Analytical Chemistry HPLC

A sensitive, accurate, robust, and precise analytical quality by design (AQbD)-based RP-HPLC (reverse-phase high-performance liquid chromatography) method has been developed for the simultaneous estimation of perindopril erbumine and moxonidine hydrochloride, both used in hypertension treatment. Initially, a three-level factorial design screened various parameters impacting chromatographic responses, followed by Central Composite Design (CCD) for optimizing the critical parameters. This cost-effective isocratic method employed a Shimpack ODS C-18 column (250 mm × 4.6 mm, 5 μm) as the stationary phase. The method, developed and validated in line with ICH Q2 (R1) guidelines, utilized a mobile phase comprising methanol: acetonitrile: phosphate buffer (34:30:36 v/v/v) adjusted to pH 3.5 with 1% ortho phosphoric acid, at a flow rate of 1.0 mL/min, a column temperature of 40 ºC, and UV detection at 214 nm, achieving efficient separation of both drugs. Linearity ranges were 25-125 μg/mL for perindopril erbumine (r 2 = 0.9996) and 1-5 μg/mL for moxonidine hydrochloride (r 2 = 0.9993). This newly developed RP-HPLC method effectively enables routine quantitative and simultaneous qualitative analysis of these compounds in bulk and synthetic mixtures. Additionally, the method's greenness and environmental safety were evaluated using eco-analytical metrics (eco analytical scale, NEMI, GAPI, and AGREE), confirming its ecological capability.

Analisis senyawa acetaminophen dalam sampel biologis dan sediaan farmasi memiliki peran penting dalam memastikan keamanan, kualitas, dan efektivitas obat. Penelitian ini membahas berbagai metode analisis yang digunakan untuk mendeteksi dan mengukur kadar acetaminophen, termasuk titrasi iodometri, kromatografi lapis tipis (TLC), spektrofotometri UV-Vis, kromatografi cair kinerja tinggi (HPLC), dan spektrometri massa. Setiap metode memiliki keunggulan dan batasannya masing-masing, tergantung pada jenis sampel dan tujuan analisis. Metode HPLC dan spektrometri massa menunjukkan sensitivitas dan spesifisitas tinggi untuk mendeteksi kadar acetaminophen pada konsentrasi rendah, menjadikannya pilihan utama dalam penelitian farmakokinetik dan toksikologi. Di sisi lain, metode sederhana seperti spektrofotometri UV-Vis dan TLC tetap relevan untuk analisis cepat dan ekonomis. Studi ini juga menyoroti pentingnya validasi metode analitik untuk memastikan keandalan hasil, terutama dalam mendeteksi residu acetaminophen dalam tubuh manusia yang dapat menyebabkan efek toksik jika digunakan berlebihan. Penelitian ini merekomendasikan pengembangan metode berbasis teknologi canggih, seperti sensor nanoteknologi, yang ramah lingkungan dan efisien untuk kebutuhan analisis modern. Selain itu, pendekatan kombinasi metode tradisional dan inovatif diharapkan dapat meningkatkan kualitas pengawasan farmasi. Penelitian lebih lanjut diperlukan untuk mengeksplorasi teknologi terbaru dalam deteksi dan analisis senyawa farmasi.
Kata Kunci: Acetaminophen, analisis farmasi, HPLC, spektrofotometri UV-Vis, toksikologi

The present study reveals an improved method for the evaluation of Adhatoda vasica extract by quantitation of major metabolite vasicine. The plant powder was extracted with methanol and evaporated in vaccuo to obtain an oily residue. The residue was diluted with aqueous solution of citric acid (1%) and was further extracted with chloroform. The preliminary screening for alkaloid was done with alkaloidal reagents and the presence of vasicine was assessed by thin layer chromatography (TLC). The extractive was then subjected to column chromatography using a mixture of ethyl acetate and methanol adjusted to pH 9.5 with liquid ammonia as mobile phase. The TLC chromatograms of the collected column fractions were developed and the fractions indicated the presences of vasicine were combined together. The residue obtained from evaporation of solvent was subjected to preparative TLC along with reference standard to get pure crystalline vasicine. The compound vasicine was ascertained by UV-Vis, FT-IR and LCMS/MS analyses. The quantitative estimation of vasicine was carried out by reversed phase HPLC method. About 99.50% of pure vasicine (647 mg) compared to reference standard was obtained from 250 gm of leaf powder (0.2588% w/w).

A simple and sensitive indirect spectrophotometric flow injection analysis (FIA) method for the determination of cefixime (CF), ceftriaxone (CFTR), cefotaxime (CFX) and cefuroxime (CFU) in pharmaceutical formulations and biological fluids has been suggested. Drugs under investigation are firstly oxidized by excess N-bromosuccinimide (NBS) in acidic medium, then the excess NBS is reacted with Rhodamine B (RB) to bleach its pinkish red color. Chemical variables and flow injection variables are all optimized to enhance reproducibility and sensitivity. The suggested procedure followed Beer's law over concentration ranges of 5 -30 μg.ml-1 for all tested drugs. The method is successfully applied to the determinations of the drugs under investigation in different pharmaceutical preparations. Results obtained by the suggested method were in excellent agreement with those obtained by the formal HPLC methods.

Cosmetics are known to contain a long list of toxic chemicals. The aim of this study was to detect and determine the toxic substance para-phenylenediamine (PPD) in some cosmetics such as Kohl and black Lipstick using Reversed Phase-High Pressure Liquid Chromatography (RP-HPLC). In this study, ten cosmetics samples were collected from some local markets in the Libyan city of El-Beida. A simple, rapid and reliable method was developed and validated for the determination of PPD in kohl and lipstick samples using solution of methanol (50%) as solvent. The method has been validated over a wide linear range of 5-25 μg/ml with correlation coefficients being consistently greater than 0.997. The lowest PPD level was observed in Hashmi black Kohl sample (0.0058 % w/w), while the Flormar Eyeliner powder sample showed the highest PPD content (0.0105 % w/w). The RP-HPLC measurments indicated that the PPD content in four samples under study (Hashmi (Kajal) Kohl, Kohl Homemade from Massa, Libya, Kohl Homemade from El-Beida, Libya and Pupa Pencil Eyeliner) were free of PPD (Not Detected). The PPD content in all samples analyzed in this study is well below the permissible limits set by the US Food and Drugs Administration, and documented values.

