This study used 1H, 13C, and 11B NMR spectroscopy to analyze the chemical structure, composition, and stability of Fruitex-B, a calcium fructoborate dietary supplement. NMR methods were developed to quantify the mono-complex, di-complex, free borate, and free fructose present in Fruitex-B. The results showed the complex is predominantly a di-ester form with borate coordinated to two fructose molecules. NMR analysis of multiple product batches demonstrated consistency in the relative concentrations of complex components. The study also examined the molecular stability of Fruitex-B when exposed to temperatures from 35-70°C.
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).
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).
UV resonance raman study of streptavidin binding of biotin and 2 iminobiotin ...John Clarkson
J. Clarkson*, D.N. Batchelder & D.A. Smith, “UV Resonance Raman Study of Streptavidin Binding of Biotin and 2-iminobiotin: Comparison with Avidin”, Biopolymers (Biospectoscopy), 62, 307-314, 2001.
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.
Complete NMR Assignment of MogrosidesII A2, II E andIII A1Isolated from Luo H...iosrphr_editor
NMR analysis allowed complete assignments of three known mogrol glycosides, Mogroside IIA2 (1),
II E (2)and IIIA1 (3), isolated from the extracts of Luo Han Guo. Herein, complete 1H and 13C NMR
assignmentsof all threemogrosidesare described based on NMR experiments (1H NMR, 13C NMR, COSY,
HSQC-DEPT, HMBC, NOESY and 1DTOCSY) and mass spectral data.
SIMONA CAVALU_EPR study of non covalent spin labeled serum albumin and haemog...Simona Cavalu
Abstract
Electron Paramagnetic Resonance (EPR) was used to investigate the Tempyo spin label (3-carbamoyl-2,2,5,5-
tetramethyl-3-pyrrolin-1-yloxy) as a report group for the interactions and the conformational changes of lyophilized
bovine serum albumin (BSA) and bovine hemoglobin (BH), as function of pH values in the range 2.5–11. The EPR
spectra are similar with those of other non-covalently spin label porphyrins in frozen solution at very low temperatures.
This behavior indicated a possible spin–spin interaction between the hemic iron and the nitroxide group. The changes
in the EPR spectra as function of the pH are discussed in terms of conformational changes of the proteins. Spectral
simulations and magnetic EPR parameters reveal the following: (i) one single paramagnetic species, with Gaussian
line shape, was used for the best fits of experimental spectra in the case of serum albumin samples; and (ii) a
weighted sum of Lorentzian and Gaussian line shape in the case of hemoglobin samples. The representation of
correlation time vs. pH, reveals a dependence of degree of immobilization of spin label on the conformational changes
of proteins in acidic and basic environment.
Probing the chemistries of flavin ring systems of p hydroxybenzoate hydroxyla...John Clarkson
J. Clarkson, B. Pulfey & P.R. Carey, “Probing the Chemistries of Flavin Ring Systems of p-Hydroxybenzoate Hydroxylase by Raman Difference Spectroscopy”, Biochemistry, 36, 12560-12566, 1997.
SIMONA CAVALU_Rotational Correlation Times of proteinsSimona Cavalu
Noncovalent spin labeled proteins (ovalbumin, bovine serum albumin, hemoglobin, and cytochrome c) were
investigated in order to follow the different type of interactions between the nitroxide radical of 3-carbamoyl-
2,2,5,5-tetramethyl-3-pyrrolin-1-yloxy spin label and functional groups of heme and nonheme proteins as
well as the pH influence on molecular motion of the label with respect to these proteins. EPR spectra were
recorded at room temperature and the computer simulation analysis of spectra was made in order to obtain
the magnetic parameters. Noncovalent labeling of proteins can give valuable information on the magnetic
interaction between the label molecule and the paramagnetic center of the proteins. The relevance of this
interaction can be obtained from line shape analysis: computer simulations for nonheme proteins assume
a Gaussian line shape, whereas for heme proteins, a weighted sum of Lorentzian and Gaussian components
is assumed. In the framework of the “moderate jump diffusion” model for rotational diffusion, the rotational
correlation time is strongly influenced by pH, because of the electrostatic interactions and hydrogen bonding.
UV resonance raman study of streptavidin binding of biotin and 2 iminobiotin ...John Clarkson
J. Clarkson*, D.N. Batchelder & D.A. Smith, “UV Resonance Raman Study of Streptavidin Binding of Biotin and 2-iminobiotin: Comparison with Avidin”, Biopolymers (Biospectoscopy), 62, 307-314, 2001.
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.
