This document describes a study that developed a method to determine cabergoline and L-dopa levels in human plasma using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The researchers were able to directly analyze deproteinized plasma samples for both cabergoline and L-dopa. Calibration curves for each drug showed good linearity over specific concentration ranges. The method was then applied to analyze plasma samples from patients taking these drugs, with coefficients of variation ranging from 2.9-10.5% indicating good precision. The LC-MS/MS method proved suitable for routine therapeutic drug monitoring of cabergoline and L-dopa.
Abstract
A small set of amphetamines has been analyzed by gas chromatography (GC) high-resolution time-of-flight mass spectrometry (TOFMS) using a microplasma photoionization (MPPI) soft-ionization source. This plasma-based, wavelength selectable ionization source enables ionization of the test compounds and their corresponding derivatives at ~8-12 eV that is a softer alternative to electron ionization at 70 eV. Three plasma gases were used in this study: Xe plasma that emits photons at resonance lines of 9.57 eV and 8.44 eV; Kr plasma at 10.63 eV and 10.02 eV, and Ar plasma at 11.82 eV and 11.61 eV. Derivatization of the test compounds with trifluoroacetic anhydride and α-methoxy-α-(trifluoromethyl)-phenylacetyl pyrazole was evaluated because the MPPI mass spectra of the underivatized amphetamines yield primarily iminium ions, which make the identification of the test compounds by GC-TOFMS inconclusive. The MPPI mass spectra of the TFA-derivatized amphetamines yield abundant molecular ions, when using Xe as plasma gas, and enough fragment ions with the Ar plasma that can help in formula generation. The structure elucidation of two "known unknowns" designer drugs using this "tunable" soft-ionization source and a high-resolution TOF mass spectrometer is presented in this study.
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.
Quantification of a Novel Peptide, CPT31 in Rat and Monkey Plasma by LC-MSCovance
ASMS 2019 -- CPT31, a novel D-peptide, is being investigated in the treatment and prevention of HIV by inhibiting the viral entry of HIV. To evaluate the properties of CPT31, an accurate highly reproducible method to quantitate CPT31 in plasma was required. To this end, a robust LC-MS assay for the quantification of CPT31 in rat and monkey plasma samples is reported here. The method follows extraction and clean-up of two plasma matrices, encompasses a range of 90.0 to 45,000 ng/mL, and completes LC-MS analysis in 7.50 minutes.
Abstract
A small set of amphetamines has been analyzed by gas chromatography (GC) high-resolution time-of-flight mass spectrometry (TOFMS) using a microplasma photoionization (MPPI) soft-ionization source. This plasma-based, wavelength selectable ionization source enables ionization of the test compounds and their corresponding derivatives at ~8-12 eV that is a softer alternative to electron ionization at 70 eV. Three plasma gases were used in this study: Xe plasma that emits photons at resonance lines of 9.57 eV and 8.44 eV; Kr plasma at 10.63 eV and 10.02 eV, and Ar plasma at 11.82 eV and 11.61 eV. Derivatization of the test compounds with trifluoroacetic anhydride and α-methoxy-α-(trifluoromethyl)-phenylacetyl pyrazole was evaluated because the MPPI mass spectra of the underivatized amphetamines yield primarily iminium ions, which make the identification of the test compounds by GC-TOFMS inconclusive. The MPPI mass spectra of the TFA-derivatized amphetamines yield abundant molecular ions, when using Xe as plasma gas, and enough fragment ions with the Ar plasma that can help in formula generation. The structure elucidation of two "known unknowns" designer drugs using this "tunable" soft-ionization source and a high-resolution TOF mass spectrometer is presented in this study.
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.
Quantification of a Novel Peptide, CPT31 in Rat and Monkey Plasma by LC-MSCovance
ASMS 2019 -- CPT31, a novel D-peptide, is being investigated in the treatment and prevention of HIV by inhibiting the viral entry of HIV. To evaluate the properties of CPT31, an accurate highly reproducible method to quantitate CPT31 in plasma was required. To this end, a robust LC-MS assay for the quantification of CPT31 in rat and monkey plasma samples is reported here. The method follows extraction and clean-up of two plasma matrices, encompasses a range of 90.0 to 45,000 ng/mL, and completes LC-MS analysis in 7.50 minutes.
Poster demonstrating the results from the development/verification project for the quantitation of pyridoxal 5-phosphate and 4-pyridoxic acid in human plasma.
Quantitative Analysis of 30 Drugs in Whole Blood by SPE and UHPLC-TOF-MSAnnex Publishers
Abstract
An Ultra-High Pressure Liquid Chromatography Time-of-Flight Mass Spectrometry (UHPLC-TOF-MS) method for quantitative analysis of 30 drugs in whole blood was developed and validated. The method was used for screening and quantification of common drugs and drugs of abuse in whole blood received from autopsy cases and living persons. The compounds included: alprazolam, amphetamine, benzoylecgonine, bromazepam, cathine, cathinone, chlordiazepoxide, cocaine, codeine, clonazepam, 7-aminoclonazepam, diazepam, nordiazepam, flunitrazepam, 7-aminoflunitrazepam, ketamine, ketobemidone, 3,4-Methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), methamphetamine, methadone, morphine, 6-monoacetylmorphine, nitrazepam, 7-aminonitrazepam, oxazepam, temazepam, tramadol, O-desmethyltramadol, and zolpidem. Blood samples (200 μL) were subjected to Solid Phase Extraction (SPE). Target drugs were quantified using a Waters ACQUITY UPLC system coupled to a Waters SYNAPT G2 TOF-MS apparatus. Extraction recoveries ranged from 41% (7-aminoclonazepam) to 111% (ketamine) and matrix effects ranged from -13% (temazepam) to 50% (7-aminonitrazepam). For all compounds, a quadratic polynomial was applied for fitting the calibration curves. Lower Limits of Quantification (LOQ) ranged from 0.005 to 0.05 mg/kg. Satisfactory precisions below 15% and accuracies within 85-115% were obtained for all compounds at concentrations exceeding the LOQ. In conclusion, we present a validated UHPLC-TOF-MS method for simultaneous quantification of 30 drugs in whole blood with a run time of 15 min using 200 μL of whole blood.