Botrytis cinerea es un hongo ubicuo que ocasiona la podredumbre gris, una de las principales enfermedades de fruta en poscosecha, siendo responsable de pérdidas económicas. Debido a la creciente preocupación por los efectos adversos del uso de pesticidas, la búsqueda de alternativas se presenta como una meta prioritaria. En este contexto, los extractos de plantas representan una rica fuente de biocompuestos con atractivas propiedades antimicrobianas. Recientemente, los solventes eutécticos naturales (NADES) se han propuesto como agentes extractantes sustentables de compuestos bioactivos a partir de plantas. En el presente estudio, se evaluó un bioextracto de L. cuneifolia basado en NADES hacia B. cinerea. Para este propósito, se usó un NADES compuesto por ácido láctico, glucosa y agua (LGH) como agente de extracción y se comparó con solventes tradicionales en términos de capacidad antioxidante y contenido fenólico total. Además, la actividad antimicrobiana del bioextracto se evaluó ...

Simple, accurate, precise, and sensitive reverse-phase high-performance liquid chromatographic (HPLC) method was developed for the simultaneous measurement of ARI and ESC in pharmaceutical products by using a C18 column with dimensions of 250 mm x 4.6 mm and a particle size of 5 μm. The mobile phase consisted of a mixture of acetonitrile, methanol, and water in the ratio of 80:05:15 v/v/v. Orthophosphoric acid was added to adjust the pH to 7.00 ± 0.1. The detection wavelength used for measuring the absorbance of the components was 246 nm, and the Mobile Phase flow rate was set at 1.0 mL/min. The retention times of ARI and ESC were 5.63 and 3.65 minutes, respectively. The ARI and ESC correlation coefficients (R 2-values) were observed at 0.9995 and 0.9997, respectively, demonstrating a robust linear relationship between the analyte concentration and the observed absorbance. The linearity range for ARI was 20 to 120 μg/mL, while ESC was 60 to 360 μg/mL. The recovery percentages for ARI and ESC ranged from 99.43% to 100.65% and 99.58% to 100.15%, respectively. Additionally, all parameter uncertainty levels were below 2. The method was validated following ICH guideline Q2 (R1). The analytical method validated will be successfully applied to the pharmaceutical dosage form.

Andrographis paniculata (Burm.f.) Nees. is a medicinal herb widely used in various traditional medicines for the treatment of various ailments particularly related to viral fever, respiratory tract infections, and other pharmacological properties. Andrographolide is a diterpene present as a major active ingredient responsible for various pharmacological activities. This study was conducted to assess the mutagenicity and clastogenicity potential of the andrographolide by Ames mutagenicity and in vitro chromosomal aberration studies, respectively. Materials and Methods: Ames Mutagenicity assay was performed using Salmonella typhimurium strains, TA100, TA1535, TA102 and TA98 and TA1537 and in vitro chromosomal aberration study was performed in CHO-K1 cell line. Both these studies were conducted following the procedure mentioned in the OECD guidelines no. 471 and 473 for Ames and in vitro chromosomal aberration studies, respectively. Results: Andrographolide was found to be non-mutagenic in Ames mutagenicity assay and nonclastogenic in the in vitro chromosomal aberration study in the CHO-K1 cell line.

Puerarin is the major bioactive ingredient in the roots of Kudzu (Radix Puerariae thomsonii), a traditional herbal that has good effects in the treatment of cardiovascular disease, antioxidants and alcohol abuse. This study was performed to develop and validate the method of quantifying puerarin in Kudzu roots; The method was applied to assay the content of puerarin in some pharmaceutical samples on the market. Chromatographic conditions were determined including Ascentis® RP C18 column (250 × 4.6mm; 5µm), detection wavelength of 250nm; 1ml/min flow rate, mobile phase methanol:water (25:75). The method was validated for the specificity, linearity ranging from 0.5 to 50µg/ml (r = 0.9998), accuracy (recovery: 96.7 to 103.65%, RSD <2%), repeatability good return (RSD <2%). The content of puerarin in roots of kudzu ranges from 0.014% to 0.762% on the dry weight.

Food safety remains a major concern for society, applications to protect milk and animal productions from chemical residues by using precise methods to detect the smaller levels of these residues. The identification of DLM and IVM residues in 80 samples (40 each sheep milk and mutton) collected from different area and slaughterhouses shop of wasit was performed. Production was monitored by using HPLC technique to determine the degree of DLM and IVE residues contamination in milk and mutton from April to July of 2018. The analysis showed that 92.5 % of the milk and 90% of mutton samples were contaminated above MRLs with DLM residues ,but all samples 100% (milk and mutton) were positive for IVM residues at the period of study .The results of milk and mutton revealed were 100% violation of the MRLs (above 0.026) according to MRLs Permissible by the WHO and FAO from DLM and show a significant difference (P<0.05) between July and April ,May and June in sheep milk and mutton (0.95 ± 0.04,0.83 ± 0.10,0.62 ± 0.08, and0.82 ± 0.16), (1.34 ± 0.11,0.36 ± 0.06, 0.62 ± 0.12 and 0.86 ± 0.10) respectively , but in IVM residues (ppm) result show found significant difference (P<0.05) in sheep milk between June and April ,May and July (0.83+0.12, 0.45+0.08, 0.69+0.12 and 0.73 + 0.10) ,and between July and April ,May and June in mutton (1.2 ± 0.08, 0.84± 0.09, 0.78±0.08 and 0.99±0.09). The results of this study confirm the need for monitoring programs for the residues of pesticides and ecto-parasites in animal products to protect the consumer health from the dangers of exposure to these remains.

The disadvantage of HTPB products processed by radical mechanism is the wide range distribution of molecular weight and molecular weight tends to be higher than predicted. To be able to obtain appropriate HTPB as fuel binder of propellant, we need a method of separation of HTPB based on molecular weight. The purpose of this research was to develop a method of separation based on differences in molecular size HTPB based filtration in porous resin column. The expected benefits of the research is obtained good separation method can improve the molecular weight of the polymer so that it meets the requirements of a propellant fuel binder. Research carried out by flush the polymer into the column containing porous resin Crosslinked PE by using a solvent toluenehydrofurane. Each faction 2 mL taken and analyzed its concentration and molecular weight by gel chromatography. Separation parameters to be searched is the size of the resin, the column length and flow rate of the solvent (elution)....

Secnidazole is rapidly absorbed after oral administration and it has a bioavailability of about 80% when given in high dose of about 2 g. The high dose of Secnidazole causes many side effects even with a single dose such as dizziness, abdominal pain, urticaria, glossitis, stomatitis, taste disturbances, paresthesias and more severe side effects include leucopenia and Secnidazole-induced acute pancreatitis. Hence the present study aims to increase the bioavailability of secnidazole by a bioenhancer, piperine, in order to reduce the therapeutic dose of secnidazole, thus decreasing its side effects. Analytical and bioanalytical methods were developed and validated for simultaneous estimation of secnidazole and piperine by HPLC. Pharmacokinetic study was carried out in rats for secnidazole alone and secnidazole-piperine combination. The results revealed that the dose of Secnidazole can be reduced up to 1/3rd of its original dose, which will lead to better patient compliance.

