Quantitative High-performance Liquid Chromatography Analysis Research Articles

Enhancement of Aggregate Formation Through Aromatic Compound Adsorption in Elastin-like Peptide (FPGVG)5 Analogs.

Elastin-like peptides (ELPs) exhibit temperature-dependent reversible self-assembly. Repetitive sequences derived from elastin, such as Val-Pro-Gly-Val-Gly (VPGVG), are essential for the self-assembly of ELPs. Previously, we developed (FPGVG)5 (F5), in which the first valine residue in the VPGVG sequence was replaced with phenylalanine, which showed strong self-aggregation ability. This suggests that interactions through the aromatic amino acid residues of ELPs could play an important role in self-assembly. In this study, we investigated the thermoresponsive behavior of F5 analogs in the presence of aromatic compounds. Turbidimetry, spectroscopy, and fluorescence measurements demonstrated that aromatic compounds interacted with F5 analogs below the transition temperature and enhanced the self-assembly ability of ELPs by stabilizing amyloid-like structures. Furthermore, quantitative high-performance liquid chromatography analyses showed that the F5 analogs could adsorb and remove hydrophobic aromatic compounds from aqueous solutions during aggregate formation. These results suggested that the F5 analogs can be applicable as scavengers of aromatic compounds.

Quantification of chondroitin sulfate, hyaluronic acid and N-glycans in synovial fluid – A technical performance study

ObjectiveTo validate a quantitative high performance liquid chromatography (HPLC) assay for chondroitin sulfate (CS) and hyaluronic acid (HA) in synovial fluid, and to analyze glycan-patterns in patient samples. DesignSynovial fluid from osteoarthritis (OA, n ​= ​25) and knee-injury (n ​= ​13) patients, a synovial fluid pool (SF-control) and purified aggrecan, were chondroitinase digested and together with CS- and HA-standards fluorophore labelled prior to quantitative HPLC analysis. N-glycan profiles of synovial fluid and aggrecan were assessed by mass spectrometry. ResultsUnsaturated uronic acid and sulfated-N-acetylgalactosamine (ΔUA-GalNAc4S and ΔUA-GalNAc6S) contributed to 95% of the total CS-signal in the SF-control sample. For HA and the CS variants in SF-control the intra- and inter-experiment coefficient of variation was between 3–12% and 11–19%, respectively; tenfold dilution gave recoveries between 74 and 122%, and biofluid stability test (room temperature storage and freeze-thaw cycles) showed recoveries between 81 and 140%. Synovial fluid concentrations of the CS variants ΔUA-GalNAc6S and ΔUA2S-GalNAc6S were three times higher in the recent injury group compared to the OA group, while HA was four times lower. Sixty-one different N-glycans were detected in the synovial fluid samples, but there were no differences in levels of N-glycan classes between patient groups. The CS-profile (levels of ΔUA-GalNAc4S and ΔUA-GalNAc6S) in synovial fluid resembled that of purified aggrecan from corresponding samples; the contribution to the N-glycan profile in synovial fluid from aggrecan was low. ConclusionsThe HPLC-assay is suitable for analyzing CS variants and HA in synovial fluid samples, and the GAG-pattern differs between OA and recently knee injured subjects.

English

There have been a growing interest since decades in the medicinal plants in terms of preventive health care and treatment of various diseases. On the other hand, polyphenolic compounds present in medicinal plants are known to have antioxidant potency. In the current work, the quantitative HPLC analysis of the flowers of four plants grown in Bolu-Turkey, Silene compacta, Digitalis lamarckii, Xeranthemum annuum L., and Epilobium hirsutum L was carried out. Prior to HPLC quantification, they were detected by TLC with reference standard compounds, identified by LC-MS and compared for their profiles of phenolic acids and some flavonoids by using high-performance liquid chromatography (HPLC) with diode array detector. The highest contents of benzoic acids were detected in the flowers of E. hirsutum L. as 159 mg in 100 g dry flowers. As for cinnamic acids, the highest content value was found in S. compacta flowers  as 292 mg in 100 g dry flowers. On the other hand, E. hirsutum L. flowers contained the highest total flavonoid value as much as 3890 mg. Total polyphenolic content was the highest (4104 mg/100 g dry plant) in E. hirsutum L., however, the lowest value was found as 492 mg/100 g dry flowers for D. lamarcki. Key words:  Polyphenols,  HPLC,  LC-MS,  quantitative determination, Silene compacta, Digitalis lamarckii, Xeranthemum annuum L., Epilobium hirsutum L.

