Structural elucidation, Identification, quantization of process related impur...IOSR Journals
Major process related unknown impurity associated with the synthesis of Hydralazine hydrochloride bulk drug was detected by high performance liquid chromatography (HPLC) and was subjected to high resolution accurate liquid chromatography mass spectroscopy (HR/AM-LCMS) for identification. The proposed impurity was isolated from Hydralazine hydrochloride active pharmaceutical ingredient (API) by preparative chromatographic method and was injected on HPLC for comparison of retention time with that of the unknown process related impurity in Hydralazine hydrochloride. The molecular ion peak of preparatively isolated impurity and that of unknown process related impurity in Hydralazine hydrochloride were compared for confirmation. The postulated structure was unambiguously confirmed with the help of HR/AM- LC MS/MS, NMR and FTIR data proposed to be 1-(2-phthalazin-1-ylhydrazino)phthalazine (Hazh Dimer). This impurity of Hydralazine hydrochloride is not been previously reported. A rapid Acquity H-class gradient method with runtime of 15.0min was developed for Quantitation on Unisphere Cyno column and validated for parameters such as accuracy, precision, linearity and range, robustness. The LOD and LOQ of method were 0081% and 0.0246% respectively.
International Journal of Engineering Research and Applications (IJERA) is an open access online peer reviewed international journal that publishes research and review articles in the fields of Computer Science, Neural Networks, Electrical Engineering, Software Engineering, Information Technology, Mechanical Engineering, Chemical Engineering, Plastic Engineering, Food Technology, Textile Engineering, Nano Technology & science, Power Electronics, Electronics & Communication Engineering, Computational mathematics, Image processing, Civil Engineering, Structural Engineering, Environmental Engineering, VLSI Testing & Low Power VLSI Design etc.
Kinetics of Ruthenium(III) Catalyzed and Uncatalyzed Oxidation of Monoethanol...Ratnakaram Venkata Nadh
Kinetics of uncatalyzed and ruthenium(III) catalyzed oxidation of monoethanolamine by N-bromosuccinimide
(NBS) has been studied in an aqueous acetic acid medium in the presence of sodium acetate
and perchloric acid, respectively. In the uncatalyzed oxidation the kinetic orders are: the first order in NBS,
a fractional order in the substrate. The rate of the reaction increased with an increase in the sodium acetate
concentration and decreased with an increase in the perchloric acid concentration. This indicates that free
amine molecules are the reactive species. Addition of halide ions results in a decrease in the kinetic rate,
which is noteworthy. Both in absence and presence of a catalyst, a decrease in the dielectric constant of the
medium decreases the kinetic rate pointing out that these are dipole—dipole reactions. A relatively higher
oxidation state of ruthenium i.e., Ru(V) was found to be the active species in Ru(III) catalyzed reactions. A
suitable mechanism consistent with the observations has been proposed and a rate law has been derived to
explain the kinetic orders.
Structural elucidation, Identification, quantization of process related impur...IOSR Journals
Major process related unknown impurity associated with the synthesis of Hydralazine hydrochloride bulk drug was detected by high performance liquid chromatography (HPLC) and was subjected to high resolution accurate liquid chromatography mass spectroscopy (HR/AM-LCMS) for identification. The proposed impurity was isolated from Hydralazine hydrochloride active pharmaceutical ingredient (API) by preparative chromatographic method and was injected on HPLC for comparison of retention time with that of the unknown process related impurity in Hydralazine hydrochloride. The molecular ion peak of preparatively isolated impurity and that of unknown process related impurity in Hydralazine hydrochloride were compared for confirmation. The postulated structure was unambiguously confirmed with the help of HR/AM- LC MS/MS, NMR and FTIR data proposed to be 1-(2-phthalazin-1-ylhydrazino)phthalazine (Hazh Dimer). This impurity of Hydralazine hydrochloride is not been previously reported. A rapid Acquity H-class gradient method with runtime of 15.0min was developed for Quantitation on Unisphere Cyno column and validated for parameters such as accuracy, precision, linearity and range, robustness. The LOD and LOQ of method were 0081% and 0.0246% respectively.
International Journal of Engineering Research and Applications (IJERA) is an open access online peer reviewed international journal that publishes research and review articles in the fields of Computer Science, Neural Networks, Electrical Engineering, Software Engineering, Information Technology, Mechanical Engineering, Chemical Engineering, Plastic Engineering, Food Technology, Textile Engineering, Nano Technology & science, Power Electronics, Electronics & Communication Engineering, Computational mathematics, Image processing, Civil Engineering, Structural Engineering, Environmental Engineering, VLSI Testing & Low Power VLSI Design etc.