Complete NMR Assignment of MogrosidesII A2, II E andIII A1Isolated from Luo H...iosrphr_editor
NMR analysis allowed complete assignments of three known mogrol glycosides, Mogroside IIA2 (1),
II E (2)and IIIA1 (3), isolated from the extracts of Luo Han Guo. Herein, complete 1H and 13C NMR
assignmentsof all threemogrosidesare described based on NMR experiments (1H NMR, 13C NMR, COSY,
HSQC-DEPT, HMBC, NOESY and 1DTOCSY) and mass spectral data.
SIMONA CAVALU_EPR study of non covalent spin labeled serum albumin and haemog...Simona Cavalu
Abstract
Electron Paramagnetic Resonance (EPR) was used to investigate the Tempyo spin label (3-carbamoyl-2,2,5,5-
tetramethyl-3-pyrrolin-1-yloxy) as a report group for the interactions and the conformational changes of lyophilized
bovine serum albumin (BSA) and bovine hemoglobin (BH), as function of pH values in the range 2.5–11. The EPR
spectra are similar with those of other non-covalently spin label porphyrins in frozen solution at very low temperatures.
This behavior indicated a possible spin–spin interaction between the hemic iron and the nitroxide group. The changes
in the EPR spectra as function of the pH are discussed in terms of conformational changes of the proteins. Spectral
simulations and magnetic EPR parameters reveal the following: (i) one single paramagnetic species, with Gaussian
line shape, was used for the best fits of experimental spectra in the case of serum albumin samples; and (ii) a
weighted sum of Lorentzian and Gaussian line shape in the case of hemoglobin samples. The representation of
correlation time vs. pH, reveals a dependence of degree of immobilization of spin label on the conformational changes
of proteins in acidic and basic environment.
Probing the chemistries of flavin ring systems of p hydroxybenzoate hydroxyla...John Clarkson
J. Clarkson, B. Pulfey & P.R. Carey, “Probing the Chemistries of Flavin Ring Systems of p-Hydroxybenzoate Hydroxylase by Raman Difference Spectroscopy”, Biochemistry, 36, 12560-12566, 1997.
SIMONA CAVALU_Rotational Correlation Times of proteinsSimona Cavalu
Noncovalent spin labeled proteins (ovalbumin, bovine serum albumin, hemoglobin, and cytochrome c) were
investigated in order to follow the different type of interactions between the nitroxide radical of 3-carbamoyl-
2,2,5,5-tetramethyl-3-pyrrolin-1-yloxy spin label and functional groups of heme and nonheme proteins as
well as the pH influence on molecular motion of the label with respect to these proteins. EPR spectra were
recorded at room temperature and the computer simulation analysis of spectra was made in order to obtain
the magnetic parameters. Noncovalent labeling of proteins can give valuable information on the magnetic
interaction between the label molecule and the paramagnetic center of the proteins. The relevance of this
interaction can be obtained from line shape analysis: computer simulations for nonheme proteins assume
a Gaussian line shape, whereas for heme proteins, a weighted sum of Lorentzian and Gaussian components
is assumed. In the framework of the “moderate jump diffusion” model for rotational diffusion, the rotational
correlation time is strongly influenced by pH, because of the electrostatic interactions and hydrogen bonding.
Penicillin, one of the first and still one of the most widely used antibiotic agents, is derived from the penicillium mold. In 1928 Scottish bacteriologist alexander fleming in a contaminated green mold penicillium notatum. He isolated the mold, grew it in a fluid medium, and found that it produced a substance capable of killing many of the common bacteria that infect humans. Australian pathologist howard florey and British biochemist ernst Boris chain isolated and purified penicillin in the late 1930s, and by 1941 an injectable form of the drug was available for therapeutic use.
Penicillin's are beta lactam antibiotics and characterized by three fundamental structural requirements
The fused beta-lactam and thiazolidine ring structure.
free carboxylic acid group.
And one or more substituted acylamino side chain.
Penam nucleus: 7-oxo-l-thia-4-azabicyclo [3.2.0] heptane
Absolute configuration: 3-S, 5-R, 6-R.
Instrumental methods of characterization:
FTIR
MASS
C13-NMR
1H-NMR
FTIR: -
Penicillin G molecule and its IR spectra in D2 O and in DMSO. Spectra are characterized by the presence of three intense bands.
β- lactam CO stretching observe at 1761 cm-1 in D2O and 1762 cm-1 in DMSO solution.
Amide group is observe at 1640 cm-1 in D2O and 1674 cm-1 in DMSO solution.