This work included external morphological study of horse fly Tabanus nemoralis Meig. 1820, which belongs to family Tabanidae order: Diptera. The study involved the most important taxonomic external characters of the: head, thorax, abdomen and their appendages which are: antenna, maxillary palp, wings, legs, spotting in coloring pattern of abdomen.

Two simple, selective, precise, and accurate chromatographic methods, namely reversed-phase highperformance liquid chromatography (RP-HPLC) method and High-performance thin-layer chromatography (HPTLC), were developed and validated for the simultaneous determination of Atorvastatin (ATO) and Perindopril (PER). The developed HPTLC method was used for the separation and quantitation of the studied drugs on silica gel 60F254 TLC plates. Dichloromethane: Methanol: Ethyl acetate: Glacial acetic acid (6:2:2:0.1, v/v/v/v) was used as a developing system and the separated bands were UV-scanned at 221 nm. Linear relationships were obtained in the range of 800-4800 ng/band for Atorvastatin and 200-1200 ng/band for Perindopril with a regression coefficient greater than 0.999. The Rf value of the drug was found to be 0.70 ± 0.02 and 0.39 ± 0.03 for Atorvastatin and Perindopril respectively. The developed RP-HPLC depended on chromatographic separation of the studied drugs on a C18 column using Acetonitrile: Methanol: Potassium dihydrogen orthophosphate (pH 3 adjusted with orthophosphoric acid) ((40:10:50), v/v/v) as a mobile phase delivered at a constant flow rate of 1 mL/min with UV detection at 240 nm. The calibration curves were linear (r2 > 0.999) over the concentration range 20-100 µg/mL for Atorvastatin and 10-50µg/mL for Perindopril. The average retention times for Atorvastatin and Perindopril were 3.42 and 8.92 min, respectively. Factors affecting the developed methods have been studied and optimized. Further, methods validation has been carried out according to ICH guidelines. The proposed methods were successfully applied for the determination of the studied drug simultaneously in bulk and synthetic mixture qualitatively and quantitatively. Statistical analysis by the F test showed no significant difference between the results obtained by the two methods. The uncertainty measurement was also carried out for the quantification of both components. The proposed HPTLC method proved to be more sensitive, while the HPLC gave more reproducible results and was time-saving.

Idiopathic pulmonary fibrosis (IPF) is an interstitial lung disease characterized by collagen deposition as a consequence of excessive lung fibroblasts and myofibroblasts proliferation. We aimed to investigate for the first time the effect of rosemary leaf extract rich with carnosic acid (CA) or rosmarinic acid (RA), after industrial elimination of essential oils, against bleomycin (BLM)-induced lung fibrosis in rats. Male Wistar rats were given a single dose of BLM (4 mg/kg, intratracheal), while CA rich extract, RA rich extract or the combination RA/ CA rich extracts (10, 75 and 150 mg/kg, intraperitoneal) were administered 3 day later and continued for 4 weeks. We reveled by HPLC an important similar amount of phenolic compounds such as pyrogallol, vanillic, gallic and ellagic acids in both rosemary extracts. BLM induced lung fibrotic foci and disturbance in superoxide dismutase, catalase and malondialdehyde levels. At 10 mg/kg, both rosemary extracts administrated alone or in combination alleviated synergistically lung fibrosis and ameliorated oxidative changes induced by BLM. In conclusion, industrial elimination of essential oils from rosemary allowed us to obtain two extracts with potent antifibrotic activities due to the large amount of RA and CA that appear much higher and effective than wild rosemary extract.

Idiopathic pulmonary fibrosis (IPF) is a chronic lung disease characterized by progressive and irreversible fibroblasts proliferation leading to significant respiratory insufficiency. This study was designed to investigate the effect of sage infusion against bleomycin (BLM)induced lung fibrosis in rats. Male Wistar rats were given a single dose of BLM (4 mg/kg, intratracheal), while sage infusion (50, 100 and 150 mg/kg, intraperitoneal) was administered 3 day later and continued for 4 weeks. We reveled by HPLC and LC-MS methods an important amount of phenolic bioactive compounds such as vanillic, gallic, ellagic, rosmarinic and carnosic acids. BLM induced collagen deposition, increased lipid peroxidation (MDA) and decreased superoxide dismutase (SOD) and catalase (CAT) activities. Only sage infusion at 150 mg/kg normalized MDA and antioxidant enzyme levels (SOD and CAT) and reduced significantly lung fibrosis. Our results showed also that this high dose have no renal or hepatic cytotoxic effect. In conclusion, sage can protect against BLM-induced murine lung fibrosis and oxidative stress due to the large content of bioactive phenolic compounds.

Background: Idiopathic Pulmonary fibrosis (IPF) is a serious, progressive lung disease characterized by accumulation of extracellular matrix (ECM), leading to functional deterioration in lung parenchyma. Objectives: to characterize the effect of rosuvastatin on bleomycin-induced pulmonary fibrosis in mice, to compare its effect with the well-established anti-fibrotic effect of pirfenidone, and the role of transforming growth factor-βeta (TGF-β). Materials & methods: induction of pulmonary fibrosis was done by intra-tracheal injection of bleomycin (2U/Kg, 50 μl) to C57BL/6 mice. Then, at the 10 th day; pirfenidone (300mg/kg/d) or rosuvastatin (10mg/kg/d) given orally. After 21 days, all mice were sacrificed. Body weight, lung weight, lung index and lung TGF-β1were measured before and after induction of pulmonary fibrosis. Histopathological examinations of lung sections were done using haematoxylin and eosin. Also, Masson's trichrome stain was used for scoring fibrosis. Results: Significant decrease in body weight and increase in lung weight, lung index, TGF-β levels were observed in bleomycin group. Also, the histopathological fibrosis score was increased in comparison to control group. Both groups of pirfenidone and rosuvastatin showed significant increase in body weight, and decrease in lung weight, lung index, TGF-β levels. Histopathological fibrosis score was decreased in comparison to bleomycin group. Conclusions: Rosuvastatin effectively reduced pulmonary fibrosis as pirfenidone. This was done by decreasing TGF-β levels. Thus, the pleiotropic actions of rosuvastatin might be applied clinically for prevention or treatment of bleomycin-induced pulmonary fibrosis. However, further studies are needed to confirm the beneficial therapeutic effects of rosuvastatin in the therapy of bleomycin-induced pulmonary fibrosis.

