Development and Validation of RP-HPLC Method for the Simultaneous Determination of Trimethoprim, Sulfadimidine Sodiumand Tylosin Tartrate in injectable solution formulation

A simple, robust and reliable reversed phase HPLC method was developed and validated for the simultaneous determination of Trimethoprim (TMP), Sulfadimidine sodium (SDMS) and Tylosin tartrate (TYT) in Nuroprim® injectable solution formulation. The desired separation was achieved on XBridge C18 column (150 4.6 mm i.d., 5m) at room temperature. The optimized mobile phase consisted of a binary solvent mixture of acetonitrile and aqueous triethylamine (TEA) solution adjusted to pH 5.7 by acetic acid. The mobile phase flow rate was fixed at 1.0 ml/min and the analytes were monitored at 287 nm using photodiode array detector. The effects of the chromatographic conditions on peaks capacity factor, USP tailing factor, column efficiency and resolution were systematically optimized. The method was validated as per International Conference of Harmonization (ICH) and United States Pharmacopeia (USP) guidelines and found to be adequate for the routine quantitative determination of TMP, SDMS and TYT in commercially available Nuroprim® injectable solution dosage form.We would like to thanks Pharmacare pharmaceutical company (Palestine) for their continuous support

Anticancer activity and phytochemical composition of wild Gundelia tournefortii

Artichoke‑like wild thistles are often used in Palestinian cuisine. One of the most commercially recognized species of these wild edible thistles is Gundelia tournefortii, a common plant in the Mediterranean region. G. tournefortii, or ‘Akoob’ in Arabic, remains uncultivated, harvested wild by local populations and considered highly valuable due to its reputed health benefits. The present study aimed to investigate the anticancer effects of G. tournefortii on the human colon carcinoma HCT‑116 cell line. Methanol and hexane extracts were identified to exert considerable antitumor activity against the HCT‑116 cancer cell line, while the aqueous extract was inactive. The phytochemical profiles of the methanol and hexane extracts were investigated using gas chromatography‑mass spectrometry. A total of 6 of the 27 natural compounds identified, including sitosterol, stigmasterol, lupeol, gitoxigenin, α‑amyrin and artemisinin, have been previously validated as being active against cancerous cells. Therefore, the presence of these phytochemicals in G. tournefortii is of importance in its role in preventing and treating cancer. according to different localities. In Palestinian traditional medicine and ethno‑botany, this plant is believed to possess nutritive and curing benefits for diabetes, epilepsy, stomach and intestinal diseases (3,4). According to the literature, it has been validated to exert antioxidant, hepatoprotective and antibacterial effects (1,5). A previous study (2) conducted among Palestinians eating Akoob on a regular basis revealed a consensus belief of its capability to prevent and cure cancer. However, an intensive search of PubMed (using the terms gundelia tournefortii and cancer; on 12th September 2016) indicates that there is no single study on the effects of G. tournefortii extracts against cancer. The present study aimed to investigate the anticancer effects of G. tournefortii on the human colon cancer HCT‑116 cell line. Gas chromatography‑mass spectrometry (GC‑MS) was utilized to explore the potential phytochemicals responsible for the anticancer activity. A total of 27 constituents were identified in G. tournefortii, of which 6 phytochemicals, including sitosterol, stigmasterol, lupeol, gitoxigenin, α‑amyrin and artemisinin have been demonstrated to exhibit anticancer activities. To the best of our knowledge, the present study was the first to investigate the potential benefits of consuming wild edible G. tournefortii for cancer, and to analyze the phytochemical contents known for their anticancer effects.The present study was supported by unrestricted grants from Al‑Qasemi Research Foundation (Baqa‑El‑Gharbia, Israel; grant no. 898002) and Ministry of Science, Technology and Space, Israel

Development and Validation of a Stability-Indicating Hydrophilic Interaction Liquid Chromatographic Method for the Determination of Sulfaquinoxaline Sodium in Water Soluble Powder Formulation