Asymmetric stretching of carboxylate group is observe at 1601 cm-1 in D20 and 1615 cm-1 in DMSO solution.
A large red shift of amide , out of the frequency window, is observed upon proton exchange in DMSO.
Collision-Induced Dissociation (CID) technique
MASS:-
A high-resolution, hybrid tandem mass spectrometer was used to obtain CID spectra. The CID spectra were acquired by:
Mass selecting the precursor ions using the first mass spectrometer.
Injecting the ions into the first quadrupole (collision cell) where they undergo CID.
Mass-analyzing the fragment ions produced using the second quadrupole.
Argon was used as the collision gas, and the pressure in the collision cell was adjusted to attenuate the precursor ion intensity to 20-50% of the original intensity. The collision energy of the ions ranged from 160 to 180 eV. The mass spectra shown abundant fragmentations at m/z 160 and m/z 176 that were reported to arise from cleavage of the β-lactam ring.
protonated benzyl penicillin exhibits abundant fragment ions at m/z 160, m/z 176, m/z 217, m/z 128, and m/z 289. The most abundant CID fragment at m/z 160 and the molecular ion peak was observed at m/z 334.
C13-NMR: -
The four sp3 ring carbons give rise to resonances in the decreasing chemical shift order C-3, C-5, C-2 and C-6.
Chemical shift for C-2 is 64.9 ppm and the substituents attached with it are α-methyl 27.0 ppm and β-methyl 31.4 ppm. Chemical shift for C-3 is 73.6 ppm and 174.5 ppm for carboxylate functions (reflecting the smaller de-shielding influence of COOH over that of COO-). The chemic shift for C-5 is 67.2 ppm. The chemic shift for C-6 is 58.4 ppm.
The lactam group shows its chemical shift at 175.0 ppm
Amino group
DFT Calculations, FT Raman, FTIR Spectra and Vibrational Assignment of 2 amin...ijtsrd
The experimental FT IR and FT Raman spectra of the molecule 2 amino 5 bromobenzoic acid has been recorded and analyzed in the region 4000-400 cm 1 and 3500-50 cm 1, respectively. 2 amino 5 bromobenzoic acid, a single crystal, belongs to the amino acid group. The observed bands were interpreted with the aid of normal coordinate analysis and force field calculations. Molecular structure, vibrational wavenumbers and complete vibrational analysis of 2 amino 5 bromobenzoic acid is performed by combining the experimental and theoretical information based on density functional theory DFT using B3LYP functional theory DFT using B3LYP functional with 6 311 G and 6 3 G basis sets. The complete assignments were performed on the basis of the potential energy distribution PED of the vibrational modes, calculated with scaled quantum mechanical SQM method. The molecular structure and vibrational frequencies, infrared intensities and Raman scattering actives have been calculated frequency showed the best agreement with experimental results. The molecule has been studied for optical properties. The quantitative analysis on the molecule had been carried out using Fourier transform infrared FTIR and Fourier transforms Raman FT Raman spectral measurements. FT IR and FT Raman analysis were carried out in an integral approach. The normalized frequencies were observed with scaled values and were compared with experimental FT IR and FT Raman spectra. Most of the modes have wavenumbers in the expected range and the error obtained was very rare in general. The optimized geometric parameters bond lengths and bond angles were given and are in agreement with the corresponding experimental values. The biological activity of 2 amino 5 bromo benzoic acid has been predicted based on these values. The Fourier transform Raman and infrared spectra of 2 amino 5 bromo benzoic acid has been recorded and analyzed2. Virender Kumar | Meenu Sharma | Lokesh Sharma "DFT Calculations, FT Raman, FTIR Spectra and Vibrational Assignment of 2-amino 5-bromobenzoic Acid" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-6 | Issue-3 , April 2022, URL: https://www.ijtsrd.com/papers/ijtsrd49568.pdf Paper URL: https://www.ijtsrd.com/chemistry/other/49568/dft-calculations-ft-raman-ftir-spectra-and-vibrational-assignment-of-2amino-5bromobenzoic-acid/virender-kumar
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.
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
Micellar Effect On Dephosphorylation Of Bis-4-Chloro-3,5-Dimethylphenylphosph...IOSR Journals
The rate enhancement depends on the hydrophobicity of the nucleophile. The micellar catalyzed reaction between bis-4-chloro-3,5-dimethylphenylphosphate ester and hydroxide or hydroperoxide anions has been examined in buffered medium (pH 8-10). First order rate constant (Kψ) for the reaction of hydroxide ion with bis-4-CDMPP go through maxima with the increasing concentration of cetyltrimethylammoniumbromide (CTABr). Micelles of CTABr very effective catalyst to the reactions of phosphate diesters. Rate constants measured with OH2- ions are approximately twice and thrice than that of OH- ions in presence of CTABr.
