Characterization of Nanoparticles in 2X, 4X and 6X Potencies of Ferrum Phosphoricum.

CONTEXT: Nanomedicine is a rapidly expanding field in which nanoparticles play an integral part. They have 2 distinct characteristics, namely a small size and large surface area, which influence a drug's permeability, solubility, and bioavailability. The trituration technique used in the preparation of certain homeopathic remedies is similar to some processes used to manufacture nanoparticles. New research has confirmed the presence of nanoparticles in homeopathic remedies. The majority of these studies use transmission electron microscopy (TEM), alone or in combination with other methods, to characterize the nanoparticles. OBJECTIVES: The study intended to evaluate the use, efficacy, and application of 5 characterization techniques for investigating the presence and size of nanoparticles in Ferrum phosphoricum (iron phosphate) in 2X, 4X and 6X potencies. DESIGN: Quantitative experimental study. SETTING: The study took place in the Department of Biotechnology and Food Technology, and the Department of Chemistry (Central Analytical facility and UJ Spectrum facility) at the University of Johannesburg in Johannesburg, South Africa. OUTCOME MEASURES: The study analyzed 2X, 4X, and 6X potencies of Ferrum phosphoricum using: (1) ultraviolet-visible spectroscopy (UV-vis), (2) Fourier transform infrared spectroscopy (FTIR), (3) dynamic light scattering (DLS), (14) scanning electron microscopy (SEM) for energy-dispersive X-ray spectroscopy (EDX), and (5) transmission electron microscopy (TEM). Data were generated through imaging and the software programs of the equipment. RESULTS: All 5 procedures confirmed the presence of Ferrum phosphoricum nanoparticles and nanofibers for all 3 potencies. Using all 5 techniques permitted characterization of different aspects of the particles, such as functional groups, elemental iron, concentration base, and size distribution. CONCLUSIONS: Although all 5 techniques used in the study showed the presence of iron phosphate nanoparticles in the 2X, 4X and 6X potencies of Ferrum phosphoricum, making them essential tools in determining the presence of nanoparticles, UV-vis, FTIR, and TEM analysis were best suited for the analysis of nanoparticles in high-dilution preparations such as homeopathic remedies, due to the limitations of DLS and EDX.

Dilution-Induced Physico-Chemical Changes of Metal Oxide Nanoparticles Due to Homeopathic Preparation Steps of Trituration and Succussion.

BACKGROUND: Although the presence of starting materials in extreme dilutions of homeopathic medicines has been established, the physico-chemical changes of these materials induced by the manufacturing steps-that is, solid-solid mixing involving grinding (trituration) and slurry mixing involving impact (succussion), followed by dilution-are still unknown. METHODS: We subjected cupric oxide and zinc oxide nanoparticles (NPs) to the homeopathic processes of trituration and succussion, followed by dilution up to 6 cH. Particle image velocimetry was employed to analyze the fluid motion during succussion and its effect on the NPs. The resulting microstructural and chemical changes at different dilution steps were determined by X-ray photoelectron spectroscopy, Fourier-transform infrared spectroscopy and transmission electron microscopy. RESULTS: The succussion triggered multi-sized bubble generation and turbulent fluid motion up to a duration of 400 ms, with maximum average velocity of 0.23 m/s. Due to 1% transfer of kinetic energy from a moving eddy with this velocity, upon collision, the rate of temperature change in a particle of size 1 µm and 1 nm was predicted to rise by approximately 102 K/s and 106 K/s respectively. During trituration, the oxide NPs reduced to metals and did not aggregate by remaining within lactose, but they converted to oxidized finer NPs after impact. Silicate chains leached from the vial cross-linked after third dilution, forming large macro-particles and encapsulating the NPs that were retained and carried at higher dilution steps. CONCLUSION: The results showed that the NPs sustained significant rate of temperature change due to energy transfer from moving eddies during succussion. Different physico-chemical changes, such as size reduction, successive reduction and oxidation of NPs, and morphological changes, were achieved through trituration and succussion. The retention of NPs within cross-linked poly-siloxane chains reveals the importance of both the borosilicate glass vial and the ethanol solution during preparation of homeopathic medicines.

Why extreme dilutions reach non-zero asymptotes: a nanoparticulate hypothesis based on froth flotation.