Objective: This study aims to develop and validate a stability-indicating hydrophilic interaction liquid chromatographic method for the determination of sulfaquinoxaline sodium (SQXS) in the presence of sulfaquinoxaline related compound A (SQXA) in a commercially available water soluble powder formulation. Methods: The analytes were separated on zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) column with a solvent mixture of 200mM ammonium acetate (NH4AC) solution and acetonitrile (ACN) (10:90; v/v) at pH adjusted to 5.7 by glacial acetic acid. The mobile phase flow was fixed at 0.5 ml/min and the analytes were monitored at 263nm at ambient temperature. Forced degradation experiments were carried out by exposing sulfaquinoxaline sodium standard and the water soluble powder formulation for thermal, photolytic, oxidative and acid-base hydrolytic stress conditions. Results: The results showed that SQXA and the other degradation products were fully resolved from sulfaquinoxaline sodium and thus the proposed ZIC-HILIC method is stability-indicating. Conclusion: The method was validated as per ICH and USP guidelines and found to be adequate for routine quantitative determination of SQXS in the presence of SQXA and other degradation impurities in commercially available water soluble powder dosage forms.The authors are grateful to Pharmacare pharmaceutical company for providing the facilities to carry out this research

Phytochemical Composition and Biological Activities of Wild Scolymus maculatus L.

The wild population of spotted golden thistle, Scolymus maculatus, which belongs to the Compositae family, is believed to be one of the multi-curative wild plants mentioned in Flora Palaestina. This study aims to disclose the phytochemical composition, antioxidant potential, and antimicrobial activity of wild S. maculatus collected from the farms of Kabul, a village in northwest Galilee, for the first time. Methods: The phytochemical components of crude S. maculatus extracts from methanol, ethyl acetate, and n-hexane solvents were separated and identified using gas chromatography-mass spectrometry (GC-MS) in the electron impact (EI) mode. The free radical scavenging of the plant extracts was measured by DPPH assay. The microdilution test was used to determine the minimum inhibitory concentrations (MICs) of di erent S. maculatus extracts and to evaluate their antimicrobial activities. Results: Thirty-two phytochemicals were found in S. maculatus extracts including stigmasterol, -sitosterol, lupeol, lupeol acetate, and -amyrin. Phytochemicals, such as 2-linoleoylglycerol, -sitosterol, -amyrin, lupeol, (3 )-12-oleanen-3-yl acetate, and lupenyl acetate, were found to dominate the methanol extract. Most of these compounds were also observed in ethyl acetate and n-hexane extracts, but at di erent levels, in addition to some other minor compounds. The various extracts were investigated for their antioxidant and antimicrobial activity. The ethanolic and the methanolic extracts were shown to exhibit the highest free radical scavenging by DPPH assay with a half-maximally e ective concentration (EC50) of 0.37 and 0.65 mg/mL respectively, while the other three extracts (aqueous, ethyl acetate and n-hexane) were less active and their EC50 (e ective concentration at which DPPH radical was scavenged by 50%) were above 1.0 mg/mL. Moreover, MICs were determined to be e ective against Staphylococcus aureus, Salmonella typhimurium, and Candida albicans microorganisms. Ethyl acetate and the ethanolic extracts are active against the three types of microorganisms at a minimum inhibitory concentration (MIC) of 0.5 mg/mL, while aqueous and the n-hexane extracts are inactive against Salmonella typhimurium. Conclusions: The results show that S. maculatus extracts are a rich source of compounds that can play an important role in human health, and in a broader context, in the treatment of various diseases, such antimicrobial and antioxidant-related ailments.Funding: Al-Qasemi Research and Development Authority as well as the Ministry of Science, Technology and Space. Acknowledgments: We would like to thank the Central Public Health Laboratory in Ramallah for facilitating the use of the GC-MS instrument

Anti-inflammatory Activities of Ethanolic Extracts of curcuma Longa (Turmeric) and cinnamon (Cinnamomum verum)

Curcuma longa and Cinnamon are used in folkloric medicine and thought to have different pharmacological activities including anti-inflammatory effects. The objective of this work is to evaluate the in vitro inhibitory effect of Curcuma longa and Cinnamon ethanolic extracts on Lipopolysacaride (LPS)-induced Interlukin-6 (IL-6) and Tumor Necrosis Factor-α (TNF-α) by polymorphonuclear Cells (PMNCs). Polymorphonuclear cells were isolated from the whole blood using Histopaque (Ficol-1077) method and then cultured in an enriched Roswell Park Memorial Institute (RBMI) medium. The concentrations of TNF-α and IL-6 in the supernatant were measured after 24 h and compared using paired-samples t test. The Curcuma longa and Cinnamon extracts have shown significant reduction in the levels of both Il-6 and TNF-α. HPLC analysis of Curcuma longa extract revealed that it contains curcumin, demethoxycurcumin, and bisdemethoxycurcumin while the major compound in the extract of cinnamon was found to be cinnamic acid. Reduction in the levels of IL-6 and TNF-α upon effect of the plants” extract is an indication of their anti-inflammatory effects. The observed anti-inflammatory effect may be due to the presence of curcuminoids and cinnamic acid from Curcuma longa and Cinnamon, respectively