Determination of Caffeine and Chlorogenic Acid (CGA) in the Methanolic Extracts Coffee (C. arabica. L) To seeds and peels (Unroasted and Roasted) Cultivars Grown in Yemen by High Performance Liquid Chromatography (HPLC)

In this study methanolic extract of one plant namely Arabic coffee, were screened for the presence of analysis Constituents and tested for their of liquid chromatographic separation (HPLC). The quantitative HPLC analysis revealed the results showed presence of Caffeine, chlorogenic acid (CGA) is the concentration of the contents Seeds, peels (unroasted) samples is higher than their contents in the Seeds, and peels (roasted) samples. The highest concentration of Caffeine was 5,334 % in unroasted Arabica coffee peels (Udaini) in Ibb (Kafr) region, while the highest value of chlorogenic acid was 68,460% in unroasted Arabica coffee peels (Tufahi) in Ibb (Kafr). The highest concentration of Caffeine was 9,948% in unroasted Arabic coffee seeds (Udaini) in Ibb (Kafr) region, while the highest concentration of chlorogenic acid was 97,280% in unroasted Arabic coffee seeds (Udaini) in Ibb (Kafr) region. The highest concentration of Caffeine was 1,964 % in roasted Arabica coffee peels (Udaini) in Ibb (Kafr) region, while the highest value of chlorogenic acid was 26,260% in roasted Arabica coffee peels (Bura’ai.) in Sana'a (Haraz). The highest concentration of Caffeine was 2,324% in roasted Arabic coffee seeds (Udaini) in Al- Mahweet (Hufash) region, while the highest concentration of chlorogenic acid was 47,09% in roasted Arabic coffee seeds (Dawairi) in Al- Mahweet (Hufash).

Determination of Vitamin C, Vitamin A and Flavonoid Levels in Garcinia cowa Roxb Fruit Flesh Extract

Garcinia cowa Roxb is a plant used as traditional medicine containing antioxidant activity. Plant extracts with antioxidant activity are often used as bioactive ingredients in anti-aging cosmetics. Studies on bioactive compounds and the use of this plant in cosmeceuticals have yet to be widely reported. This study aims to determine and analyze the levels of flavonoids, vitamins C, and vitamin A in G. cowa fruit flesh extract. These three compounds are bioactive ingredients that are beneficial in reducing the symptoms of photo-aging on the skin. The G. cowa used in this study was obtained from Tampunik Kambang, Pesisir Selatan Regency, and was extracted by maceration method using 96% ethanol. The testing of vitamin C levels in the extract of G. cowa fruit flesh was carried out through High-Performance Liquid Chromatography (HPLC) method using a UV detector with a wavelength of 264 nm. Quantitative HPLC analysis was carried out based on the resulting peak area data from the chromatogram. Vitamin A and flavonoid levels were measured using ultraviolet-visible spectrophotometry (UV-VIS) at maximum wavelengths of 286 and 429 nm. The results show that the levels of vitamin C and vitamin A in G. cowa was 151.4 mg/100g extract and 1.23 g 100g extract, while the flavonoid level was 69.5 mg/100g extract. The level of vitamins C, vitamin A, and flavonoids in G. cowa can contribute to natural bioactive substances and can be developed further into anti-aging cosmeceutical products.