Extreme dilutions, especially homeopathic remedies of 30c, 200c, and higher potencies, are prepared by a process of serial dilution of 1:100 per step. As a result, dilution factors of 10(60), 10(400), or even greater are achieved. Therefore, both the presence of any active ingredient and the therapeutic efficacy of these medicines have been contentious because the existence of even traces of the starting raw materials in them is inconceivable. However, physicochemical studies of these solutions have unequivocally established the presence of the starting raw materials in nanoparticulate form even in these extreme (super-Avogadro, >10(23)) dilutions. In this article, we propose and validate a hypothesis to explain how nanoparticles are retained even at such enormous dilution levels. We show that once the bulk concentration is below a threshold level of a few nanograms/milliliter (ng/mL), at the end of each dilution step, all of the nanoparticles levitate to the surface and are accommodated as a monolayer at the top. This dominant population at the air-liquid interface is preserved and carried to the subsequent step, thereby forming an asymptotic concentration. Thus, all dilutions are only apparent and not real in terms of the concentrations of the starting raw materials.

[Preparation and preliminary characterization of chitosan from Catharsius molossus discards].

OBJECTIVE: To study preparation and characterization of chitosan from Catharsius Molossus processing discards and explore the feasibility of utilization of insects residue. METHODS: Preparation technique of chitosan was studied by orthogonal design,and properties were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), etc. RESULTS: Preparation techniques were as follows: demineralizing: soaked for 30 min at 80 degrees C with 1.3 mol/L HCl, then kept for 12 h under room temperature. Deproteinization and delipidation: treated for 6 h at 90 degrees C with 4 mol/L NaOH. Decolorizing: soaked at room temperature with 3% KM-nO4, then treated with 2% oxalic acid at 70 degrees C. Deacetylating:treated for 5 h at 110 degrees C with 14 mol/L NaOH. CONCLUSION: The technique is stable and feasible. The result also preliminarily showes that chitosan from Catharsius Molossus is better than shrimp's. It will be widely applied in biomedical and other industrial areas with such exciting properties.

Application of mid-infrared spectroscopy in the quality control of traditional Chinese medicines.

Chinese herbal medicines are often referred to as Chinese materia medica (CMM). Composite formulae containing mixtures of CMM are prescribed for treatment and prevention of diseases in the practice of traditional Chinese medicine (TCM). Some of the well-known CMM formulae (Fufang in Chinese) are manufactured and marketed as proprietary Chinese medicines (PCM). Quality assessment and assurance of these products are difficult; they are a challenging task. Mid-infrared spectroscopy, a classic molecular structure analysis method, has been innovatively applied in the quality control of TCM, and has gained significant impact and advancement in analytical fields. Infrared fingerprinting features appear particularly suitable for the identification of multicomponent matrices in samples whose chemical integrity has not been altered or destroyed because no extraction procedure is needed. This review summarizes and gives an overall view on the application of mid-infrared and two-dimensional correlation infrared (2D-IR) spectroscopy as well as chemometric techniques in the identification of CMM, investigation of TCM processing procedures, and analysis of herb extracts and preparations.

Bioinspired morphology-controlled silver nanoparticles for antimicrobial application.

Phytochemicals sources have been extensively used as reducing and capping agents for synthesis of nanoparticles (NPs). However, morphology-controlled synthesis and shape/size dependent applications of these NPs still need to be explored further, and there is a need to develop a way in which particular and optimized phytochemicals result in the desired NPs in lesser time and cost with higher reproducibility rate. The present study is focused on morphology-controlled synthesis and shape/size dependent application of silver NPs based on the fractionated phytochemicals of Elaeagnus umbellata extract (EU). Unlike other approaches, in this study the reaction parameters such as time, temperature, pH, stirring speed and concentration of the precursor solutions were not altered during the optimization process. The fractionated phytochemicals were used separately for the synthesis of AgNPs, and the synthesized NPs were characterized by UV-visible, FT-IR, atomic force microscopy (AFM) and scanning electron microscopy (SEM). Our findings suggested that the constituents of the extract fractions varied with the selection of the extraction solvent, and the shape/size, bactericidal properties and toxicity of the NPs have a strong correlation with the phytochemicals of the plant extract. The fractionated phytochemicals present in the water fractions (EUW) resulted in monodispersed spherical AgNPs in the size about 40 nm. The NPs have significant stability in physiological conditions (i.e. temperature, pH and salt), have good antibacterial activity, and were found to be non-toxic. Furthermore, AFM and SEM analysis exposed that the NPs killed the bacteria by disturbing the cellular morphology and releasing the cellular matrix. Our results justify the use of different fractions of plant extract to obtain detail implications on shape, size, antibacterial potential and toxicity of AgNPs. This is the first step in a controllable, easy and cheap approach for the synthesis of highly stable, uniform, non-toxic and bactericidal AgNPs using five fractions of EU. The findings suggested that the synthesized NPs, particularly from EUW, could be used in pharmaceutical and homeopathic industry for the development of antibacterial medications.