Keywords: Drugs of abuse, UHPLC-TOF-MS, Whole blood, SPE, Quantification
Drug discovery library design by biomimetic hplcKlara Valko
The slides explain the early drug discovery process and how the measurements of biomimetic properties can help to design the best ADME properties of compound libraries.
Poster demonstrating the results from the development/verification project for the quantitation of all- trans retinol and alpha tocopherol in human serum.
Spectroscopic and Thermal Characterization of Charge-Transfer Complexes Forme...IJMER
The spectrophotometric characteristics of the solid charge-transfer molecular complexes
(CT) formed in the reaction of the electron donor 2-amino-6-ethylpyridine (2A6EPy) with the π-
acceptors tetracyanoethylene (TCNE), 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and 2,4,4,6-
tetrabromo-2,5-cyclohexadienone (TBCHD) have been studied in chloroform at 25 0C. These were
investigated through electronic, infrared, mass spectra and thermal studies as well as elemental
analysis. The results show that the formed solid CT- complexes have the formulas [(2A6EPy)(TCNE)2],
[(2A6EPy)2(DDQ)], [(2A6EPy)4(TBCHD)] for 2-amino-6-ethylpyridine in full agreement with the
known reaction stoichiometries in solution as well as the elemental measurements. The formation
constant kCT, molar extinction coefficient εCT, free energy change ∆G
0
and CT energy ECT have been
calculated for the CT- complexes [(2A6EPy)(TCNE)2], [(2A6EPy)2(DDQ)].
This is nice presentation given by Vishal Goyani, an Analytical student of National Institute Of Pharmaceutical education and research-INDIA.
mail your query at - vngoyanii@gmail.com
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).
Nanobiosensors can be built via functionalization of atomic force microscopy (AFM) tips with
biomolecules capable of interacting with the analyte on a substrate, and the detection being
performed by measuring the force between the immobilized biomolecule and the analyte.
The optimization of such sensors may require multiple experiments to determine suitable
experimental conditions for the immobilization and detection. In this study we employ molecular
modeling techniques to assist in the design of nanobiosensors to detect herbicides. As a proof
of principle, the properties of acetyl co-enzyme A carboxylase (ACC) were obtained with
molecular dynamics simulations, from which the dimeric form in an aqueous solution was
found to be more suitable for immobilization owing to a smaller structural fluctuation than
the monomeric form. Upon solving the nonlinear Poisson–Boltzmann equation using a
finite-difference procedure, we found that the active sites of ACC exhibited a positive surface
potential while the remainder of the ACC surface was negatively charged. Therefore, optimized
biosensors should be prepared with electrostatic adsorption of ACC onto an AFM tip
functionalized with positively charged groups, leaving the active sites exposed to the analyte.
The preferential orientation for the herbicides diclofop and atrazine with the ACC active site
was determined by molecular docking calculations which displayed an inhibition coefficient
of 0.168 mM for diclofop, and 44.11 mM for atrazine. This binding selectivity for the herbicide
family of diclofop was confirmed by semiempirical PM6 quantum chemical calculations which
revealed that ACC interacts more strongly with the herbicide diclofop than with atrazine,
showing binding energies of 119.04 and +8.40 kcal mol1, respectively.
Poster demonstrating the results from the development/verification project for the quantitation of pyridoxal 5-phosphate and 4-pyridoxic acid in human plasma.
Quantitative Analysis of 30 Drugs in Whole Blood by SPE and UHPLC-TOF-MSAnnex Publishers
Abstract
An Ultra-High Pressure Liquid Chromatography Time-of-Flight Mass Spectrometry (UHPLC-TOF-MS) method for quantitative analysis of 30 drugs in whole blood was developed and validated. The method was used for screening and quantification of common drugs and drugs of abuse in whole blood received from autopsy cases and living persons. The compounds included: alprazolam, amphetamine, benzoylecgonine, bromazepam, cathine, cathinone, chlordiazepoxide, cocaine, codeine, clonazepam, 7-aminoclonazepam, diazepam, nordiazepam, flunitrazepam, 7-aminoflunitrazepam, ketamine, ketobemidone, 3,4-Methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), methamphetamine, methadone, morphine, 6-monoacetylmorphine, nitrazepam, 7-aminonitrazepam, oxazepam, temazepam, tramadol, O-desmethyltramadol, and zolpidem. Blood samples (200 μL) were subjected to Solid Phase Extraction (SPE). Target drugs were quantified using a Waters ACQUITY UPLC system coupled to a Waters SYNAPT G2 TOF-MS apparatus. Extraction recoveries ranged from 41% (7-aminoclonazepam) to 111% (ketamine) and matrix effects ranged from -13% (temazepam) to 50% (7-aminonitrazepam). For all compounds, a quadratic polynomial was applied for fitting the calibration curves. Lower Limits of Quantification (LOQ) ranged from 0.005 to 0.05 mg/kg. Satisfactory precisions below 15% and accuracies within 85-115% were obtained for all compounds at concentrations exceeding the LOQ. In conclusion, we present a validated UHPLC-TOF-MS method for simultaneous quantification of 30 drugs in whole blood with a run time of 15 min using 200 μL of whole blood.
