Abstract: Substandard and counterfeit antimalarials is a major problem in malaria endemic areas. The availability of counterfeit/ substandard medicines is not only decreasing the efficacy in patients, but it is also one of the contributing factors for developing antimalarial drug resistance. Owing to this, a pilot study was conducted to survey quality of drugs collected from different malaria endemic areas of India. Artesunate+Sulphadoxine-Pyrimethamine (AS+SP), Artemether-Lumefantrine (AL), Chloroquine (CQ) tablets were randomly picked from public health facilities in selected states of India. The quality of antimalarial drugs from these areas was assessed by using Global Pharma Health Fund Minilab test kit. This includes physical/visual inspection and disintegration test. Thin-layer chromatography (TLC) was carried out for semi-quantitative assessment of active pharmaceutical ingredients. A total of 45 brands, out of which 21 were for CQ, 14 for AL and 10 for AS+SP were tested from Uttar Pradesh (U.P.), Mizoram, Meghalaya and Gujrat states. One out of 45 samples showed variable disintegration and retension factor. The variable disintegration and retention factor which would have been due to substandard quality or other factors including storage. However, HPLC analysis confirms standard active pharmaceutical ingredient, but may be due to humid temperature and moisture in storage may account for the observed result.
Abstract: Anogeissus leiocarpus (Combretaceae) is well known
for its medicinal uses in African traditional medicine, for treating
many human diseases mainly skin diseases and infections. Mycetoma
disease is a fungal and/ or bacterial skininfection, mainly cause by
Madurella mycetomatis fungus. This study was carried out in vitro to
investigate the antifungal activity of Anogeissus leiocarpus leaf
extracts against the isolated pathogenic Madurella mycetomatis, by
using the NCCLS modified method compared to Ketoconazole
standard drug, and MTT assay. The bioactive fraction was subjected
to chemical analysis implementing different chromatographic
analytical methods (TLC, HPLC, and LC-MS/MS). The results
showed significance antifungal activity of A. leiocarpus leaf extracts
against the isolated pathogenic M. mycetomatis, compared to negative
and positive controls. The chloroform fraction showed the highest
antifungal activity. The chromatographic analysis of the chloroform
fraction with the highest activity showed the presence of important
bioactive compounds such as ellagic and flavellagic acids derivatives,
flavonoids and stilbenoid, which are well known for their antifungal
activity.
Abstract: High Performance Liquid Chromatography (HPLC)
method was developed and validated for simultaneous estimation of
6-Gingerol(6G) and 6-Shogaol(6S) in joint pain relief gel containing
ginger extract. The chromatographic separation was achieved by
using C18 column, 150 x 4.6mm i.d., 5μ Luna, mobile phase
containing acetonitrile and water (gradient elution). The flow rate
was 1.0 ml/min and the absorbance was monitored at 282 nm. The
proposed method was validated in terms of the analytical parameters
such as specificity, accuracy, precision, linearity, range, limit of
detection (LOD), limit of quantification (LOQ), and determined
based on the International Conference on Harmonization (ICH)
guidelines. The linearity ranges of 6G and 6S were obtained over 20-
60 and 6-18 μg/ml respectively. Good linearity was observed over the
above-mentioned range with linear regression equation Y= 11016x-
23778 for 6G and Y = 19276x-19604 for 6S (x is concentration of
analytes in μg/ml and Y is peak area). The value of correlation
coefficient was found to be 0.9994 for both markers. The limit of
detection (LOD) and limit of quantification (LOQ) for 6G were
0.8567 and 2.8555 μg/ml and for 6S were 0.3672 and 1.2238 μg/ml
respectively. The recovery range for 6G and 6S were found to be
91.57 to 102.36 % and 84.73 to 92.85 % for all three spiked levels.
The RSD values from repeated extractions for 6G and 6S were 3.43
and 3.09% respectively. The validation of developed method on
precision, accuracy, specificity, linearity, and range were also
performed with well-accepted results.
