spectrophotometric method has been developed and validated for the determination of cinitapride hydrogen tartrate (CHT) in bulk and solid pharmaceutical dosage form. Drug absorption was measured in different analytical mediums however; maximum absorption was seen in 0.1 N HCl at wavelength (λmax) of 266 nm. The calibration curve was found to be linear over the concentration range from 6 to14µg/mL with the correlation coefficient value (r) of 0.999. The LOD and LOQ were estimated to be 0.1019µg/mL and 0.309µg/mL respectively. The accuracy was evaluated by determining the percent drug recovery, performed at three different levels of 50%, 100% and 150%. The% recovery was found to be in the range of 99.96-100.64%. The precision of the method was determined by inter-day and intra-day variations. The % RSD value <0.5 indicates the underlying method is precise and accurate as well. The developed method was applied to characterize in vitro assay content of few brands of cinitapride (1 mg) available in local market. No interference of the formulation excipients with the drug absorption was observed during assay. Drug substance and drug product were exposed to various stressed conditions (acid, base, oxidative, thermal and photolysis). Forced degradation testing of drug product showed that the oxidation (20%) was found to be the major degradation pathway of the cinitapride. However; drug estimation was not influenced in presence of degradation moieties formed during acid, base, oxidation, thermal and photolytic breakdown. Overall, the investigated technique is robust and specific that would be successfully used to quantify the cinitapride hydrogen tartarate in pharmaceutical dosage and bulk form in future.
Subject(s)
Benzamides/analysis , Spectrophotometry, Ultraviolet , Technology, Pharmaceutical/methods , Calibration , Drug Stability , Limit of Detection , Reference Standards , Reproducibility of Results , Spectrophotometry, Ultraviolet/standards , Tablets , Technology, Pharmaceutical/standardsABSTRACT
The European Pharmacopoeia (Ph. Eur.) general chapter 5.14. Gene transfer medicinal products for human use suggests the use of absorbance measurements at 260 nm to determine the DNA concentration of plasmid vectors used for the preparation of gene therapy products for human use. An international collaborative study was organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM) to confirm the suitability of UV spectrophotometry for the quantification of plasmid vectors used in gene therapy (GT). Three Official Medicine Control Laboratories (OMCLs of the European OMCL Network) and members of the OMCL Working Group for GT products took part in the study, in which various types of spectrophotometers were assessed using common test samples. Results of the study demonstrated that UV spectrophotometry can be considered suitable for the quantification of plasmid DNA in GT products regardless of the instrument used.
Subject(s)
Genetic Therapy/methods , Genetic Vectors/analysis , Plasmids/analysis , Spectrophotometry, Ultraviolet , Calibration , Europe , Genetic Therapy/standards , Genetic Vectors/genetics , Genetic Vectors/standards , Humans , Linear Models , Observer Variation , Plasmids/genetics , Plasmids/standards , Quality Control , Reference Standards , Reproducibility of Results , Spectrophotometry, Ultraviolet/standardsABSTRACT
A high performance liquid chromatography (HPLC) method for simultaneous analysis of venlafaxine and its major metabolite 0-desmethylvenlafaxine and vilazodone and its methabolite M10 have been devel- oped and validated. Chromatography was performed on the Phenyl-Hexyl column with mobile phase containing methanol, acetate buffer at pH 3.5 and diethylamine. The application of stationary phase with 7r-7c moieties and mobile phase containing diethylamine as silanol blocker lets to obtain double protection against silanols and thus very high theoretical plate numbers were obtained. The good separation selectivity, good peaks' symmetry and very high systems efficiency for all investigated compounds were obtained in applied chromatographic system. The method is very efficient and suitable for the analysis of investigated drugs and their metabolites in human serum for patients' pharmacotherapy control.
Subject(s)
Antidepressive Agents/blood , Chromatography, High Pressure Liquid , Cyclohexanols/blood , Spectrometry, Mass, Electrospray Ionization , Spectrophotometry, Ultraviolet , Venlafaxine Hydrochloride/blood , Vilazodone Hydrochloride/blood , Biotransformation , Calibration , Chromatography, High Pressure Liquid/standards , Humans , Limit of Detection , Reference Standards , Reproducibility of Results , Spectrometry, Mass, Electrospray Ionization/standards , Spectrophotometry, Ultraviolet/standardsABSTRACT
A collaborative study was conducted to evaluate an HPLC method for determining phenolic compounds in Echinacea spp. raw materials, powdered extracts, and tinctures. Eleven collaborating laboratories received three practice samples representing each matrix type, phenolic reference standards, eight test samples as blind duplicates, the validated analytical method, and instructions. Test samples included two raw materials, four extracts (including one in combination with astragalus and reishi), one ethanolic tincture in combination with goldenseal, and one glycerite tincture. Each material was extracted with a 60% methanol aqueous solution, separated on a C18 column, and detected at 330 nm. Results reported by laboratories for total phenolics in Echinacea roots, aerial parts, and extracts ranged from 9.5 to 62.9 mg/g with RSDR ranging from 3.64 and 7.95% and Horwitz ratio (HorRat) values ranging from 1.06 to 2.01. Total phenolics in the ethanolic tincture ranged from 4837 to 5962 µg/mL, with an RSDR of 6.35% and a HorRat value of 1.45. The glycerite tincture showed poor interlaboratory precision with a HorRat value of 3.32, an RSDR of 21.8%, and reported total phenolic values ranging from 257 to 539 µg/mL.