Kinetics of Ruthenium(III) Catalyzed and Uncatalyzed Oxidation of Monoethanol...Ratnakaram Venkata Nadh
Kinetics of uncatalyzed and ruthenium(III) catalyzed oxidation of monoethanolamine by N-bromosuccinimide
(NBS) has been studied in an aqueous acetic acid medium in the presence of sodium acetate
and perchloric acid, respectively. In the uncatalyzed oxidation the kinetic orders are: the first order in NBS,
a fractional order in the substrate. The rate of the reaction increased with an increase in the sodium acetate
concentration and decreased with an increase in the perchloric acid concentration. This indicates that free
amine molecules are the reactive species. Addition of halide ions results in a decrease in the kinetic rate,
which is noteworthy. Both in absence and presence of a catalyst, a decrease in the dielectric constant of the
medium decreases the kinetic rate pointing out that these are dipole—dipole reactions. A relatively higher
oxidation state of ruthenium i.e., Ru(V) was found to be the active species in Ru(III) catalyzed reactions. A
suitable mechanism consistent with the observations has been proposed and a rate law has been derived to
explain the kinetic orders.
A STUDY ON FORMATION OF SALYCILIC ACID FORMALDEHYDE POLYMER SAMPLEEDITOR IJCRCPS
Condensation of salicylic acid (0.02 mole) with formaldehyde (0.016 mole) in presence of aqueous 40% H2SO4.
Keywords: pipette,thermometer,spectro-photometer,conicalflakk,waterbath.
Spectroscopic, Thermal, Magnetic and conductimetric studies on some 7-hydroxy...IOSR Journals
7-hydroxy-4-methyl-8-(p-methylphenylazo)coumarin (L1) and 7-hydroxy-4-methyl-8-(p-methoxyphenylazo)coumarin (L2) have been prepared and characterized by elemental analysis, infrared (IR), proton nuclear magnetic resonance (1H NMR) and mass spectra. The important infrared (IR) spectral bands corresponding to the active groups in the two ligands and the solid complexes under investigation were studied. Also the important fragments in the ligands and complexes were done using mass spectra and the main peaks were corresponding to the molecular weights of the ligands and complexes. The solid complexes have been synthesized and characterized by elemental and thermal analyses (TG and DTA) as well as by IR, 1H NMR, magnetic measurements, electronic transition, molar conductance, mass and electron spin resonance (ESR) spectra. The proposed steriochemical structures for the investigated metal complexes suggest octahedral geometry with respect to Mn, Co, Ni, Cu and Zn divalent metal ions with the investigated ligands 1:1 and 1:2 complexes and all of the formed complexes contain coordinated and hydrated water molecules. All of the prepared solid complexes behave as non-electrolytes in chloroform.
DETERMINATION OF MIANSERIN USING TROPAEOLIN-OOO BY ION PAIR FORMATIONRatnakaram Venkata Nadh
Objective: The present study was aimed at the development of a simple visible spectrophotometric method for the assay of mianserin, a drug used for the treatment of depression.
Methods: The method was developed using tropaeolin-ooo (TPooo) as an ion associative complex forming a chromophore. Developed the chromophore by sequential mixing of aqueous solutions of mianserin, hydrochloric acid, and TPooo. Chromophore was extracted into an organic solvent (chloroform) and absorbance values of organic layers were measured. As per the existing guidelines of an international conference on harmonization (ICH), various parameters of the method were tested for validation.
Results: At the optimized reaction conditions, the formed chromophore (λ max
Conclusion: Due to lack of pre-treatment process for this method, it was simple. All the tested parameters of the method were validated as per ICH guidelines.
Mannich Synthesis Under Ionic Liquid [Et3NH][HSO4] CatalysisIOSRJAC
Ionic liquid [Et3NH][HSO4] was found to be a particularly efficient catalyst for the synthesis of β- amino carbonyl pyrimidines through the Mannich condensation reaction of substituted pyrimidin-2(1H)-ones, cyclohexanone and 4-fluro/chlorobenzaldehyde under ultrasonic irradiation at room temperature. The present methodology offers several advantages such as excellent yields, simple procedure and mild conditions.
Complexation, Spectroscopic, Thermal, Magnetic And Conductimetric Studies On ...IOSR Journals
7-hydroxy-4-methyl-8-(phenylazo) coumarin (L1)and 7-hydroxy-4-methyl-8-(o-carboxyphenylazo) coumarin (L2) have been prepared and characterized by elemental analysis, infrared (IR), proton nuclear magnetic resonance (1H NMR) and Mass spectra. The important infrared (IR) spectral bands corresponding to the active groups in the two ligands and the solid complexes under investigation were studied. Also the important fragments in the ligands and the complexes were done using mass spectra and the main peaks were corresponding to the molecular weights of the ligands and complexes. The solid complexes have been synthesized and studied by elemental and thermal analyses (TG and DTA) as well as by IR, 1H NMR, magnetic measurements, electronic transition, molar conductance, mass spectra and electron spin resonance (ESR) spectra. The proposed steriochemical structures for the investigated metal complexes suggest octahedral geometry with respect to Mn, Co, Ni, Cu and Zn metal ions and all of the formed complexes contain coordinated and hydrated water molecules. All of the prepared solid complexes behave as non-electrolytes in chloroform.
