Present study deals with the preliminary study of Glycine Max Ethanol Extract (GMEE). GMEE stacks more macro and micro nutrients with many pharmacological and nutraceutical standards. GMEE was preliminary screened by simple test methods and instrumentation methods such as RP-HPLC, IR and GC-MS. The obtained results from IR predicted the presence of different functional groups such as OH, CH2, C=O, C-O and cyclic ring. While, the RP-HPLC and GC-MS profiles of GMEE predicted the presence of lipids, polyphenols, alkaloids and flavonoids in the extract.
Comparative Assessment of Total Polyphenols and Antioxidant Activity of Comme...AnuragSingh1049
Green Tea, made from Camellia sinensis plant leaves, is one of the most popular drinks in the world. For the past decades, scientists have studied this plant in terms of potential health benefits. Research has shown that green tea helps prevent stroke, malignancy and infections. In this paper, antioxidant activity and total phenol content of 4 samples of green tea from local Tuzla stores were investigated, of which two were of foreign origin. The antioxidant activity of the samples was analyzed using FRAP and DPPH methods. The obtained results show that the highest content of total phenols and the largest antioxidant capacity has a sample of foreign origin. The content of total phenols in the samples ranges from 60.01 to 79.34 mg GAE/g. The highest FRAP value is 3.34 mmol/g. The antioxidant capacity was also confirmed by the DPPH method. The IC50 value ranges from 0.014 to 0.030 mg/mL.
VALIDATED LIQUID CHROMATOGRAPHY/TANDEM MASS SPECTROMETRY METHOD FOR DETERMINA...Manik Ghosh
A simple, highly sensitive and rapid LC-MS/MS method has been developed and validated for the quantification of metolazone in rat plasma using irbesartan as internal standard (IS). After simple protein precipitation extraction by acetonitrile, the analyte and IS were extracted from 50 μL plasma sample on an Agilent Poroshell 120, EC- C18 (50 mm × 4.6 mm, i.d., 2.7 μm) column using 5μL injection volume with a total run time of 2 min. Acidified methanol/water mixture was used as a mobile phase. The parent/product ion transitions for metolazone (m/z 366.1/258.9) and IS (m/z 429.2/207.0) were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring and positive ion mode. The method was found to be linear in the range of 0.05 – 200 metolazone. The method was validated with respect to selectivity, linearity, accuracy, precision, recovery and stability according to accepted regulatory guidelines. The described method was successfully applied to preclinical pharmacokinetic studies of analytes after an oral administration of metolazone (1 mg/kg) in rats.
Characterization of intact antibodies by pre-fractionation using gel electrop...Expedeon
Antibodies represent an important class of proteins due to their central role in the immune response. Moreover, there is an increasing interest in the use of recombinant antibodies as novel drug therapies.
Qualitative tests of proteins, color reaction of proteins,biuret's test, colo...ShwetaMishra115
Qualitative tests of proteins
color reaction of proteins
biuret's test, color reaction of proteins, millon's test, ninhydrin test, qualitative tests of proteins, sakaguchi test, sodium nitroprusside test, xanthoproteic test
Present study deals with the preliminary study of Glycine Max Ethanol Extract (GMEE). GMEE stacks more macro and micro nutrients with many pharmacological and nutraceutical standards. GMEE was preliminary screened by simple test methods and instrumentation methods such as RP-HPLC, IR and GC-MS. The obtained results from IR predicted the presence of different functional groups such as OH, CH2, C=O, C-O and cyclic ring. While, the RP-HPLC and GC-MS profiles of GMEE predicted the presence of lipids, polyphenols, alkaloids and flavonoids in the extract.
