Detection and Extraction of Heparin from Camel Lungs

2019 ◽  
Vol 20 (6) ◽  
pp. 476-482
Author(s):  
Ahmad Almeman ◽  
Kasem Abdulmajed ◽  
Eltayeb E. Eid
Keyword(s):  
Lung Tissue ◽  
Method Development ◽  
Factor Xa ◽  
Extraction Methods ◽  
Raw Material ◽  
Activity Measurement ◽  
Business Opportunity ◽  
Heparin Activity ◽  
Heparin Preparation ◽  
Viable Business

Background: Heparin is an essential drug used as an anticoagulant. Access to raw material suitable for heparin extraction is critical for creating a viable business opportunity. In Saudi Arabia, large amounts of raw material with potential for heparin extraction are wasted. Objective: To extract heparin and low-molecular-weight heparin (LMWH) from the camel lung, and measure its potency and activity. Methods: Heparin preparation included three steps: extraction, electrophoretic identification, and activity measurement. Fresh lung tissue (100 g) was minced and homogenized in a blender. Crude heparin extracts were prepared using Charles’s or Volpi’s method with slight modifications. Heparin was purified by electrophoresis using high-purity agarose gels in barium acetate buffer. The heparin activity of purified samples was assayed spectrophotometrically using commercial heparin kits. Results: Charles’s and Volpi’s extraction methods were simple and easy to establish. The yield was 90 mg crude heparin per 100 g of camel lung tissue following Volpi’s extraction protocol, whereas Charles’s method did not yield any heparin. The separation of heparin and LMWH by gel electrophoresis resulted in sharp and clear product bands using material prepared according to Volpi’s method. The heparin preparation had an anti-factor Xa activity of 37 IU/mg, indicating weak potency. Conclusion: Preparation of active heparin from camel lung tissue is a technology applicable in manufacturing. Further method development is needed to increase heparin purity and potency.

Anthocyanins: Chemical Properties and Health Benefits: A Review

2021 ◽  
Vol 17 ◽  
Author(s):  
Siraj Salman Mohammad ◽  
Renata Oliveira Santos ◽  
Maria Ivone Barbosa ◽  
José Lucena Barbosa Junior
Keyword(s):  
Raw Materials ◽  
Health Benefits ◽  
Chemical Properties ◽  
Extraction Methods ◽  
The Body ◽  
Raw Material ◽  
Extraction Properties ◽  
Therapeutic Properties ◽  
Food Processes ◽  
Shed Light

: Anthocyanins are widely spread in different kinds of food, especially fruits and floral tissues, there is an extensive range of anthocyanin compounds reach more than 600 exist in nature. Anthocyanins can be used as antioxidants and raw material for several applications in food and pharmaceutical industry. Consequently, a plenty of studies about anthocyanins sources and extraction methods were reported. Furthermore, many studies about their stability, bioactive and therapeutic properties have been done. According to the body of work, we firstly worked to shed light on anthocyanin properties including chemical, antioxidant and extraction properties. Secondly, we reported the applications and health benefits of anthocyanin including the applications in food processes and anthocyanin characteristics as therapeutic and prophylactic compounds. We reviewed anticancer, anti-diabetic, anti-fatness, oxidative Stress and lipid decreasing and vasoprotective effects of anthocyanins. In conclusion, because the importance of phytochemicals and bioactive compounds the research is still continuing to find new anthocyanins from natural sources and invest them as raw materials in the pharmaceutical and nutrition applications.

scholarly journals Obtaining Bioactive Compounds from the Coffee Husk (Coffea arabica L.) Using Different Extraction Methods

Molecules ◽  
2020 ◽  
Vol 26 (1) ◽  
pp. 46
Author(s):  
Mariana de Oliveira Silva ◽  
John Nonvignon Bossis Honfoga ◽  
Lorena Lucena de Medeiros ◽  
Marta Suely Madruga ◽  
Taliana Kênia Alencar Bezerra
Keyword(s):  
Phenolic Compounds ◽  
Bioactive Compounds ◽  
Extraction Methods ◽  
Water Bath ◽  
Raw Material ◽  
Total Phenolic ◽  
Total Phenolic Compounds ◽  
Ethanol Mixture ◽  
Coffee Husk ◽  
Coffea Arabica L