Objective: In the present work, rapid and sensitive isocratic RP-UPLC method was established and comprehensive validation study for the estimation of Tolperisone HCl and Paracetamol was carried out according to international conference on harmonization (ICH) guidelines. Methods: Simultaneous estimation was chromatographed using 0.1% o-phosphoric acid in water and acetonitrile (70: 30 v/v) as a mobile phase at a flow rate of 0.20 ml/min with 35 ºC column temperature. Chromatographic separation accomplished isocratically on Acquity UPLC BEH C18 (50 mm ×2.1 mm, particle size 1.7 µm) and detection utilizing photodiode array detector at 254 nm. Injection volume was 2.0 µl. Results: The calibration curve was linear over the wide concentration range of 6.0µg/ml to 54.0µg/ml and 20.0µg/ml to 180.0µg/ml for Tolperisone HCl and Paracetamol, respectively. The retention time of Tolperisone HCl and Paracetamol was 1.396 and 2.625 min, respectively and the total analysis time was 5.0 min. Based on the results, the validated method was effectively applied for the estimation of Tolperisone HCl and Paracetamol in combined dosage form and in single pharmaceutical formulations with a new generation instrument, ultra performance liquid chromatography (UPLC). Moreover, the method helps to get better quality control and to pledge therapeutic efficacy. Conclusion: The method is simple, less time consuming and comparatively cost effective than existing methods.

Objective: To develop an accurate, precise and linear Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method and validate as per ICH guidelines for the quantitative estimation of Ribavirin (200mg) in tablets. Methods: The optimized method uses a reverse phase column, Enable Make KromasilC18 (250 X 4.6 mm; 5μ), a mobile phase of phosphate buffer (pH 4.2): acetonitrile in the proportion of 85:15 v/v, flow rate of 1.0 ml/min and a detection wavelength of 215 nm using a PDA detector. Results: The developed method resulted in Ribavirin eluting at 2.606 min. Ribavirin exhibited linear in the range 25-150μg/ml. The precision is exemplified by the relative standard deviation of 0.4%. Percentage Mean recovery was found to be in the range of 98-102, during accuracy studies. The limit of detection (LOD) and limit of quantitiation (LOQ) was found to be 0.24ng/ml and 0.73ng/ml respectively. Conclusion: An accurate, precise and linear RP-HPLC method was developed and validated for the quantitative estimation of Ribavirin in VIRAZIDE (200mg) tablets as per ICH guidelines and hence it can be used for the routine analysis in various pharmaceutical industries.

Background: Aflatoxin B1 (AFB1) is a carcinogenic metabolite produced by Aspergillus falvus which cause several diseases in humans. This study aimed to determinate the role of polymerase chain reaction (PCR) and high-performance liquid chromatography (HPLC) techniques in discrimination between aflatoxigenic and non-aflatoxigenic producer isolates of A. flavus and molecular expression for aflatoxin encoding genes (aflM ver-1, aflD, and aflR). Furthermore, to assess the cytotoxic effect of AFB on isolated hematopoietic stem cells (HSCs) obtained from the human umbilical cord. Materials and Methods: The extracted aflatoxin was detected by HPLC. Fungal DNA was extracted and submitted to PCR for amplifying AFB encoding genes (nor1 aflD, aflM, and aflR). The collection of umbilical cord blood samples was accomplished using the system close method. CD34 were used for immunophenotypic analysis of HSC which were obtained using buffy coat method. An experimental study was conducted in vitro to assess the cytotoxicity effects of AFB with certain concentrations on the viability of AFB treated-HSC. Results: Out of 15 A. flavus, the genes (AflR, nor1 aflD) were founded in 11 (73%) of A. flavus strains, while the (ver aflM) was appeared in 10 (67%) of strains. HPLC technique had been detected efficiently AFB1-producing isolates in 10 (66.7%) out of 15 isolates of A. flavus with rates ranged from 0.78 to 45.03 ppm. It was yielded that the inhibition rate of isolated HSC was increased seriously with an increase of aflatoxins concentration leading to decrease in viability of AFB-treated cells. Conclusions: Aflatoxin production was directly associated with the appearance and expression of genes (aflM ver-1, aflD, and aflR) by PCR. Furthermore, HPLC is a superior technique in identifying and analyzing aflatoxin with high sensitivity and accuracy. Further, in a cytotoxicity assay, whenever the aflatoxin concentration was high, the rate of HSC growth inhibition increases and viability of AFB-treated cells decreased.

The IUPAC name of Cfx is (6R,7R)-7-[[(2Z)-2-(2-amino-1,3-thiazol-4-yl)-2(carboxymethoxyimino)acetyl] amino]-3-ethenyl-8-oxo-5-thia-1-azabicyc-ABSTRACT Cefixime (Cfx) is a member of the third generation of Cephalosporin antibiotics. It used on a wide scale in prescribed antibiotic drugs as anti-infection for Gram-positive and Gram-negative microorganisms. The present study aimed to develop an HPLC method of Cfx analysis enjoyed highly linearity, repeatability, robustness, ruggedness, selectivity, rapidly, and economical to use. The chromatographic system depends on the RP-BDS column (250 mm x 4.6 mm x 5 μm). The mobile phase was prepared by mixing Methanol: Phosphate buffer (3:7, v/v) at flow rate 1.0 ml/min with wavelength detection at 254 nm, the temperature at 30° C with injection volume 20 μL. The method revealed that satisfied linearity regression R 2 (0.9996) with repeatability (0.94%) with DL and QL; 59.3 ng/ml and 179.8 ng/ml respectively. The method showed successful and satisfying results for Cfx in bulk and pharmaceutical formulations and urine samples at low levels.

New, simple and sensitive batch and Flow-injecton spectrophotometric methods for the determination of Thymol in pure form and in mouth wash preparations have been proposed in this study. These methods were based on a diazotization and coupling reaction between Thymol and diazotized procaine HCl in alkaline medium to form an intense orange-red water-soluble dye that is stable and has a maximum absorption at 474 nm. A graphs of absorbance versus concentration show that Beer's law is obeyed over the concentration range of 0.4-4.8 and 4-80 µg.ml-1 of Thymol, with detection limits of 0.072 and 1.807 µg.ml-1 of Thymol for batch and FIA methods respectively. The FIA procedure sample throughput was 80 h-1. All different chemical and physical experimental parameters that affecting on the development and stability of the colored product were carefully studied and the proposed methods were successfully applied to the determination of Thymol in mouth wash preparations.

One of the long-known separation methods is chromatography, such as high performance liquid chromatography (HPLC). This separation essentially uses column containing particle-packed which generates high flow-resistance and low mass transfer. As matter of fact, it has an impact to high amount mobile phase usage. Meanwhile, organic polymer monolithic column has been widely used as alternative due to its high mass transfer. In addition, the resulting column has stability over wide pH range, high temperature, shrinkage and swelling of the reservoir. The aim of this research was to produce monolithic column which able to utilized for separation with low amount of mobile phase usage. Preparation of monolithic column begins with pretreatment of polyetereterketone tubing column inner wall. It conducted by activating the inner wall with H 2 SO 4 49% (v/v), followed by vinylization with glycidyl methacrylate. Furthermore, pretreated column filled by polymer mixture consisted of 30 wt% of monomers glycidyl methacrylate:trimetilolpropane trimethacrylate 4:1 (w/v), 70 wt% of pore-forming agents (1-propanol/1,4-butanediol/water 7:4:1 w/v), and azoisobutyronitrile 1 wt% of total monomers amount. The polymerization conducted at 60°C for 12 h. Produced column connected to HPLC then applied to separate toluene and amylbenzene. The result shows both compounds successfully n flow rate of acetonitrile:water (75:25) at 84 and 132 min respectively.