Synthesis and characterization of some metal complexes of 2- Phenyl-3,4-dihyd...IOSRJAC
2-Phenyl-3,4-dihydro-quinazolin-4-yloxy)-acetic acid (L1) metal complexes with Mn2+ , Co2+, Ni2+ Cu2+ , and Zn2+ ions were studied and the structure of the complexes were elucidated using elemental analyses, infrared (IR), 1H nuclear magnetic resonance (NMR), magnetic moment and thermal analysis measurements. Besides the characterization of complexes by physicochemical technique, Biological activities of the synthesized complexes were examined against some microbial strains for evaluation of antibacterial and antifungal activities.
Volumetric Analysis
Types of titration
Acid- Base Theory
Reaction, End Point & Indicators
Acid- Base titration
Titration curve
Non- Aqueous Titration
Precipitation Titration
Complexometric Titration
Oxidation- Reduction Titration,
Calculation. Errors
General Informations,
Synthesis, Characterization and Antibacterial Activity of New Complexes of So...IOSR Journals
Complexes of some lanthanide picrates (Ln3+ = Pr3+, Nd3+ and Dy3+) with benzo-18-crown-6 and 221-cryptand were synthesized and characterized by elemental analysis, FTIR, and UV-Visible. Spectrophotometric methods, thermal analysis (TGA & DTG), melting point, magnetic susceptibility and molar conductance. Also an in-vitro study on gram positive (Staphylococcus aureus) and gram negative bacteria (Escherichia coli, Salmonella and pseudomonas aeruginosa) was performed and the results were compared to those of the broad spectrum antibiotic Chloramphinicol. The benzo-18-crown-6 complexes have the general formula of [Ln.L.(Pic)2]Pic.nH2O , where; (Ln3+ = Pr3+, Nd3+, and Dy3+) , (L = Benzo-18-crown-6) , (Pic = Picrate anion) , (n = 1-2). In these complexes two picrate anions are coordinated to the metal ion through the phenolic oxygen and oxygen of the ortho nitro group, thus, the metal ions in these complexes have a coordination number of (10). The complexes of 221-cryptand have the general formula of [Ln.L.(Pic)]Pic2.nH2O where; (Ln3+ = Pr3+, Nd3+, and Dy3+), (L = 221-cryptand), (Pic = Picrate anion), (n = 1,2 or 7). In these complexes one picrate anion is coordinated to the metal ion, also through the phenolic oxygen and the oxygen from the ortho nitro group, thus the metal ions in the cryptand complexes have a coordination number of (9).
Titrimetric analysis is a method of analysis in which a solution of the substance being determined is treated with a solution of a suitable reagent of exactly known concentration. The reagent is added to the substance until the amount added is equivalent to the amount of substance to be determined.
A STUDY ON FORMATION OF SALYCILIC ACID FORMALDEHYDE POLYMER SAMPLEEDITOR IJCRCPS
Condensation of salicylic acid (0.02 mole) with formaldehyde (0.016 mole) in presence of aqueous 40% H2SO4.
Keywords: pipette,thermometer,spectro-photometer,conicalflakk,waterbath.
Spectroscopic, Thermal, Magnetic and conductimetric studies on some 7-hydroxy...IOSR Journals
7-hydroxy-4-methyl-8-(p-methylphenylazo)coumarin (L1) and 7-hydroxy-4-methyl-8-(p-methoxyphenylazo)coumarin (L2) have been prepared and characterized by elemental analysis, infrared (IR), proton nuclear magnetic resonance (1H NMR) and mass spectra. The important infrared (IR) spectral bands corresponding to the active groups in the two ligands and the solid complexes under investigation were studied. Also the important fragments in the ligands and complexes were done using mass spectra and the main peaks were corresponding to the molecular weights of the ligands and complexes. The solid complexes have been synthesized and characterized by elemental and thermal analyses (TG and DTA) as well as by IR, 1H NMR, magnetic measurements, electronic transition, molar conductance, mass and electron spin resonance (ESR) spectra. The proposed steriochemical structures for the investigated metal complexes suggest octahedral geometry with respect to Mn, Co, Ni, Cu and Zn divalent metal ions with the investigated ligands 1:1 and 1:2 complexes and all of the formed complexes contain coordinated and hydrated water molecules. All of the prepared solid complexes behave as non-electrolytes in chloroform.