Characterization of entrapment behavior of polyphenols in nanostructured lipid carriers and its effect on their antioxidative activity

Polyphenols are widely used as antioxidant agents to protect human health. Resveratrol, kaempferol, and quercetin have been reported to have potent antioxidant activity; however, these compounds have many problems related to their practical application, such as instability and insolubility. Thus, a nanostructured lipid carrier (NLC) was utilized as a drug delivery system (DDS) to overcome these limitations. This study investigated the particle stability, drug loading performance, and antioxidant activity of polyphenols-incorporated NLCs. The particle size and distribution were suitable for DDS applications, and all the samples demonstrated good stability after 2 months of storage. Based on Raman spectroscopy analysis, polyphenols were successfully encapsulated in NLCs. Quantitative high-performance liquid chromatography analysis indicated that NLCs could load resveratrol more than kaempferol and quercetin. In addition, NLCs have successfully improved all the antioxidant activity per unit concentration of polyphenol (specific antioxidant activity) compared to the free polyphenols. Quercetin-incorporated NLCs showed the highest specific antioxidant activity. This result is the opposite of entrapment efficiency and actual antioxidant activity, most likely influenced by the location of entrapped polyphenol molecules. As it was performed, NLCs are highly recommended to be applied as an antioxidant delivery system.

Development of the technique for quality control of 1,3-bis(3,4-dicyanophenoxy)benzene by HPLC

Objectives. Determination of target products and byproducts is necessary for the quality control of phthalonitrile monomer synthesis as well as production scaling and performing related kinetic studies. High-performance liquid chromatography (HPLC) is a simple and affordable method for quantitative chemical analysis, which also verifies the quality of raw materials. The objective of this study was to develop an HPLC technique for determining the composition of the reaction mixture in the synthesis of 1,3-bis(3,4-dicyanophenoxy)benzene (DPB).Methods. Reversed-phase HPLC was used to quantitatively analyze the reaction mixture.Results. A simple and rapid method for the quantitative HPLC analysis of phthalonitrile monomers and their mixtures with reagents was developed. Reaction times and the accumulation of byproducts were also studied.Conclusions. The successful performance of the developed technique allows us to recommend it for practical applications. The results obtained for reactors of different sizes have good convergence, and DPB synthesis was successfully scaled up to intermediate scale equipment.

Identification of Chemopreventive Components from Halophytes Belonging to Aizoaceae and Cactaceae Through LC/MS-Bioassay Guided Approach.

Six halophytes, namely, Aptenia cordifolia var. variegata, Glottiphyllum linguiforme, Carpobrotus edulis, Ferocactus glaucescens, F. pottsii and F. herrerae were investigated for chemopreventive effect. Prioritization of most promising plant for further investigation was carried out through an integrated liquid chromatography-high resolution electrospray ionization mass spectrometry profiling-bioassay guided approach. NAD(P)H: quinone oxidoreductase-1 (NQO-1) induction in cultured murine hepatoma cells (Hepa-1c1c7) and inhibition of nitric oxide (NO) production in lipopolysaccharide-activated macrophages (RAW 264.7) were carried out to investigate chemopreventive effect. Bioassay data revealed that F. herrerae, A. cordifolia, C. edulis and F. glaucescens were the most active with 2-, 1.7-, 1.6- and 1.5-folds induction of NQO-1 activity. Only F. glaucescens exhibited >50% inhibition of NO release. LCMS profiling of the F. glaucescens revealed its high content of flavonoids, a known micheal acceptor with possible NQO-1 induction, as proved by quantitative high-performance liquid chromatography analysis. Thus, the extract of F. glaucescens was subjected to chromatographic fractionation leading to the isolation of four compounds including (i) 2S-naringenin, (ii) trans-dihydrokaempferol (aromadendrin), (iii) 2S-naringenin-7-O-β-d-glucopyranoside and (iv) kaempferol-7-O-β-d-glucopyranoside (populnin). The current study through an LCMS dereplication along with bio guided approach reported the activity of populnin as NO inhibitor and NQO-1 inducer with promising chemopreventive potential.