Keywords: Drugs of abuse, UHPLC-TOF-MS, Whole blood, SPE, Quantification
Drug discovery library design by biomimetic hplcKlara Valko
The slides explain the early drug discovery process and how the measurements of biomimetic properties can help to design the best ADME properties of compound libraries.
Poster demonstrating the results from the development/verification project for the quantitation of all- trans retinol and alpha tocopherol in human serum.
Spectroscopic and Thermal Characterization of Charge-Transfer Complexes Forme...IJMER
The spectrophotometric characteristics of the solid charge-transfer molecular complexes
(CT) formed in the reaction of the electron donor 2-amino-6-ethylpyridine (2A6EPy) with the π-
acceptors tetracyanoethylene (TCNE), 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) and 2,4,4,6-
tetrabromo-2,5-cyclohexadienone (TBCHD) have been studied in chloroform at 25 0C. These were
investigated through electronic, infrared, mass spectra and thermal studies as well as elemental
analysis. The results show that the formed solid CT- complexes have the formulas [(2A6EPy)(TCNE)2],
[(2A6EPy)2(DDQ)], [(2A6EPy)4(TBCHD)] for 2-amino-6-ethylpyridine in full agreement with the
known reaction stoichiometries in solution as well as the elemental measurements. The formation
constant kCT, molar extinction coefficient εCT, free energy change ∆G
0
and CT energy ECT have been
calculated for the CT- complexes [(2A6EPy)(TCNE)2], [(2A6EPy)2(DDQ)].
This is nice presentation given by Vishal Goyani, an Analytical student of National Institute Of Pharmaceutical education and research-INDIA.
mail your query at - vngoyanii@gmail.com
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).
Nanobiosensors can be built via functionalization of atomic force microscopy (AFM) tips with
biomolecules capable of interacting with the analyte on a substrate, and the detection being
performed by measuring the force between the immobilized biomolecule and the analyte.
The optimization of such sensors may require multiple experiments to determine suitable
experimental conditions for the immobilization and detection. In this study we employ molecular
modeling techniques to assist in the design of nanobiosensors to detect herbicides. As a proof
of principle, the properties of acetyl co-enzyme A carboxylase (ACC) were obtained with
molecular dynamics simulations, from which the dimeric form in an aqueous solution was
found to be more suitable for immobilization owing to a smaller structural fluctuation than
the monomeric form. Upon solving the nonlinear Poisson–Boltzmann equation using a
finite-difference procedure, we found that the active sites of ACC exhibited a positive surface
potential while the remainder of the ACC surface was negatively charged. Therefore, optimized
biosensors should be prepared with electrostatic adsorption of ACC onto an AFM tip
functionalized with positively charged groups, leaving the active sites exposed to the analyte.
The preferential orientation for the herbicides diclofop and atrazine with the ACC active site
was determined by molecular docking calculations which displayed an inhibition coefficient
of 0.168 mM for diclofop, and 44.11 mM for atrazine. This binding selectivity for the herbicide
family of diclofop was confirmed by semiempirical PM6 quantum chemical calculations which
revealed that ACC interacts more strongly with the herbicide diclofop than with atrazine,
showing binding energies of 119.04 and +8.40 kcal mol1, respectively.
CAIRO, EGYPT--(Marketwire - Apr 26, 2012) -
OCI Formally Signs US$ 363 Million Contract to Build a 1,000 MW Gas Turbine Power Plant in Iraq
Orascom Construction Industries (OCI) announced today that it formally signed the US$ 363 million contract to build a 1,000 MW gas turbine power plant in Baija, Iraq. The Iraqi cabinet approved the project in January 2012.
The power plant was tendered and awarded by the Iraqi Ministry of Electricity, which was established in 2003. The project is expected to be completed in 21 months.
Retete de la GoodFood Romania BBQ Party #2Bogdan Epure
A doua editie a Good Food BBQ Party a strans in jur de 100 de persoane in gradina La Seratta, inclusiv cei mai buni bloggeri de food din tara, chef Joseph Hadad, chef Cezar Munteanu, Marius Tudosiei de la emisiunea “Sanatatea in bucate” si cititori fideli ai revistei Good Food.
LC-MS/MS method for the quantification of carbinoxamine in human plasmaIOSR Journals
A simple, reverse-phase high performance liquid chromatographic method with mass spectrometric detection (HPLC-MS/MS) was developed for determination of carbinoxamine in human plasma using pargeverine HCl as an internal standard. The procedure involves a simple protein precipitation technique using BDS HYPERSIL C8 (100 x 4.6mm) column. The mobile phase used was acetonitrile: buffer (25mm ammonium formate solution) (80:20). Precipitation was done using acetonitrile and detection was done in MRM mode, using an Electro Spray positive ionization. The ion transition monitored was (m/z) carbinoxamine (Q1 Mass: 291.2; Q3 Mass: 167.1), Internal standard (Q1 Mass: 338.1; Q3 Mass; 167.0). The retention time of carbinoxamine and internal Standard were 1.61 and 1.75 respectively. Method was evaluated in terms of linearity, accuracy, precision, recovery, sensitivity. The simple extraction procedure and short chromatographic runtime make the method suitable for therapeutic drug monitoring studies.