DETERMINATION OF MIANSERIN USING TROPAEOLIN-OOO BY ION PAIR FORMATIONRatnakaram Venkata Nadh
Objective: The present study was aimed at the development of a simple visible spectrophotometric method for the assay of mianserin, a drug used for the treatment of depression.
Methods: The method was developed using tropaeolin-ooo (TPooo) as an ion associative complex forming a chromophore. Developed the chromophore by sequential mixing of aqueous solutions of mianserin, hydrochloric acid, and TPooo. Chromophore was extracted into an organic solvent (chloroform) and absorbance values of organic layers were measured. As per the existing guidelines of an international conference on harmonization (ICH), various parameters of the method were tested for validation.
Results: At the optimized reaction conditions, the formed chromophore (λ max
Conclusion: Due to lack of pre-treatment process for this method, it was simple. All the tested parameters of the method were validated as per ICH guidelines.
Mannich Synthesis Under Ionic Liquid [Et3NH][HSO4] CatalysisIOSRJAC
Ionic liquid [Et3NH][HSO4] was found to be a particularly efficient catalyst for the synthesis of β- amino carbonyl pyrimidines through the Mannich condensation reaction of substituted pyrimidin-2(1H)-ones, cyclohexanone and 4-fluro/chlorobenzaldehyde under ultrasonic irradiation at room temperature. The present methodology offers several advantages such as excellent yields, simple procedure and mild conditions.
Complexation, Spectroscopic, Thermal, Magnetic And Conductimetric Studies On ...IOSR Journals
7-hydroxy-4-methyl-8-(phenylazo) coumarin (L1)and 7-hydroxy-4-methyl-8-(o-carboxyphenylazo) coumarin (L2) have been prepared and characterized by elemental analysis, infrared (IR), proton nuclear magnetic resonance (1H NMR) and Mass spectra. The important infrared (IR) spectral bands corresponding to the active groups in the two ligands and the solid complexes under investigation were studied. Also the important fragments in the ligands and the complexes were done using mass spectra and the main peaks were corresponding to the molecular weights of the ligands and complexes. The solid complexes have been synthesized and studied by elemental and thermal analyses (TG and DTA) as well as by IR, 1H NMR, magnetic measurements, electronic transition, molar conductance, mass spectra and electron spin resonance (ESR) spectra. The proposed steriochemical structures for the investigated metal complexes suggest octahedral geometry with respect to Mn, Co, Ni, Cu and Zn metal ions and all of the formed complexes contain coordinated and hydrated water molecules. All of the prepared solid complexes behave as non-electrolytes in chloroform.
Synthesis and characterization of some metal complexes of 2- Phenyl-3,4-dihyd...IOSRJAC
2-Phenyl-3,4-dihydro-quinazolin-4-yloxy)-acetic acid (L1) metal complexes with Mn2+ , Co2+, Ni2+ Cu2+ , and Zn2+ ions were studied and the structure of the complexes were elucidated using elemental analyses, infrared (IR), 1H nuclear magnetic resonance (NMR), magnetic moment and thermal analysis measurements. Besides the characterization of complexes by physicochemical technique, Biological activities of the synthesized complexes were examined against some microbial strains for evaluation of antibacterial and antifungal activities.
Volumetric Analysis
Types of titration
Acid- Base Theory
Reaction, End Point & Indicators
Acid- Base titration
Titration curve
Non- Aqueous Titration
Precipitation Titration
Complexometric Titration
Oxidation- Reduction Titration,
Calculation. Errors
General Informations,
Synthesis, Characterization and Antibacterial Activity of New Complexes of So...IOSR Journals
Complexes of some lanthanide picrates (Ln3+ = Pr3+, Nd3+ and Dy3+) with benzo-18-crown-6 and 221-cryptand were synthesized and characterized by elemental analysis, FTIR, and UV-Visible. Spectrophotometric methods, thermal analysis (TGA & DTG), melting point, magnetic susceptibility and molar conductance. Also an in-vitro study on gram positive (Staphylococcus aureus) and gram negative bacteria (Escherichia coli, Salmonella and pseudomonas aeruginosa) was performed and the results were compared to those of the broad spectrum antibiotic Chloramphinicol. The benzo-18-crown-6 complexes have the general formula of [Ln.L.(Pic)2]Pic.nH2O , where; (Ln3+ = Pr3+, Nd3+, and Dy3+) , (L = Benzo-18-crown-6) , (Pic = Picrate anion) , (n = 1-2). In these complexes two picrate anions are coordinated to the metal ion through the phenolic oxygen and oxygen of the ortho nitro group, thus, the metal ions in these complexes have a coordination number of (10). The complexes of 221-cryptand have the general formula of [Ln.L.(Pic)]Pic2.nH2O where; (Ln3+ = Pr3+, Nd3+, and Dy3+), (L = 221-cryptand), (Pic = Picrate anion), (n = 1,2 or 7). In these complexes one picrate anion is coordinated to the metal ion, also through the phenolic oxygen and the oxygen from the ortho nitro group, thus the metal ions in the cryptand complexes have a coordination number of (9).