In-vitro propagation, callus culture and bioactive lignan production in Phyllanthus tenellus Roxb: a new source of phyllanthin, hypophyllanthin and phyltetralin

This is the first report on identification and quantification of important hepatoprotective and anticancer polyphenolic lignans such as phyllanthin (PH), hypophyllanthin (HPH), niranthin (NH) and phyltetralin (PT) in natural plant and in vitro cultures of Phyllanthus tenellus Roxb. The identification of lignans was carried out by Liquid Chromatography–High Resolution Mass Spectrometry (LC–HRMS) and quantified using High-Performance Liquid Chromatography (HPLC). In addition, an efficient protocol has been developed for multiple shoot induction in nodal explants of in vitro derived shoots of P. tenellus. Maximum number of shoot regeneration (7.83 ± 0.15) was achieved on medium incorporated with 1.0 mg/l 6-Benzylaminopurine (BAP). The medium containing Indole-3-acetic acid (IAA) 2 mg/l was superior for induction of rooting in in vitro raised shoots. The plantlets were acclimatized to the field condition with 100% survival. The quantitative HPLC analysis showed that the lignan content was variable with the auxins and cytokinins incorporated in the medium. The lignan content was higher in callus grown on Murashige and Skoog (MS) medium + 2.0 mg/l Naphthaleneacetic acid (NAA). The reported protocol can be used for mass propagation and application of biotechnological approaches for improvement of P. tenellus. The results indicate intriguing possibilities for the utilization of P. tenellus plant parts as an alternative source and of callus culture to scale up bioactive lignan production for pharmaceutical applications.

Inhibitory Effects of Raw-Extract Centella asiatica (RECA) on Acetylcholinesterase, Inflammations, and Oxidative Stress Activities via In Vitro and In Vivo.

Centella asiatica (C. asiatica) is one of the medicinal plants that has been reported to exert comprehensive neuroprotection in vitro and in vivo. In view of this, the present study was performed to investigate the effect of ethanolic extract of C. asiatica, designated as raw-extract of C. asiatica (RECA) in reducing the acetylcholinesterase (AChE), inflammations, and oxidative stress activities via both in vitro (SH-SY5Y and RAW 264.7 cells) and in vivo (Sprague Dawley rats). Quantitative high-performance liquid chromatography analysis reveals that RECA contains a significantly high proportion of glycosides than the aglycones with madecassoside as the highest component, followed by asiaticoside. Treatment of SH-SY5Y cells with RECA significantly reduced the AChE activity in a concentration-dependent manner with an IC50 value of 31.09 ± 10.07 µg/mL. Furthermore, the anti-inflammatory and antioxidant effects of RECA were evaluated by lipopolysaccharides (LPS)-stimulated RAW 264.7 cells. Our results elucidated that treatment with RECA significantly suppressed the level of pro-inflammatory cytokine/mediators and oxidative stress released in a concentration-dependent manner. Interestingly, these patterns of inhibition were consistent as observed in the LPS-induced neuroinflammation Sprague Dawley rats’ model. The highest concentration used in the two models presented the most significant results. Herein, our findings strongly suggest that RECA may offer therapeutic potential for the treatment of Alzheimer’s disease through inhibiting the AChE, inflammation, and oxidative stress activities.

Quantitative HPLC analysis of flavonoid in three different solvent extracts in leaves of Gynura procumbens

In Malaysia, traditional folks call Gynura procumbens as Sambung nyawa or translated as longevity life. The name given might be because of the claim made by them that the leaves extracts seems to be valuable and possesses high theuraphatic potential treatment of various diseases. Extraction of the leaves have been conducted by many researchers to identify the properties for antidiabetic/antihyperglygemic, antihypertensive, antimicrobial, antioxidant, antiimflammatory, antiulcer, anticancer, antiherpes and immunomodulatory. Extracts have been obtained using different solvent. Four alcoholic extracts and one water extract were studied separately. Soxhlet extraction was conducted in the following manner: plant material (3.0 g) was placed in the Soxhlet apparatus. Extraction process was carried out for three hours using 99.9% ethanol, methanol and water as a solvent (250 mL) until the solvent discoloration (3 hours). Yield of extraction techniques was determined by evaporating the solvent under vacuum using rotary evaporator. Previous research on solvent extraction of Gynura procumbens were also compared to whereby the methanol extract obtained by other researcher was 12.2%, ethanol extract with 14.73% and 27.83%. There is also one study extracting Gynura procumbens extracts via water, however the yield is not mentioned. High performance liquid chromatography (HPLC) coupled with ultra violet (UV) detector has been used for the separation and quantification of kaempferol from the Gynura procumbens leaves extract. Reversed phase high performance liquid chromatography separation was performed, using 0.1% phosphoric acid:acetonitrile as eluent in mobile phase. Our results confirm previous reports concerning the presence of several flavonoids.