Micellar Effect On Dephosphorylation Of Bis-4-Chloro-3,5-Dimethylphenylphosph...IOSR Journals
The rate enhancement depends on the hydrophobicity of the nucleophile. The micellar catalyzed reaction between bis-4-chloro-3,5-dimethylphenylphosphate ester and hydroxide or hydroperoxide anions has been examined in buffered medium (pH 8-10). First order rate constant (Kψ) for the reaction of hydroxide ion with bis-4-CDMPP go through maxima with the increasing concentration of cetyltrimethylammoniumbromide (CTABr). Micelles of CTABr very effective catalyst to the reactions of phosphate diesters. Rate constants measured with OH2- ions are approximately twice and thrice than that of OH- ions in presence of CTABr.
ACETYLATION OF BENZYLIC ALCOHOLS OVER BiFeO3 (BFO), Bi0.86Sm0.07Eu0.07FeO3 (B...EDITOR IJCRCPS
BiFeO3 (BFO), Bi0.86Sm0.07Eu0.07FeO3 (BSEFO), and Bi0.86Sm0.07Cd0.07FeO3 (BSCFO) nanopowders were prepared by the sol-gel
combustion method and the catalytic performances were evaluated in acetylation reaction of benzyl alcohol. The physical chemical
properties of catalysts were characterized by using XRD, FT-IR, scanning electron microscope (SEM), EDX and BET surface.
Efficient acetylation of benzyl alcohol was carried out over all the nano powders using acetyl chloride/ acetonitrile at room
temperature. Among the nanopowders, BSCFO showed the highest catalytic performance and the yield of benzyl acetate was 89,
45, and 69 percent over BSCFO, BFO, and BSEFO, respectively. Partial substitution of Sm-Eu and Sm-Cd in bismuth ferrite
improved the catalytic performance and increased the specific surface area of the catalysts. A direct relationship was resulted
between catalytic performance and surface of catalysts, where BSCFO with the highest surface area (111m2/g) exhibited the
superior catalytic performance. The quantitative yield for acetate product was also resulted for acetylation of p-methyl benzyl
alcohol, p-nitro benzyl alcohol and p-chloro benzyl alcohol on BSCFO. The catalysts showed good reusability in the process. The
study confirmed the catalysts could be promising catalyst for acetylation of alcohols.
Keywords: Europium, Samarium, Bismuth ferrites, nano perovskite, doping, Acetylation, benzylic alcohols.
Similar to Multinuclear liquid and solid-state NMR of Fructoborate complex (20)
1H qNMR of EPA and DHA Omega-3 Fatty Acid: PLS-Regression Models Obtained at ...John Edwards
Application of 60 and 300 MHz 1H NMR with PLS-Regression to correlate 1H NMR spectra variability with GC derived EPA and DHA Content in a fish oil nutritional supplement manufacturing process.
Benchtop NMR of Adulterants in Sexual Enhancement and Weight-Loss Supplements...John Edwards
NMR utilization as screening tool for illegal adulteration of herbal supplements with pharmaceutical doses of viagra, tadalafil and their various analogs.
Presented at SMASH 2014, Atlanta, September 2014
Hamdard Laboratories (India), is a Unani pharmaceutical company in India (following the independence of India from Britain, "Hamdard" Unani branches were established in Bangladesh (erstwhile East Pakistan) and Pakistan). It was established in 1906 by Hakeem Hafiz Abdul Majeed in Delhi, and became
a waqf (non-profitable trust) in 1948. It is associated with Hamdard Foundation, a charitable educational trust.
Hamdard' is a compound word derived from Persian, which combines the words 'hum' (used in the sense of 'companion') and 'dard' (meaning 'pain'). 'Hamdard' thus means 'a companion in pain' and 'sympathizer in suffering'.
The goals of Hamdard were lofty; easing the suffering of the sick with healing herbs. With a simple tenet that no one has ever become poor by giving, Hakeem Abdul Majeed let the whole world find compassion in him.
They had always maintained that working in old, traditional ways would not be entirely fruitful. A broader outlook was essential for a continued and meaningful existence. their effective team at Hamdard helped the system gain its pride of place and thus they made an entry into an expansive world of discovery and research.