Coffee husks (Coffea arabica L.) are characterized by exhibiting secondary metabolites such as phenolic compounds, which can be used as raw material for obtaining bioactive compounds of interest in food. The objective of this study is to evaluate different methods for obtaining the raw material and extracting solutions of bioactive compounds from coffee husks. Water bath and ultrasound-assisted extraction methods were used, using water (100%) or ethanol (100%) or a mixture of both (1:1) as extracting solutions and the form of the raw material was in natura and dehydrated. The extracts were evaluated by their antioxidant potential using DPPH radicals, ABTS, and iron reduction (ferric reducing antioxidant power (FRAP)), and later total phenolic compounds, total flavonoids, and condensed tannins were quantified the phenolic majority compounds were identified. It was verified that the mixture of water and ethanol (1:1) showed better extraction capacity of the compounds with antioxidant activity and that both conventional (water bath) or unconventional (ultrasound) methods showed satisfactory results. Finally, a satisfactory amount of bioactive compounds was observed in evaluating the chemical composition (total phenolic compounds, total flavonoids, condensed tannins, as well as the analysis of the phenolic profile) of these extracts. Corroborating with the results of the antioxidant activities, the best extracting solution was generally the water and ethanol mixture (1:1) using a dehydrated husk and water bath as the best method, presenting higher levels of the bioactive compounds in question, with an emphasis on chlorogenic acid. Thus, it can be concluded that the use of coffee husk as raw material to obtain extracts of bioactive compounds is promising. Last, the conventional method (water bath) and the water and ethanol mixture (1:1) stood out among the methods and extracting solutions used for the dehydrated coffee husk.

scholarly journals Chromogenic anti-Xa test: the ratio between heparin activity units and concentration of apixaban and rivaroxaban

2020 ◽  
pp. 96-104
Author(s):  
E. V. Titaeva ◽  
A. B. Dobrovolsky
Keyword(s):  
Oral Anticoagulants ◽  
Direct Oral Anticoagulants ◽  
Plasma Concentrations ◽  
Factor Xa ◽  
Activity Test ◽  
Formation Dynamics ◽  
Test Version ◽  
Heparin Activity ◽  
The Relationship ◽  
Thrombin Formation

Introduction. The direct oral anticoagulants (DOC) therapy does not require alaboratory control; however, it may be required to determine the anticoagulationlevel to choose a treatment strategy if alarge bleeding is developing or emergency surgery is needed.The objective of this experimental study was to investigate the relationship between the residual factor Xa (FXa) activity, anti-Xa activity units oflow molecular weight heparins (LMWH), and the apixaban and rivaroxaban plasma concentrations in a chromogenic anti-Xa assay.Material and methods. Concentrated DOC solutions were prepared by extracting apixaban and rivaroxaban from crushed tablets using methanol and dimethyl sulfoxide, respectively. The resulting solutions were added to the donor plasma pool until final inhibitor concentrations are achieved in the range from 10 to 100 ng/ml plasma. Anti-Xa activity was determined using an STA-compact analyser and the Liquid anti-Xa reagent kit, an analysis protocol, and calibrators designed to control the LMWH therapy. The effect on the thrombin formation dynamics was investigated using the thrombin generation test (TGT) and the PPR reagent as a trigger (final concentrations of tissue factor are 5 pM, and those of phospholipids are 4 μM). TGT curves were analysed using the Thrombinoscope program.Results. It was shown that in the anti-Xa activity test version designed to control the LMWH therapy, there is a high correlation (R2 > 0.98) between thelogarithm of the residual factor Xa activity and the content of apixaban and rivaroxaban in the range from 10 to 80 ng/ml. Rivaroxaban shows about 1.5 times more anti-Xa activity than apixaban at equal concentrations. It was also shown that apixaban and rivaroxaban at doses equal both in concentration and in anti-Xa activity differ in their effect on the thrombin formation dynamics and thrombin inactivation in the TGT.Conclusion. In the LMWH anti-Xa activity test version, the measured range of apixaban and rivaroxaban includes 30 ng/ml and 50 ng/ ml concentrations taken as “cut-off points” to determine the treatment tactics in emergency cases. However, thelack of certified DOC calibratorslimits the use of this test in clinical practice.

scholarly journals EXTRACTION AND PURIFICATION OF VEGETABLE PEROXIDASE: A REVIEW

2019 ◽  
Vol 16 (31) ◽  
pp. 692-703
Author(s):  
Aline HAAS ◽  
Cleiton VAZ ◽  
Aniela Pinto KEMPKA
Keyword(s):  
Enzymatic Activity ◽  
Specific Activity ◽  
Extraction Methods ◽  
Raw Material ◽  
Partial Purification ◽  
Buffer Solution ◽  
Degradation Of Dyes ◽  
Extraction And Purification ◽  
Wide Range ◽  
Purification Methods