Titrimetric analysis is a method of analysis in which a solution of the substance being determined is treated with a solution of a suitable reagent of exactly known concentration. The reagent is added to the substance until the amount added is equivalent to the amount of substance to be determined.
في اعتقادي فان سورية بلد غني بموارده و ثرواته، ولذلك فهو مرشح لان يكون بلداً منتجاً لغذاء سليم وآمن وخال من الملوثات المختلفة، نظراً للسياسات الحكيمة التي تتبعها الحكومة في ظل قيادة السيد الرئيس الدكتور بشار الأسد حفظه الله، والمتمثلة بجملة من التشريعات والقوانين الهادفة الى حماية المستهلك والبيئة والثروات الغذائية ومصادر المياه، إضافة إلى نشر الوعي عن طريق وسائل الإعلام المختلفة و الحرص على توفير الكوادر في الإعلام التخصصي الغذائي و البيئي.
In view of the medical and social problem caused by the increasing number of drug addicts the determination of opium alka-loids is of special importance. Morphine is known as a highly addictive and potent narcotic but drug users prefer heroin because of its more intense immediate effect. As heroin is hydrolysed in the organism to morphine the knowledge of the morphine content of biological fluids and tissues is indispensable for forensic and therapeutic purposes. The methods recently used for the determination of morphine in urine are gas-liquid chromatography 1, high-pressure liquid chromatography with fluorimetric determination 2 and gas chromatography mass spectrometry 3. In this paper a simple and inexpensive kinetic method is presented , based on the decomposition of the coloured compound formed by the reaction of hydrogen peroxide with cobalt(II) and morphine, in the presence of carbonate buffer. There is a definite concentration range over which the decomposition rate of the compound mentioned is a linear function of the morphine concentration.
Similar to Microchimica Acta Volume 84 issue 5-6 1984 [doi 10.1007_bf01197162] G. A. Milovanovic; M. A. Sekheta -- Kinetic determination of morphine in urine
Austin Journal of Bioorganic & Organic Chemistry is a peer reviewed, open acc...Austin Publishing Group
Austin Journal of Bioorganic & Organic Chemistry is a peer reviewed, open access journal publishes manuscripts in the following areas but not limited to structures, synthesis, kinetics, organic synthesis, physical organic chemistry, supramolecular chemistry and chemical biology.
Austin Journal of Bioorganic & Organic Chemistry accepts original research articles, review articles, commentaries, Letters, perspectives, and rapid communication on all the aspects of Bioorganic & Organic Chemistry.
Metabolomic Profiling of Spent Biomass Of Marine Microalgae, Chlorella vulgarispriyanka raviraj
OBJECTIVE:
To evaluate the presence of any high value added compounds in the spent biomass of C. vulgaris
To identify the biological activity of the extracted compounds
To evaluate the structure and nature of the compounds using Nuclear Magnetic Resonance Spectroscopy and other analytical techniques.
Development of economically viable methodologies for the simultaneous extraction of by-products from a single set of biomass.
biological activities performed -Total antioxidant capacity, Anti bacterial activity, Anti-tuberculosis activity, Anti proliferative assay
After a pilot scale production of a rhamnolipidic biosurfactant, it is purified using two methods, chemical separation and chromatography. We use UPLC-MS/MS for the characterization of the chemical species.
IOSR Journal of Pharmacy (IOSRPHR), www.iosrphr.org, call for paper, research...iosrphr_editor
IOSR Journal of Pharmacy (IOSRPHR), www.iosrphr.org, call for paper, research paper publishing, where to publish research paper, journal publishing, how to publish research paper, Call for research paper, international journal, publishing a paper, call for paper 2012, journal of pharmacy, how to get a research paper published, publishing a paper, publishing of journal, research and review articles, Pharmacy journal, International Journal of Pharmacy, hard copy of journal, hard copy of certificates, online Submission, where to publish research paper, journal publishing, international journal, publishing a paper
DETERMINATION OF MIANSERIN USING TROPAEOLIN-OOO BY ION PAIR FORMATIONRatnakaram Venkata Nadh
Objective: The present study was aimed at the development of a simple visible spectrophotometric method for the assay of mianserin, a drug used for the treatment of depression.
Methods: The method was developed using tropaeolin-ooo (TPooo) as an ion associative complex forming a chromophore. Developed the chromophore by sequential mixing of aqueous solutions of mianserin, hydrochloric acid, and TPooo. Chromophore was extracted into an organic solvent (chloroform) and absorbance values of organic layers were measured. As per the existing guidelines of an international conference on harmonization (ICH), various parameters of the method were tested for validation.
