Dr. Charbell Miguel Haddad Kury
Professor, Department of Pediatrics and Biochemistry, Medicine School of Campos Dos Goytacazes, Universidade Federal do Rio de Janeiro, Brazil.

ISBN 978-93-90888-00-9 (Print)
ISBN 978-93-90768-97-4 (eBook)
DOI: 10.9734/bpi/cacb/v4

This book covers key areas of chemistry and biochemistry. The contributions by the authors include monomers, multistage chemical reaction, low frequency mechanical action, digestibility, gut microflora, prebiotic assay, thermolysin, antioxidative, antihypertensive, protein hydrolysate, gas chromatography, mass spectrometers, ion mobility spectrometers, miniaturization, mobile instruments, absorbance ratio, iso?absorptive point, regression coefficient, anticonvulsant activity, strychnine induced model, thiosemicarbazide induced model, 4-amino pyridine induced model, neurotoxicity screening, insulin resistance, metabolic syndrome, deuterium-depleted water, deuterium depletion, endemic plants, polysaccharides, saponification, medicinal- plant -based drugs, phytochemicals, inbred lines, hybrids, acid soil, analgesic activity, anti-inflammatory activity, antioxidant, anticandidal activity, blood ionized calcium, urinary tract infections, uropathogenic Escherichia coli, A-type proanthocyanidins, phenolic metabolites, antiadhesive activity, probiotics, oxidative stability, nuclear magnetic resonance, direct immersion SPMEfree fatty acids, hydroperoxides, monohydroxy-conjugated dienes, epoxides, di- or poly-hydroxy-compounds. This book contains various materials suitable for students, researchers and academicians in the field of chemistry and biochemistry.


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The effect of periodic mechanical action on the kinetics and the mechanism of liquid-phase reactions were studied. This is the first stage of extensive research in which mathematical modeling of simple kinetic schemes taking into account the formation of associates is planned to carry out. In case of a multistage reaction existence of several quasi stationary states and oscillations of concentration of reagent is possible.

At a research the method of mathematical model operation of temporary instability of reaction found two types of oscillations of concentration of intermediate (harmonic and relaxation) characterized significantly by the differing times With increasing frequency and amplitude of low-frequency sonic waves (up to some critical value), we observed the disappearance of trimers and concomitant cessation of the reaction. Model operation of process showed a possibility of speed control of reaction at external lowfrequency acoustic influence.

Evaluation of Glycan Compound from Swiftlet’s Edible Nest (Aerodramus fuciphagus) as Potential Prebiotic Material

Nur ‘Aliah Daud, Salma Mohamad Yusop, Seng Joe Lim, Abdul Salam Babji

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 7-15

Swiftlet’s edible nest, the white nest which produced by Aerodramus fuciphagus species has been consumed for centuries and is well-known as one of the nutritious and medicinal delicacy in curing many illnesses. Among the benefits of consuming the bird nest, the benefit towards the human gut health has not yet been well explored. A part of the major factor shaping the composition and physiology of gut microbiota is the availability of glycans, most of which cannot be digested by enzymes encoded in the human genome. Gut microbiota can digest the indigestible glycans and further produce short-chain fatty acids, which serve as nutrients for colonocytes and other gut epithelial cells. The improvement of growth of the gut microbiota may contribute to strengthening immune system of the host. Mucus glycoproteins of the swiftlet’s edible nest are mainly constituted of numerous glycans which attached to the bird nest protein backbone. In this study, the glycans from the edible bird nest (EBN) were extracted from the native glycoprotein by the reaction of alkaline hydrolysis. The extracted glycan was purified using gel chromatography and the glycan’s purity was tested using FTIR spectroscopy. Molecular weight of the glycan was determined using gel electrophoresis. The extracted glycan was further tested for its ability to resist human digestion system to be a potential prebiotic compound. Results showed that the glycan of EBN with molecular weight in the range of 21.2 kDa to 66.6 kDa can resist human digestion, thus have a potential to be delivered into the gut environment and act as prebiotic. The potential prebiotic activity can be further study using a mixed culture of bacteria and/or selected probiotic strains.