Peroxidases are enzymes that catalyze the oxidation of various substrates, maintaining their enzymatic activity in wide ranges of pH and temperatures. These enzymes are used in processes for the degradation of dyes and phenolic compounds. Peroxidases are present in the tissues of several plants, and the search for new sources of this enzyme is necessary. This literature review aims to compile information about the extraction and/or purification of peroxidases contained in different plant tissues, presenting extraction methods, purification processes, enzymatic activities and their increments, according to the chemical and physical processes applied. Several plant sources can be raw material to obtain these enzymes, through different forms of extraction, where the processes of comminution predominate in the presence of buffer solution. For partial purification, are used precipitation with solvents (acetone and ethanol) and salts (ammonium sulfate) and centrifugation. For purification, chromatographic processes are used, in which molecular exclusion and affinity chromatography are prominent. It is concluded that there is a wide range of possibilities for obtaining the enzyme peroxidase from plants, with variability in the enzymatic activity when different extraction methods are applied. The purification methods used provide increases in the specific activity of the peroxidases.

scholarly journals WEEE collection and CRM recovery trials: piloting a holistic approach for Scotland

2018 ◽  
Vol 20 (4) ◽  
pp. 712-718
Keyword(s):  
Raw Materials ◽  
Scale Up ◽  
Holistic Approach ◽  
Extraction Methods ◽  
Social Enterprises ◽  
Raw Material ◽  
Household Waste ◽  
Sustainable Supply Chain ◽  
Gold Silver ◽  
Critical Raw Materials

<p>Re-Tek UK and its partners, Enscape Consulting and the University of West of Scotland commenced trials for the collection and recovery of critical raw materials from waste electrical and electronic (WEEE) products in July 2016. Sponsored by the EU LIFE funded project ‘Critical Raw Material Closed Loop Recovery’ coordinated by WRAP with EARN, ERP UK Ltd, KTN Ltd and Wuppertal Institute as beneficiaries. The trials are aimed at boosting the recovery of critical raw materials (CRMs) from household waste electrical and electronic products (WEEE) and Information Communications Technology (ICT) in particular, after functioning equipment is separated out for re-use. The new collection models provided residents with the opportunity to drop-off unwanted electrical and electronic appliances at a time and place that suits them, through a collaborative approach which encourages local authorities, educational establishments, businesses, and Social Enterprises, etc to act as hub sites. Hubs were designed to minimize product damage and encourage drop-off, rather than hoarding. Extraction methods developed after the collection phase of the trial looked at the opportunity to recover cobalt, gold and silver from ICT products, with the potential to inform how a more sustainable supply chain could be developed in Scotland. The elements studied were selected to demonstrate financial opportunity (gold/silver) and a strategic priority material (cobalt) for long term supply. These are based on bioleaching and electrochemical recovery using novel carbon based electrode systems, and chemical processing methods using extraction techniques with an assessment of pilot performance and scale up challenges. Our report is on the state of progress towards practical solutions to WEEE and CRM recovery.</p>

A Comparison of Anticoagulation Activity of a Potent Heparin Preparation in Different Test

1979 ◽  
Author(s):  
A.S. Bhargava ◽  
J. Heinick ◽  
Chr. Schöbel ◽  
P. Günzel
Keyword(s):  
Blood Clotting ◽  
Anticoagulant Activity ◽  
Factor Xa ◽  
Thrombin Time ◽  
Beagle Dogs ◽  
Clotting Time ◽  
Relative Potencies ◽  
Heparin Preparation

The anticoagulant effect of a new potent heparin preparation was compared with a commercially available heparin in vivo after intravenous application in beagle dogs. The anticoagulant activity was determined using thrombin time, activated partial thromboplastin time and whole blood clotting time after 5, 10 and 30 minutes of application. The relative potency of the new heparin preparation (Scherinq) was found to be 1.62 to 2.52 times higher than heparin used for comparison (150 USP units/mg, Dio-synth). The anticoagulant properties of both preparations were also studied in vitro using dog and human plasma. The relative potencies in vitro correlated well with those obtained in vivo. Further characterization with amidolytic method using chromogenic substrate for factor Xa and thrombin (S-2222 and S-2238 from KABI, Stockholm) showed that heparin (Schering) contains 243 to 378 USP units/raq depending upon the test systems used to assay the anticoagulation activity and in addition, proves the validity of the amidolytic method.

Dextran sulfate included in factor Xa assay reagent overestimates heparin activity in patients after heparin reversal by protamine

2003 ◽  
Vol 111 (4-5) ◽  
pp. 273-279 ◽  
Author(s):  
Christine Mouton ◽  
Joachim Calderon ◽  
Gérard Janvier ◽  
Marie-Christine Vergnes
Keyword(s):  
Dextran Sulfate ◽  
Factor Xa ◽  
Heparin Activity

The Use of Anti-Xa Assay to Monitor Intravenous Unfractionated Heparin Therapy

2010 ◽  
Vol 23 (3) ◽  
pp. 210-216 ◽  
Author(s):  
Amy Fann Rosenberg ◽  
Marc Zumberg ◽  
Lisa Taylor ◽  
Aimée LeClaire ◽  
Neil Harris
Keyword(s):  
Clinical Trials ◽  
Unfractionated Heparin ◽  
Pediatric Population ◽  
Factor Xa ◽  
Heparin Therapy ◽  
Activated Partial Thromboplastin Time ◽  
Controlled Clinical Trials ◽  
Randomized Controlled Clinical Trials ◽  
Randomized Controlled ◽  
Heparin Activity