Haemolysis effect of Mefenamic Acid 250 mg Capsule in Bio analysis by liquid ...IOSR Journals
A rapid, simple and specific method for estimation of Mefenamic acid in human plasma was validated using Indomethacin as internal standard. The analyte and internal standard were extracted from plasma using simple solid phase extraction. The compound were separated on a reverse-phase column with an isocratic mobile phase consisting of 2 mM Ammonium Acetate in Water and acetonitrile (20:80, v/v) and detected by tandem mass spectrometry in negative ion mode. The ion transition recorded in multiple reaction monitoring mode were m/z 240.1 196.0 for Mefenamic acid and m/z 356.1312.0 for internal standard. Linearity in plasma was observed over the concentration range 35.000 – 7000.000 ng/mL for Mefenamic acid. The cv of the assay was 4.89 % to 5.98 % and accuracy was 99.36 to 102.20 % Intra and Interday respectively at LLOQ level. The validated method was applied to bioequivalence study of 250 mg Mefenamic acid in 28 healthy human volunteers. Total 50 samples from individual volunteers identified as Haemolyzed which were analyze initial and repeat again to cross check the method reproducibity for Haeamolysis effect and compared which found acceptable range
Analytical Method Development and Validation for the Estimation of Zolmitript...ijtsrd
In this work the authors have proposed a simple, specific, economic and accurate reverse phase liquid chromatographic method for the estimation of Zolmitriptan as an active pharmaceutical ingredient and in pharmaceutical formulation. The main objective of the current research paper is to To develop simple, precise and accurate RP HPLC method for Zolmitriptan also to validate the developed method as per ICH guideline Q2R1 and to explore the applicability of the method in finished product formulation for estimation of Zolmitriptan during its lifecycle. The objective was achieved by optimized condition with Phonemenex C18 column 150mm×4.6mm , 5µm. And mobile phase Phosphate buffer pH 3.5 85 Methanol 15. The separation was done with a flow rate of 0.9ml min, detection with 224nm. The retention was found to be 3.57 minute. LOD and LOQ were found to be 2.45 and 7.42 respectively. So in order to obtain the correct results various validations methods are performed to get the results. The results obtained from those validation methods are plotted in the form of the charts as well as the different curves. Mr. Rahul M. Sagde | Mr. Pawan N. Karwa | Mr. Vivek M. Thorat | Sanjay S. Jadhav "Analytical Method Development and Validation for the Estimation of Zolmitriptan by RP HPLC Method" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-5 , August 2019, URL: https://www.ijtsrd.com/papers/ijtsrd26474.pdfPaper URL: https://www.ijtsrd.com/pharmacy/analytical-chemistry/26474/analytical-method-development-and-validation-for-the-estimation-of-zolmitriptan-by-rp-hplc-method/mr-rahul-m-sagde
Development and validation of HPLC method for the estimation of Escitalopram ...SriramNagarajan15
A simple, specific, robust, accurate and precise isocratic HPLC method has been developed and subsequently validated for simultaneous determination of escitalopram (ESP) in pharmaceutical dosage forms. Kromosil (250x4.6)mm 5µ with flow rate of 1ml/ min by using JASCO PU-1580 and UV/VIS JASCO UV-1570 at 238 nm. The separation was carried out using a mobile phase consisting of acetonitrile, methanol and 5mM ammonium acetate buffer (pH 3.0) in the ratio 30:20:50 respectively. The retention time for escitaloparm was found to be 5.36 minutes respectively. The correlation coefficient was found to be 0.9997 (ESP). The mean percentage recovery was found to be 101.86 respectively. The % estimation of the drugs was found near to 100 % representing the accuracy in the method. The proposed method was also validated and applied for the analysis of drugs in tablet formulation.
Elevating Tactical DDD Patterns Through Object CalisthenicsDorra BARTAGUIZ
After immersing yourself in the blue book and its red counterpart, attending DDD-focused conferences, and applying tactical patterns, you're left with a crucial question: How do I ensure my design is effective? Tactical patterns within Domain-Driven Design (DDD) serve as guiding principles for creating clear and manageable domain models. However, achieving success with these patterns requires additional guidance. Interestingly, we've observed that a set of constraints initially designed for training purposes remarkably aligns with effective pattern implementation, offering a more ‘mechanical’ approach. Let's explore together how Object Calisthenics can elevate the design of your tactical DDD patterns, offering concrete help for those venturing into DDD for the first time!
Smart TV Buyer Insights Survey 2024 by 91mobiles.pdf91mobiles
91mobiles recently conducted a Smart TV Buyer Insights Survey in which we asked over 3,000 respondents about the TV they own, aspects they look at on a new TV, and their TV buying preferences.
GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using Deplo...James Anderson
Effective Application Security in Software Delivery lifecycle using Deployment Firewall and DBOM
The modern software delivery process (or the CI/CD process) includes many tools, distributed teams, open-source code, and cloud platforms. Constant focus on speed to release software to market, along with the traditional slow and manual security checks has caused gaps in continuous security as an important piece in the software supply chain. Today organizations feel more susceptible to external and internal cyber threats due to the vast attack surface in their applications supply chain and the lack of end-to-end governance and risk management.
The software team must secure its software delivery process to avoid vulnerability and security breaches. This needs to be achieved with existing tool chains and without extensive rework of the delivery processes. This talk will present strategies and techniques for providing visibility into the true risk of the existing vulnerabilities, preventing the introduction of security issues in the software, resolving vulnerabilities in production environments quickly, and capturing the deployment bill of materials (DBOM).
Speakers:
Bob Boule
Robert Boule is a technology enthusiast with PASSION for technology and making things work along with a knack for helping others understand how things work. He comes with around 20 years of solution engineering experience in application security, software continuous delivery, and SaaS platforms. He is known for his dynamic presentations in CI/CD and application security integrated in software delivery lifecycle.
