scholarly journals Development, Validation and Application of a UHPLC-MS Method for the Quantification of Chios Mastic Gum Triterpenoids in Human Plasma

Planta Medica ◽  
2021 ◽  
Author(s):  
Vincent Brieudes ◽  
Eleni V. Mikropoulou ◽  
Errikos Kallergis ◽  
Andriana C. Kaliora ◽  
Efstathia Papada ◽  
...  
Keyword(s):  
Human Plasma ◽  
Biological Fluids ◽  
Structural Characteristics ◽  
Biological Properties ◽  
Lipid Lowering ◽  
High Yield ◽  
Limit Of Quantification ◽  
Isolation And Characterization ◽  
Starting Point ◽  
Chios Mastic Gum

AbstractChios mastic gum is the resinous secretion obtained from the barks of the shrub Pistacia lentiscus var. Chia, which is endemic to the Greek island of Chios. Since antiquity, Chios mastic gum has found several uses as a phytotherapeutic remedy, primarily for the treatment of gastrointestinal disorders while recently, Chios mastic gum was also recognized by EMA as an herbal medicinal product with specific indications. Chios mastic gumʼs biological properties are attributed to triterpenes which comprise the major chemical group (approx. 70%) and notably isomasticadienonic acid and masticadienonic acid. However, due to their structural characteristics, the isolation thereof in high yield and purity is challenging and since they are not commercially available, pharmacological studies aiming to assess their biological properties are limited. In the present work, masticʼs phytochemical investigation by UPLC-HRMS is followed by the isolation and characterization of isomasticadienonic acid and masticadienonic acid to be used as analytical standards for their accurate and reliable quantification in human plasma. A UHPLC-tQ-MS method that was developed and validated (in terms of specificity, linearity, limit of quantification, accuracy and precision), for the direct quantification of the targeted compounds in the low ng/mL range of concentration, was subsequently implemented on plasma samples of healthy volunteers thus demonstrating its fitness for purpose. The results presented herein might provide insight to the understanding of this traditional natural product consumed notably for its anti-inflammatory, antioxidant and lipid lowering properties. Moreover, this method might serve as a starting point for any study aiming to monitor bioactive triterpenes in biological fluids.

scholarly journals Micellar Enhanced Spectrofluorimetric Method for the Determination of Ponatinib in Human Plasma and Urine via Cremophor RH 40 as Sensing Agent

2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Hany W. Darwish ◽  
Ahmed H. Bakheit ◽  
Ali Saber Abdelhameed ◽  
Amer S. AlKhairallah
Keyword(s):  
Human Plasma ◽  
Fluorescence Intensity ◽  
Limit Of Detection ◽  
Biological Fluids ◽  
Great Success ◽  
Limit Of Quantification ◽  
Human Plasma And Urine ◽  
Spectrofluorimetric Method ◽  
Fluorescence Characteristics

An impressively simple and precise spectrofluorimetric procedure was established and validated for ponatinib (PTB) quantitation in biological fluids such as human plasma and human urine. This method depends on examining the fluorescence characteristics of PTB in a micellar system of Cremophor RH 40 (Cr RH 40). Cr RH 40 enhanced the intrinsic fluorescence of PTB distinctly in aqueous water. The fluorescence spectra of PTB was recorded at 457 nm following its excitation at 305 nm. Maximum fluorescence intensity was attained by addition of 0.7 mL of Cr RH 40 and one mL of phosphate buffer to PTB aliquots and then dilution with distilled water. There is a linear relationship between the fluorescence intensity of PTB and its concentration over the range 5–120 ngmL−1, with limit of detection and limit of quantification equal to 0.905 ngmL−1and 2.742 ngmL−1, respectively. The accuracy and the precisions of the proposed method were checked and gave adequate results. The adopted method was applied with a great success for PTB quantitation in different biological matrices (spiked human plasma and urine) giving high recovery values.

scholarly journals Development and Validation of Valganciclovir and its Active Metabolite Ganciclovir Determination in Human Plasma by HPLC-MS/MS Method

2021 ◽  
Vol 10 (1) ◽  
pp. 120-128
Author(s):  
T. N. Komarov ◽  
I. E. Shohin ◽  
M. A. Tokareva ◽  
O. A. Archakova ◽  
D. S. Bogdanova ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
High Performance ◽  
Biological Fluids ◽  
Detection Methods ◽  
Pharmacokinetic Studies ◽  
Limit Of Quantification ◽  
Carry Over