Validated HPLC Method to Simultaneously Determine Amprolium Hydrochloride, Sulfaquinoxaline Sodium and Vitamin K3 in A.S.K Powder on ZIC-HILIC Column

A new HPLC method that is based on zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) coupled with ultraviolet detection was developed, optimized and validated for the simultaneous determination of amprolium hydrochloride, sulfaquinoxaline sodium, and Vitamin K3 (as menadione sodium bisulfite) in A.S.K powder. The separation was carried out using ZIC-HILIC column (250 mm × 4.6 mm, 5 mm) and a mobile phase of 0.2 M Ammonium acetate (NH4AC) buffer and acetonitrile (15:85; v/v) with pH adjusted to 5.7 by glacial acetic acid at a flow rate of 0.5 ml/min. The analytes were monitored by UV detection at 263 nm. The effects of the operational chromatographic conditions on retention and resolution were tested. Different concentrations of the organic solvent in the mobile phase, the ionic strength of the NH4AC buffer and pH of the mobile phase were investigated. The optimized method was subjected to validation by examining specificity, accuracy, precision, linearity, range, ruggedness and robustness. The results were evaluated as per the International Conference on Harmonization (ICH) and United States Pharmacopoeia (USP33/NF28) guidelines and it fulfilled the validation criteria. The method is sensitive, specific, fast, accurate, and requires minimum sample manipulation. It was applied on commercial A.S.K batches, to which all the active ingredients were separated from their excipients.We would like to thank Pharmacare pharmaceutical company (Palestine) for their support of equipments, materials, columns needed to finish this study

Bicarbonate In-Vitro Effect on Beta-Hematin Inhibition by Artemisia sieberi Aqueous Infusion

Malaria is still considered the most threatening disease in Africa. Plasmodium; the malaria parasite, forms during its intra-erythrocytic stage a pigment called hemozoin, which acts as a protection shield against oxygen radical-mediated stress that leads to parasite’s death. Many drugs targeting hemozoin formation such as chloroquine and amodiaquine, but recently strains of Plasmodium have gained resistance to such drugs. Artemisia sieberi stem and leaf water infusion extract compared with A. sieberi bicarbonate aqueous infusion were tested using a semi-quantitative in-vitro method based on the inhibition of ferriprotoporphyrin IX (FP) bio- mineralization developed by Deharo et al. to reveal the differences in antimalarial activity. Reversed phase preparative liquid chromatography coupled to Photo Diode Array (HPLC-PDA) detector was also used to explain this dissimilarity in antimalarial activity. We found that A. sieberi bicarbonate aqueous infusion inhibits the formation of β-hematin better than standard water infusion. The bicarbonate addition increases the extraction of more compounds as the chromatographic HPLC results revealed. Other Artemisia plants (A. vulgaris and A. herba alba) were also tested to explore any inhibition effects

Correlation between Antibacterial Activity and Free-Radical Scavenging: In-Vitro Evaluation of Polar/Non-Polar Extracts from 25 Plants