Gopinath Rebala
Gopinath Rebala is the CTO of OpsMx, where he has overall responsibility for the machine learning and data processing architectures for Secure Software Delivery. Gopi also has a strong connection with our customers, leading design and architecture for strategic implementations. Gopi is a frequent speaker and well-known leader in continuous delivery and integrating security into software delivery.
Dev Dives: Train smarter, not harder – active learning and UiPath LLMs for do...UiPathCommunity
💥 Speed, accuracy, and scaling – discover the superpowers of GenAI in action with UiPath Document Understanding and Communications Mining™:
See how to accelerate model training and optimize model performance with active learning
Learn about the latest enhancements to out-of-the-box document processing – with little to no training required
Get an exclusive demo of the new family of UiPath LLMs – GenAI models specialized for processing different types of documents and messages
This is a hands-on session specifically designed for automation developers and AI enthusiasts seeking to enhance their knowledge in leveraging the latest intelligent document processing capabilities offered by UiPath.
Speakers:
👨🏫 Andras Palfi, Senior Product Manager, UiPath
👩🏫 Lenka Dulovicova, Product Program Manager, UiPath
Builder.ai Founder Sachin Dev Duggal's Strategic Approach to Create an Innova...Ramesh Iyer
In today's fast-changing business world, Companies that adapt and embrace new ideas often need help to keep up with the competition. However, fostering a culture of innovation takes much work. It takes vision, leadership and willingness to take risks in the right proportion. Sachin Dev Duggal, co-founder of Builder.ai, has perfected the art of this balance, creating a company culture where creativity and growth are nurtured at each stage.
UiPath Test Automation using UiPath Test Suite series, part 3DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 3. In this session, we will cover desktop automation along with UI automation.
Topics covered:
UI automation Introduction,
UI automation Sample
Desktop automation flow
Pradeep Chinnala, Senior Consultant Automation Developer @WonderBotz and UiPath MVP
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
Accelerate your Kubernetes clusters with Varnish CachingThijs Feryn
A presentation about the usage and availability of Varnish on Kubernetes. This talk explores the capabilities of Varnish caching and shows how to use the Varnish Helm chart to deploy it to Kubernetes.
This presentation was delivered at K8SUG Singapore. See https://feryn.eu/presentations/accelerate-your-kubernetes-clusters-with-varnish-caching-k8sug-singapore-28-2024 for more details.
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.
DevOps and Testing slides at DASA ConnectKari Kakkonen
My and Rik Marselis slides at 30.5.2024 DASA Connect conference. We discuss about what is testing, then what is agile testing and finally what is Testing in DevOps. Finally we had lovely workshop with the participants trying to find out different ways to think about quality and testing in different parts of the DevOps infinity loop.
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.
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
UiPath Test Automation using UiPath Test Suite series, part 4
1 s2.0-s1570023203002794-main
1. Journal of Chromatography B, 792 (2003) 55–61
www.elsevier.com/locate/chromb
Determination of cabergoline and L-dopa in human plasma
using liquid chromatography–tandem mass spectrometry
a , a a b b
*Kazuo Igarashi , Koichiro Hotta , Fumiyo Kasuya , Kazuo Abe , Saburo Sakoda
a
Laboratory of Biochemical Toxicology, Faculty of Pharmaceutical Sciences and High Technology Research Center,
Kobe Gakuin University, Ikawadanicho, Nishi-ku, Kobe 651-2180, Japan
b
Department of Neurology, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan
Received 8 November 2002; received in revised form 31 January 2003; accepted 17 March 2003
Abstract
We determined cabergoline and L-dopa in human plasma using liquid chromatography–mass spectrometry with tandem
mass spectrometry (LC–MS–MS). The deproteinized plasma samples with organic solvent or acid were analyzed directly by
reversed-phase liquid chromatography. Using multiple reaction monitoring (MRM, product ions m/z 381 of m/z 452 for
cabergoline and m/z 152 of m/z 198 for L-dopa) on LC–MS–MS with electrospray ionization (ESI), cabergoline and L-dopa
in human plasma were determined. Calibration curves of the method showed a good linearity in the range 5–250 pg/ml for
cabergoline and 1–200 ng/ml for L-dopa, respectively. The limit of determination was estimated to be approximately
2 pg/ml for cabergoline and approximately 0.1 ng/ml for L-dopa, respectively. The method was applied to the analysis of
cabergoline and L-dopa in plasma samples from patients treated with these drugs. The precision of analysis showed
coefficients of variation ranging from 3.8% to 10.5% at cabergoline concentration of 13.8–26.2 pg/ml and from 2.9% to
8.9% at an L-dopa concentration of 302.5–522.1 ng/ml in patient plasma. As a result, the procedure proved to be very
suitable for routine analysis.
2003 Elsevier Science B.V. All rights reserved.
Keywords: L-Dopa; Cabergoline
1. Introduction patients [3]. Moreover, it has been reported that
cabergoline improves Parkinson symptoms in MPTP-
Cabergoline, N-[3-(dimethylamino)propyl]-N- treated monkeys [4] and is effective in the treatment
(ethylamino)carbonyl - 6 - (2-propenyl) - ergoline - 8b- of Parkinson’s disease [5,6].
carboxamide (Fig. 1) is an ergot alkaloid derivative An oral dose of cabergoline lower than 1 mg/day
with dopamine agonist activity [1]. It has been is usually used for the treatment of Parkinson’s
shown to induce long-lasting inhibition of prolactin disease, and it has been reported that the plasma
secretion in rats [2] and in hyperprolactinemia levels of cabergoline in healthy volunteers who took
a single oral dose of 0.6 mg is in the range 80–800
pg/ml using the radioimmunoassay method [7]. The
*Corresponding author. Tel.: 181-78-974-1551; fax: 181-78-
very low dosages in humans has required the de-974-5689.
velopment of a more sensitive and selective methodE-mail address: igarashi@pharm.kobegakuin.ac.jp (K.