Introduction. Currently, physicochemical methods of quantification are actively used to determine the content of drugs in biological fluids. High-performance liquid chromatography with various detection methods is particularly widespread. One of the most difficult practical tasks is the chromatographic separation of so-called poorly retained compounds – drug substances poorly retained on the chromatographic column. Valganciclovir and Ganciclovir are among such substances. Aim. The aim of this study is to develop a method for valganciclovir and ganciclovir in human plasma by high performance liquid chromatography with tandem mass-spectrometry (HPLC-MS/MS) for pharmacokinetic studies.Materials and methods. Determination of valganciclovir and ganciclovir in plasma by HPLC-MS/MS. The samples were processed by acetonitrile protein precipitation.Results and discussion. This method was validated by next parameters: selectivity, matrix effect, calibration curve, accuracy, precision, recovery, lower limit of quantification, carry-over and stability.Conclusion. The method of the determination of valganciclovir and ganciclovir in human plasma was developed and validated by HPLC-MS/MS. The linearity in plasma sample was achieved in the concentration range of 5.00–1000.00 ng/ml for valganciclovir and 50.00–10000.00 ng/ml for ganciclovir. Method could be applied to valganciclovir and ganciclovir determination in plasma for PK and BE studies.

scholarly journals ADVANCED BIOMEDICAL HYDROGELS: MOLECULAR ARCHITECTURE AND ITS IMPACT ON MEDICAL APPLICATIONS

2021 ◽  
Author(s):  
Jonathan T Peters ◽  
Marissa Wechsler ◽  
Nicholas A Peppas
Keyword(s):  
Biological Fluids ◽  
Structural Characteristics ◽  
Biological Properties ◽  
Molecular Characteristics ◽  
Medical Applications ◽  
Biological Applications ◽  
Molecular Architecture ◽  
Polymeric Networks ◽  
Wide Range ◽  
External Stimuli

Abstract Hydrogels are crosslinked polymeric networks swollen in water, physiological aqueous solutions, or biological fluids. They are synthesized by a wide range of polymerization methods that allow for the introduction of linear and branched units with specific molecular characteristics. In addition, they can be tuned to exhibit desirable chemical characteristics including hydrophilicity or hydrophobicity. The synthesized hydrogels can be anionic, cationic, or amphiphilic, and can contain multifunctional crosslinks, junctions or tie points. Beyond these characteristics, hydrogels exhibit compatibility with biological systems, and can be synthesized to render systems that swell or collapse in response to external stimuli. This versatility and compatibility have led to better understanding of how the hydrogel’s molecular architecture will affect their physicochemical, mechanical, and biological properties. We present a critical summary of the main methods to synthesize hydrogels which define their architecture, and advanced structural characteristics for macromolecular/biological applications.

Purification of Urokinase from Complex Mixtures Using Immobilized Monoclonal Antibody Against Urokinase Light Chain

1983 ◽  
Vol 49 (01) ◽  
pp. 024-027 ◽  
Author(s):  
David Vetterlein ◽  
Gary J Calton
Keyword(s):  
Monoclonal Antibody ◽  
Light Chain ◽  
Culture Media ◽  
Biological Fluids ◽  
Single Step ◽  
High Yield ◽  
Step Method ◽  
Commercial Preparation ◽  
E Coli ◽  
Tissue Culture Media

SummaryThe preparation of a monoclonal antibody (MAB) against high molecular weight (HMW) urokinase light chain (20,000 Mr) is described. This MAB was immobilized and the resulting immunosorbent was used to isolate urokinase starting with an impure commercial preparation, fresh urine, spent tissue culture media, or E. coli broth without preliminary dialysis or concentration steps. Monospecific antibodies appear to provide a rapid single step method of purifying urokinase, in high yield, from a variety of biological fluids.

Entrepreneurship Management at the Local Government Level: The Experience of the Cities for Entrepreneurs Program

2018 ◽  
Vol 9 (5) ◽  
pp. 388-407
Author(s):  
Patricio Gigli ◽  
◽  
Donatela Orsi ◽  
Marisel Martín Aramburú ◽  
◽  
...  
Keyword(s):  
Human Capital ◽  
Local Government ◽  
Local Governments ◽  
Structural Characteristics ◽  
National Level ◽  
Development Issue ◽  
Starting Point ◽  
Local Government Level ◽  
Micro Level ◽  
Government Level

This paper aims at describing the experience of the Cities for Entrepreneurs Program (Ciudades para Emprender or CPE) of the National Directorate of Community and Human Capital (which belongs to the SEPYME), National Ministry of Production. This paper starts from the premise that entrepreneurship takes place at the most micro level of the offer and, therefore, is a concept associated with the characteristics of the environment closest to that offer: the local territory. However, there is little history in the country of public policies relating the issue of entrepreneurship with the local management. That is why we take as a starting point the conceptualization of the chosen framework: local governments and the development issue, seen from the perspective of entrepreneurships. Moreover, an overview is given on the structural characteristics of municipalities in Argentina. In addition, some international experiences and attempts to promote entrepreneurship at a national level are analyzed. Finally, the Cities for Entrepreneurs Program (CPE) is outlined, based on a summary of the diagnoses of the Entrepreneurial Ecosystems of the selected cities and the tools used and their execution status at the time of publication of this paper.