Hamdard Laboratories was founded in 1906 in Delhi by Hakeem Hafiz Abdul Majeed and Ansarullah Tabani, a Unani practitioner. The name Hamdard means "companion in suffering" in Urdu language.(itself borrowed from Persian) Hakim Hafiz Abdul Majeed was born in Pilibhit City UP, India in 1883 to Sheikh Rahim Bakhsh. He is said to have learnt the complete Quran Sharif by heart. He also studied the origin of Urdu and Persian languages. Subsequently, he acquired the highest degree in the unani system of medicine.
Hakim Hafiz Abdul Majeed got in touch with Hakim Zamal Khan, who had a keen interest in herbs and was famous for identifying medicinal plants. Having consulted with his wife, Abdul Majeed set up a herbal shop at Hauz Qazi in Delhi in 1906 and started to produce herbal medicine there. In 1920 the small herbal shop turned into a full-fledged production house.
Hamdard Foundation was created in 1964 to disburse the profits of the company to promote the interests of the society. All the profits of the company go to the foundation.
After Abdul Majeed's death, his son Hakeem Abdul Hameed took over the administration of Hamdard Laboratories at the age of fourteen.
Even with humble beginnings, the goals of Hamdard were lofty; easing the suffering of the sick with healing herbs. With a simple tenet that no one has ever become poor by giving, Hakeem Abdul Majeed let the whole world find compassion in him. Unfortunately, he passed away quite early but his wife, Rabia Begum, with the support of her son, Hakeem Abdul Hameed, not only kept the institution in existence but also expanded it. As he grew up, Hakeem Abdul Hameed took on all responsibilities. After helping with his younger brother's upbringing and education, he included him in running the institution. Both brothers Hakeem Abdul Hameed and Hakim Mohammed
Hotel management involves overseeing all aspects of a hotel's operations to ensure smooth functioning and exceptional guest experiences. This multifaceted role includes tasks such as managing staff, handling reservations, maintaining facilities, overseeing finances, and implementing marketing strategies to attract guests. Effective hotel management requires strong leadership, communication, organizational, and problem-solving skills to navigate the complexities of the hospitality industry and ensure guest satisfaction while maximizing profitability.
Vietnam Mushroom Market Growth, Demand and Challenges of the Key Industry Pla...IMARC Group
The Vietnam mushroom market size is projected to exhibit a growth rate (CAGR) of 6.52% during 2024-2032.
More Info:- https://www.imarcgroup.com/vietnam-mushroom-market
Vietnam Mushroom Market Growth, Demand and Challenges of the Key Industry Pla...
Multinuclear liquid and solid-state NMR of Fructoborate complex
1. Solid-State 1H, 13C and 11B qNMR Analysis of Fruitex-B® – A Calcium Fructoborate
Complex: Chemical Structure, Identification, Quantitative Analysis and Stability Study
Boris Nemzer1,3, John Hunter1, John C. Edwards2
1. VDF Futureceuticals Inc., Momence, IL USA 2. Process NMR Associates LLC, Danbury, CT USA
3. University of Illinois at Urbana-Champaign, Urbana, IL USA
Overview of Study
Fruitex-B® (FrxB) is a patented plant mineral complex that is marketed as a nutritional
supplement with potential health benefits for conditions linked to inflammation such as bone,
joint and cardiovascular conditions. The product is a calcium fructoborate complex formed by
the reaction of boric acid with fructose and calcium carbonate. Liquid and solid-state 13C and
11B NMR was utilized to establish a baseline for product quality and to establish a robust testing
method for both identification and quantification of the mono-complex and di-complex present
in the product, as well as free borate and free fructose that is present in the finished product. A
quantitative 11B NMR method was developed to quantify the amount of FrxB present in
products where it was tableted or capsuled with magnesium stearate or maltodextrin. A
quantitative 13C NMR method was developed to quantify free fructose content in the complex.
Finally, an NMR based product stability study was performed to monitor molecular level
stability of the complex at temperature ranging from 35-70oC with exposure lasting from 2-18
hours.