Igarashi). for measuring plasma levels of cabergoline. Al-
1570-0232/03/$ – see front matter 2003 Elsevier Science B.V. All rights reserved.
doi:10.1016/S1570-0232(03)00279-4
2. 56 K. Igarashi et al. / J. Chromatogr. B 792 (2003) 55–61
2. Experimental
2.1. Chemicals and reagents
Cabergoline and deuterated internal standard (CG-
095, I.S.) (Fig. 1) were provided by Kissei Pharma-
ceutical Co. (Matsumoto, Japan). L-Dopa and a-
methyldopa (I.S.) were obtained from Sigma Chemi-
cal Co. (St. Louis, USA). Acetonitrile (HPLC grade),
methanol (HPLC grade), high purity ammonium
formate and formic acid were obtained from
NACALAI Tesque Inc. (Kyoto, Japan). All other
chemicals and reagents were of analytical grade from
commercial sources.
Fig. 1. Chemical structures of cabergoline, deuterated cabergoline
2.2. Sample collection
(I.S.), L-dopa and a-methyldopa (I.S.).
Four schizophrenic patients (41–63 years old)
though an RIA method was developed to quantify volunteered to take part in this study; informed
cabergoline in plasma samples, it is thought that its consent was obtained from each subject. Blood
method cannot give a satisfactory result because its samples were taken 1 h after oral administration of
major metabolite, N-demethyl metabolite, cross- the drug (cabergoline 0.25–1.0 mg or L-dopa 200
reacts with the antibody. For quantitative analysis, a mg) and placed in heparinized tubes. The plasma
liquid chromatography–mass spectrometry (LC– was separated by centrifugation and stored at 240 8C
MS) method has recently become a powerful tech- until analysis.
nique. In addition, the application of tandem mass Plasma samples from healthy volunteers were also
spectrometry (MS–MS) has improved both the sen- used as control for development of extraction pro-
sitivity and selectivity of the determination. A large cedure for cabergoline and L-dopa.
number of studies have been performed using liquid
chromatography combined with tandem mass spec- 2.3. Sample preparation for LC–MS
trometry (LC–MS–MS) for the determination of
drugs in biological fluids [8–13]. This method has For determination of cabergoline, plasma samples
recently been used for the determination of caber- (100 ml) were mixed with 50 ml of acetonitrile, 20
goline in human plasma [14]. In the present study, mM ammonium formate solution (90:10, v/v) and
we also developed the modified method to determine 50 ml of I.S. solution (CG-095, 1 ng/ml in acetoni-
plasma levels of cabergoline using LC–MS–MS. trile) in the microcentrifuge tubes. It was then
Combination treatment of L-dopa with cabergoline centrifuged at 10 000 g for 10 min and the superna-
has been used for the treatment of Parkinson’s tant was filtered with a 0.45-mm filter. The filtrate
disease. In this case, it is important that the plasma was transferred to the autosampler vial insert, and 30
concentration of L-dopa is determined in the patients ml was injected into the LC–MS–MS system.
administered with cabergoline. Although many meth- For determination of L-dopa, plasma samples (50
ods have been developed for the determination of ml) were added to 60 ml of 0.4 M perchloric acid and
L-dopa in plasma, reversed-phase high performance 40 ml of I.S. solution (a-methyldopa, 100 ng/ml in
liquid chromatography (HPLC) with electrochemical 0.4 M perchloric acid) in the microcentrifuge tubes.
detection (ECD) [15–18] is currently used. In the After shaking the tube for 1 min, it was centrifuged
present study, the analytical procedure using LC– at 10 000 g for 10 min and the supernatant was
MS–MS was applied to the analysis of L-dopa in filtered with a 0.45-mm filter. The filtrate was diluted
human plasma samples and developed. with 100 ml of the HPLC mobile phase solution, and
3. K. Igarashi et al. / J. Chromatogr. B 792 (2003) 55–61 57
was then transferred to the autosampler vial insert Ionization conditions for LC–MS–MS were as fol-
where 20 ml was injected into the LC–MS–MS lows: for cabergoline, capillary voltage 2.7 kV, cone
system. voltage 35 V, collision energy 18 eV, source tempera-
ture 80 8C, desolvation temperature 200 8C; for L-
2.4. LC–MS–MS dopa, capillary voltage 3.2 kV, cone voltage 15 V,
collision energy 14 eV, source temperature 80 8C,
LC–MS–MS was performed using a Quattro desolvation temperature 300 8C, and argon was used
Ultima triple quadrupole tandem mass spectrometer as collision gas.