BOTANICAL COMPOSITION AND PRODUCTIVITY OF PERENNIAL GRASSES AGROPHYTOCENOSES WITH TETRAPLOID MEADOW CLOVER

1970 ◽  
pp. 59-62
Author(s):  
N. I. Kasatkina ◽  
Zh. S. Nelyubina
Keyword(s):  
Dry Weight ◽  
Single Species ◽  
Lotus Corniculatus ◽  
Specific Weight ◽  
Biological Properties ◽  
Phleum Pratense ◽  
Perennial Grasses ◽  
High Yield ◽  
Botanical Composition ◽  
The Third

The biological properties of plants, their mutual relations under different growth conditions and at different periods of their life, must be known for obtaining highly productive agrophytocenoses with participation of a meadow clover (Trifolium pratense L.). Botanical composition and fodder productivity of perennial grasses in agrocenoses with participation of meadow tetraploid clover Kudesnik were studied in 2014-2017. It was revealed that in the first and second years of use the agrophytocenosis, the yield of green mass was formed due to meadow tetraploid clover, the share of its participation in the first mowing was at level of 71-87% and 64-97% respectively. Specific weight of clover in multispecies agrocenoses considerably decreased by the third year of use: in the first mowing up to 32-68%, in the second - up to 8-52%. At the same time, the percentage of long-term herbaceous grasses increased: meadow timothy (Phleum pratense L.) - up to 34-54%, eastern galega (Galéga orientális Lam.) - up to 33%, changeable alfalfa (Medicago x varia Martyn) - up to 22-54%, lotus corniculatus (Lotus corniculatus L.) - up to 14-19%. The proportion of weed admixture in single-species clover planting was 12%, in agrocenoses - 2-14%. The grass mixtures clover + timothy and clover + alfalfa + timothy were less infested by weeds. High yield of dry weight of single-species sowing of meadow tetraploid clover was obtained in the first two years of use - 7.8 and 6.5 tons / ha, respectively. By the third year of use, the productivity of clover has decreased to 2.9 t / ha. On average, for three years of use, the highest yield (6.2-6.3 t / ha) was formed by agrocenoses meadow tetraploid clover + meadow timothy and meadow tetraploid clover + changeable alfalfa + meadow timothy.

PERSPECTIVES OF APPLICATION OF ANTIBODIES AGAINST ENDOGLIN (CD105) FOR VISUALIZATION AND ANTI-ANGIOGENE THERAPY OF TUMORS

2018 ◽  
Vol 64 (4) ◽  
pp. 504-507
Author(s):  
Vladimir Klimovich ◽  
Natalya Vartanyan ◽  
Anastasiya Stolbovaya ◽  
Lidiya Terekhina ◽  
Olga Shashkova ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Monoclonal Antibodies ◽  
Human Plasma ◽  
Vascular Endothelium ◽  
Immune Complexes ◽  
Targeted Delivery ◽  
Biological Fluids ◽  
Therapeutic Drugs ◽  
Cultured Endothelial Cells ◽  
Hybridoma Technology

During last years monoclonal antibodies (MAB) directed against vascular endothelium markers demonstrated their efficiency for visualization and targeted delivery of therapeutic drugs to tumors. Endoglin (CD105) which serves as a key element that determines endothelial cells quiescence or activation is one of such markers. Endoglin is highly expressed on the vascular endothelium of growing tumors. A first panel of MAB against endoglin in our country was produced at the hybridoma technology laboratory of RRC RST named after A.M. Granov. On the basis of these MAB ELISA was created allowing detection of endoglin in human plasma and other biological fluids. Several MAB had been shown to bind endoglin on the membrane of the cultured endothelial cells and to persist there for several hours. During the first 30 min after binding some of the immune complexes “endoglin-MAB” were internalized into the cytoplasm and were found included in the endosomes. In future these MAB can be used to create the reagents for the addressed delivery of isotope tags both on the membrane and into the cytoplasm of endothelial cells.