Results: At the optimized reaction conditions, the formed chromophore (λ max
Conclusion: Due to lack of pre-treatment process for this method, it was simple. All the tested parameters of the method were validated as per ICH guidelines.
Similar to Microchimica Acta Volume 84 issue 5-6 1984 [doi 10.1007_bf01197162] G. A. Milovanovic; M. A. Sekheta -- Kinetic determination of morphine in urine (20)
اليوم الأول دورة تدريبية في معايير السلامة في المختبرات والمعامل الكيميائية [...Sekheta Bros Company
دورة تمهيدية من ثلاثة أيام يخضع لها السيدات والسادة مدراء الفروع والأقسام والعاملين المعنيين بأمور الأمن والصحة والسلامة في مؤسسات القطاع العام، تهتم بتوضيح أهم المفاهيم السائدة اليوم حول سلامة وأمن المختبرات والمنشآت وصحة وسلامة الأفراد العاملين في مجال التحليل الكيميائي والبحث والتطوير ومراقبة الجودة وتشمل:
الإداريين ومساعديهم
إدارة المخاطر والأقسام التابعة لها (في الإدارات المتطورة)
الموارد البشرية العاملة في مختبرات البحث والتطوير ومراقبة الإنتاج والجودة.
العاملين في مجال تخزين ونقل المواد الكيميائية.
العاملين في أقسام الطوارئ وأعمال التمديدات الكهربائية والتركيب والصيانة والإصلاح
Dr mohamad azzam sekheta review in hazards in haccp and iso 1st int symp hama...
Microchimica Acta Volume 84 issue 5-6 1984 [doi 10.1007_bf01197162] G. A. Milovanovic; M. A. Sekheta -- Kinetic determination of morphine in urine
1. MikrochimicaActa [Wien] 1984III, 477--483
9 by Springer-Verlag1984
Chemical Institute, Faculty of Sciences, University of Belgrade
Kinetic Determination of Morphine in Urine""
By
G. A. Milovanovic and M. A. Sekheta
With 1Figure
(Received November 27, 1984)
In view of the medical and social problem caused by the in-
creasing number of drug addicts the determination of opium alka-
loids is of special importance.
Morphine is known as a highly addictive and potent narcotic
but drug users prefer heroin because of its more intense immediate
effect. As heroin is hydrolysed in the organism to morphine the
knowledge of the morphine content of biological fluids and tissues
is indispensable for forensic and therapeutic purposes.
The methods recently used for the determination of morphine
in urine are gas-liquid chromatography1, high-pressure liquid chro-
matography with fluorimetric determination2 and gas chromatog-
raphy-mass spectrometry3.
In this paper a simple and inexpensive kinetic method is pre-
sented, based on the decomposition of the coloured compound
formed by the reaction of hydrogen peroxide with cobalt(II) and
morphine, in the presence of carbonate buffer. There is a definite
concentration range over which the decomposition rate of the com-
pound mentioned is a linear function of the morphine concentration.
* Presented at the first International Symposium on Kinetics in Ana-
lytical Chemistry, C6rdoba, September 27--30, 1983.
31 Mikrochim. Acta 1984 1II/5-6
2. 478 G.A. Milovanovicand M. A.Sekheta:
Experimental
Apparatus
The reaction rate was followed photometrically with a Zeiss Specol by
measuring the solution absorbance at 350 nm in a 5-cm cell, every 15 sec
during the first 3 rain of the reaction. The temperature was kept constant
with an "Ultrathermostat nach HSppler", type NBE (VEB Prfifger~ite-
Werk, Medingen). The pH-measurements were performed by means of a
Radiometer 4C pH-meter.
Reagents
Analytical grade chemicals and redistilled water were used. Concen-
trations of the stock solutions were: hydrogen peroxide (Merck), 9.8 M;
cobalt(II) sulphate (Merck), 1• 10-~ M; morphine hydrochloride (Alkaloid),
1x 10-2 M. Carbonate buffer solution was made by mixing 1M sodium
bicarbonate and 1M sodium carbonate (Merck).
Procedure
The reaction was performed in a special vessel with three com-
partments. The solution of cobalt(II) was measured into one com-
partment, buffer and morphine into the second, and hydrogen per-
oxide and enough water to make a total volume of 25 ml into the
third compartment of the vessel. The vessel was brought to
25_+0.1o C in the thermostat and the reaction was started by mix-
ing all the solutions in the vessel.
Extraction of Free Morphine from Urine Samples
The urine sample (25 ml) was extracted with chloroform-iso-
propyl alcohol (95 : 5) mixture at pH 8.9 (carbonate buffer). After
extraction the mixture was centrifuged and the organic layer was
separated and evaporated. The residue was dissolved in 5 ml of
redistilled water, and analysed for morphine as just described.