(Micromass Inc., Manchester, UK) equipped with
electrospray ionization (ESI). The HPLC system
consisted of a Waters Alliance 2690 pump equipped 3. Results and discussion
with an autosampler (Waters Co., Milford, MA,
USA) and the reversed-phase COSMOSIL 5C8-MS The ESI mass spectra of cabergoline and CG-095
column (2 mm I.D.3150 mm, 5-mm particle diam- (I.S.) are shown in Fig. 2. Cabergoline and CG-095
1
eter, NACALAI Tesque Inc., Kyoto, Japan) for showed a protonated molecule ion (M1H) of m/z
1
cabergoline analysis or the reversed-phase COS- 452 and 456, respectively. These (M1H) ions were
MOSIL 5C -MS column (2 mm I.D.3150 mm, predominant on the ESI mass spectra. Investigation18
5-mm particle diameter, NACALAI Tesque Inc., of the MS/MS behavior of cabergoline indicated that
Kyoto, Japan) for L-dopa analysis. The flow-rate of the product ion m/z 381 of cabergoline (m/z 452)
mobile phase for cabergoline analysis was set at 0.2 was the most suitable ion for quantitative determi-
ml/min, with gradient elution starting at 20% nation by multiple reaction monitoring (MRM) (Fig.
acetonitrile–80% water (containing 20 mM ammo- 3). Moreover, the product ion m/z 385 was chosen
nium formate) for 3 min, followed by a linear as the most adequate ion of deuterated cabergoline
increase to 80% acetonitrile in 1 min, and held there (I.S.). Fig. 4a shows the MRM chromatograms of
for 10 min before rapidly returning to the initial cabergoline (125 pg/ml) and I.S. standard solutions
conditions. For L-dopa analysis, a mobile phase using the product ions with m/z 381 and 385,
consisting of 10% methanol–90% water (containing respectively. The MRM chromatograms of plasma
05% formic acid) was delivered at a flow-rate of 0.2 obtained from patients who were treated with caber-
ml/min. In both case, sample introduction to the goline are shown in Fig. 4b. It was a clean chromato-
mass spectrometer was stopped for 0–6 min for gram free from interfering contaminants.
cabergoline and for 0–2 min for L-dopa using a For the determination of cabergoline in human
switching valve after injection into the system. plasma, liquid–liquid extraction was first used as the
Fig. 2. LC–ESI mass spectra of cabergoline (a) and deuterated cabergoline (I.S.) (b).
4. 58 K. Igarashi et al. / J. Chromatogr. B 792 (2003) 55–61
1
Fig. 3. Product ion spectra on LC–MS–MS of the [M1H] ion of cabergoline (a) and deutrated cabergoline (I.S.) (b).
pretreatment of sample according to the previous observed over the concentration range examined (5–
method [14,19]. However, the extraction efficiency 250 pg/ml for standard solution, y50.0051x2
2
of cabergoline to organic solvent was not as much 0.0036, r 50.9974). The calibration curve showed
(about 52% at 20 pg/ml), and its method could not little day-to-day variability in the slopes and the
give good results on the precision of the measure- intercepts [coefficient of variation (C.V.), ,8%]. The
ments (data not shown). Therefore, we modified the lower limit of detection was approximately 2 pg/ml
preparation method of plasma samples to inject into (S/N54) for standard solution.
the LC–MS–MS system. The plasma samples were Experiments with spiked samples resulted in a
added with acetonitrile and 20 mM ammonium recovery of 94.866.4% at a concentration of 20
formate solution (90:10, v/v) and after mixing and pg/ml of control plasma sample (data not shown).
centrifugation, the supernatants were directly in- Table 1 gives the intra-day and inter-day precision
jected into the LC–MS–MS system. The calibration (C.V.%) of the method using plasma samples of
curve for cabergoline using MRM was obtained by patients treated with cabergoline. The intra-day
plotting the peak area ratio of cabergoline (product precision ranged from 3.8 to 8.7%, and the inter-day
ion m/z 381) to I.S. (product ion m/z 385) versus precision ranged from 6.4 to 10.5%. These results
the amount of cabergoline. Good linearity was indicate good reproducibility between experiments.
Fig. 4. MRM chromatograms on LC–MS–MS of standard solution (a) and human plasma sample. The standard solution (a) is 125 pg/ml
and plasma sample obtained from patient is 15.5 pg/ml.
5. K. Igarashi et al. / J. Chromatogr. B 792 (2003) 55–61 59
Table 1 methyldopa (I.S.) showed a protonated molecule ion
1Intra-day and inter-day precision of LC–MS–MS method for the
(M1H) of m/z 198 and 212, respectively (Fig. 5).
determination of cabergoline in human plasma
Moreover, ESI-MS–MS mass spectra are shown in
Patient Plasma Precision (C.V.%) Fig. 6. It indicates that the product ions m/z 152 of
concentration
Intra-day Intra-day L-dopa (m/z 198) and m/z 139 of I.S. (m/z 212) are
(pg/ml)
(n55) (n52, 4 days) the most adequate ions for MRM of LC–MS–MS.
We examined the quantitative determination of L-K (female, 63) 19.561.7 8.7 9.1
S (female, 58) 26.261.0 3.8 6.4 dopa using MRM with these ions. Without the use of
Y (male, 59) 13.860.7 5.1 10.5 liquid–liquid extraction, the plasma samples were
N (female, 41) 17.561.4 8.0 9.8
simply treated with perchloric acid solution. After
Plasma samples were obtained from patients 1 h after oral removing the plasma protein, it was diluted with
treatment with cabergoline (0.5 mg). Plasma concentration repre-
HPLC mobile and then injected directly into the
sents mean6SD of five experiments.
LC–MS–MS system. As shown in Fig. 7, the MRM
We also determined L-dopa in human plasma using chromatogram was free of interference at the re-
the same LC–MS–MS method as that of caber- tention times of interest. The calibration curve
goline. The ESI mass spectra of L-dopa and a- plotted the peak area ratio of L-dopa (product ion
Fig. 5. LC–ESI mass spectra of L-dopa (a) and a-methyldopa (I.S.) (b).