Background
Boron is naturally occurring and essential element for plant and animal life. There are many
different biological compounds that can form complexes with boron. Compounds capable of
complexing with boric acid include sugar alcohols, pyranose and furanose sugars or their
derivatives, organic acids. Boric acid forms esters and complexes with a wide variety of mono-,
di-, and polyhydroxy compounds (Woods, 1996). One of the most stable esters of boric acid are
complexes where boric acid is a bridge between two carbohydrate molecules, e.g. fructose-boron-
fructose. The examination of boron complexation in plants and plant extracts by 11B
NMR demonstrated the majority of the boron was associated with a diester complexes of diols
and hydroxycarboxylic acids in radish and apple respectively (Matsunaga & Nagata, 1995). The
authors made conclusion that fructose is the most significant boron complexing molecule. Later
these hypotheses were verified (Brown & Shelp, 1997 and Hu, et al., 1997) after successful
isolation and full characterization of soluble boron complexes from higher plans. Calcium
fructoborate (CF) is most commonly found in fresh fruits and vegetables. As a dietary
supplement it is manufacture by VDF FutureCeuticals, Inc under the commercial name Fruitex-
B®(FrxB) based on the US patent 5.962.049 (Miljkovic, 1999). The characterization of this
complex has been reported previously (Rotaru et al., 2010) using thermal analysis, X-rays
diffraction, ICP-MS, Raman spectrometry techniques.
In this study we investigate molecular composition, stability and identification of FrxB used as a
dietary supplement for human nutrition (Dinca & Scorei, 2013, and Reyes-Izquierdo et al., 2012
) using liquid- and solid-state 11B and 13C NMR.
Materials and Method
Materials
Fruitex-B ® calcium fructobotrate (FrxB) was manufactured and provided by Futureceuticals,
Momence IL, USA according to the Miljkovic patent (US 5, 962,049).
All NMR analysis was performed in D2O or H2O/D2O. D2O (99.9%D) was obtained from
Cambridge Isotopes Laboratories, Tewksbury MA, USA.
Samples were observed directly after they were received, after they had been thermally
treated in a Duratech TCON dry bath system (capable of holding temperatures to +/- 0.1 oC), or
as calibration standards which were made by mixing accurately weighed samples of FrxB with
magnesium stearate or maltodextrin. Samples for solid-state NMR were weighed to the nearest
0.1 mg on a Sartorius GD-503-NTEP microbalance after they were packed into the MAS rotor.
NMR Spectroscopy
Liquid-state 11B, 13C, and 1H NMR was performed on a Varian Mercury 300MVX NMR spectrometer
equipped with a 5mm Varian ATB Probe at a resonance frequencies of 96.14 MHz (11B), 75.36 MHz
(13C) and 299.67 MHz (1H), respectively. 11B spectra were acquired with a 45 degree tip angle pulse
width, a relaxation delays of 0.2 seconds, an acquisition time of 80 ms with 8K points acquired with a
spectral width of 100 kHz, and 1024 pulses were averaged. The data was zero filled to 65K points. The
13C NMR was acquired with a 30 degree tip angle pulse width, a 5 seconds relaxation delay, 0.96
second acquisition time, with 24K points acquired with a spectral width of 25 kHz, and 10-12,000
pulses were averaged. The data was zero filled to 131K points. The 1H NMR spectra were obtained
with a 30 degree pulse angle, a 2 second relaxation delay, a 4.448 second acquisition time, with 32K
points acquired over a spectral width of 7.2 kHz, 128 pulses were averaged. The data was zero-filled
to 131K points. The data was acquired in a quantitative manner with inverse gated decoupling of
protons during the acquisition of the 11B and 13C experiments. All samples were dissolved in D2O
(Cambridge Isotope Laboratories). No pH adjustments were performed on the samples after
dissolution.
Solid-State 13C (50.30 MHz) and 11B (64.17 MHz) NMR spectra were obtained on a Varian UnityPlus-
200 NMR spectrometer equipped with a Doty Scientific 7mm Supersonic CP-MAS probe. Magic angle
spinning (MAS) speeds of around 6 kHz were employed. The 13C NMR data was acquired using cross
polarization which prepares the magnetization on the protons initially and then transfers the spin
locked magnetization to the 13C nuclei. The advantage of this experiment is the fact that the
experiment is performed at the spin-lattice relaxation rate (T1) of protons in the sample which is
considerably shorter than the T1 of 13C nuclei in the same sample. Thus, one obtains a significant
enhancement of the 13C signal from the polarization transfer and can pulse at a shorter pulse-repetition
rate. The 13C experiment on calcium fructoborate complex were acquired with an 8 second
relaxation rate, and acquisition time of 25.6 ms, with 1K points being acquired over a spectral with of
40 kHz, and 4096 pulses were averaged. The exception to these acquisition parameters were those
used for pure crystalline fructose. The 11B NMR spectra were acquired with MAS and with the sample
remaining static in the NMR probe. The experiments were acquired with a central transition selective
pulse width, a 0.2 second relaxation time, with 1K points being acquired in an acquisition time of 10.2
ms, and with a spectral width of 100 kHz.