Comparative Assessment of Total Polyphenols and Antioxidant Activity of Comme...AnuragSingh1049
Green Tea, made from Camellia sinensis plant leaves, is one of the most popular drinks in the world. For the past decades, scientists have studied this plant in terms of potential health benefits. Research has shown that green tea helps prevent stroke, malignancy and infections. In this paper, antioxidant activity and total phenol content of 4 samples of green tea from local Tuzla stores were investigated, of which two were of foreign origin. The antioxidant activity of the samples was analyzed using FRAP and DPPH methods. The obtained results show that the highest content of total phenols and the largest antioxidant capacity has a sample of foreign origin. The content of total phenols in the samples ranges from 60.01 to 79.34 mg GAE/g. The highest FRAP value is 3.34 mmol/g. The antioxidant capacity was also confirmed by the DPPH method. The IC50 value ranges from 0.014 to 0.030 mg/mL.
VALIDATED LIQUID CHROMATOGRAPHY/TANDEM MASS SPECTROMETRY METHOD FOR DETERMINA...Manik Ghosh
A simple, highly sensitive and rapid LC-MS/MS method has been developed and validated for the quantification of metolazone in rat plasma using irbesartan as internal standard (IS). After simple protein precipitation extraction by acetonitrile, the analyte and IS were extracted from 50 μL plasma sample on an Agilent Poroshell 120, EC- C18 (50 mm × 4.6 mm, i.d., 2.7 μm) column using 5μL injection volume with a total run time of 2 min. Acidified methanol/water mixture was used as a mobile phase. The parent/product ion transitions for metolazone (m/z 366.1/258.9) and IS (m/z 429.2/207.0) were monitored on a triple quadrupole mass spectrometer, operating in the multiple reaction monitoring and positive ion mode. The method was found to be linear in the range of 0.05 – 200 metolazone. The method was validated with respect to selectivity, linearity, accuracy, precision, recovery and stability according to accepted regulatory guidelines. The described method was successfully applied to preclinical pharmacokinetic studies of analytes after an oral administration of metolazone (1 mg/kg) in rats.
Characterization of intact antibodies by pre-fractionation using gel electrop...Expedeon
Antibodies represent an important class of proteins due to their central role in the immune response. Moreover, there is an increasing interest in the use of recombinant antibodies as novel drug therapies.
Qualitative tests of proteins, color reaction of proteins,biuret's test, colo...ShwetaMishra115
Qualitative tests of proteins
color reaction of proteins
biuret's test, color reaction of proteins, millon's test, ninhydrin test, qualitative tests of proteins, sakaguchi test, sodium nitroprusside test, xanthoproteic test
Study on Fructooligosaccharide (Fos) Production By Enzyme Pectinex Ultra Sp-l...IJAEMSJORNAL
Fructooligosaccharide (FOS) is a new alternative sweetener with its characteristics such as low energy and safety for people with diabetes. Pectinex Ultra SP-L which has fructosyltransferase, catalyzes the reaction to produce short chain fructooligosaccharides. The research was conducted to enhance the high FOS content in process of FOS production by immobilized enzyme. Results achieved by Empirical planning Design Expert 7.0 - central composite method (CCD) identified at optimum conditions for process of immobilized enzyme with alginate 3.3 (%), ratio of enzyme: alginate was 0.79 (w/w), CaCl2 3.75 (%). Efficiency of loading protein reached 73.32 (%). Reaction conditions: 60 (oC), shaking velocity 90 (rpm), the initial sucrose concentrations of 50 (%), pH 5.75. When produced in 20 (h), the reaction obtained the highest level of FOS with column reaction system. The FOS for producted by immobilized enzyme achieved 47.87 (%), of which 1-kestose obtained 37.06 (%), the remaining was 10.81 (%) including nystose and fructofuranosylnystose. Efficiency of producing FOS by using immobilized enzyme compared to the free enzyme was high up to 89.49 (%).
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
Evaluation of antioxidant properties of pomegranate peel extract in compariso...Pritam Kolge
Evaluation of antioxidant properties of pomegranate peel extract in comparison with pomegranate pulp extract.....
This is Journal Club activity Presentation with the reference of various research papers.
This Presentation Contain following...
#Info about Paper
#Highlights
#Introduction
#Materials
#Methods
#Results
#Discussion
#Conclusion
#References
Important Methods used
1)Antioxidants extraction
2)Determination of total phenolics content
3)FRAP Assay
4)Determination of content of phenolic compounds in the
extract
5)Superoxide radical scavenging activity (DPPH)
6)Hydroxyl radical (OH) prevention activity
Journal Club Presentation at Bharati Vidyapeeth College of Pharmacy, Kolhapur.