Chemical Composition, Gastroprotective, and Antioxidant Activities of Schrebera arborea Fruits

ABSTRACTThe chemical composition, gastroprotective, and antioxidant activities of Schrebera arborea fruits were investigated. The solvent extracts were analyzed for fatty acid and polyphenolic content using gas chromatography-mass spectrometry (GC–MS) and high performance liquid chromatography (HPLC), respectively. Acute toxicity study was done by oral administration at 2,000 mg kg–1. Gastroprotective effect was evaluated at 50, 100, or 150 mg kg–1 in ethanol and indomethacin-induced ulcer models in rats. The antioxidant activities of the extract were also evaluated. Sitosterol (41.55%) was the major component identified by GC–MS. Quantitative HPLC analysis for phenolic compounds indicated the presence of gallic acid (0.19 mg g–1), caffeic acid (0.13 mg g–1), rutin (0.30 mg g–1), apigenin-7-glucoside (1.21 mg g–1), catechin (1.84 mg g–1), and kaempferol (6.53 mg g–1), respectively. The ethanolic extract of the fruit produced 63.5% and 80.8% inhibition of ethanol and indomethacin-induced ulcer at 100 mg kg–1, respectively. The extract at 100 mg kg–1 reduced MDA level and increased glutathione, catalase, and superoxide dismutase activities in rat stomach tissue. Significant radical scavenging antioxidant activity was recorded in vitro.

Phenolic profile, antioxidant capacity and antimicrobial activity of Calligonum arich L., desert endemic plant in Tunisia

Calligonum arich L. is an endemic plant belonging to the Polygonaceae family and growing in North Africa, particularly in Tunisia. This study aims to determine the phenolic profile and to evaluate the antioxidant and antibacterial activities of C. arich grown in Southern Tunisia environmental conditions. The samples were extracted using methanol solvent. Spectrophotometric methods were used to assess the total polyphenol content (TPC) and total flavonoid content (TFC) using respective standards (gallic acid and quercetin). Individual phenolic compounds were identified using high performance liquid chromatography (HPLC). The in vitro antioxidant capacity was studied adopting three different tests including total antioxidant capacity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-Azino-bis(3-ethylbenothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities. Results showed that total polyphenol content ranged from 32.02 mg GAE/g DW in roots to 34.27 mg GAE/g DW in leaves. Total flavonoid content ranged from 22.79 to 26.41 mg QE/g DW, respectively in stem and leaves. Qualitative and quantitative HPLC analysis revealed the presence of 14 phenolic compounds that varied significantly with plant parts. Interestingly, the antimicrobial test showed that C. arich leaves extract exhibited the highest antibacterial activity against five microorganisms (two Gram-positive and three Gram-negative bacteria). Moreover, both leaves and roots present a valuable source of phenolic compounds providing to C. arich a high antioxidant capacity with significant variability. This plant could be exploited as a potential source of natural bioactive molecules which confers on this species a greater nutritional value replacing synthetic antioxidants.

Lipid Nanocarriers for Oral Delivery of Serenoa repens CO2 Extract: A Study of Microemulsion and Self-Microemulsifying Drug Delivery Systems.