Structure of Fruitex-B Fructoborate Complex
1H NMR
The spectrum above shows the comparison of the 1H NMR spectra of pure D-fructose and
Fruitex-B fructoborate complex (FrxB). Free fructose is observed as well as the mono-ester/
di-ester complex in the FrxB sample, but the overall spectrum is complicated and no
assignments have been made due to its complexity. However, the 1H NMR spectrum can be
used to quantify the presence of FrxB in maltodextrin and other soluble adulterants.
Figure 3: 13C NMR of D-Fructose compared to several production lots of Fruitex-B
Figure 4: 13C NMR of Fruitext-B Fructoborate complex with assignments
Figure 2: 13C NMR spectrum of D-fructose with assignments (Consonni and Cagliani, 2008,
Mazzoni et al, 1997).
Fructose
Tautomer Mole%
b-FP 71.1
b-FF 22.4
a-FF 6.5
D-Fructose Tautomer Type
13C NMR - Anomeric Region
Fructose Tautomer
Lot 1
(mole%)
Lot 2
(mole%)
Lot 3
(mole%)
b-FP (Free Fructose 15.5 13.9 15.7
b-FF (Free Fructose) 7.9 7.1 8.5
a-FF (Free Fructose) 1.6 2.0 1.8
a-FF-B-Complex 48.3 50.5 47.6
b-FF-B-Complex 17.0 16.3 16.3
b-FP-B-Complex 9.6 10.2 10.0
Relative Molar Concentrations found in Different Lots of
Calcium Fructoborate Complex
Unreacted fructose (free fructose) is observed in the complex mixture and a 3-7 ppm
downfield shift of the fructose resonances is observed for the carbons coordinated to
borate in the fructoborate complex. The change of relative signal intensities in the
regions of the spectrum that are associated with furanose tautomers, indicates that the
tautomer distribution of the complex strongly favors the reaction of borate with the
fructofuranose (FF) form.
We have also used 13C NMR of the anomeric (C-2) region to determine the consistency
of the FrxB product from one batch to the next (see Figure 4).
The fact that the FrxB complex peaks are multi-component in all cases leads to the
conclusion that the borate reacts with multiple hydroxyls with OH condensation
reactions occurring predominantly on the C-1/C-2 as well as on the C-3/C-4 of the FF
forms. It is expected from the mole ratios utilized in the synthesis of the FrxB complex
that the complex is predominantly the di-ester form BL2
- form (one borate coordinated
to two fructose molecules) with the minor constituent being the monoester form (BL-),
as well as some free/unreacted borate.
13C NMR
Figure 1
Figure 5: 11B NMR of boric acid and 3 lots of Fruitex-B
11B NMR: Liquid-state 11B NMR has been utilized often in the study of biomedical
applications of boron (Bendel, 2005). In this study liquid-state 11B NMR was obtained
on order to observe the FrxB complex from the perspective of the boron chemistry.
Previous research has identified that three basic types of boron are observed in
aqueous solutions of FrxB. Free boric acid is observed at 0 ppm, the di-ester complex
(BL2
-) is observed at -9 ppm, and the mono-ester (BL-) complex is observed at -13
ppm (Makkee et al., 1985, Reyes-Izquierdo et al., 2012, and Smith et al., 1998). The
relative molar concentrations of these three types of boron were found to be
approximately 5%, 85%, and 10%, respectively. Figure 5 shows the liquid-state 11B
NMR spectra of boric acid and 3 batches of FrxB.
Fructose
BL-BL2
-
The above structures represent the mixture of component fructoborate
Species present in the Fruitex-B product
Liquid-state 13C NMR Thermal Stability of Fruitex-B Product
Solid-state 13C NMR
Liquid-state 11B NMR
Solid-state 11B NMR
Static Powder Lineshape
The thermal stability of the Fruitex-B product was tested
by exposing the product to temperatures of 35, 50 and
70oC for between 2 and 18 hours. These figures show the
results obtained by liquid and solid-state NMR experiments.
The samples showed no observable changes over the
course of the stability test. Free borate, BL- and BL2- were
calculated from the 11B NMR and the 13C NMR was utilized
to calculate free fructose and the a-FF/b-ff/b-FP
component complex concentrations.