Simple, accurate, precise, and sensitive reverse-phase high-performance liquid chromatographic (HPLC) method was developed for the simultaneous measurement of ARI and ESC in pharmaceutical products by using a C18 column with dimensions of 250 mm x 4.6 mm and a particle size of 5 μm. The mobile phase consisted of a mixture of acetonitrile, methanol, and water in the ratio of 80:05:15 v/v/v. Orthophosphoric acid was added to adjust the pH to 7.00 ± 0.1. The detection wavelength used for measuring the absorbance of the components was 246 nm, and the Mobile Phase flow rate was set at 1.0 mL/min. The retention times of ARI and ESC were 5.63 and 3.65 minutes, respectively. The ARI and ESC correlation coefficients (R 2-values) were observed at 0.9995 and 0.9997, respectively, demonstrating a robust linear relationship between the analyte concentration and the observed absorbance. The linearity range for ARI was 20 to 120 μg/mL, while ESC was 60 to 360 μg/mL. The recovery percentages for ARI and ESC ranged from 99.43% to 100.65% and 99.58% to 100.15%, respectively. Additionally, all parameter uncertainty levels were below 2. The method was validated following ICH guideline Q2 (R1). The analytical method validated will be successfully applied to the pharmaceutical dosage form.

Two simple, selective, precise, and accurate chromatographic methods, namely reversed-phase highperformance liquid chromatography (RP-HPLC) method and High-performance thin-layer chromatography (HPTLC), were developed and validated for the simultaneous determination of Atorvastatin (ATO) and Perindopril (PER). The developed HPTLC method was used for the separation and quantitation of the studied drugs on silica gel 60F254 TLC plates. Dichloromethane: Methanol: Ethyl acetate: Glacial acetic acid (6:2:2:0.1, v/v/v/v) was used as a developing system and the separated bands were UV-scanned at 221 nm. Linear relationships were obtained in the range of 800-4800 ng/band for Atorvastatin and 200-1200 ng/band for Perindopril with a regression coefficient greater than 0.999. The Rf value of the drug was found to be 0.70 ± 0.02 and 0.39 ± 0.03 for Atorvastatin and Perindopril respectively. The developed RP-HPLC depended on chromatographic separation of the studied drugs on a C18 column using Acetonitrile: Methanol: Potassium dihydrogen orthophosphate (pH 3 adjusted with orthophosphoric acid) ((40:10:50), v/v/v) as a mobile phase delivered at a constant flow rate of 1 mL/min with UV detection at 240 nm. The calibration curves were linear (r2 > 0.999) over the concentration range 20-100 µg/mL for Atorvastatin and 10-50µg/mL for Perindopril. The average retention times for Atorvastatin and Perindopril were 3.42 and 8.92 min, respectively. Factors affecting the developed methods have been studied and optimized. Further, methods validation has been carried out according to ICH guidelines. The proposed methods were successfully applied for the determination of the studied drug simultaneously in bulk and synthetic mixture qualitatively and quantitatively. Statistical analysis by the F test showed no significant difference between the results obtained by the two methods. The uncertainty measurement was also carried out for the quantification of both components. The proposed HPTLC method proved to be more sensitive, while the HPLC gave more reproducible results and was time-saving.

Clinical trials have demonstrated that combining fimasartan potassium trihydrate and atorvastatin calcium produces better results for essential hypertension and dyslipidemia with fewer side effects than individual drugs. However, no analytical method is available to accurately determine both drugs' qualitative and quantitative amounts when used together. Therefore, we are interested in developing and validating three analytical methods for simultaneously estimating fimasartan potassium trihydrate and atorvastatin calcium in laboratory-prepared tablets. These methods include a novel, simple, precise, sensitive, robust HPLC technique, and two UV spectrophotometric methods. The HPLC method used a reversed-phase Shimpack ODS C18 column as a stationary phase with an isocratic procedure. A mobile phase consisting of a mixture of acetonitrile and 0.01 M ammonium acetate buffer, (pH adjusted to 3.7 using formic acid) at a ratio of 70:30 v/v, at a constant flow rate of 1 mL/min and chromatograms were recorded at 234 nm for the two analytes. Vierordt's simultaneous equation method used two wavelengths (247 nm and 263 nm) to estimate the drugs' amounts and derive a simultaneous equation for FIMA and ATOR. Meanwhile, the first-order derivative method used zero-crossing points of 224nm and 261nm to estimate the concentrations of ATOR and FIMA. A statistical analysis using an analysis of variance test revealed no significant differences among the three methods regarding accuracy and precision. Moreover, the three developed methods underwent rigorous statistical validation by the ICH Q2R1 guidelines, demonstrating their successful application in the concurrent analysis of the two drugs in bulk and tablets prepared within the laboratory. To assess the ecological impact of the new UV spectrophotometric technique and HPLC method, the greenness profile was evaluated utilizing Analytical eco scale (AES), National environmental method index (NEMI), the Green analytical procedure index (GAPI), and the software-based Analytical greenness metric (AGREE) tools. This evaluation substantiated the method's environmental friendliness regarding solvent usage, chemical substances, energy consumption, and waste generation.

Objective Review and assess pharmaceutical and clinical characteristics of chloroquine including high-performance liquid chromatography (HPLC)-based methods used to quantify the drug in pharmaceutical products and biological samples. Evidence acquisition A literature review was undertaken on the PubMed, Science Direct, and Scielo databases using the following keywords related to the investigated subject: 'chloroquine', 'analytical methods', and 'HPLC'. Results For more than seven decades, chloroquine has been used to treat malaria and some autoimmune diseases, such as lupus erythematosus and rheumatoid arthritis. There is growing interest in chloroquine as a therapeutic alternative in the treatment of HIV, Q fever, Whipple's disease, fungal, Zika, Chikungunya infections, Sjogren's syndrome, porphyria, chronic ulcerative stomatitis, polymorphic light eruption, and different types of cancer. HPLC coupled to UV detectors is the most employed method to quantify chloroquine in pharmaceutical products and biological samples. The main chromatographic conditions used to identify and quantify chloroquine from tablets and injections, degradation products, and metabolites are presented and discussed. Conclusion Research findings reported in this article may facilitate the repositioning, quality control, and biological monitoring of chloroquine in modern pharmaceutical dosage forms and treatments.