Results and Discussion
To establish the optimum conditions for the determination of
morphine, the kinetics of the decomposition of the unstable product
formed between hydrogen peroxide, cobalt(II) and morphine has
been studied.
It was found that the rate of decomposition increases with in-
creasing concentration of morphine and of cobalt(II), and is maximal
when the two components are in 1 : 1 molar ratio; on this basis
3. Kinetic Determination of Morphine in Urine 479
it was concluded that the decomposition rate depends on the cata-
lytically active 1:1 complex which cobalt forms with morphine.
From the results obtained the kinetic equation of the reaction has
been postulated and the apparent rate constant calculated:
da
dt = ~ [Co-morphine] [H20211/4
/~= (0.51 • 0.03) mole-1/4.11/4. min-1.
This kinetic equation is valid for hydrogen peroxide concentra-
tions ranging from 1.2 x 10-~ M to 19.6 x 10-3 M, buffer concentra-
tions ranging from 1.6 x 10-2 M to 5.6 x 10-2 M, and pH=8.9;
T=25 +_0.10C.
From the rate of the reaction at various temperatures in the
range 20--35 o C the activation energy and thermodynamic param-
eters have been calculated and are given in Table L The activation
Table I. The Activation Energy and Thermodynamic Parameters for the Decom-
position of the Compound Formed Between Hydrogen Peroxide, Cobalt(II) and
Morphine
E A H ++ AS ++ A G ++ pK++
(kJ/mole) (kJ/mole) (J.K-l.mole-i) (kJ/mole)
37.8 35.3 - 131 74.5 13.1
energy is lower than that for the decomposition of hydrogen per-
oxide by inorganic ions, which is about 3 46 kJ/mole. It may be con-
cluded that a catalytically active complex, with catalyse-like action
in the decomposition of hydrogen peroxide, is probably formed
between cobalt(II) and morphine.
Table II. Kinetic Determination of Morphine (5 Determinations)
No. Taken Found Relative
standard
(~g/mI) (#g/ml) deviation (%)
5 1.50 1.66 + 0.13 7.8
5 7.50 7.16 __+0.17 2.4
5 12.3 12.07__+0.36 3.0
From the kinetic investigations the optimum conditions for the
determination of morphine have been established: hydrogen per=
oxide, 19.6 x 10 '3 M; carbonate buffer, 4.0 x 10-2 M (pH 8.9),
31"
4. 480 G.A. Milovanovic and M. A. Sekheta:
cobalt(II) sulphate, 4.0 x 10 .5 M. Determination of morphine by
the differential tangent method gives a calibration graph of the
type shown in Fig. 1. The results obtained are given in Table II.
I /t~. j
2
88 "'13" I'2 lg 20 Nor],pg/mt~
0 I 2 3 4 5 [Mor], M- 105
Fig. 1. Calibration graph for the determination of morphine
Initial concentrations: hydrogen peroxide, 19.6x10-aM; carbonate buffer
(pH 8.9), 4.0 x 10-~ M; cobalt(II) sulphate, 4.0 x 10-5 M
Morphine was determined in concentrations ranging from 1.5 to
12.3/~g/ml, with relative standard deviation up to 7.8%.
A study was made of possible interference by the related com-
pounds of the opium alkaloid group as well as by some medical
Table III. Tolerance Ratio for Foreign Substances in the Determination of
Morphine (2 x 10-~ M)
Narcotine interferes
Codeine 1 : I
Thebaine 5 : 1
Dionin 5 : 1
Papaverine 1 : 1
Methadone 10 : 1
Benzoctamine 10 : 1
Valium 10 : 1
Lasdol 2 : 1
drugs. Results presented in Table III show that narcotine interferes
with the determination, whereas papaverine and codeine (when
present at the same concentration as morphine) do not affect the
5. Kinetic Determination of Morphine in Urine 481
accuracy of the determination. Lasdol (lysine-acetylsalicylate +gly-
cine, 9 : 1) does not interfere when present at twice the concentra-
tion of morphine, and thebaine and dionin can be tolerated at
five times the morphine concentration. Other compounds examin-
ed do not interfere even when present at ten times the concentra-
tion of morphine. It may be concluded that other alkaloids of this
group under the same experimental conditions may also form cata-
lytically active complexes with cobalt. Table IV shows how dif-
Table IV. Relative Molar Coefficient of Catalytic Activity, Given as F Values*
of Complexesof Morphine and Its Derivativeswith Cobalt(II)
Compound R1 R2 F value
Morphine - OH - OH 1.000
Codeine - OCH~ - OH 0.065
Dionin - OC2Ha - OH 0.051
Thebaine - OCH~ - OCH3 0.032
* The F value is the quotient of the molar concentrations of a reference
catalyst (in this case the morphine complex) and of the complex considered,
which have the same catalytic activity under identical conditions5.
ferent substituents affect the catalytic activity of the complexes
formed. The catalytic activity decreases on the replacement of the
phenolic hydroxyl group of morphine by a methoxy group in codeine
or an ethoxy group in dionin, and thebaine, which contains two
methoxy groups, exerts the lowest catalytic activity.