Thanks for Help and Guidance of Dr. P. B. Choudhari (Assistant Professor, Pharmaceutical Chemistry) and Mr. D. P. Mali Assistant Professor, Pharmaceutical Chemistry
Analysis of Sugars in Honey Using the PerkinElmer Altus HPLC System with RI D...PerkinElmer, Inc.
Honey consumption has grown significantly during the last few decades due to its high nutritional value and unique flavor. The price of natural bee honey is much higher than other sweeteners making it susceptible to adulteration with cheaper sweeteners, primarily sucrose. Besides lower levels of nonsugar ingredients, natural honey primarily consists of glucose and fructose and may contain low levels of sucrose and/or maltose. However, according to the international regulations, any commercially available “pure”-labeled honey products that are found to have in excess of 5% by weight of sucrose or maltose are considered to be adulterated. With the focus on possible honey adulteration, this application highlights the LC separation of various sugars found in honey and the analysis of these components in four store-bought honey samples. Method conditions and performance data, including linearity and repeatability, are presented.
Validation of Factor IIa for heparin sodium or heparin injectionkrishgen
A chromogenic assay intended for the quantitative determination in purified solutions by measurement of factor IIa or Xa inhibition activity. The kit can be used for 100 test reactions as per microtiter plate protocol.
Validation of factor xa assay for heparin sodium heparin injectionkrishgen
chromogenic assay intended for the quantitative determination in purified solutions by measurement of factor IIa or Xa inhibition activity. The kit can be used for 100 test reactions as per microtiter plate protocol.
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
Study on Fructooligosaccharide (Fos) Production By Enzyme Pectinex Ultra Sp-l...IJAEMSJORNAL
Fructooligosaccharide (FOS) is a new alternative sweetener with its characteristics such as low energy and safety for people with diabetes. Pectinex Ultra SP-L which has fructosyltransferase, catalyzes the reaction to produce short chain fructooligosaccharides. The research was conducted to enhance the high FOS content in process of FOS production by immobilized enzyme. Results achieved by Empirical planning Design Expert 7.0 - central composite method (CCD) identified at optimum conditions for process of immobilized enzyme with alginate 3.3 (%), ratio of enzyme: alginate was 0.79 (w/w), CaCl2 3.75 (%). Efficiency of loading protein reached 73.32 (%). Reaction conditions: 60 (oC), shaking velocity 90 (rpm), the initial sucrose concentrations of 50 (%), pH 5.75. When produced in 20 (h), the reaction obtained the highest level of FOS with column reaction system. The FOS for producted by immobilized enzyme achieved 47.87 (%), of which 1-kestose obtained 37.06 (%), the remaining was 10.81 (%) including nystose and fructofuranosylnystose. Efficiency of producing FOS by using immobilized enzyme compared to the free enzyme was high up to 89.49 (%).
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
Evaluation of antioxidant properties of pomegranate peel extract in compariso...Pritam Kolge
Evaluation of antioxidant properties of pomegranate peel extract in comparison with pomegranate pulp extract.....
This is Journal Club activity Presentation with the reference of various research papers.
This Presentation Contain following...
#Info about Paper
#Highlights
#Introduction
#Materials
#Methods
#Results
#Discussion
#Conclusion
#References
Important Methods used
1)Antioxidants extraction
2)Determination of total phenolics content
3)FRAP Assay
4)Determination of content of phenolic compounds in the
extract
5)Superoxide radical scavenging activity (DPPH)
6)Hydroxyl radical (OH) prevention activity
Journal Club Presentation at Bharati Vidyapeeth College of Pharmacy, Kolhapur.
Thanks for Help and Guidance of Dr. P. B. Choudhari (Assistant Professor, Pharmaceutical Chemistry) and Mr. D. P. Mali Assistant Professor, Pharmaceutical Chemistry
Analysis of Sugars in Honey Using the PerkinElmer Altus HPLC System with RI D...PerkinElmer, Inc.