The aim of this study was the development and characterization of lipid nanocarriers using food grade components for oral delivery of Serenoa repens CO2 extract, namely microemulsions (MEs) and self-microemulsifying drug delivery systems (SMEDDSs) to improve the oral absorption. A commercial blend (CB) containing 320 of S. repens CO2 extract plus the aqueous soluble extracts of nettle root and pineapple stem was formulated in two MEs and two SMEDDSs. The optimized ME loaded with the CB (CBM2) had a very low content of water (only 17.3%). The drug delivery systems were characterized by dynamic light scattering, transmission electron microscopy, and high-performance liquid chromatography (HPLC) with a diode-array detector analyses in order to evaluate the size, the homogeneity, the morphology, and the encapsulation efficiency. β-carotene was selected as marker for the quantitative HPLC analysis. Additionally, physical and chemical stabilities were acceptable during 3 wk at 4 °C. Stability of these nanocarriers in simulated stomach and intestinal conditions was proved. Finally, the improvement of oral absorption of S. repens was studied in vitro using parallel artificial membrane permeability assay. An enhancement of oral permeation was found in both CBM2 and CBS2 nanoformulations comparing with the CB and S. repens CO2 extract. The best performance was obtained by the CBM2 nanoformulation (~ 17%) predicting a 30 - 70% passive oral human absorption in vivo.

Altered Expression of Ganglioside Metabolizing Enzymes Results in GM3 Ganglioside Accumulation in Cerebellar Cells of a Mouse Model of Juvenile Neuronal Ceroid Lipofuscinosis.

Juvenile neuronal ceroid lipofuscinosis (JNCL) is caused by mutations in the CLN3 gene. Most JNCL patients exhibit a 1.02 kb genomic deletion removing exons 7 and 8 of this gene, which results in a truncated CLN3 protein carrying an aberrant C-terminus. A genetically accurate mouse model (Cln3Δex7/8 mice) for this deletion has been generated. Using cerebellar precursor cell lines generated from wildtype and Cln3Δex7/8 mice, we have here analyzed the consequences of the CLN3 deletion on levels of cellular gangliosides, particularly GM3, GM2, GM1a and GD1a. The levels of GM1a and GD1a were found to be significantly reduced by both biochemical and cytochemical methods. However, quantitative high-performance liquid chromatography analysis revealed a highly significant increase in GM3, suggesting a metabolic blockade in the conversion of GM3 to more complex gangliosides. Quantitative real-time PCR analysis revealed a significant reduction in the transcripts of the interconverting enzymes, especially of β-1,4-N-acetyl-galactosaminyl transferase 1 (GM2 synthase), which is the enzyme converting GM3 to GM2. Thus, our data suggest that the complex a-series gangliosides are reduced in Cln3Δex7/8 mouse cerebellar precursor cells due to impaired transcription of the genes responsible for their synthesis.

Validation of the Best Procurement Season of Arjuna Bark [Terminalia arjuna (Roxb. ex DC.) Wight & Arn.] by Comparative HPLC and Pharmacognostical Studies

Aim: The present study was undertaken to establish the best procurement time for Arjuna (Terminalia arjuna (Roxb. ex DC.) Wight & Arn.) by analyzing the seasonal variation in bioactive secondary metabolite in the bark with quantitative high performance liquid chromatography (HPLC) as well as comparative pharmacognosy by taking consideration of Ayurvedic literature. In Ayurveda, it is mentioned that for all the herbs, the best-suited procurement time is Sharad Ritu, especially for bark.1,2 Therefore, to validate the time of collection, the present study has been planned. Materials and methods: Arjuna bark from the same habitat in all six seasons described in Ayurveda, i.e., Shishir (January–February), Vasant (March–April), Grishma (May-June), Varsha (July–August), Sharad (September–October), and Hemant (November–December), has been collected. Authentication of the source of the collected plant material was done and the accession number was given by the herbarium of NVARI, Jhansi. Identification and comparative pharmacognosy in each season at the macroscopic and microscopic levels along with powder microscopy of the useful part of the plant per the standard procedures was done at NVARI Jhansi. The extraction of the economically important plant bark and the quantitative HPLC analysis of the extracted material in all six seasons and comparison of HPLC analysis of different seasons were carried out at Captain Srinivasa Murthy Research Institute of Ayurveda and Siddha Drug Development (CSMRIASDD), Chennai, Tamil Nadu, India. Results: The present study showed that internal microscopical features remained the same throughout the year. A selected marker compound, ellagic acid, was quantified in each season by HPLC. The extractive value was found to be maximum, e.g., 1.7698 g of hydro-alcoholic extract in the Sharad Ritu sample. HPLC estimation showed that the abundance of ellagic acid is more in the Sharad Ritu sample, e.g., 0.0826 to 0.1103. Results indicated that a maximum concentration of bioactive secondary metabolite, i.e., ellagic acid, is in the Sharad Ritu sample. Conclusion: Findings suggest that if Arjuna [Terminalia arjuna (Roxb. ex DC.) Wight & Arn.] barks are collected in Sharad Ritu, then it will provide better therapeutic results.