Abstract: The consumption of food contaminated with molds
(microscopic filamentous fungi) and their toxic metabolites results in
the development of food-borne mycotoxicosis. The spores of molds
are ubiquitously spread in the environment and can be detected
everywhere. Ochratoxin A is a toxic and potentially carcinogenic
fungal toxin found in a variety of food commodities. In this study, the
mycological quality of various ready-to-eat local and imported pork
meat and meat byproducts sold in Egyptian markets were assessed
and the presence of various molds was determined in pork used as a
raw material, edible organs as liver and kidney as well as in
fermented raw meat by-products. The study assessed the mycological
quality of pork raw meat and their by-products sold in commercial
shops in Cairo, Egypt. Mycological analysis was conducted on
(n=110) samples which included pig’s livers and kidneys from
Egyptian Bassatin slaughter house; local and imported processed
pork meat by-products from Egyptian pork markets. The isolates
were identified using traditional mycological and biochemical tests.
All kidney and liver samples were positive to molds growth while all
byproducts were negative. Ochratoxin A levels were quantitatively
analyzed using the high performance liquid chromatography (HPLC)
and the highest results were present in kidney 7.51 part per billion
(ppb) followed by minced meat 6.19 ppb generally the local samples
showed higher levels than the imported ones. To the best of our
knowledge, this is the first report on mycotoxins detection and
quantification from pork by-products in Egypt.
Abstract: Phthalates are ubiquitous environmental pollutants
well known because of their endocrine disrupting activity in human
organism. The aim of our study was, by biological monitoring,
investigate exposure to phthalates of Roma ethnicity group i.e.
children and adults from 5 families (n=29, average age 11.8 ± 7.6
years) living in western Slovakia. Additionally, we analysed some
associations between anthropometric measures, questionnaire data
i.e. socio-economic status, eating and drinking habits, practise of
personal care products and household conditions in comparison with
concentrations of phthalate metabolites. We used for analysis of urine
samples high performance liquid chromatography and tandem mass
spectrometry (HPLC-MS/MS) to determine concentrations of
phthalate metabolites monoethyl phthalate (MEP), mono-n-butyl
phthalate (MnBP), mono-iso-butyl phthalate (MiBP), mono(2-ethyl-
5-hydroxyhexyl) phthalate (5OH-MEHP), mono(2-ethyl-5-oxohexyl)
phthalate (5oxo-MEHP) and mono(2-etylhexyl) phthalate (MEHP).
Our results indicate that ethnicity, lower socioeconomic status and
different housing conditions in Roma population can affect urinary
concentration of phthalate metabolites.
Abstract: The potential neuroprotective effect of Phyllantus
nuriri against Fe2+ and sodium nitroprusside (SNP) induced oxidative
stress in mitochondria of rats brain was evaluated. Cellular viability
was assessed by MTT reduction, reactive oxygen species (ROS)
generation was measured using the probe 2,7-dichlorofluoresce
indiacetate (DCFH-DA). Glutathione content was measured using
dithionitrobenzoic acid (DTNB). Fe2+ (10μM) and SNP (5μM)
significantly decreased mitochondrial activity, assessed by MTT
reduction assay, in a dose-dependent manner, this occurred in parallel
with increased glutathione oxidation, ROS production and lipid
peroxidation end-products (thiobarbituric acid reactive substances,
TBARS). The co-incubation with methanolic extract of Phyllantus
nuriri (10-200 μg/ml) reduced the disruption of mitochondrial
activity, gluthathione oxidation, ROS production as well as the
increase in TBARS levels caused by both Fe2+ and SNP in a dose
dependent manner. HPLC analysis of the extract revealed the
presence of gallic acid (20.540.01), caffeic acid (7.930.02), rutin
(25.310.05), quercetin (31.280.03) and kaemferol (14.360.01).