Quantitation of Fruitex-B in the Presence of Excipients and Adulterants
y = 20525x - 44667
R² = 0.9966
0
500000
1000000
1500000
2000000
2500000
0 20 40 60 80 100
11B Signal Intensity
FruitexB Wt%
11B NMR - Total 11B Signal Vs
FruitexB Wt%
y = 19.759x - 12.505
R² = 0.9971
0
20
40
60
80
100
0.0 1.0 2.0 3.0 4.0 5.0 6.0
Calcium Fructoborate Content (Wt%)
Di-Ester/Mono-Ester Ratio
Change in Di-Complex/Mono-Complex
Ratio with Increasing Adulteration
with Maltodextrin
Liquid-state 11B qNMR – Calibration of Fruitex-B in Maltodextrin
Calibration of Wt% Fruitex-B from
absolute 11B NMR signal intensity
Calibration of Wt% Fruitex-B from
Change in Di-ester/mono-ester ration
With changing maltodextrin content
Solid-State 11B MAS NMR – Fruitex-B Wt% in Maltodextrin and Magnesium Stearate
Stearate Fruitex-B
Stearate
Liquid-state 1H NMR – Fruitex-B content in Maltodextrin
Calibration obtained from
reduction of maltodextrin
anomeric proton peak with
increasing Fruitex-B content.
Quantitation is possible by 1H, 13C, 11B NMR with either liquid or solid-state experiments where appropriate.
References (Contd)
Makkee, M., Keibook, A.P.G., van Bekkum, H. (1985). Studies on borate esters III. Borate esters of D-mannitol, D-glucitol, D-fructose, D-glucose in water. Recl. Trav. Chim. Pays-Bas, 104, 230-
235.
Matsunaga, T., and Nagata, T. (1995). In vivo 11B NMR observation of plant tissue. Anal. Sci. 11, 889-892.
Mazzoni, V., Bradesi, P., Tomi, F., Casanova, J. (1997). Direct qualitative and quantitative analysis of carbohydrate mixtures using 13C NMR spectroscopy: application to honey. Magn. Reson.
Chem. 35, S81-90.
Miljkovic, D. (1999) US patent 5.962.049 (issued October 5, 1999)
Pauli, G.F., Gödecke, T., Jaki, B.U., Lankin, D.C. (2012). Quantitative 1H NMR. Development and Potential of an Analytical Method: An Update. J. Nat. Prod., 75(4), 834–851.
Reyes-Izquierdo, T., Nemzer, B., Gonzalez, A.E., Zhou, Q., Argumedo, R., Shu, C., Pietrzkowski, Z. (2012). Short-term intake of Calcium Fructoborate improves WOMAC and McGill scores and
beneficially
modulates biomarkers associated with knee osteoarthritis: A pilot clinical double-blinded placebo-controlled study. Am. J. Biomed. Sci., 4(2) 111-122.
Rotaru, P., Scorei, R., Harabor, A., Dumitru, M. (2010) Thermal analysis of calcium fructoborate sample. Thermochem. Acta, 506, 1-2, 8-13.
Smith, B.M., Owens, J.L., Bowman, C.N., Todd, P. (1998). Thermodynamics of borate ester formation by three readily grafted carbohydrates. Carbohydr. Res., 308, 173-179.
Wagner, C.C., Ferraresi Curotto, V., Pis Diez, R., Baran, E.J. (2008). Experimental and Theoretical Studies of Calcium Fructoborate. Biol. Trace Elem. Res., 122, 64-72.
Woods, W.G. (1996). Review of possible boron speciation relating to its essentiality. J. Trace Elem Exp. Med. 9, 153-163.
References
Bendel, P. (2005). Biomedical applications of 10B and 11B NMR. NMR Biomed., 19, 74-82.
Brown, P.H. and Shelp, B.J. (1997). Boron mobility in plants. Plant Soil 193, 85-101.
Caytan, E., Botosoa, E.P., Silvestre, V., Robins, R.J., Akoka, S., Remaud, G.S. (2007). Accurate Quantitative 13C NMR
Spectroscopy: Repeatability over Time of Site-Specific 13C Isotope Ratio Determination. Anal. Chem., 79 (21), 8266–
8269.
Consonni, R., Cagliani, L.R. (2008). Geographical Characterization of Polyfloral and Acacia Honeys by Nuclear Magnetic
Resonance and Chemometrics. J. Agric. Food Chem., 56, 6873-6880.
Dinca, L., Scorei, R. (2013). Boron in Human Nutrition and its Regulations Use. J. Nutr. Ther., 2, 22-29.
Hu, H., Penn, S.C., Lebrilla, C.B. and Brown, P.H. (1997). Isolation and characterization of soluble boron complexes in
higher plants. Plant Physiol. 113, 649-655.
Conclusion
Multinuclear liquid and solid-state NMR spectroscopy was capable
of defining the fructoborate structure, and allowed identification
of the pure complex as well as Quantitation of Fruitex-B product in
the presence of excipient and adulterants.