Open-label uncontrolled pilot study on antipsoriatic activity of Rosa hemisphaerica

Background and objective: Rosa hemisphearica (Rosaceae) is highly cultivated worldwide as an ornamental plant and medicinally have an antibacterial and antioxidant effect. This study aimed to investigate clinically and histopathologically the anti-inflammatory activity of Rosa hemisphearica in psoriasis and to correlate the anti-inflammatory activity with the active constituents in the plant by the phytochemical study. Methods: In this open-label uncontrolled pilot study, a topical preparation of R. hemisphearica stem extract was used by 20 patients suffering from psoriasis for three weeks. Three histopathological slides for each patient, one each week for three weeks was done for anti-inflammatory response evaluation. Subsequently, total phenolic content was measured followed by thin layer chromatography (TLC) and high-performance liquid chromatography (HPLC) analysis for the identification of phenolic acid content in the stem. Results: By histopathological examination revealed that both Munro’s and Kogoj’s abscesses were gradually disappeared, the granular layer gradually restored to normal and the inflammatory cells gradually reduced in number prominently polymorph (neutrophils) and eventually histocytes disappeared. Total phenolic contents in R. hemisphearica stem extract were determined as 205.625 µg gallic acid equivalent each gm of the sample extract, TLC, and HPLC results showed the presence of rosmarinic acid in the stem extract. On quantitative HPLC analysis, the percentage of rosmarinic acid in the stem extract was 0.024 with a retention time (Rt) of 3.4 min. The proposed HPLC method was found to be linear, accurate and precise. Conclusion: The organic solvent extract of R. hemisphearica stem showed a significant reduction in an inflammatory cell among psoriatic patients under study. Rosmarinic acid was identified by TLC and HPLC as important phenolic acid constituents.

Paving the Way for Dose Banding of Chemotherapy: An Analytical Approach.

Background: In an interdepartmental cooperation, we investigated the feasibility and benefits of implementing dose banding of chemotherapy at our medical center. Based on this concept, chemotherapy doses are clustered into bands of similar dosage levels, thereby allowing the preproduction of frequently used standard doses of drugs, with sufficient physicochemical stability. Although established practice in the United Kingdom, there is little published evidence of its introduction elsewhere. Methods: We performed an analysis of local prescribing practice (22,310 chemotherapies) and identified gemcitabine, 5-fluorouracil, and carboplatin, among various others, as cytotoxic drugs suitable for dose banding. Results: First, we determined the physicochemical stability of the selected chemotherapy drugs during 12-weeks' storage by performing pH analysis and visual examination for color change or particles. No relevant changes were identified. Gemcitabine was selected for quantitative high-performance liquid chromatography analysis and we were able to show that ≥95% remained after 12 weeks' storage, in accordance with international guidelines. To simulate a worst case scenario, we performed microbiological stability testing of simulated cytotoxic compounding by replacing the cytotoxic drug with liquid media. Samples were incubated over defined storage time points (3, 6, and 12 weeks) and evaluated using the direct inoculation method. For the container integrity test, we deposited the samples into highly contaminated broth for 1 hour. Microbiological stability was demonstrated in both tests for the full storage period. Conclusions: Our data show that 12-weeks' storage of selected cytotoxic products is feasible from a microbiological perspective. Sterility of prepared products was maintained under extreme storage conditions. Gemcitabine content was in accordance with international guidelines after 12-weeks' storage. These results support the introduction of dose-banded gemcitabine products with the predicted advantages of optimized pharmacy workflow and reduced patient waiting times. We highlight the need for further research and consensus on the performance of purity analyses in dose-banded drug products.