Characterization and synergistic antibacterial potential of green synthesized silver nanoparticles using aqueous root extracts of important medicinal plants of Pakistan.

In the past few years, biologically synthesized silver nanoparticles (AgNPs) have been standout amongst the most utilized nanoparticles both in the field of therapeutics and clinical practices. Therefore, the current study aimed to synthesize AgNPs for the first time using aqueous root extracts of important plants of Pakistan i.e. Bergenia ciliata, Bergenia stracheyi, Rumex dantatus and Rumex hastatus and characterize them. In addition, antibacterial activity of synthesized AgNPs at 30-150 µg/well was assessed using well diffusion method against Staphylococcus aureus, Staphylococcus haemolyticus, Bacillus cereus, Escherichia coli, Salmonella typhi and Pseudomonas aeruginosa bacterial strains that are considered most harmful bacteria for human beings. The characterization of synthesized AgNPs showed the absorption maxima ranged from 434 to 451 nm and XRD confirmed the crystalline nature of AgNPs as well as FTIR elucidated the involvement of biomolecules for reduction and capping of AgNPs. SEM determined the average size of AgNPs ranging from 25 to 73 nm and strong signals of silver were captured in EDX images. The result of antibacterial activity showed that only aqueous root extracts of all selected plants were inactive against all the tested bacterial strains. However, importantly, direct relationship between zone of inhibition of S. aureus, S. typhi and P. aeruginosa was found with increasing concentration of AgNPs of each selected plant. Moreover, S. haemolyticus was only inhibited by R. hastatus based AgNPs at only high concentrations and E. coli was inhibited by R. dantatus and R. hastatus based AgNPs. However, B. cereus was not inhibited by any AgNPs except R. hastatus and R. hastatus based AgNPs have greater antibacterial potential among all the synthesized AgNPs. These results suggest that synthesized AgNPs have improved antibacterial potential of root extracts of each selected plant and these synthesized AgNPs could be used in pharmaceutical and homeopathic industry for the cure of human diseases.

Molecularly Imprinted Polymer of Colocynthin, An Effective Tool for Quality Control of Citrullus colocynthis Extracts.

BACKGROUND: Different parts of Colocynth, Citrullus colocynthis (L.) Schrad., are used in traditional phytotherapy and homeopathy. OBJECTIVE: In our new approach, a molecularly imprinted polymer was synthesized to absorb colocynthin, the major plant marker, and its capability was evaluated using HPLC-UV. METHOD: A new method was considered to achieve optimal conditions. FT-infrared, N2 adsorption porosimetry, fluorescent and scanning electron microscopy and thermo gravimetric profile of the polymers were studied. The imprinted polymer was applied as molecularly imprinted solid phase extraction sorbent to enrich colocynthin from colocynth oil extract, a traditional medicine dosage form. RESULTS: The imprinted polymer showed high capacity and affinity toward colocynthin. Physical assessments demonstrated no major differences between imprinted and nonimprinted polymers. The imprinted polymer was able to absorb colocynthin more efficiently than non-imprinted and control simple solvent extraction from the real sample. CONCLUSION: In conclusion, this polymer is capable of being applied as a promising adsorbent for analysis of colocynth traditional medicine products.

Variation in Fourier transform infrared spectra of some homeopathic potencies and their diluent media.

OBJECTIVE: The aim of this study was to determine whether potentized homeopathic drugs and their diluent media differ from each other with respect to their Fourier transform infrared (FTIR) spectra. DESIGN: FTIR spectra of Nux vomica 30C, Lycopodium 30C, Santonin 30C, Cina 30C, Cina 206C, Cina 1006C, and their diluent media 90% ethanol and Ethanol 30C were obtained in the wave number range of 2000-1000 cm1 at 20 degrees C. Potassium bromide powder soaked with the potencies, pressed into pellets, and air dried were used to measure the spectra. Because water structures in homeopathic potencies are thought to carry specific information on drug molecules and because O-H bending vibrational band (v2) exclusively belongs to water, the study was restricted to the bands in that wave number region. Alcohol has no absorption in the O-H bending region. RESULTS: The potencies were found to differ from each other and their diluent media in the number of v2 bands, their wave number (cm1), shape, and half-width (cm1) of the bands. CONCLUSIONS: The number and other characteristics of the v2 band represent the number of hydrogen-bonded water species and their hydrogen-bonding strength, respectively. The potencies and their diluent media therefore differ from each other in the number of hydrogen-bonded water species and their hydrogen-bonding strength. The observation that KBr pellets soaked with a potentized drug retains its specific spectral absorption properties simply confirms that medicated sucrose globules, used in homeopathic dispensing, are capable of retaining the therapeutic properties of the drug.