Honey consumption has grown significantly during the last few decades due to its high nutritional value and unique flavor. The price of natural bee honey is much higher than other sweeteners making it susceptible to adulteration with cheaper sweeteners, primarily sucrose. Besides lower levels of nonsugar ingredients, natural honey primarily consists of glucose and fructose and may contain low levels of sucrose and/or maltose. However, according to the international regulations, any commercially available “pure”-labeled honey products that are found to have in excess of 5% by weight of sucrose or maltose are considered to be adulterated. With the focus on possible honey adulteration, this application highlights the LC separation of various sugars found in honey and the analysis of these components in four store-bought honey samples. Method conditions and performance data, including linearity and repeatability, are presented.
Validation of Factor IIa for heparin sodium or heparin injectionkrishgen
A chromogenic assay intended for the quantitative determination in purified solutions by measurement of factor IIa or Xa inhibition activity. The kit can be used for 100 test reactions as per microtiter plate protocol.
Validation of factor xa assay for heparin sodium heparin injectionkrishgen
chromogenic assay intended for the quantitative determination in purified solutions by measurement of factor IIa or Xa inhibition activity. The kit can be used for 100 test reactions as per microtiter plate protocol.
The IOSR Journal of Pharmacy (IOSRPHR) is an open access online & offline peer reviewed international journal, which publishes innovative research papers, reviews, mini-reviews, short communications and notes dealing with Pharmaceutical Sciences( Pharmaceutical Technology, Pharmaceutics, Biopharmaceutics, Pharmacokinetics, Pharmaceutical/Medicinal Chemistry, Computational Chemistry and Molecular Drug Design, Pharmacognosy & Phytochemistry, Pharmacology, Pharmaceutical Analysis, Pharmacy Practice, Clinical and Hospital Pharmacy, Cell Biology, Genomics and Proteomics, Pharmacogenomics, Bioinformatics and Biotechnology of Pharmaceutical Interest........more details on Aim & Scope).
All manuscripts are subject to rapid peer review. Those of high quality (not previously published and not under consideration for publication in another journal) will be published without delay.
Determination of Total Protein Using the LAMBDA UV/Vis SpectrophotometerPerkinElmer, Inc.
The Lowry and Biuret methods are standard methods for protein quantification. Though the latter is more sensitive and is used for investigative work, it is limited by (1) poor stability of the combined reagent, (2) non-reproducibility of color, especially at low protein concentrations, and (3) a non-linear chromogenic response with protein concentration. Ohnishi and Barr1 modified and simplified the Biuret combined reagent for the Lowry procedure and at the same time improved its stability. This application note describes the modified Lowry procedure for protein analysis.
Protein quantification is divided into "total quantification method" of whole protein and "individual quantification method" of specific protein according to its purpose. It is an indispensable part of biological experiments.
The presentation is about the chemical residues that cloud be seen in the milk. It includes the chemical residues like the antibiotic residues, pesticides, detergents and heavy metals.
the presentation contain ways used to estimate proteins, this presentation prepared by TONNYBITE, a student from KILIMANJARO CHRISTIAN MEDICAL UNIVERSITY COLLEGE, TANZANIA
The Art of the Pitch: WordPress Relationships and SalesLaura Byrne
Clients don’t know what they don’t know. What web solutions are right for them? How does WordPress come into the picture? How do you make sure you understand scope and timeline? What do you do if sometime changes?
All these questions and more will be explored as we talk about matching clients’ needs with what your agency offers without pulling teeth or pulling your hair out. Practical tips, and strategies for successful relationship building that leads to closing the deal.
LF Energy Webinar: Electrical Grid Modelling and Simulation Through PowSyBl -...DanBrown980551
Do you want to learn how to model and simulate an electrical network from scratch in under an hour?
Then welcome to this PowSyBl workshop, hosted by Rte, the French Transmission System Operator (TSO)!
During the webinar, you will discover the PowSyBl ecosystem as well as handle and study an electrical network through an interactive Python notebook.