This result suggests that these phytochemicals account for the
protective actions of P. niruri against Fe2+ and SNP -induced
oxidative stress. Our results show that P. nuriri consist important
bioactive molecules in the search for an improved therapy against the
deleterious effects of Fe2+, an intrinsic producer of reactive oxygen
species (ROS), that leads to neuronal oxidative stress and
neurodegeneration.
Abstract: The work allowed gaining knowledge about redox and
speciation changes of As, Cr and Sb ionic forms in Klodnica River
water. This kind of studies never has been conducted in this region of
Poland. In study optimized and validated previously HPLC-ICP-MS
methods for determination of As, Sb and Cr was used. Separation
step was done using high-performance liquid chromatograph
equipped with ion-exchange column followed by ICP-MS
spectrometer detector. Preliminary studies included determination of
the total concentration of As, Sb and Cr, pH, Eh, temperature and
conductivity of the water samples. The study was conducted monthly
from March to August 2014, at six points on the Klodnica River. The
results indicate that exceeded at acceptable concentration of total Cr
and Sb was observed in Klodnica River and we should qualify
Klodnica River waters below the second purity class. In Klodnica
River waters dominates oxidized antimony and arsenic forms, as well
as the two forms of chromium Cr(VI) and Cr(III). Studies have also
shown the methyl derivative of arsenic's presence.
Abstract: The influences of cell-free solutions (CFSs) of lactic
acid bacteria (LAB) on cadaverine and other biogenic amines
production by Listeria monocytogenes and Staphylococcus aureus
were investigated in lysine decarboxylase broth (LDB) using HPLC.
Cell free solutions were prepared from Lactococcus lactis subsp.
lactis, Leuconostoc mesenteroides subsp. cremoris, Pediococcus
acidilactici and Streptococcus thermophiles. Two different
concentrations that were 50% and 25% CFS and the control without
CFSs were prepared. Significant variations on biogenic amine
production were observed in the presence of L. monocytogenes and S.
aureus (P < 0.05). The function of CFS on biogenic amine production
by foodborne pathogens varied depending on strains and specific
amine. Cadaverine formation by L. monocytogenes and S. aureus in
control were 500.9 and 948.1 mg/L, respectively while the CFSs of
LAB induced 4-fold lower cadaverine production by L.
monocytogenes and 7-fold lower cadaverine production by S. aureus.
The CFSs resulted in strong decreases in cadaverine and putrescine
production by L. monocytogenes and S. aureus, although remarkable
increases were observed for histamine, spermidine, spermine,
serotonin, dopamine, tyramine and agmatine in the presence of LAB
in lysine decarboxylase broth.
Abstract: Formation of histamine, tryptamine, phenylethylamine and tyramine (vasoactive amines) in dry fermented sausage Petrovská klobása during drying and ripening in traditional room (B1) and industrial ripening chamber (B3) were investigated. Dansyl chloride derivatized vasoactive amines were determined using HPLC-DAD on Eclipse XDB-C18 column.
Histamine, the most important amine from food safety point of view, was not detected in any analyzed sample. Unlike most of the other fermented sausages, where tyramine is reported as the most abundant amine, in Petrovská klobása tryptamine was the most abundant vasoactive amine in both groups of sausages even though concentrations of tryptamine and tyramine in B3 sausages at the end of ripening were nearly the same (39.8 versus 39.6mg/kg). Sum of vasoactive amines in samples varied from not detected ND (B3) to 176 mg/kg (B1), with concentration of 36.1 (B3) and 73.6 (B1) mg/kg at the end of drying and 96 (B3) and 176 (B1) mg/kg at the end of ripening period. Although the sum of vasoactive amines has increased from the end of drying (45. and 90. day) to the end of ripening period (120. day), during whole production period these values did not exceed 200 mg/kg proposed as possible indicator of hygienic conditions and GMP in the sausage production.