The concept of implementing e-government systems is growing widely all around the world and becoming an interest to all governments. However, governments are still seeking for effective ways to implement e-government systems properly and successfully. As services of e-government increased and citizens' demands expand, the e-government systems become costlier to satisfy the growing needs. The cloud computing is a technique that has been discussed lately as a solution to overcome some problems that an e-government implementation or expansion is going through. This paper is a proposal of a new model for egovernment on basis of cloud computing. E-Government Public Cloud Model EGPCM, for egovernment is related to public cloud computing.

Context: Isoflavones may act as selective regulators in the prevention of various diseases. The most important source of isoflavones is the soy, from which different phytotherapeutics are elaborated of use in Ecuadorian population. However, its concentration varies depending on several factors, therefore quality assessment need to be carried out through out several analytical methods. Aims: To validate an analytical method by high precision liquid chromatography (HPLC) to quantify total isoflavones in herbal medicine. Methods: To quantify isoflavones, it was used a brand liquid chromatography with UV/VIS detector at 260 nm, C-18 column using isocratic method. The mobile phase was composed of 2% acetic acid: acetonitrile (75:25). The quantification was performed against reference standard. The parameters for the validation followed the established in the USP 33. Results: The chromatogram presented six peaks with elution between 1.557 and 18.913 min. The linearity of the system and th...

This paper introduces an original, rapid, efficient and reliable HPLC method for the accurate and simultaneous quantification (g/L) of chloride in samples containing sugars, organic acids and alcohols. Separation was achieved using a HI-Plex H column at 35 C, with H 2 SO 4 (0.005 N) as the mobile phase at a flow rate of 0.4 mL/min. The column effluent was monitored by a Refractive Index (RI) detector. A linear response was achieved over NaCl concentrations of 0.25-2.5 g/L and 5-40 g/L. The analytical method inter-and intra-run accuracy and precision were better than AE10.0%. Investigating the mechanism of detection using different chloride and sodium s reviled that this method can be used for determining the total concentration of chloride salts when in suspension. This method was successfully applied to 15 samples of commercial food products and the salt content obtained from this method was compared with 3 other methods for salt determination. The (HI-Plex H) column was designed for determining the concentrations of sugars, organic acids and alcohols when in solution. Hence, application of our new methodology would allow the determination of sugars, alcohols and organic acids in samples derived from seawater-based fermentation media as well as samples from salty food and dairy products.

RESUMO: Separações cromatográficas são importantes no fim de processos que se utilizam de rotas químicas, visando a recuperação e o aumento da pureza do produto de interesse. O piperonal, obtido por síntese, apresenta pureza cerca de 85 %. Devido à sua relevância para a indústria de cosméticos é necessário purezas cada vez maiores. O objetivo deste trabalho é o de separar o piperonal em escala semipreparativa em batelada. Fizeram-se injeções em escala analítica para obter os parâmetros seletividade e resolução. Com tais parâmetros, foram obtidos o tempo de ciclo de cada injeção, o tempo de coleta e a concentração da solução de alimentação. O sistema de separação consistiu de uma coluna analítica e uma semipreparativa preenchidas com fase estacionária C 18 , tendo como fase móvel etanol/água, na proporção 70/30 (v/v) à 35 o C. Com a cromatografia semipreparativa sequencial foi possível obter o piperonal com pureza elevada, na ordem de 99 %.

The issue of sustainability is a growing concern and has led to many environmentally friendly chemical productions through a great intensification of the use of biomass conversion processes. Thermal conversion of biomass is one of the most attractive tools currently used, and pyrolytic treatments represent the most flexible approach to biomass conversion. In this scenario, microwave-assisted pyrolysis could be a solid choice for the production of multi-chemical mixtures known as bio-oils. Bio-oils could represent a promising new source of high-value species ranging from bioactive chemicals to green solvents. In this review, we have summarized the most recent developments regarding bio-oil production through microwave-induced pyrolytic degradation of biomasses.

The issue of sustainability is a growing concern and has led to many environmentally friendly chemical productions through a great intensification of the use of biomass conversion processes. Thermal conversion of biomass is one of the most attractive tools currently used, and pyrolytic treatments represent the most flexible approach to biomass conversion. In this scenario, microwave-assisted pyrolysis could be a solid choice for the production of multi-chemical mixtures known as bio-oils. Bio-oils could represent a promising new source of high-value species ranging from bioactive chemicals to green solvents. In this review, we have summarized the most recent developments regarding bio-oil production through microwave-induced pyrolytic degradation of biomasses.

Porous material in contact with vapor tends to adsorb fluid in the pores. The amount of adsorbed fluid depends on the vapor pressure, but depends in some parameter ranges as well on the history of the system. When raising the vapor pressure the adsorption may be smaller than while lowering the pressure. Thus, the measured adsorption isotherm consists of two branches that appear stable on the time scale of experiments. The phenomenon is known as adsorption hysteresis and has been extensively discussed in the literature as it is in distinct contrast to the expectations of thermodynamics:the system has more than one answer to one set of boundary conditions (here: grand canonical boundary conditions). The common explanation offered in the literature introduces the concept of metastable states, conceiving either or both branches of the isothermas being metastable. Even though the concept of metastability cannot be separated from the concept of a lifetime against decay into the correspond...

The monument to the Actium victory raised by Octavian Augustus on the northern outskirts of Nicopolis, in the area where the Actian games were taking place, consists of an enormous podium built in ashlar masonry on the slopes of a hill sacred to Apollo. On the upper part of the terrace created by the podium, a porticus triplex was surrounding a courtyard. The area of the courtyard was occupied by a monumental altar and by two pedestals carrying bronze statues of unknown figures. The walls of the porticus and of an annex on its north wing were decorated with frescos fragments which were preserved in situ, while several other fragments were found scattered throughout the monument. In this article, we present the results of the identification of the organic binding medium, as well as the isolation of microorganisms from the wall painting fragments using HPLC and classical microbiological isolation techniques OM and SEM, respectively. Ten samples were selected for HPLC-FD analysis and compared with two reference mixtures of egg yolk with gelatin 3:1 w/w and 5:1 w/w. Six of the ten samples gave proteinaceous profile results, yielding an amino acid profile like the mixture of egg yolk/gelatin 3:1 w/w, which is relatively poor in gelatin content. The fungal species isolated were Aspergillus sp., Penicillium sp., unidentified strains of mold and fungi and Bacillus sp. This study aims to assess the present-day condition of the wall paintings for future investigation. It is well known that the metabolism of organic bindings in the substrate, like animal glue loses their binding effect when they are contaminated or when they have been in use for an extended time and consolidation is required. During a microbial infestation, these organic bindings decompose thus losing their consolidation effect. The metabolism of organic bindings is restricted to natural organic bindings and affects synthetic polymers applied for consolidation purposes.