Finally, the application of the method to the determination of
morphine in urine has been investigated. The calibration graph was
obtained by spiking blank urine samples with known amounts of
morphine to give concentrations from 0.0 to i5.0 ,ug/ml. It was
found to be linear (-tane=l.808 x 10-9' x Cmo~phin~+0.008) with a
linear regression correlation coefficient of 0.997.
The efficiency of the extraction was checked by extracting
known concentrations of morphine from quintreplicate urine sam-
ples. The mean recovery of morphine was 85%.
The precision and reproducibility were obtained by determining
morphine at three concentration levels (five replicates) (Table V).
The proposed method was also applied to the analysis of five
urine samples from persons suspected of having taken morphine or
heroin. As morphine is present in urine predominantly as the glu-
curonide, for determination of its total morphine content the urine
was hydrolysed before extraction.
6. 482 G.A. Milovanovicand M. A.Sekheta:
To 25 mI of urine 2 ml of concentrated hydrochloric acid were
added and the solution was digested in a boiling water-bath for
30 rain. After cooling, the solution was adjusted to pH 9 with
ammonia solution, then buffered and extracted as described for the
free morphine in urine.
Two of the samples investigated were found to be negative
and were eliminated from further testing.
Table V. ReproducibilityData for Morphine in Urine Samples
(5 Determinations)
Taken Found Relative standard
(,t~g/ml) (#g/ml) deviation (%)
1.50 1.70+0.13 7.6
7.51 8.20+__0.62 7.6
13.53 13.47__+0.57 4.2
In the other samples the free morphine determined ranged from
2.0 to 3.5 #g/ml and the total morphine from 5 to 9#g/ml. The
results obtained were compared with those of HPLC determina-
tions by the method of Jane and Taylorz. The correlation coefficient
of 0.94 for the two sets of data appeared to indicate satisfactory
overall agreement.
If codeine is present it could be expected partly to undergo
O-demethylation to form morphine. However, the main metabolic
pathway is conjugation with glucuronic acid, and free codeine inter-
feres with the determination only when present in concentrations
greater than that of morphine. Other alkaloids, which under the
experimental conditions react like morphine, do not interfere at
low concentrations.
Acknowledgement
The authors are grateful to the Serbian Republic Research Fund
for financial support.
Summary
Kinetic Determination o[ Morphine in Urine
A kinetic method for the determination of morphine in urine is
presented. It is based on the decomposition of the coloured com-
pound formed by the reaction between hydrogen peroxide, cobalt(II)
7. Kinetic Determination of Morphine in Urine 483
and morphine, in alkaline media. The decomposition rate of the
coloured reaction product was followed photometrically. Morphine
was determined in concentrations ranging from 1.5 to 13.5/~g/ml,
and the relative standard deviation was not higher than 8%. The
determination was performed by the differential tangent method.
The effect of related substances from the opium alkaloid group was
also examined. The method determines the free morphine present.
For total morphine the glucuronides have to be hydrolysed by acid
digestion first.
Zusammenfassung
Kinetische Bestimmung yon Morphin in Harn
Die beschriebene Methode beruht auf der Zersetzung des geffirbten
Produktes, das sich aus Morphin, Wasserstoffperoxid und Kobalt(II) in
alkalischer L6sung bildet. Die Geschwindigkeit dieser Zersetzung wurde
photometrisch verfolgt. Morphin wurde im Konzentrationsbereich 1,50--
13,53/~g/ml bestimmt. Die relative Standardabweichung war nicht h6her
als 8%. Die Bestimmung wurde mit der differentiellen Variante der Tan-
gentenmethode ausgefiihrt. Der Einflut~ verwandter Substanzen aus der
Gruppe der Opiumalkaloide wurde untersucht.
References
1 G. R. Nakamura and E. L. Way, Analyt. Chemistry 47, 775 (1975).
2 I. Jane and J. F. Taylor, J. Chromatography 109, 37 (1975).
8 E. J. Cone, C. W. Gorodetzky, S. Y. Yeh, W. D. Darwin, and W. F.
Buchwald, J. Chromatography 230, 57 (1982).
4 N. I. Kobozev, Chosen Works (Russian), Moscow (1978).
5 S. Pantel, Analyt. Chim. Acta 141, 353 (1982).
Correspondence and reprints: Dr. G.A. Milovanovid, Institute of
Chemistry, University of Belgrade, Studentski trg 16, P. O. Box 550, 11001
Belgrade, Yugoslavia.