PowSyBl is an open source project hosted by LF Energy, which offers a comprehensive set of features for electrical grid modelling and simulation. Among other advanced features, PowSyBl provides:
- A fully editable and extendable library for grid component modelling;
- Visualization tools to display your network;
- Grid simulation tools, such as power flows, security analyses (with or without remedial actions) and sensitivity analyses;
The framework is mostly written in Java, with a Python binding so that Python developers can access PowSyBl functionalities as well.
What you will learn during the webinar:
- For beginners: discover PowSyBl's functionalities through a quick general presentation and the notebook, without needing any expert coding skills;
- For advanced developers: master the skills to efficiently apply PowSyBl functionalities to your real-world scenarios.
Generating a custom Ruby SDK for your web service or Rails API using Smithyg2nightmarescribd
Have you ever wanted a Ruby client API to communicate with your web service? Smithy is a protocol-agnostic language for defining services and SDKs. Smithy Ruby is an implementation of Smithy that generates a Ruby SDK using a Smithy model. In this talk, we will explore Smithy and Smithy Ruby to learn how to generate custom feature-rich SDKs that can communicate with any web service, such as a Rails JSON API.
Neuro-symbolic is not enough, we need neuro-*semantic*Frank van Harmelen
Neuro-symbolic (NeSy) AI is on the rise. However, simply machine learning on just any symbolic structure is not sufficient to really harvest the gains of NeSy. These will only be gained when the symbolic structures have an actual semantics. I give an operational definition of semantics as “predictable inference”.
All of this illustrated with link prediction over knowledge graphs, but the argument is general.
Software Delivery At the Speed of AI: Inflectra Invests In AI-Powered QualityInflectra
In this insightful webinar, Inflectra explores how artificial intelligence (AI) is transforming software development and testing. Discover how AI-powered tools are revolutionizing every stage of the software development lifecycle (SDLC), from design and prototyping to testing, deployment, and monitoring.
Learn about:
• The Future of Testing: How AI is shifting testing towards verification, analysis, and higher-level skills, while reducing repetitive tasks.
• Test Automation: How AI-powered test case generation, optimization, and self-healing tests are making testing more efficient and effective.
• Visual Testing: Explore the emerging capabilities of AI in visual testing and how it's set to revolutionize UI verification.
• Inflectra's AI Solutions: See demonstrations of Inflectra's cutting-edge AI tools like the ChatGPT plugin and Azure Open AI platform, designed to streamline your testing process.
Whether you're a developer, tester, or QA professional, this webinar will give you valuable insights into how AI is shaping the future of software delivery.
Essentials of Automations: Optimizing FME Workflows with ParametersSafe Software
Are you looking to streamline your workflows and boost your projects’ efficiency? Do you find yourself searching for ways to add flexibility and control over your FME workflows? If so, you’re in the right place.
Join us for an insightful dive into the world of FME parameters, a critical element in optimizing workflow efficiency. This webinar marks the beginning of our three-part “Essentials of Automation” series. This first webinar is designed to equip you with the knowledge and skills to utilize parameters effectively: enhancing the flexibility, maintainability, and user control of your FME projects.
Here’s what you’ll gain:
- Essentials of FME Parameters: Understand the pivotal role of parameters, including Reader/Writer, Transformer, User, and FME Flow categories. Discover how they are the key to unlocking automation and optimization within your workflows.
- Practical Applications in FME Form: Delve into key user parameter types including choice, connections, and file URLs. Allow users to control how a workflow runs, making your workflows more reusable. Learn to import values and deliver the best user experience for your workflows while enhancing accuracy.
- Optimization Strategies in FME Flow: Explore the creation and strategic deployment of parameters in FME Flow, including the use of deployment and geometry parameters, to maximize workflow efficiency.
- Pro Tips for Success: Gain insights on parameterizing connections and leveraging new features like Conditional Visibility for clarity and simplicity.
We’ll wrap up with a glimpse into future webinars, followed by a Q&A session to address your specific questions surrounding this topic.
Don’t miss this opportunity to elevate your FME expertise and drive your projects to new heights of efficiency.