This present review submitted high-performance liquid chromatography (HPLC) methods for determining the presence of meloxicam used in clinical treatment within various fields of application, including biological samples and pharmaceutical formulations, between 1990 and 2011. Meloxicam is a cyclooxygenase-2 selective nonsteroidal anti-inflammatory drug (NSAIDs), which is the group most often used in human health care. We compared the methods from the standpoint of sensitivity. HPLC methods with MS/MS detection have become the method of choice in bioanalytical applications.

Resumen Los copolímeros de estireno-divinilbenceno (EST-DVB) tienen un amplio rango de aplicaciones como resinas de intercambio iónico y absorbentes. La síntesis de estos copolímeros ha sido objeto de estudios durante mucho tiempo debido a las diversas variables que se pueden ajustar o modificar, como, por ejemplo, porógeno, tamaño, tiempo de agitación, velocidad de agitación, agente sulfonante, entre otros, con la finalidad de obtener un soporte polimérico con características físico-químicas adecuadas, tales como, inercia, estabilidad térmica, y la capacidad de ser reutilizables y de baja toxicidad. En este trabajo se plantea la variación de parámetros de síntesis en suspensión de estas resinas, como la cantidad de porógeno a utilizar, la velocidad de agitación y la cantidad de agente entrecruzante divinilbenceno (DVB), para determinar su influencia en la capacidad de intercambio. Se pudo obtener tres resinas con tamaño de partículas entre 55-75 µm (Resina 1) y 90-200 µm (Resinas 2...

The aim of this experiment was to analyze a mixture of 3 polycyclic aromatic hydrocarbons (PAHs), namely naphthalene, acenaphthene and phenanthrene by reversed-phase liquid chromatography, coupled with UV and FLD detectors. A soil sample was extracted, purified and injected into the instrument to separate the PAHs.

The gain in sensitivity in fluorescence compared to UV detection is associated more with the absence of background noise and the possibility of selecting both an excitation and emission wavelength and which can be optimized by systematically testing the signal-to-noise ratio as a function of the detector gain setting. This is all the more true given that
• the coefficients of variation for the readings of the enriched standards are practically the same in both UV and FLD and that,
• the recovery rates are around 80% for naphthalene and acenaphthene both in UV and FLD and that,
• the limits of detection (LOD), for the 3 PAHs, are around 0.1 in FLD and 0.2 mg/L in UV.
This greater sensitivity of fluorescence for our 3 PAHs does not translate into a lower variation in dispersion – since the standard deviation also increases. However, a coefficient of variation of fluorescence data lower than 10% could suggest more grouped data and a more homogeneous distribution, but this is not the case.

The greater precision of acenaphthene in UV and a low relative standard deviation for triplicates thereof explains the low average coefficient of variation for the 3 products. And the fact that this relative standard deviation for acenaphthene differs greatly from those observed for naphthalene and phenanthrene in UV does not allow a meaningful comparison to be made between the two detectors if it is based solely on precision, repeatability and variation in dispersion.
The slight increase of 1.2 seconds in fluorescence retention times is due to the longer tube after column for this detector. And this difference cannot be explained by the simple fact that the UV detector, on the same line at the exit of the column, precedes the fluorescence detection – a difference of 1.2 seconds would not be enough to justify such an increase in the dispersion.
The detection limits calculated in Table 18 are probably not the lowest PAH concentrations that can be analyzed by our method. Similar to detection, the limits of quantification calculated and displayed in this table are unlikely to represent a signal-to-noise ratio of 10 and should be evaluated on the merit of linearity. The reproducibilities in Table 17 have no intrinsic comparative significance except proportionality to the standard deviation, since they all originate from 2 measurements and the t (Student) value is the same for each.

Although it has not been confirmed, this HPLC-FLD-UV technique is surely more sensitive than the GC-MS method seen in experiment 4 for the identification of essential oils. Thus, the HPLC–FLD-UV method was used to monitor both two levels of enrichment and solid phase extraction (SPE) using commercial Restek cartridges and which were also evaluated for the recovery of PAH. Recoveries between 75% and 90% have been achieved using these SPE cartridges. Considering the non-polar nature of PAHs, it would have been interesting to verify whether better results could have been obtained using a liquid-liquid extraction technique for the analysis of real water or soil samples. We could thus have compared the limits obtained from Table 18, the reproducibilities from Table 17 and especially the recovery rates from Table 16 which, all in all, had reasonable values except for phenanthrene in UV.

We determined the PAH concentrations in the unknown B1. The two detectors indicate similar values for the 3 PAHs in the unknown. The concentrations of naphthalene, acenaphthene and phenanthrene by FLD are 4.50, 11.37 and 5.39 mg/L while by UV, they are 4.3, 10.72 and 5.55 mg /L respectively. The dispersion variation is 25% lower with the UV detector with average standard deviations of 0.06 in absorbance (compared to 0.08 in fluorescence) – therefore better repeatability for UV, the lowest LOD of the naphthalene for UV (0.074 mg/L) and better resolutions in UV too.

Numerous medicinal plants are found to have a potent transcriptional activity for the genes involved in cellular function. This study is aimed at determining the effects of Eruca sativa seeds extract (SE) on PTEN tumor suppressor gene, which is a candidate gene inactivated in a number of different tumor types including breast cancer. Effect of SE on the activation of PTEN gene expression level was investigated in rat mammary gland carcinogenesis induced by 7,12-dimethylbenz(α)anthracene (DMBA). Total RNA was extracted to evaluate the expression level of PTEN by quantitative real time PCR (qRT-PCR). The results showed that the expression level of PTEN gene was significantly increased by about 938% compared to the cancer group (DMBA) as a result of SE treatment. This study indicates that the SE has a significant inhibitory effect in targeting breast cancer by activating the PTEN gene expression.

En este trabajo se desarrollan y validan las vitaminas A y D mediante dos métodos analíticos por Cromatografía Líquida de Alta Resolución (CLAR), aplicables al control de la calidad del Aceite de Hígado de Tiburón. Ambos métodos se basan en la separación de las vitaminas a través una columna cromatográfica Lichrosorb RP - 18 (5 µm, 25 cm x 4 mm), empleando una fase móvil compuesta por metanol-agua (98:2) y la cuantificación de las mismas frente a muestras de referencia empleando el método del estándar externo. Las longitudes de ondas empleadas fueron 325 y 254 nm, para la Vitamina A y D, respectivamente. Los métodos analíticos desarrollados resultaron lineales, precisos, específicos y exactos en el rango de concentraciones estudiadas.In this work two analytic, easy and economic methods to determine vitamins A and D by High Performance Liquid Chromatography (HPLC) applicable to the quality control of the Shark Liver Oil Pool were developed and validated. Both methods are based in the...