Continuous infusion unfractionated heparin (UH) has traditionally been monitored using the activated partial thromboplastin time (aPTT). The use of this test to monitor heparin therapy is not based on randomized controlled clinical trials, and the test is associated with significant intra- and inter-patient variability that is not related to circulating blood heparin activity. Due to these and other limitations, the use of aPTT alone to monitor UF has been questioned. Many laboratories are now transitioning to monitoring actual heparin activity (by anti-factor Xa analysis). In this review, we discuss the limitations of using the aPTT to monitor UH therapy and additionally the limitations of solely using heparin activity to monitor therapy. We also include a discussion of the challenges with monitoring heparin therapy in the pediatric population.

A DEFINITION OF HEPARIN ANTICOAGULANT POTENCY APPLICABLE TO ALL HEPARINS AND HEPARIN-LIKE SUBSTANCES AND ITS PRACTICAL APPLICATION IN ASSAYING HEPARIN

1987 ◽  
Author(s):  
Craig M Jackson
Keyword(s):  
Rate Constant ◽  
Anticoagulant Activity ◽  
Factor Xa ◽  
Order Rate Constant ◽  
Red Cross ◽  
Order Kinetic ◽  
High Affinity ◽  
General Terms ◽  
The Individual ◽  
Heparin Preparation

Heparins increase the rate of inactivation of proteinases by antithrombin without being consumed in the inactivation reaction. The anticoagulant activity of any heparin or heparin preparation is thus determined by the increase in the inactivaton rate which it produces. This rate increase is dependent on the concentration of the heparin in the sample and on some now well known structural properties of the individual heparin molecules that produce high affinity for antithrombin . All proteinases are not inactivated by antithrombin equally rapidly in the absence of heparin, nor are heparins and heparin derivatives of different molecular weight equally effective in the inactivation of the same proteinase. Under appropriate conditions, the observed rate constant (kObs) for the heparin catalyzed proteinase inactivation reaction is simply related to the intrinsic potencies and concentrations of the individual high affinity heparin molecules in the sample. The intrinsic potency of a high affinity heparin molecule is the efficiency with which it catalyzes the inactivation of the particular proteinase, e.g. Factor Xa or thrombin, i.e., it is a second order rate constant, (designated k*) . After k* has been determined from kobs for a known heparin or heparin preparation and a particular proteinase, the concentration of heparin in an unknown sample can be calculated from the equation[H] = [HAT] = kobs/k* In general terms, the appropriate conditions, i.e.,the antithrombin and proteinase concentrations, the pH, and ionic strength, required for this equation to be used are those conditions for which all of the high affinity heparin is bound to the antithrombin and pseudo first order kinetic behavior occurs. At very low heparin concentrations, a correction for the inactivation of the proteinase by antithrombin alone is necessary, but is easily made.Supported by Organon Teknika Corporation and an Established Investigator Award from the American National Red Cross

scholarly journals The Activity of Heparin Preparations Studied by Amidolytic and Clotting Methods

1977 ◽  
Author(s):  
A.N. Teien ◽  
U. Abildgaard ◽  
M. Höök ◽  
U. Lindahl
Keyword(s):  
Antithrombin Iii ◽  
Specific Activity ◽  
Factor Xa ◽  
Test System ◽  
Assay Method ◽  
Thrombin Time ◽  
Assay Methods ◽  
Specific Activities ◽  
The Difference ◽  
Heparin Preparation

Two heparin standards, heparin isolated from human mastocytoma tissue, four commercial heparins and two heparin preparations separated by affinity chromatography (“High affinity heparin”=HAH and “Low affinity heparin”=LAH) were assayed by the activated partial thromboplastin time method (APTT), the calcium thrombin time method (CaTT) and two amidolytic methods (measuring the accelerating effect of heparin on the inactivation of thrombin or factor Xa by antithrombin III), with and without plasma in the test system. The specific activities of the various heparins were expressed relative to that of the 3rd. Int. Standard (=100). Found specific activities ranged 3 - 198 (LAH and HAH, respectively). In all assay systems HAH had the highest specific activity, followed by one of the commercial preparations and the 3rd Int. Standard. LAH and human heparin had very low specific activities, except in the APTT test system, an assay method which in addition mirrors other anticoagulant effects of heparin than the acceleration of antithrombin III. Apart from the higher effect of LAH and human heparin on the APTT, the difference in specific activities found for each individual heparin preparation with these various assay methods was slight.In view of the reproducibility and simplicity of the amidolytic methods, it is suggested that they be adapted for heparin standardization.

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