JMeter webinar - integration with InfluxDB and GrafanaRTTS
Watch this recorded webinar about real-time monitoring of application performance. See how to integrate Apache JMeter, the open-source leader in performance testing, with InfluxDB, the open-source time-series database, and Grafana, the open-source analytics and visualization application.
In this webinar, we will review the benefits of leveraging InfluxDB and Grafana when executing load tests and demonstrate how these tools are used to visualize performance metrics.
Length: 30 minutes
Session Overview
-------------------------------------------
During this webinar, we will cover the following topics while demonstrating the integrations of JMeter, InfluxDB and Grafana:
- What out-of-the-box solutions are available for real-time monitoring JMeter tests?
- What are the benefits of integrating InfluxDB and Grafana into the load testing stack?
- Which features are provided by Grafana?
- Demonstration of InfluxDB and Grafana using a practice web application
To view the webinar recording, go to:
https://www.rttsweb.com/jmeter-integration-webinar
De-mystifying Zero to One: Design Informed Techniques for Greenfield Innovati...
Lowry vs biuret final (1)
1. UNIVERSITI PENDIDIKAN SULTAN IDRIS
35900 TANJONG MALIM, PERAK DARUL RIDZUAN
FAKULTI :
SAINS DAN TEKNOLOGI
PRINCIPLE IN BIOCHEMISTRY (SBK3013)
EXPERIMENT 2:
PROTEIN ANALYSIS
PREPARED BY :
UMI ABIBAH BT SULAIMAN D20091034811
SITI RAHAYU BT MOHAMED NOOR D20091034855
AZMA AMIRA MOHAMAD D20091034859
NUR AFIQAH BT MUHAMAD APANDI D20091034872
AMEERA BT YAHYA D20091034814
SEMESTER 8
SESI 2009/2010
LAB SESSION
THURSDAY(10.30 A.M. - 1.30 P.M.)
LAB INSTRUCTOR
DR ROSMILA MISNAN
2. PROTEIN EXPERIMENT
Experiment 2: The determination of three protein samples using Biuret and Lowry Assay.
Objective:
1. To learn the principles of protein assays.
2. To determine protein concentrations of protein samples using the Biuret Protein Assay
and Lowry assay.
Introduction:
The determination of protein concentration is an essential technique in all aspects of
protein studies.This lab activity is designed to teach students the principles behind a common
protein estimation assay known as the BiuretProtein Assay and Lowry Protein
Assay.Although there are a wide variety of protein assays available none of the assays can be
used without first consideringtheir suitability for the application. Each method has its own
advantages and limitations and often it is necessary to obtainmore than one type of protein
assay for research applications.
In the copper ion based protein assays, protein solutions are mixed with an alkaline
solution of copper salt, cupricions (Cu2+). The protein assay is based on the interaction of
cupric ions with protein in an alkaline solution and is commonlyreferred to as the Biuret
assay. The interaction of cupric ions (Cu2+) with protein results in a purple color that can be
read at 540nm. The amount of color produced is proportional to protein concentration.
The Lowry protein assay method for protein concentration determination is one of the
most venerable and widely-used protein assays. Hydrolysis is probably the most accurate
method of determining protein concentration followed by amino acid analysis. Most other
methods are sensitive to the amino acid composition of the protein, and absolute
concentrations cannot be obtained. The Lowry procedure is sensitive, and is moderately
constant from protein to protein. The Lowry protein estimation has been so widely used that
it is a completely acceptable alternative to a rigorous absolute determination in almost all
circumstances in which protein mixtures or crude extracts are involved.