Abstract: Background and objectives: Most of the agricultural products are processed by blanching. Blanching can increase antioxidant activity in white saffron products. The objective of this research were to determine antioxidant activity, to identify, and to measure changes in phenolic substances of fresh and blanched white saffron rhizomes (Curcuma mangga Val.). Methods: White saffron rhizomes were peeled, washed and blanched in boiling water containing 0% or 0.05% citric acid solution for 5 and 10 minutes. Samples were extracted using methanol, rotaevaporated, and freezedried. Dried extract was determined antioxidant activity by DPPH method, identified and quantified for the phenolic substances by High Performance Liquid Chromatography (HPLC) equipped with coloumn C18 and Photodiode-array detector (PAD). Result: This research showed that the quantity of the 6 phenolic substances identified in blanched white saffron in citric acid solution increased significantly compared to that of the non-blanched. Blanching white saffron in 0.05% citric acid media for 5 minutes increased its antioxidant activity, and total phenolic content. Conclusions: The identified phenolic substances of white saffron were Gallic Acid (GA), Catechin (C), Epicatechin (EC), Epigallocatechin (EGC), Epigallocatechingallat (EGCG) and Gallocatechingallat (GCG). The blanched white saffron contained C and EGCG significantly higher than that of fresh rhizomes.
Abstract: Quercetin and (+)-catechin are metabolites present in Phyllanthus niruri plant, have potential in medicinal uses as anticancer and antioxidant agents. Studies on production of quercetin and (+)-catechin from P. niruri callus culture via in vitro technique were carried out and the results were compared to the intact plant. P. niruri explants were cultured on Murashige and Skoog (MS) solidified media supplemented with several phytohormone combinations for one month. The metabolites were extracted from P. niruri callus and intact plant by using carbon dioxide supercritical fluid extraction (SFE) with ethanol as modifier and solvent extraction techniques. The extracts were analyzed by means of HPLC method. Results showed that P. niruri callus culture was successfully established. The highest content of quercetin (1.72%) was found from P. niruri callus grown in media supplemented with 0.8mg/L kinetin and 0.2mg/L 2,4-dicholophenoxyacetic acid (2,4-D), which was 1.2 fold higher than intact plant. Meanwhile, the highest amounts of (+)-catechin (0.63%) was found from P. niruri callus grown in media with addition of 0.2mg/L 1-naphthalene acetic acid (NAA) and 0.8mg/L 2,4-D. The SFE condition in this study showed better extraction efficiency when higher contents of selected metabolites were found in all SFE extracts compared to the common solvent extracts.
Abstract: A new and cost effective RP-HPLC method was
developed and validated for simultaneous analysis of non steroidal
anti inflammatory dugs Diclofenac sodium (DFS), Flurbiprofen
(FLP) and an opioid analgesic Tramadol (TMD) in advanced drug
delivery systems (Liposome and Microcapsules), marketed brands
and human plasma. Isocratic system was employed for the flow of
mobile phase consisting of 10 mM sodium dihydrogen phosphate
buffer and acetonitrile in molar ratio of 67: 33 with adjusted pH of
3.2. The stationary phase was hypersil ODS column (C18, 250×4.6
mm i.d., 5 μm) with controlled temperature of 30 C°. DFS in
liposomes, microcapsules and marketed drug products was
determined in range of 99.76-99.84%. FLP and TMD in
microcapsules and brands formulation were 99.78 - 99.94 % and
99.80 - 99.82 %, respectively. Single step liquid-liquid extraction
procedure using combination of acetonitrile and trichloroacetic acid
(TCA) as protein precipitating agent was employed. The detection
limits (at S/N ratio 3) of quality control solutions and plasma samples
were 10, 20, and 20 ng/ml for DFS, FLP and TMD, respectively.