Determination of Antioxidative and ACE-inhibitory Activities from Red Tilapia Protein Hydrolysates Generated by Alcalase and Thermolysin

Nur ‘Aliah Daud, Abdul Salam Babji, Salma Mohamad Yusop

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 16-23

Production of fish protein hydrolysates has been widely studied in order to explore the potential of fish protein and industrial potential of fish wastes as functional bioactive peptides. In this research, the activities of antioxidative and angiotensin converting enzyme (ACE)-inhibitory peptides derived from freshwater fish of Red Tilapia protein (Oreochromis niloticus) using alcalase and thermolysin enzymes were evaluated. The enzymatic hydrolysis process was performed for 4 hours at 37°C, pH 7.4. After two hours of hydrolysis with thermolysin and alcalase enzymes resulted in degree of hydrolysis of 76.29% and 63.49%, respectively. Hydrolysates obtained after 1 hour and 2 hours hydrolysis were chosen for further study on the bioactive activities. The result showed that the thermolysin enzyme yielded higher antioxidant activities than alcalase enzyme based on ABTS radical scavenging activity and reducing power assays. For ACE-inhibitory assay, thermolysin enzyme yielded higher ACE-inhibitory activities after 1 hour hydrolysis while alcalase enzyme yielded higher inhibition activities after 2 hours hydrolysis. Chosen cut-off interval of hydrolysates showed that the thermolysin hydrolysates have strong inhibition effects towards ACE enzyme than alcalase hydrolysates. This study had demonstrated that the hydrolysates produced from Red Tilapia protein may contribute as a health promoting ingredient, to reduce free radical in human system as well as improving the shelf-life of functional food products. The hydrolysates may also be used as an ingredient which have the antihypertensive effects towards the mild hypertension patients.

Developments in Miniaturization of Gas Chromatography

Waldemar Wardencki, Z. Witkiewicz

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 24-42

The tendency towards miniaturization is visible in many areas of the wider word. The downsizing of many everyday used items (e.g. wristwatches, i-phones) is possible mainly due to the avaailability of newly developed technology. The same trend is also observed in many fields of analytical chemistry. It is related with the contemporary movement in analytics to perform analysis at the place where analytes are present. The miniaturization of conventional analytical instruments can provide many attractive features, e. g. reduced space requirements, better flexibility and faster results, without sacrifice in performance, and perhaps, with improved performance.

The chapter provides a general overview of the progress and current status of miniaturization of one of the most widely used analytical technique – gas chromatography (GC). First, the general characteristics of miniaturized gas chromatographs and a brief history of the miniaturized GC systems is presented. Next, the technologies used for fabrication of miniaturized gas chromatographs and the developments of a key components of micro gas chromatographs (?GC) systems, i.e. injectors/preconcentrators, columns and detectors, based on microelectromechanical technology (MEMS), are discussed. In the last section, the hybrid miniaturized systems, with emphasis on coupling of gas chromatographs with mass and ion mobility spectrometers (MS and IMS), are discussed. Finally, the examples of selected applications of gas chromatographs with MS (GC-MS) and IMS (GC-IMS) are given.

Two simple, sensitive, accurate, precise, rapid and economical methods were developed for the estimation of Ofloxacin and Tinidazole from combined tablet dosage form. First method is based simultaneous equation and second method is based on ?analysis (absorbance ratio method). Ofloxacin and Tinidazole show  at 300 nm and 275 nm in 0.  Methanolic?HCI (20:80) respectively. The linearity was obtained in the concentration ranges of  for Ofloxacin and  for Tinidazole with Regression Coefficient (R2) greater than 0.999. In the first method concentration and subsequently amount of drug determined by using simultaneous equations and in second method concentration and amount of drug determined by using ratio of absorbance at Iso?absorptive point (which was found to be 286 nm) and at  of one of the drugs. The results of analysis have been validated statistically and subsequently by assay and recovery studies.

Research on Synthesis and Anti Convulsant Activity of Novel Oxadiazole Substituted Phenothiazine Derivatives

N. S. Dighe, A. A. Bankar, D. S. Musmade, S. A. Nirmal

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 54-60

The purpose of this research is directed towards the synthesis and evaluation of novel agents for the treatment of a variety of neurological disorders. In the past, the phenothiazine nucleus has been thoroughly investigated for various biological activities. The oxadiazole substituted phenothiazine has piqued interest as a potential treatment for a variety of neurological disorders. In light of these findings, researchers attempted to synthesise substituted phenothiazine and test them for anticonvulsant activity. Strychnine and 4-aminopyridine mediated models were used to test the anti-convulsant efficacy of synthesised compounds. These compounds can be used for ligand based drug designing purpose by which exact molecular mechanism responsible for showing anti-convulsant effect can be explored in future.