3. Result:
Biuret Assay
Table 1.1: Biuret assay
Concentration (mg/ml) Absorbance (nm)
Blank 0.00
1 0.138
2 0.139
3 0.190
4 0.192
5 0.194
6 0.211
Table 1.2: Determination using Biuret assay
Samples Absorbance
before
dilution (nm)
Dilution Absorbance
after dilution
(nm)
Concentration
(mg/ml)
Chicken egg 0.936 1mL of sample + 9 mL
of distilled water
0.106 2.0 x 10=20
Quail egg 0.870 1mL of sample + 9 mL
of distilled water
0.090 1.6 x 10= 16
Duck egg 0.764 1mL of sample + 9 mL
of distilled water
0.066 1.0 x 10= 10
Chicken egg
(ayam
kampung)
0.230
1mL of sample + 9 mL
of distilled water
0.195 1.8 x 10= 18
Omega egg 0.256 1mL of sample + 9 mL
of distilled water
0.197 1.9 x 10= 19
4. Graph of absorbance value (nm) versus gelatin concentration (mg/ml) for Biuret assay
0
0.05
0.1
0.15
0.2
0.25
0 1 2 3 4 5 6 7
absorbancevalue(nm)
Gelatin concentration (mg/ml)
Y-Values
5. Lowry Assay
Table 2.1: Lowry assay
Concentration Absorbance (nm)
Blank 0.00
0.1mg/mL 0.053
0.2 mg/mL 0.133
0.3mg/mL 0.143
0.4 mg/mL 0.207
0.5mg/mL 0.185
0.6 mg/mL 0.166
Table 2.2: Determination using Lowry assay
Samples Dilution Absorbance
after dilution
(nm)
Concentration
(mg/ml)
Chicken egg 1mL of sample + 9 mL
of distilled water
0.072 0.16 x 10=1.6
Quail egg 1mL of sample + 9 mL
of distilled water
0.134 0.3x 10= 3.0
Duck egg 1mL of sample + 9 mL
of distilled water
0.185 0.56 x 10= 5.6
Chicken egg
(ayam
kampung)
1mL of sample + 9 mL
of distilled water
0.173 0.41 x 10= 4.1
Omega egg 1mL of sample + 9 mL
of distilled water
0.131 0.25 x 10= 2.5
6. Graph of absorbance value (nm) versus gelatin concentration (mg/ml) for Lowry assay
0
0.05
0.1
0.15
0.2
0.25
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7
Absorbancevalue(nm)
Gelatin concerntration (mg/ml)
Y-Values
7. RESULT SUMMARY
Table 3.1 Protein concentration obtained from Biuret assay and Lowry assay
Type of sample Concentration (mg/ml)
Biuret Assay Lowry Assay
Chicken egg 20 1.6
Quail egg 16 3.0
Duck egg 10 5.6
Chicken egg
(ayam kampung)
18 4.1
Omega egg 19 2.5
Discussion:
In this experiment, we had used Biuret assay and Lowry assay to determine protein
concentration in five protein food samples which are chicken egg, quail egg, duck egg,
chicken egg (ayam kampung) and omega egg.
Biuret test indicate the presence of peptide bond between the amino group and the
carboxyl acid group on adjacent amino acids in a protein. The violet colour is a positive test
for the presence of protein. The greater the violet colour intense, the greater the number of
peptide bonds that reacts.
Biuret test also has its limitations and not very sensitive which are the Biuret only test
whether peptide bonds in protein are present in a sample, it will not determine how much
there is unless we compare our sample of unknown concentration with a standard of known
protein concentration.
However biuret test will not detect free amino acids and the characteristic purple
colour will not show up if we only have small peptides, since the number of peptide bonds
may not be sufficient to yield colour. The Biuret assay is not much good for protein
concentrations below about 5 mg/ml. By using the Folin‐Ciocalteu reagent to detect reduced
copper makes the Lowry assay nearly 100 times more sensitive than the Biuret reaction alone.
Based on the result obtained we had plotted the standard curve by using Biuret assays,
and we compare with the five type of albumin sample. We found that the absorbance value of
8. these five types of albumin sample is higher than the standard curve. Then the five must do
the dilution to lower the reading from the standard curve. For dilution, the ratio used is 1 ml
of protein samples to 9.0 ml of distilled water. After dilution, we get the results which are the
protein concentration in chicken egg is the highest which is 20 mg/mL, the second higher of
protein concentration by using Biuret assays is omega egg which is 19 mg/mL and the third
higher protein concentration using Biuret assays is chicken egg(ayam kampung) which is
18mg/mL. Quail egg show the protein concentration 16mg/mL and the lowest protein
concentration by using Biuret assays is duck egg which is 10 mg/mL.