The Assay was acceptable in linear dynamic range. All other
validation parameters were found in limits of FDA and ICH method
validation guidelines. The proposed method is sensitive, accurate and
precise and could be applicable for routine analysis in
pharmaceutical industry as well as in human plasma samples for
bioequivalence and pharmacokinetics studies.
Abstract: A new reverse phase-high performance liquid chromatography (RP-HPLC) method with fluorescent detector (FLD) was developed and optimized for Norfloxacin determination in human plasma. Mobile phase specifications, extraction method and excitation and emission wavelengths were varied for optimization. HPLC system contained a reverse phase C18 (5 μm, 4.6 mm×150 mm) column with FLD operated at excitation 330 nm and emission 440 nm. The optimized mobile phase consisted of 14% acetonitrile in buffer solution. The aqueous phase was prepared by mixing 2g of citric acid, 2g sodium acetate and 1 ml of triethylamine in 1 L of Milli-Q water was run at a flow rate of 1.2 mL/min. The standard curve was linear for the range tested (0.156–20 μg/mL) and the coefficient of determination was 0.9978. Aceclofenac sodium was used as internal standard. A detection limit of 0.078 μg/mL was achieved. Run time was set at 10 minutes because retention time of norfloxacin was 0.99 min. which shows the rapidness of this method of analysis. The present assay showed good accuracy, precision and sensitivity for Norfloxacin determination in human plasma with a new internal standard and can be applied pharmacokinetic evaluation of Norfloxacin tablets after oral administration in human.
Abstract: Pomegranate and pomegranate juices (PJs) have taken
great attention for their health benefits in the last years. As there is an
increasing concern about potential health benefits of ellagic acid, it is
of great interest to evaluate alterations in ellagic acid concentration of
commercial PJs. The purpose of this study is to analyze total
phenolic, free and total ellagic acid content of six commercial PJs
sold in Turkish markets using HPLC.
The results showed that some commercial PJs had markedly high
total phenolic and ellagic acid content. Total phenolic substances of
commercial PJs range from 796.71 to 4608.91 mg GAE/l. Free
amount of ellagic acid in commercial PJs range from 27.64 to 111.78
mg/l. Samples are hydrolyzed with concentrated HCl at 93oC for 2
and 24 hour and influences of temperature and time parameters on
hydrolization were investigated. Thermal processing for
pasteurization increased ellagic acid via ellagitannins hydrolysis.
Abstract: Commercially available lipases (Candida antarctica lipase B, Novozyme 435, Thermomyces lanuginosus lipase, and Lipozyme TL IM), as well as sol-gel immobilized lipases, have been screened for their ability to acylate regioselectively xylitol, sorbitol, and mannitol with a phenolic ester in a binary mixture of t-butanol and dimethylsulfoxide. HPLC and MALDI-TOF MS analysis revealed the exclusive formation of monoesters for all studied sugar alcohols. The lipases immobilized by the sol-gel entrapment method proved to be efficient catalysts, leading to high conversions (up to 60%) in the investigated acylation reactions. From a sequence of silane precursors with different nonhydrolyzable groups in their structure, the presence of octyl and i-butyl group was most beneficial for the catalytic activity of sol-gel entrapped lipases in the studied process.
Abstract: The aim of the present study was to develop and
validate an inexpensive and simple high performance liquid
chromatographic (HPLC) method for the determination of colistin
sulfate. Separation of colistin sulfate was achieved on a ZORBAX
Eclipse XDB-C18 column using UV detection at λ=215 nm. The
mobile phase was 30 mM sulfate buffer (pH 2.5):acetonitrile(76:24).
An excellent linearity (r2=0.998) was found in the concentration
range of 25 - 400 μg/mL. Intra- day and inter-day precisions of
method (%RSD, n=3) were less than 7.9%.The developed and
validated method was applied to determination of the content of
colistin sulfate in medicated premix and animal feed sample.The
recovery of colistin from animal feed was satisfactorily ranged from
90.92 to 93.77%. The results demonstrated that the HPLC method
developed in this work is appropriate for direct determination of
colistin sulfate in commercial medicated premixes and animal feed.