Preliminary Study on the Effect of Systemic Subnormal Deuterium Level on Metabolic Syndrome-Related and Other Blood Parameters in Humans

Gábor Somlyai, Ildikó Somlyai, István Fórizs, György Czuppon, András Papp, Miklós Molnár

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 61-71

The effects of deuterium depletion on the human organism have been, except for the antitumor action, seldom investigated by now and the available data are scarce. In oncological patients who also suffered from diabetes and were treated with deuterium-depleted water (DDW), an improvement of glucose metabolism was observed, and rat studies also proved the efficacy of DDW to reduce blood sugar level. In the present work, 30 volunteers with pre- or manifest diabetes were enrolled to a clinical study. The patients received 1.5 L of water with reduced deuterium content (104 ppm instead of 145 ppm, equivalent 12 mmol/L in human) daily for 90 days. The effects on fasting glucose and insulin level, on peripheral glucose disposal, and other metabolic parameters were investigated. The effects of DDW on other physiological parameters like qualitative blood count was also investigated.  Fasting insulin and glucose decreased, and insulin reaction on glucose load improved, in 15 subjects, while in the other 15 the changes were opposite. Peripheral glucose disposal was improved in 11 of the subjects. In the majority of the subjects, substantial increase of serum high-density lipoprotein (HDL) cholesterol and significant decrease of serum Na+ concentration were also seen-the latter possibly due to activation of a Na+/H+ antiporter by the decreased intracellular deuterium level. The results support the possible beneficial role of DDW in disorders of glucose metabolism but leave questions open, requiring further studies.

In vitro Studies on Proliferation and Production of Cytokine and IgG by Human PBMCs Stimulated with Polysaccharide Extract from Plants Endemic to Gabon

Line Edwige Mengome, Aline Voxeur, Jean Paul Akue, Patrice Lerouge

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 72-85

Polysaccharides were extracted from seven plants endemic to Gabon to study their potential immunological activities. Peripheral blood mononuclear cell (PBMC) (5 × 105 cells/mL) proliferation, cytokine and immunoglobulin G (IgG) assays were performed after stimulation with different concentrations of polysaccharide fractions compared with lipopolysaccharides (LPS) and concanavalin A (ConA) from healthy volunteers. The culture supernatants were used for cytokine and IgG detection by enzyme-linked immunosorbent assay (ELISA). The results show that pectin and hemicellulose extracts from Uvaria klainei, Petersianthus macrocarpus, Trichoscypha addonii, Aphanocalyx microphyllus, Librevillea klaineana, Neochevalierodendron stephanii and Scorodophloeus zenkeri induced production levels that were variable from one individual to another for IL-12 (3–40 pg/mL), IL-10 (6–443 pg/mL), IL-6 (7–370 pg/mL), GM-CSF (3–170 pg/mL) and IFN-? (5–80 pg/mL). Only hemicelluloses from Aphanocalyx microphyllus produce a small amount of IgG (OD = 0.034), while the proliferation of cells stimulated with these polysaccharides increased up to 318% above the proliferation of unstimulated cells. However, this proliferation of PBMCs was abolished when the pectin of some of these plants was treated with endopolygalacturonase (p < 0.05), but the trend of cytokine synthesis remained the same, both before and after enzymatic treatment or saponification. This study suggests that these polysaccharides stimulate cells in a structure-dependent manner. The rhamnogalacturonan-I (RGI) fragment alone was not able to induce the proliferation of PBMC. This study confirms the immunostimulatory properties of polysaccharides.