1
Fig. 6. Product ion spectra on LC–MS–MS of the [M1H] ion of L-dopa (a) and a-methyldopa (I.S.) (b).
6. 60 K. Igarashi et al. / J. Chromatogr. B 792 (2003) 55–61
Fig. 7. MRM chromatograms on LC–MS–MS of standard solution (a) and human plasma sample. The standard solution (a) is 800 ng/ml
and plasma sample obtained from the patient is 285.5 ng/ml.
m/z 152) to I.S. (product ion m/z 139) against the tem. In the previous methods, sample preparations
L-dopa concentration (ng/ml) showed a good lineari- for LC, for example, liquid–liquid extraction for
ty over the range examined (1–200 ng/ml for cabergoline [14,19] and purification by alumina for
2
standard solution) (y50.1328x10.0207, r 5 L-dopa [18] were complex and took a lot of time, and
0.9981). The lower limit of detection was approxi- good reproducibility could not be obtained. In order
mately 0.1 ng/ml (S/N54) for standard solution. to improve these problems, we examined the direct
Furthermore, the intra-day and inter-day precision analysis by MRM method on LC–MS–MS without
(C.V.%) of the method using plasma samples of liquid–liquid extraction. After injection of the sam-
patients treated with L-dopa are shown in Table 2. ple into the system, the delay of sample introduction
The intra-day precision ranged from 2.9 to 6.4%, and to the mass spectrometer by use of a switching valve
the inter-day precision ranged from 6.5 to 8.1%. could be protected from the interference of ionization
These results indicate good reproducibility between and increase the ion intensity. The present procedure
experiments. demonstrated that this technique was simple, had
We have developed a simple determination for good reproducibility and was very useful for routine
cabergoline and L-dopa using an LC–MS–MS sys- analysis of cabergoline and L-dopa.
Table 2
Intra-day and inter-day precision of LC–MS–MS method for the determination of L-dopa in human plasma
Sample Plasma concentration Precision (C.V.%)
(pg/ml)
Intra-day Intra-day
(n55) (n52, 4 days)
S (female, age 58)
(CG-105, 0.25 mg) 405.2111.7 2.9 6.5
(CG-105, 0.5 mg) 522.1132.9 6.3 8.1
(CG-105, 1.0 mg) 302.5119.4 6.4 8.9
Plasma samples obtained from patients 1 h after oral treatment with L-dopa (200 mg) and cabergoline (CG-105, 0.25–1.0 mg) plasma
concentration represents mean6S.D of five experiments.
7. K. Igarashi et al. / J. Chromatogr. B 792 (2003) 55–61 61
[7] S. Persiani, E. Pianezzola, F. Broutin, G. Fonte, M.S.Acknowledgements
Benedetti, J. Immunoassay 13 (1992) 457.
[8] W.M. Muck, J. Chromatogr. A 712 (1995) 45.
This study was partly supported by a Research [9] B. Lansecker, G. Hopfgartner, J. Chromatogr. B 658 (1995)
Grant from Kissei Pharmaceutical Co. Ltd. and 281.
Pharmacia and Upjohn Inc. in Japan. We thank Dr [10] T.V. Constanzer, C.M. Chavez, B.K. Matuszewski, J. Pharm.
Biomed. Anal. 12 (1994) 705.M. Nishiyama for providing the internal standard
[11] A.T. Murphy, P.L. Bonatte, S.C. Kasper, T.A. Gillespie, A.F.
(CG-095).
Delong, Biol. Mass Spectrom. 23 (1994) 621.
[12] M. Rittner, F. Pragst, W. Bork, J. Neumann, J. Anal. Toxicol.
25 (2001) 115.
References [13] D. Fluchard, S. Kiebooms, M. Dubois, P. Delahaut, J.
Chromatogr. B 744 (2000) 281.
[14] C. Allievi, P. Dostert, Rapid Commun. Mass Spectrom. 12
[1] M.S. Benedetti, P. Dostert, D. Barone, C. Ethymiopoulos, G.
(1998) 33.
Peretti, R. Roncucci, Eur. J. Pharmacol. 187 (1990) 399.
[15] S. Dethy, M.A. Laute, N.V. Blercom, P. Damhaut, S. Gol-
[2] E.D. Salle, G. Ornati, G. Briatico, J. Endocrinol. Invest. 5
dman, J. Hilderbrand, Clin. Chem. 43 (1997) 740.
(1982) 45.
[16] F. Blandini, E. Martignoni, C. Pacchetti, S. Desideri, D.
[3] C. Ferrari, C. Barbieri, R. Caldara, M. Mucci, F. Codecasa,
Rivellini, G. Nappi, J. Chromatogr. B 700 (1997) 278.
A. Paracchi et al., J. Clin. Endocrinol. Metab. 63 (1986) 941.
[17] I. Rondelli, D. Acerbi, F. Mariotti, P.Ventura, J. Chromatogr.
[4] N. Arai, M. Isaji, H. Miyata, J. Fukuyama, E. Mizuta, S.
B 653 (1994) 17.
Kuno, J. Neural. Transm. 10 (1995) 55.
[18] T. Wikberg, J. Pharm. Biomed. Anal. 9 (1991) 167.
[5] A. Lieberman, S. Imke, M. Muenter, K. Wheeler, J.E.
[19] E. Pianezzola, V. Bellotti, R.L. Croix, M. Benedetti, J.
Ahiskog, J.Y. Matsumoto et al., Neurology 43 (1993) 1981.
Chromatogr. 574 (1992) 170.
[6] M. Jori, A. Dubin, Mov. Disord. 5 (1990) 62.