Abstract: In the last decade, carbohydrates have attracted great
attention as renewable resources for the chemical industry.
Carbohydrates are abundantly found in nature in the form of
monomers, oligomers and polymers, or as components of
biopolymers and other naturally occurring substances. As natural
products, they play important roles in conferring certain physical,
chemical, and biological properties to their carrier molecules.The
synthesis of this particular carbohydrate glycomonomer is part of our
work to obtain biodegradable polymers. Our current paper describes
the synthesis and characterization of a novel carbohydrate
glycomonomer starting from D-glucose, in several synthesis steps,
that involve the protection/deprotection of the D-glucose ring via
acetylation, tritylation, then selective deprotection of the aromaticaliphatic
protective group, in order to obtain 1,2,3,4-tetra-O-acetyl-
6-O-allyl-β-D-glucopyranose. The glycomonomer was then obtained
by the allylation in drastic conditions of 1,2,3,4-tetra-O-acetyl-6-Oallyl-
β-D-glucopyranose with allylic alcohol in the presence of
stannic chloride, in methylene chloride, at room temperature. The
proposed structure of the glycomonomer, 2,3,4-tri-O-acetyl-1,6-di-
O-allyl-β-D-glucopyranose, was confirmed by FTIR, NMR and
HPLC-MS spectrometry. This glycomonomer will be further
submitted to copolymerization with certain acrylic or methacrylic
monomers in order to obtain competitive plastic materials for
applications in the biomedical field.
Abstract: The strawberry jam is rich in bioactive compounds. It
is economically and commercially important and widely consumed.
Different strawberries cultivars can be used for its preparation,
however, a careful selection should be performed to guarantee the
preservation of bioactive compounds during jam storage. Two
strawberry cultivars (Camarosa and American 13) were analyzed by
HPLC, three anthocyanins: cyanidin-3-glucoside, pelargonidin-3-
glucoside and pelargonidin-3-rutinoside were quantified. Camarosa
strawberries presented significantly higher concentration of
anthocyanins (p
Abstract: Agricultural waste is mainly composed of cellulose
and hemicelluloses which can be converted to sugars. The
inexpensive reducing sugar from durian peel was obtained by
hydrolysis with HCl concentration at 0.5-2.0% (v/v). The hydrolysis
range of time was for 15-60 min when the mixture was autoclaved at
121 °C. The result showed that acid hydrolysis efficiency (AHE)
highest to 80.99% at condition is 2.0%concentration for 15 min.
Reducing sugar highest to 56.07 g/litre at condition is 2.0%
concentration for 45min. Total sugar highest to 59.83 g/litre at
condition is 2.0%concentration for 45min, which was not significant
(p < 0.05) with condition 2.0% concentration for 30 min and 1.5 %
concentration for 45 and 60 min. The increase in concentration
increased AHE, reducing sugar and total sugar. The hydrolysis time
had no effect on AHE, reducing sugar and total sugar. The maximum
reducing sugars of each concentration were at hydrolysis time 45
min .The hydrolysated were analysis by HPLC, the results revealed
that the principle of sugar were glucose, fructose and xylose.
Abstract: The field of polymeric biomaterials is very important
from the socio-economical viewpoint. Synthetic carbohydrate
polymers are being increasingly investigated as biodegradable,
biocompatible and biorenewable materials. The aim of this study was
to synthesize and characterize some derivatives based on D-mannose.
D-mannose was chemically modified to obtain 1-O-allyl-2,3:5,6-di-
O-isopropylidene-D-mannofuranose and 1-O-(2-,3--epoxy-propyl)-
2,3:5,6-di-O-isopropylidene-D-mannofuranose.
The chemical structure of the resulting compounds was
characterized by FT-IR and NMR spectroscopy, and by HPLC-MS.