Objectives: The current study aimed to measure the antioxidant and antibacterial activities of 25 wild Palestinian edible plants, which were subjected to extraction by polar and non-polar solvents. Correlations between free radical scavenging activity and antibacterial activity of the extracts were assessed for both polar and non-polar fractions. Materials: Twenty-five wild edible plant species that are frequently consumed by people in Palestine (mainly in a rural area) were examined. Among them, 10 plant species were among those with the highest mean cultural importance values, according to an ethnobotanical survey that was conducted in the West Bank, Palestine, a few years ago. Method: The protocol of the DPPH assay for testing free-radical scavenging was utilized for determining EC50 values, while microdilution tests were conducted to determine the 50% inhibitory concentration (IC50) of the extracts for the microorganism Staphylococcus mutans. Results and Discussion: Eight extracts (non-polar fractions) were found to possess an antibacterial IC50 of less than 20 ppm, such as Foeniculum vulgare, Salvia palaestinafruticose, Micromeria fruticose, Trigonella foenum-graecum, Cichorium pumilum jacq, Salvia hierosolymitana boiss, Ruta chalepensis, and Chrysanthemum coronarium. The polar fractions possess higher antioxidant activity, while non-polar fraction possess higher antibacterial activity. Looking at all the results together can deceive and lead to the conclusion that there is no correlation between antibacterial activity against S. mutans and free radical scavenging (R2 equals 0.0538). However, in-depth analysis revealed that non-polar plant extracts with an EC50 of free radical scavenging 100 ppm have a four-fold order of enrichment toward more activity against S. mutans. These findings are of high importance for screening projects. A four-fold order of enrichment could save plenty of time and many in screening projects. The antibacterial active extracts marked by low-medium free radical scavenging might act through a mechanism of action other than that of highly active, free radical scavenging extracts. Conclusion: The screening of antioxidant and antimicrobial activity performed on 25 selected wild plant extracts revealed a satisfactory free radical scavenging and antimicrobial potential that could be of value in the management of oxidative stress. Further studies are recommended to explore novel and highly active natural antibacterial products.The Al-Qasemi Research Authority, and the Faculty of Medicine of Al Najah University, supported this work. We declare that the funders had no role in the study design, data collection, analysis, decision to publish, or preparation of the manuscript

Separation and identification of phenolics and flavonoids from wild Pistacia palaestina extract and its antioxidant activity

An in-vitro evaluation of the antioxidant activities of wild Palestinian Pistacia palaestina extracts was done. In parallel, the total phenolic content (TPC) and the total flavonoids content (TFC) were measured. The antioxidant activities were determined spectrophotometrically by DPPH, FRAP, CUPRAC and the ABTS methods. The phenolic and flavonoid contents were separated and identified using LC-PDA-MS. The P. palaestina extract was found to contain many phenolic and flavonoids that enhance its reducing activity and free radical scavenging ability. Total phenolic content, and total flavonoids contents were found to be 66.5 ± 2.2 mg Gallic acid/g, and 20.3 ± 1.1 mg catechin/g, respectively. Antioxidant activity represented as FRAP, CUPRAC, DPPH and ABTS was found to be 23.5± 1.2 mmol Fe+2/g, 4562 ± 63 μmol Trolox/g, 344 ± 11 μmol/g, 53.1 ± 6.6 μmol/g, respectively. The aim of the study is therefore to employ different antioxidant tests to evaluate the antioxidant activities of crude ethanol leaf extracts of P. palaestina, and to determine its phenolic and flavonoids content

Analysis of phenolic and flavonoids of wild ephedra alata plant extractsby lc/pda and lc/ms and their antioxidant activity

Background: Ephedra is among Palestinian medicinal plants that are traditionally used in folkloric medicine for treating many diseases. Ephedra is known to have antibacterial and antioxidant effects. The goal of this study is to evaluate the antioxidant activity of different extracts from the Ephedra alata plant growing wild in Palestine, and to analyze their phenolic and flavonoid constituents by HPLC/PDA and HPLC/MS. Materials and Methods: Samples of the Ephedra alata plant grown wild in Palestine were extracted with three different solvents namely, 100% water, 80% ethanol, and 100% ethanol. The extracts were analyzed for their total phenolic content (TPC), total flavonoid content (TFC), antioxidant activity (AA), as well as phenolic and flavonoids content by HPLC/PDA/MS. Results: The results revealed that the polarity of the extraction solvent affects the TPC, TFC, and AA of extracts. It was found that both TPC and AA are highest for plant extracted with 80% ethanol, followed by 100% ethanol, and finally with 100% water. TFC however was highest in the following order: 100% ethanol > 80% ethanol > water. Pearson correlation indicated that there is a significant correlation between AA and TPC, but there is no correlation between AA and TFC. Simultaneous HPLC-PDA and UHPLC-MS analysis of the ethanolic plant extracts revealed the presence of Luteolin-7-O-glucuronide flavone, Myricetin 3-rhamnoside and some other major polyphenolic compounds that share myricetin skeleton. Conclusion Ephedra alata extract is rich in potent falvonoid glycosidic compounds as revealed by their similar overlaid UV-Vis spectra and UHPLC-MS results. On the basis of these findings, it is concluded that Ephedra alata constitutes a natural source of potent antioxidants that may prevent many diseases and could be potentially used in food, cosmetics, and pharmaceutical products