Subject(s)
Dietary Supplements/analysis , Echinacea/chemistry , Phenols/analysis , Chromatography, High Pressure Liquid/standards , Laboratories/standards , Spectrophotometry, Ultraviolet/standardsABSTRACT
The use of mycelia as biocatalysts has technical and economic advantages. However, there are several difficulties in obtaining accurate results in mycelium-catalysed reactions. Firstly, sample extraction, indispensable because of the presence of mycelia, can bring into the extract components with a similar structure to that of the analyte of interest; secondly, mycelia can influence the recovery of the analyte. We prepared calibration standards of 3-phenoxy-1,2-propanediol (PPD) in the pure solvent and in the presence of mycelia (spiked before or after extraction) from five fungi (Aspergillus niger, Aspergillus tubingensis, Penicillium aurantiogriseum, Penicillium sp. and Aspergillus terreus). The quantification of PPD was carried out by HPLC-UV and UV-vis spectrophotometry. The manuscript shows that the last method is as accurate as the HPLC method. However, the colorimetric method led to a higher data throughput, which allowed the study of more samples in a shorter time. Matrix effects were evaluated visually from the plotted calibration data and statistically by simultaneously comparing the intercept and slope of calibration curves performed with solvent, post-extraction spiked standards and pre-extraction spiked standards. Significant differences were found between the post- and pre-extraction spiked matrix-matched functions. Pre-extraction spiked matrix-matched functions based on A. tubingensis mycelia, selected as the reference, were validated and used to compensate for low recoveries. These validated functions were successfully applied to the quantification of PPD achieved during the hydrolysis of glycidyl phenyl ether by mycelium-bound epoxide hydrolases and equivalent hydrolysis yields were determined by HPLC-UV and UV-vis spectrophotometry. This study may serve as starting point to implement matrix effects evaluation when mycelium-bound epoxide hydrolases are studied.
Subject(s)
Epoxide Hydrolases/metabolism , Fungal Proteins/metabolism , Fungi/metabolism , Aspergillus/metabolism , Aspergillus niger/metabolism , Biotechnology , Calibration , Catalysis , Chromatography, High Pressure Liquid/standards , Glycerol/analogs & derivatives , Glycerol/metabolism , Mycelium/metabolism , Penicillium/metabolism , Phenyl Ethers/metabolism , Reference Standards , Solvents , Spectrophotometry, Ultraviolet/standardsABSTRACT
L-asparaginase is an effective anti-tumor agent for acute lymphoblastic leukemia. This work presents the development of an activity determination of L-ASNase preparations for pharmaceutical quality control purposes, in accordance with analytical Quality by Design principles. Critical method attributes, the absorbance at 450 nm (A450) of the Nessler product as well as its variability, were evaluated as a function of critical method variables, by using experimental designs. The design space of the enzyme activity assay was defined (Nessler method: C(KI)/C(HgI2) of 1.90-1.95, C(NaOH)/C(HgI2) of 17.0-18.0, C(HgI2final) of 20-40 mM and time of 10-40 min; enzyme activity conditions: temperature range of 36.6-37.4 °C, pH range of the KH2PO4 buffer from 7.1 to 7.7, KH2PO4 buffer concentration: 0.18-0.22 M and L-Asn concentration of 18-22 mM), leading to a final enzyme activity assay method. A control strategy was ultimately implemented using system suitability tests.
Subject(s)
Asparaginase/analysis , Asparaginase/metabolism , Chemistry, Pharmaceutical/methods , Chemistry, Pharmaceutical/standards , Enzyme Activation/physiology , Protein Structure, Secondary , Spectrophotometry, Ultraviolet/methods , Spectrophotometry, Ultraviolet/standardsABSTRACT
Acetylcholinesterase reactivators (oximes) are compounds used for antidotal treatment in case of organophosphorus poisoning. The dissociation constants (pK(a1)) of ten standard or promising acetylcholinesterase reactivators were determined by ultraviolet absorption spectrometry. Two methods of spectra measurement (UV-vis spectrometry, FIA/UV-vis) were applied and compared. The soft and hard models for calculation of pK(a1) values were performed. The pK(a1) values were recommended in the range 7.00-8.35, where at least 10% of oximate anion is available for organophosphate reactivation. All tested oximes were found to have pK(a1) in this range. The FIA/UV-vis method provided rapid sample throughput, low sample consumption, high sensitivity and precision compared to standard UV-vis method. The hard calculation model was proposed as more accurate for pK(a1) calculation.