Chemometrics meets homeopathy--an exploratory analysis of infrared spectra of homeopathic granules.

10 homeopathic remedies commercially available (each in 3 dilutions) as sugar granules, where half of them were of organic (and half inorganic) origin were subjected to solid-state infrared spectroscopy, both in middle infrared (ATR-FTIR) and near infrared (NIR) range. Measurements were repeated six times (six days, each sample was measured once in the same day, samples were measured in random order). The obtained spectra was subjected to unsupervised (PCA) and supervised (PLS-DA) chemometric techniques to check any visible differnces in spectral data between homeopathic remedies, including also feature selection approaches. It can be concluded that the only one information encoded in this dataset is the atmospheric drift of spectra between consecutive measurement days. This proves that homeopathy is not "infrared visible" in the case of proper experimental design. These results can be useful in further investigations of possible mechanisms of action of homeopathy (if they exist).

[Analysis of Lapis Micae Aureus and Lapis Chloriti by IR spectra].

OBJECTIVE: To develop a new method of discriminating Lapis Micae Aureus and Lapis Chloriti. METHODS: Fourier transform infrared (FTIR) absorption spectrometry. RESULTS: There were obvious differences between the FTIR spectrometry of Lapis Micae Aureus and Lapis Chloriti from different sources. CONCLUSION: FTIR technique provided a method for the quality control and discrimination of Lapis Micae Aureus and Lapis Chloriti.

[To identify human and sheep placentas by FT-IR and RP-HPLC techniques].

Placenta has a long history of being Chinese traditional medicine. The quality control methods used nowadays in the markets were mainly based on the amino acid content of the placenta. However, it is hard to distinguish human placenta from sheep placenta with these methods. In this paper, a novel method was established employing FT-IR and RP-HPLC techniques. This method can distinguish human placenta from sheep placenta and may help to improve quality control of placenta in the markets.

Variation in free and bound water molecules indifferent homeopathic potencies as revealed by their Fourier Transform Infrared Spectroscopy (FTIR)

Homeopathic potencies 12cH and above cross the Avogadro number, and as such do not containany original drug molecules in their aqueous ethanol medium. It is thought H-bonded waterstructures preserved by ethanol carry the information of initial drug molecules. Potentized drugs show some differences with respect to their infrared (IR) absorption spectra. In a water-ethanol solution, free water molecules vary according to the concentration of ethanol. In the present study the concentration of ethanol has been kept constant at 0.03 molar fractions in 6 differenthomeopathic potencies. Aim: to see whether different homeopathic potencies having fixed ethanolcontent show variation in FTIR spectra and also free water molecules. Two potencies like 8cH and32cH of three homeopathic drugs Natrum mur, Cantharis and Nux vomica were used in thestudy, and their ethanol concentration was kept fixed at 0.03 molar fraction. The control wasconsidered to be aqueous ethanol at the same concentration. Spectrum of reference water wasalso taken. Fourier transform infrared (FTIR) absorption spectra were obtained in the wavenumber region of 4000 – 2800 cm-1. The half-width at half-maximum was measured for eachspectrum. The intensity of each spectrum was normalized at 3410 cm-1 close to the peak. The difference spectrum (absorbance of drug solution – absorbance of reference water) for each drugand the control was obtained. FTIR spectra showed variation in absorbance intensity on both thehigh and low frequency side of the O-H stretching band in different drugs as well as the control. The C-H stretching band of 2977 cm-1 also showed variation in intensity in different drugs...

Variation in free and bound water molecules indifferent homeopathic potencies as revealed by their Fourier Transform Infrared Spectroscopy (FTIR)