Development and Validation of a New Method for the Determination of Anti-hepatitis C Agent Simeprevir in Human Plasma using HPLC with Fluorescence Detection

2020 ◽  
Vol 16 (4) ◽  
pp. 428-435
Author(s):  
Ahmed F.A. Youssef ◽  
Yousry M. Issa ◽  
Kareem M. Nabil
Keyword(s):  
Hepatitis C ◽  
Human Plasma ◽  
Fluorescence Detection ◽  
Internal Standard ◽  
Protein Precipitation ◽  
Assay Method ◽  
Fluorescence Detector ◽  
Limit Of Quantification ◽  
Relative Standard

Background: Simeprevir is one of the recently discovered drugs for treating hepatitis C which is one of the major diseases across the globe. Objective: The present study involves the development of a new and unique High-Performance Liquid Chromatography (HPLC) method using fluorescence detection for the determination of simeprevir (SIM) in human plasma. Methods: Two methods of extractions were tested, protein precipitation using acetonitrile and liquidliquid extraction. A 25 mM dipotassium hydrogen orthophosphate (pH 7.0)/ACN (50/50; v/v), was used as mobile phase and C18 reversed phase column as the stationary phase. The chromatographic conditions were optimized and the concentration of simeprevir was determined by using the fluorescence detector. Cyclobenzaprine was used as an internal standard. Results: Recovery of the assay method based on protein precipitation was up to 100%. Intra-day and inter-day accuracies range from 92.30 to 107.80%, with Relative Standard Deviation (RSD) range 1.65-8.02%. The present method was successfully applied to a pharmacokinetic study where SIM was administered as a single dose of 150 mg SIM/capsule (Olysio®) to healthy individuals. Conclusion: This method exhibits high sensitivity with a low limit of quantification 10 ng mL-1, good selectivity using fluorescence detection, wide linear application range 10-3000 ng mL-1, good recovery and highly precise and validation results. The developed method can be applied in routine analysis for real samples.

scholarly journals Analysis of Pembrolizumab in Human Plasma by LC-MS/HRMS. Method Validation and Comparison with Elisa

Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 621
Author(s):  
Aurélien Millet ◽  
Nihel Khoudour ◽  
Jérôme Guitton ◽  
Dorothée Lebert ◽  
François Goldwasser ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Human Plasma ◽  
High Resolution Mass Spectrometry ◽  
Internal Standard ◽  
Therapeutic Drug ◽  
Limit Of Quantification ◽  
Immunoglobulin G4 ◽  
Positive Mode ◽  
Surrogate Peptide ◽  
First Time

Pembrolizumab is a humanized immunoglobulin G4-kappa anti-PD1 antibody used in the treatment of different solid tumors or haematological malignancies. A liquid chromatography coupled with a high resolution mass spectrometry (orbitrap technology) method was fully developed, optimized, and validated for quantitative analysis of pembrolizumab in human plasma. A mass spectrometry assay was used for the first time a full-length stable isotope-labelled pembrolizumab-like (Arginine 13C6-15N4 and Lysine 13C6-15N2) as an internal standard; the sample preparation was based on albumin depletion and trypsin digestion and, finally, one surrogate peptide was quantified in positive mode. The assay showed good linearity over the range of 1–100 μg/mL, a limit of quantification at 1 μg/mL, excellent accuracy from 4.4% to 5.1%, and also a between-day precision below 20% at the limit of quantification. In parallel, an in-house ELISA was developed with a linearity range from 2.5 to 50 µg/mL. Then, results were obtained from 70 plasma samples of cancer patients that were treated with pembrolizumab and quantified with both methods were compared using the Passing-Bablok regression analysis and Bland-Altman plotting. The LC-MS/HRMS method is easy to implement in the laboratory for use in the context of PK/PD studies, clinical trials, or therapeutic drug monitoring.

scholarly journals Isolation and Characterization of Phages Active against Paenibacillus larvae Causing American Foulbrood in Honeybees in Poland

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1217
Author(s):  
Ewa Jończyk-Matysiak ◽  
Barbara Owczarek ◽  
Ewa Popiela ◽  
Kinga Świtała-Jeleń ◽  
Paweł Migdał ◽  
...  
Keyword(s):  
Biological Properties ◽  
Field Application ◽  
Paenibacillus Larvae ◽  
American Foulbrood ◽  
Isolation And Characterization ◽  
Safety Studies ◽  
Spreading Disease ◽  
Intended Use ◽  
Antibacterial Potential

The aim of this study was the isolation and characterization, including the phage effect on honeybees in laboratory conditions, of phages active against Paenibacillus larvae, the causative agent of American Foulbrood—a highly infective and easily spreading disease occurring in honeybee larva, and subsequently the development of a preparation to prevent and treat this dangerous disease. From the tested material (over 2500 samples) 35 Paenibacillus spp. strains were obtained and used to search for phages. Five phages specific to Paenibacillus were isolated and characterized (ultrastructure, morphology, biological properties, storage stability, and genome sequence). The characteristics were performed to obtain knowledge of their lytic potential and compose the final phage cocktail with high antibacterial potential and intended use of future field application. Preliminary safety studies have also been carried out on healthy bees, which suggest that the phage preparation administered is harmless.

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