Ajuga chamaecistus subsp. scoparia (Boiss.) Rech.f.: A new source of phytochemicals for antidiabetic, skin-care, and neuroprotective uses

The genus Ajuga is used traditionally as food and medicine. The aim of this study was to investigate the antidiabetic, anti-Alzheimer’s diseases, skin-protective, and antioxidant activities of essential oil and extracts of Ajuga chamaecistus subsp. scoparia (Boiss.) Rech.f., together with the identification of their chemical composition. The 1,1-diphenyl-2-picrylhydrazyl radical and 2,2-azino-bis (3-ethylbenzothiazloine-6-sulfonic acid) radical cation scavenging activity, metal chelating activity, reducing power, content of total bioactive compounds, general toxicity, and enzyme inhibitory potential of essential oil, ethanol extract, and water decoction of the plant were evaluated. Phenolics profile of the extracts was analyzed using the reverse-phase high-performance liquid chromatography with diode array detector and the chemical composition of the essential oil was determined using the gas chromatography and gas chromatography–mass spectroscopy techniques. The essential oil could significantly inhibit the activity of α-glucosidase (4.3mmol Acarbose equivalents/g), α-amylase (2.8mmol Acarbose equivalents/g), acetylcholinesterase (1.96mg gallic acid equivalents/g), butyrylcholinesterase (2.2mg gallic acid equivalents/g), and tyrosinase (36mg kojic acid equivalents/g) while ethanol and water extracts showed moderate enzyme inhibitory activity. Furthermore, essential oil had the strongest antioxidant capacity in the 1,1-diphenyl-2-picrylhydrazyl scavenging, 2,2-azino-bis (3-ethylbenzothiazloine-6-sulfonic acid) scavenging, cupric reducing antioxidant capacity, ferric reducing antioxidant power, and metal chelating assays. Spathulenol (18.0%), thymol (15.1%), octen-3-ol (14.3%), and linalool oxide (11.2%) were identified as major constituents in the essential oil. Quantitative high-performance liquid chromatography analysis indicated that p-coumaric acid, gallic acid, and ferulic acid are the main phenolic compounds in this plant. The results suggested that Ajuga chamaecistus could be useful for several applications as functional foods, cosmetics, and pharmaceuticals.

Forsythia suspensa fruit extracts and the constituent matairesinol confer anti-allergic effects in an allergic dermatitis mouse model

Ethnopharmacological relevanceForsythia suspensa is used in traditional medicine to treat inflammation. To clarify the anti-inflammatory and anti-allergic effects of F. suspensa fruits, we determined the therapeutic effects of crude extract, fractions, and a constituent from F. suspensa fruits on a murine atopic dermatitis (AD) model. Materials and methodsWe investigated the inhibitory effects of F. suspensa extract (FSE), extract fractions, and the constituent matairesinol on histamine release from MC/9 mast cells activated by compound 48/80 and the development of AD-like skin lesions and symptoms in NC/Nga mice exposed to Dermatophagoides farinae (mite) extract. High performance liquid chromatography (HPLC) analysis of FSE and its fractions were evaluated using matairesinol standard. ResultsFSE, FSE methylene chloride fraction (FSE-MC), and FSE water fraction (FSE-water) inhibited compound 48/80-induced histamine release from MC/9 mast cells. Topical application of FSE or FSE-MC to NC/Nga mice exposed to Dermatophagoides farinae suppressed the development of AD-like skin lesions. Quantitative HPLC analysis of FSE and FSE-MC identified the presence of matairesinol. Topical application of matairesinol to NC/Nga mice effectively reduced AD symptoms, inhibited inflammatory cell infiltration, and lowered immunoglobulin E levels in serum. Further study demonstrated that DfE-induced changes in IL-4 and IFN-γ mRNA expression in the ears of NC/Nga mice were reversed by matairesinol application. ConclusionsThese results indicate that the F. suspensa and its constituent matairesinol might be a therapeutic candidate for treating allergic inflammatory disorders such as AD.