Homeopathic potencies 12cH and above cross the Avogadro number, and as such do not containany original drug molecules in their aqueous ethanol medium. It is thought H-bonded waterstructures preserved by ethanol carry the information of initial drug molecules. Potentized drugs show some differences with respect to their infrared (IR) absorption spectra. In a water-ethanol solution, free water molecules vary according to the concentration of ethanol. In the present study the concentration of ethanol has been kept constant at 0.03 molar fractions in 6 differenthomeopathic potencies. Aim: to see whether different homeopathic potencies having fixed ethanolcontent show variation in FTIR spectra and also free water molecules. Two potencies like 8cH and32cH of three homeopathic drugs Natrum mur, Cantharis and Nux vomica were used in thestudy, and their ethanol concentration was kept fixed at 0.03 molar fraction. The control wasconsidered to be aqueous ethanol at the same concentration. Spectrum of reference water wasalso taken. Fourier transform infrared (FTIR) absorption spectra were obtained in the wavenumber region of 4000 – 2800 cm-1. The half-width at half-maximum was measured for eachspectrum. The intensity of each spectrum was normalized at 3410 cm-1 close to the peak. The difference spectrum (absorbance of drug solution – absorbance of reference water) for each drugand the control was obtained. FTIR spectra showed variation in absorbance intensity on both thehigh and low frequency side of the O-H stretching band in different drugs as well as the control. The C-H stretching band of 2977 cm-1 also showed variation in intensity in different drugs...(AU)

Aplicações da espectroscopia no infravermelho no controle de qualidade de medicamentos contendo diclofenaco de potássio. Parte I: dosagem por regressão multivariada / Infrared spectroscopy applied to the drugs quality control. Part I: multivariate regression for the quantification of potassium diclofenac

No presente trabalho são combinadas as potencialidades das ferramentas quimiométricas e as facilidades na obtenção de espectros pela técnica de espectroscopia por refletância difusa no infravermelho médio com transformada de Fourier (DRIFTS), visando à dosagem do fármaco diclofenaco de potássio. Foram formuladas 20 diferentes concentrações do princípio ativo em excipiente amido entre 0,15 e 0,50 mg/g, com variação de aproximadamente 0,02 mg/g na concentração entre cada amostra. Das 20 amostras, 14 foram empregadas para a calibração e 6 para validação. Os espectros foram adquiridos em um espectrômetro NICOLET Magna 550, em duplicata das amostras. Para a construção dos modelos de regressão multivariada, as informações espectrais foram processadas no aplicativo Pirouette® 2.7 da Infometrix, utilizando-se as regiões 3.715-3.088 cm-1 e 1.634 -1.191 cm-1. A calibração foi realizada pelo método de regressão dos mínimos quadrados parciais (PLS), utilizando pré-processamento autoescalado, e tratamento com correção do espalhamento de luz (MSC). Foram selecionados os melhores modelos, considerando os valores do coeficiente de correlação (R²) e da raiz quadrada dos erros médios quadráticos de validação (RMSEV). Os resultados permitem concluir que modelos de calibração, empregando dados DRIFTS e regressão PLS, podem ser empregados como alternativa na dosagem de diclofenaco de potássio, agregando como vantagens frente aos métodos usuais, a não geração de resíduos, a fácil realização da análise e o baixo custo operacional.

In this work the alternative method for quantification of potassium diclofenac was studied, using diffuse reflectance infrared Fourier transform spectrocopy (DRIFTS), together with partial least squares regression (PLS). Two spectra of 20 samples with different concentrations (0.15-0.50 mg/g) of potassium diclofenac in starch had been colleted in Nicolet Magna 550 spectrophotometer, with resolution of the 4 cm-1 and 32 scans. For multivariate regression 14 samples had been used for calibration set and 6 samples had been used for validation set. For the PLS modeling, the spectral information (3715-3088 cm-1 and 1634-1191 cm-1) after multiplicative scatter correction (MSC) and autoscaling preprocess, had been processed in Pirouette® 2.7. The best models with better coefficient regression (R²), these were improved using root mean square error of validation (RMSEV). The new technique propose using DRIFTS/PLS shows an excellent choices for the quality control of the productive process of factories and drugstores that produce or handle these materials in large scale, presenting low time of analysis, no sample destruction and no wastes production.

Análise exploratória aplicada no estudo de medicamentos contendo piroxicam / Exploratory analysis applied in study of pharmaceutical formulations with piroxicam

A identificação de diferentes formulações de medicamentos manipulados contendo piroxicam foi estudada, empregando epectros de reflexão difusa no infravermelho médio com transformada de Fourier (DRIFTS), em associação com a técnica de análise por agrupamentos hierárquicos (AAH). Os espectros de amostras, de 5 diferentes farmácias de manipulação, contendo piroxicam (10 mg e 20 mg) e seus respectivos excipientes, foram adquiridos em um espectrofotômetro NICOLET Magna 550, obtendo-se duas réplicas de cada amostra. Para a análise multivariada, as informações espectrais foram tratadas no programa Pirouette® 2.7 da Infometrix, utilizando-se as regiões espectrais...