Medicinal- plant -based drugs have the advantage of being simple, effective and have broad spectrum activity. An ethno-botanical and ubiquitous plant serves as rich resources of natural drugs for research development. Ocimum gratissimum (fam. Lamiaceae) ‘Scent leaf’ and Vernonia amygdalina (fam. Asteraceae) ‘Bitter leaf’ are commonly consumed vegetables in Southern Nigeria. O. gratissimum is used by the ‘Igbos’ of South East Nigeria to flavor soups and stews while it is popular with the ‘Yorubas’ of South West Nigeria for treatment of stomach disorders. V. amygdalina is a well known vegetable in West Africa used as both food and for its pharmaceutical properties. The enormous use of these vegetables in the diet motivated the present study whereby the food compositions, mineral constituents and the phytochemicals of the two vegetables were determined. Qualitative analysis, of the aqueous extracts, revealed the presence of saponins, terpenoids, tannins, flavonoids, steroids, alkaloids, phenolics and anthraquinones in both vegetables. Generally V. amygdalina contained higher quantities of secondary metabolites, saponin which was reported as the highest amount was 5.71±0.12 mg/g and 3.52±0.01 mg/g followed by terpenoids 5.64±0.11 mg/g and 3.40±0.11 mg/g in V. amygdalina and O. gratissimum respectively. There is however a minute amount of anthraquinone in the two vegetables. Proximate analysis and macro elements were determined using their various standard methods. The results of proximate analysis revealed the presence of moisture (12.28 ± 0.02% and 10.01± 0.01%) protein (35.37±0.11% and 22.20±0.02%) and total ash (6.00±0.20% and 5.75±0.10%) each in V. amygdalina and O. gratisimum respectively. The following minerals were present calcium, magnesium, potassium, sodium, iron, copper and zinc, with zinc being in the highest concentration (46.80± 0.03 mg/kg and 58.00±0.12 mg/kg) followed by copper (9.45±0.11 mg/kg and 16.60 ± 0.03 mg/kg) each in V. amygdalina and O. gratissimum respectively. There are higher values of phytochemicals and food components in V. amygdalina while O. gratissimum had higher mineral contents. These two vegetables are potential sources of components for complementary medicine.

Line x Tester Analysis and Potential for Aluminum and Manganese Tolerance in an International Collection of Maize

Honoré Tekeu, Liliane N. Tandzi, Eddy E. M. Ngonkeu, Pierre F. Djocgoué

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 96-110

Soil acidity is a limiting factor that has a significant impact on maize growth and yield. The aim of this study was to determine the breeding value of inbred lines and to evaluate the hybrid maize test-cross output under acid soil conditions. During the 2012 cropping season in Cameroon, a line x tester analysis involving 63 test-crosses created by crossing 20 maize inbred lines with 3 testers and between testers themselves, as well as 1 standard check, was conducted in two contrasted regions with aluminium and manganese toxicity.. Both acid and non-acid soil treatments with dolomitic lime were tested in a randomised full block design with 3 replications.  Seven hybrids producing high grain yield (4.27 to 9.88 t/ha), with high specific combining ability (SCA) and slow rate of yield loss were discovered such as tolerant for both types of acidic soils. Likewise, four inbred lines (ATP-46, 87036, and Cam Inb gp117, C4SRRA7) with high general combining ability (GCA) were retained as good progenitors. The GCA and SCA effects revealed that additive effects of genes mediated resistance to aluminium toxicity, while non-additive effects of genes dominated on acid soil with manganese toxicity. However, for a potential extension, a molecular characterization of the selected varieties will be needed.

Aiming to synthesize new anti-inflammatory 1,7-dihydropyrazolo[3,4-d] imidazo[1,2-f] pyrimidine 5 from aminocyanopyrazole, a forthright method has been developed. It is for their anti-inflammatory, gastroprotective, analgesic, antioxidant and anticandidal activities that these compounds were screened. The investigation of the structure-activity relationships have brought about the choice of compound 6-(4-methoxyphenyl)-3-methyl-1,7-dihydropyrazolo[3,4-d]imidazo[1,2-f]pyrimidine, 5a, which revealed the most powerful activities. All novel compounds’ structures were expounded using IR, 1H NMR, 13C NMR and HRMS.

Research on Stable Ionized Calcium Concentration in Uncapped Plasma Samples

Victoria Richardson, Quinnita Reid, William A. Anong

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 125-131

Background: Blood ionized calcium (iCa) concentration has been shown to be pH dependent. In the clinical laboratory, plasma specimen for iCa measurement is routinely rejected when exposed to air prior to analysis. A preanalytical variable such as specimen exposure to air is believed to alter the pH and consequently the iCa concentration. Specimen exposure leads to the loss of carbon dioxide resulting in an increase pH and decrease iCa concentration. As the most active form physiologically, iCa measurement represents the best indicator of a patient’s calcium status. The purpose of this study is to investigate the rate at which these changes affect iCa concentration. We hypothesize that the changes are slow and insignificant to warrant the rejection of plasma specimen.