The aim of the present work was to develop and vali date a simple, effective and economical HPLC method for the analysis of Torsemide in bulk, dosage forms and in issolution samples. A Zorbax C 18 reverse phase column (150 × 3.0mm, 3.5μm) and mobile phase containing 5mM ammon iu acetate: acetonitrile (75:25 v/v) were used in isocratic mode. Quantification was achieved at 287n m. The retention time of Torsemide was 2.37 min and showed a good linearity in the concentration range of 10-50μ g/mL with a correlation coefficient of 0.998. The v alidation parameters included specificity, linearity, and lim it of detection, limit of quantification, precision , robustness and stability which fulfilled regulatory requirements in all the cases. The percent recoveries were in be twe n 98-102 (RSD &lt; 2). The method was successfully applied for the analysis of Torsemide in bulk, pharmaceutical d osage forms and in dissolution samples.

A new simple, accurate and cost-effective spectrophotometric method has been developed for the analysis of some cephalosporins (ceftriaxone, ceftazidime, cefixime, cefotaxime and cefuroxime) in bulk samples and pharmaceutical dosage forms. The reaction involves a two-step process of diazotization of the cephalosporins with acidified NaNO 2 at 0-5°C and coupling with acidified p-dimethylaminobenzaldehyde (DMAB). Optimal temperature and time for coupling were established at 50 and 60°C with coupling time ranging from 15 to 20 min. All the cephalosporins gave azo adducts with DMAB that absorbed light optimally at 400-430 nm. The reaction with DMAB occurred at a stoichiometric ratio of 1:1. Optimization of DMAB concentration revealed the superiority of using 0.3% DMAB in 0.0625 M H 2 SO 4 with the best diluting solvent being methanol. Beer's law was obeyed at concentrations ranging from 5 to 60 lg/mL with correlation coefficients >0.9980 in all cases. Overall recoveries were of the order of 95-103% with errors generally less than 6%. The method was successfully applied to the determination of the cephalosporins in dosage forms and it was found to be equivalent accuracy to the official (USP and BP) HPLC assay procedures for these drugs. There was no interference from commonly adopted excipients. The method could find application as a rapid and cost-effective alternative for the quality control of these cephalosporins, especially in preliminary studies.

The sedimentation procedure has been adopted in production of ground silica monolith particles to improve chromatographic separation efficiency of the resultant phase. First, silica monolith particles have been successfully prepared in a large scale by a sol-gel process followed by grinding. The particles after calcination were separated by sedimentation into three zones using an Imhoff sedimentation cone. The particles of the bottom zone were derivatized with a C18 ligand and end-capped. The sedimentation process was found to not only eliminate troublesome minute particles but also narrow down the particle size distribution. The resultant phase was packed in glass lined stainless steel micro-columns. The average number of theoretical plates (N) of the columns for a test mixture was 47,000 and 29,300 for the 300 and 150mm columns (1mm internal diameter), corresponding to 157,000/m and 195,000/m, respectively.

Porous silica monolith particles have been successfully prepared in an enhanced scale (80-85 g/batch) and derivatized with some ligands and packed in micro-columns. The separation efficiencies as high as 127 000 and 175 000 plates /m were achieved with 30 and 15 cm columns packed with C18 modified particles, respectively. The average particle size and particle size distribution were reduced by scale-up and subtle change of the formulation of reaction mixture. This phase also has shown some encouraging possibility for fast analysis. Another phase derivatized with mixed ligands of C8 and urea functionalities showed promising feasibility for proteomic analysis.

Objective: The combination of hydrocodone and ibuprofen is prescribed to temporarily relieve severe pain Methods: The analysis of hydrocodone and ibuprofen was executed on phase Zodiac C18 (250 × 4.6 mm, 5 μm) column with sodium dihydrogen phosphate (0.1M, pH5.0) and methanol (60:40, v: v) as a mobile phase with a flow rate of 1.0 ml/min. The detection and measurement were done at 234 nm. The stability of hydrocodone and ibuprofen was examined under different conditions recommended by ICH, including alkaline, acidic, neutral, oxidative, photolytic and thermal.. A new selective, precise and accurate stability indicating reverse-phase high-performance liquid chromatography (RP-HPLC) technique was developed for identification and estimation of hydrocodone and ibuprofen in tablets. Results: Hydrocodone and ibuprofen are eluted at 4.019 min and 4.999 min, respectively. The assay method was linear (range: 3.75-11.25 µg/ml and 100-300 μg/ml) with R 2 Conclusion: This method can be opted in the routine quality control test of fixed-dose tablet combination comprising of hydrocodone and ibuprofen. values of 0.9996 and 0.9999 for hydrocodone and ibuprofen, respectively. Method was accurate (recovery: 99.65% and 99.37% for hydrocodone and ibuprofen, respectively), precise (RSD: 0.262% and 0.261% for hydrocodone and ibuprofen, respectively), selective and robust. In degradation studies, peaks of degradants did not interfere with the peaks of hydrocodone and ibuprofen. This validated method was applied to assay the content of hydrocodone and ibuprofen in tablets.

Botrytis cinerea is a ubiquitous fungus causing gray mold, the main postharvest disease in fruit, which implies important economic losses in agriculture. With growing concern over health and environmental effects of pesticides, the search for eco-friendly alternatives is a clear priority. Plant extracts represent a rich source of biocompounds with attractive antimicrobial properties. In the last decade, Natural Deep Eutectic Solvents (NADES) has emerged as an auspicious green extraction media to achieve bioextract for a sustainable postharvest control. In the present study, a novel L. cuneifolia NADES-based bioextract was evaluated against B. cinerea. To this purpose, a NADES composed by lactic acid, glucose and water (LGH) was used as extracting agent and compared with traditional solvents in terms of antioxidant capacity and total phenolic content. Furthermore, the bioextract antifungal activity was tested in vitro and also in vivo on artificially inoculated grapes, in order to obtain preliminary data about the efficacy on gray mold development. The antimicrobial activity of the bioextract was assessed using agar diffusion method against B. cinerea, inhibition of 92% was achieved with the bioextract at 2%. Notably, L. cuneifolia bioextract showed an excellent performance for gray mold control on grapes, supporting their potential as alternative green fungicide. Keywords natural deep eutectic solvents • biocompounds • antimicrobial activity • medicinal plant • postharvest control J. Boiteux et al.

The sedimentation procedure has been adopted in production of ground silica monolith particles to improve chromatographic separation efficiency of the resultant phase. First, silica monolith particles have been successfully prepared in a large scale by a sol-gel process followed by grinding. The particles after calcination were separated by sedimentation into three zones using an Imhoff sedimentation cone. The particles of the bottom zone were derivatized with a C18 ligand and end-capped. The sedimentation process was found to not only eliminate troublesome minute particles but also narrow down the particle size distribution. The resultant phase was packed in glass lined stainless steel micro-columns. The average number of theoretical plates (N) of the columns for a test mixture was 47,000 and 29,300 for the 300 and 150mm columns (1mm internal diameter), corresponding to 157,000/m and 195,000/m, respectively.