Next we had carried out Lowry assay to determine the protein concentration in three
protein samples. The Lowry method is more sensitive since it combines the reactions of
copper ions with the peptide bonds under alkaline conditions with the oxidation of
aromatic protein residues. The Lowry method is best used with protein concentrations of 0.01-
1.0 mg/mL and is based on the reaction of Cu+
, produced by the oxidation of peptide bonds,
with Folin-Ciocalteu reagent.
The following substances are known to interfere with the Lowry assay are detergents,
carbohydrates, glycerol, potassium compounds, sulfhydryl compounds, most phenols, uric
acid, guanine, and calcium. Many of these interfering substances are commonly used in
buffers for preparing proteins or in cell extracts.
From the result of Lawry assays we also had been plotted standard curve graph, and
we compare with the five type of albumin sample. For dilution, the ratio used is 1 ml of
protein samples to 9.0 ml of distilled water. After dilution, we get the results which are the
protein concentration in duck egg is the highest among the other four which is 5.6 mg/mL, the
second higher of protein concentration by using Lowry assays is chicken egg (ayam
kampung) egg which is 4.1 mg/mL and the third highest protein concentration using Lowry
assays is quail egg which is 3.0 mg/mL. Omega egg shows the protein concentration
2.5mg/mL and the lowest protein concentration by using Lowry assays is chicken egg which
is 1.6 mg/mL.
When compare the result of Lowry assay with biuret essay, there is some differences.
This is might be because of some errors that occur while we conduct our experiment. For
instance we just estimate the volume of the sample when we poured into the vial. It should an
9. actual volume that is same for all protein. Second, there might be error when we dilute the
protein, caused by parallax error when reading the measuring cylinder.
As the conclusion, Lowry assays is best method in determining the protein
concentration because it is best used with protein concentrations of 0.01-1.0 mg/mL which is
suitable when test with 0.25 mL of protein sample of albumin of chicken, duck, quail, ayam
kampong and omega which is in the range of 0.01-1.0 mg/mL. Meanwhile, Biuret assay is not
very efficient in determining the protein concentration as it is not much good for protein
concentrations below about 5 mg/ml. Thus, albumin of duck has the highest protein
concentration, albumin of ayam kampong is the second highest in protein concentration,
albumin of quail has the third highest in protein concentration, omega chicken has the fourth
highest in protein concentration and the last one is albumin of chicken.
Instead of these two method, there are several method can be used in determining
protein concentration. First is The Bradford assay, a colorimetric protein assay, is based on
an absorbance shift of the dye Coomassie Brilliant Blue G-250 in which under acidic conditions the
red form of the dye is converted into its bluer form to bind to the protein being assayed. It also has
disadvantages which is the Bradford assay is linear over a short range, typically from 0 µg/ml to
2000 µg/ml, often making dilutions of a sample necessary before analysis. It is also inhibited by the
presence of detergents.
10. Conclusion
1. Protein concentration can be determined using Biuret protein assay and Lowry protein
assay.
2. The protein concentration in chicken egg is 20mg/mL , omega egg is 19 mg/mL,
chicken egg(ayam kampong) is 18mg/mL, quail egg is 16mg/mL and duck egg is
10mg/mL based on Biuret method.
3. The protein concentration in duck egg is 5.6mg/mL , chicken egg(ayam kampong) is
4.1 mg/mL, quail egg is 3.0mg/mL, omega egg is 2.5mg/mL and chicken egg is
1.6mg/mL based on Lowry method.
4. Protein concentration can be found out from the standard curve.
References
1. www.gbiosciences.com/.../633453707995878750.pdf
2. wolfson.huji.ac.il/purification/PDF/Protein.../PIERCE_BIURET.pdf
3. biochemistry.musc.edu/.../Lowry%20Protein%20Assay... - United States
4. www.molecularstation.com/protein/lowry-protein-assay/