Methods: Appropriately collected whole blood specimen were centrifuged and analyzed for iCa concentration per laboratory procedure on AVL by Roche. The first, at time zero and subsequent measurements were made at various time intervals of sample exposure to air. The pH of plasma and whole blood samples exposed to air at various intervals were also measured to evaluate for changes.

Results: The iCa concentrations remained stable in plasma samples exposed to air for a considerable period of time. Exposed plasma pH measurements were equally stable (for up to ninety minutes), compared to that of whole blood specimen. The average changes seen in whole blood pH (~0.5 units) were ten times greater than that of plasma pH (~0.05 units) for the same length of time.

Conclusion: Plasma samples for iCa that are accidentally exposed to air should not be summarily rejected because both the pH and iCa level are stable for almost eighty minutes. The changes in whole blood pH are due to the respiring red cells. Red cells contain carbonic anhydrase (not found in plasma) that catalyzes the reversible interconversion of carbon dioxide and bicarbonate ion. Unnecessary specimen rejection/redraw request is costly. Besides, repeat injection is painful, exposes the patient to infections and hematoma at the injection site.

Cranberry Polyphenols and Prevention against Urinary Tract Infections: A Brief Review

Dolores González de Llano, M. Victoria Moreno-Arribas, Begoña Bartolomé

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 132-148

Cranberry (Vaccinium macrocarpon) is a distinctive source of polyphenols as flavonoids and phenolic acids that has been described to display beneficial effects against urinary tract infections (UTIs), the second most common type of infections worldwide. UTIs can lead to significant morbidity, especially in healthy females due to high rates of recurrence and antibiotic resistance. Strategies and therapeutic alternatives to antibiotics for prophylaxis and treatment against UTIs are continuously being sought after. Different to cranberry, which have been widely recommended in traditional medicine for UTIs prophylaxis, probiotics have emerged as a new alternative to the use of antibiotics against these infections and are the subject of new research in this area. Besides uropathogenic Escherichia coli (UPEC), the most common bacteria causing uncomplicated UTIs, other etiological agents, such as Klebsiella pneumoniae or Gram-positive bacteria of Enterococcus and Staphylococcus genera, seem to be more widespread than previously appreciated. Considerable current effort is also devoted to the still-unraveled mechanisms that are behind the UTI-protective effects of cranberry, probiotics and their new combined formulations. All these current topics in the understanding of the protective effects of cranberry against UTIs are reviewed in this paper. Finally, pieces of evidence regarding treatment against UTIs with probiotics and their combinations with cranberry are too limited to draw any conclusions but they seem to indeed be a promising strategy for future treatments.

A Rapid Analysis of Soybean Oil Minor Component Profile as a Tool to Predict Its Oxidative Stability: A Comprehensive View

A. S. Martin-Rubio, P. Sopelana, María D. Guillén

Current Advances in Chemistry and Biochemistry Vol. 4, 5 May 2021, Page 149-169

The minor components of vegetable oils are important for their oxidative stability. In order to know to what extent they can influence oil behaviour under oxidative conditions, two commercial soybean oils, one virgin and the other refined, both with very similar compositions in acyl groups but differing in their minor component profiles, were subjected to accelerated storage conditions. They were characterized by 1H nuclear magnetic resonance (NMR) and direct immersion solid-phase microextraction coupled to gas chromatography/mass spectrometry (DI-SPME-GC/MS), while oil oxidation was monitored by 1H-NMR. The lower levels of tocols and sterols in the virgin oil, together with its higher free fatty acid content when compared to the refined one, result in a lower oxidative stability. This is deduced from faster degradation of acyl groups and earlier generation of oxidation products in the virgin oil. These latter include a broad variety of compounds, some of them detected for the first time in the oxidation process of soybean oil, like mono- and di-hydroperoxides, monohydroxy-conjugated dienes, epoxides, monoketo-conjugated dienes, di-or poly-hydroxy-compounds, formic acid, formates or poly-formates, compounds with 5-alkyl-(5H)-furan-2-one structure and aldehydes. These findings reveal that commercial virgin soybean oil quality is not necessarily higher than that of the refined type, and that a simple and rapid analysis of oil minor components by DI-SPME-GC/MS would enable one to establish quality levels within oils originating from the same plant species and similar unsaturation level regarding composition in potentially bioactive compounds and oxidative stability.