Spectrophotometric Quantification Research Articles

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The purpose of the work is to determine the amount of flavonoids in the grass Vicia faba using spectrophotometry and assess the validity of the method. The raw materials of the Vicia faba species used for research were delivered in July 2022 from the Guba region of the Republic of Azerbaijan during the flowering phase of the plant. The validation parameters of the method for spectrophotometric quantification of flavonoids in the Vicia faba herb were assessed in accordance with the requirements of international documents. The amount of flavonoids in the Vicia faba herb was determined using a spectrophotometric method and it was found that the amount of flavonoids was ± 2.5%. A validation assessment of the spectrophotometric method for the quantitative determination of flavonoids in Vicia faba raw materials was carried out, the reliability interval of the method accuracy indicator was 0.09; linearity index in the concentration range of 0.02-0.04 mg/ml (R2=0.998); coefficient of variation of the accuracy indicator - 0.14% on the first day, 0.32% on the second day; The limit of determination of the method was found to be 0.001 mg/ml, and the limit of determination of quantity was found to be 0.004 mg/ml. The conducted studies show that the herb Vicia faba is quite rich in flavonoids, which lays the foundation for expanding the local medicinal plant raw material base in the future.

Supramolecular Solvent-Based Liquid–Liquid Microextraction and Preconcentration of Rhodamine B, Methyl Violet, and Eriochrome Black T in Water, Cosmetics, and Food with Spectrophotometric Quantification

Dyes are complex organic compounds that are utilized in many industries. However, excessive exposure to dyes increases susceptibility to various diseases. Their concentrations are below the detection level of many available analytical techniques. Therefore, preconcentration methodology is highly needed prior to their quantification. Herein, a new supramolecular solvent decanol-tetrahydrofuran-based liquid-liquid microextraction method has been established for the first time for the determination of Rhodamine B (Rh B), methyl violet (MV), and Eriochrome black T (EBT) in commercial spices, cosmetics, and water before spectrophotometric quantification. 556, 590, and 530 nm were used for the measurement of Rhodamine B (Rh B), methyl violet (MV), and Eriochrome black T (EBT), respectively. The optimization experiments revealed the best performance at pH 7, 0.9 mL of decanol, 1.3 mL of tetrahydrofuran (THF), and 5 min of vortex time. Under optimized conditions, the limits of detection (LOD) (0.480–0.511 µg L−1), limits of quantification (LOQ) (1.455–1.549 µg L−1), preconcentration factor (PF) (15–20), enhancement factor (EF) (15–20), and relative standard deviation (RSD) (0.5–0.8%) were evaluated. The validation of the method was carried out by standard addition experiments for food, cosmetic, and environmental water samples. The recovery results from 95 ± 6 to 100 ± 4% demonstrate that the method can be successfully applied to practical analysis.

Spectrophotometric determination of celecoxib and tramadol in the new approved formulated dosage form using principle component regression assistive model

Celecoxib and tramadol have been combined in a novel FDA-approved medication to address acute pain disorders requiring opioid treatment when other analgesics proved either intolerable or ineffective. The absorbance spectra of celecoxib and tramadol exhibit significant overlap, posing challenges for their individual quantification. This study introduces a spectrophotometric quantification approach for celecoxib and tramadol using a principle component regression assistive model to assist resolving the overlapped spectra and quantifying both drugs in their binary mixture. The model was constructed by establishing calibration and validation sets for the celecoxib and tramadol mixture, employing a five-level, two-factor experimental design, resulting in 25 samples. Spectral data from these mixtures were measured and preprocessed to eliminate noise in the 200–210 nm range and zero absorbance values in the 290–400 nm range. Consequently, the dataset was streamlined to 81 variables. The predicted concentrations were compared with the known concentrations of celecoxib and tramadol, and the errors in the predictions were evidenced calculating root mean square error of cross-validation and root mean square error of prediction. Validation results demonstrate the efficacy of the models in predicting outcomes; recovery rates approaching 100 % are demonstrated with relative root mean square error of prediction (RRMSEP) values of 0.052 and 0.164 for tramadol and celecoxib, respectively. The selectivity was further evaluated by quantifying celecoxib and tramadol in the presence of potentially interfering drugs. The model demonstrated success in quantifying celecoxib and tramadol in laboratory-prepared tablets, producing metrics consistent with those reported in previously established spectrophotometric methods.

Achyrocline satureioides infusion, popularly prepared and consumed, has an in vitro protective effect on human neural cells exposed to rotenone

Ethnopharmacological relevanceTraditional harvest of Achyrocline satureioides (AS) occurs at dawn on Good Friday in some South American countries. Inflorescences are traditionally used as infusions for several disorders, including neuropsychiatric disorders. Pillows and cushions are popularly filled with AS to attenuate the symptoms of depression, anxiety, and sleep disturbances. However, evidence for the potential beneficial effects of AS on human neural cells remains unclear. Aim of the studyAn in vitro model of SH-SY5Y human neural cells was applied to evaluate the effect of AS infusion, prepared as commonly used, on cells exposed to rotenone and to investigate its potential for neuropsychiatric disorders. Materials and methodsA hot aqueous extract was obtained from a traditionally prepared AS inflorescence infusion and chemically characterized by high-resolution mass spectrometry and spectrophotometric quantification of total polyphenols, tannins, and flavonoids. The SH-SY5Y cell cultures were treated with AS extract at concentrations of 1, 3, 5, 10, 50, 100, and 300 μL/mL to determine the potential cyto- and genotoxic effects of AS on neural cells using MTT, Neutral Red, and GEMO assays. Apoptosis modulation was assessed using flow cytometry and apoptosis-modulating genes were evaluated by qRT-PCR. The protective effect of AS on the neurotoxicity triggered by rotenone exposure (30 nM) was determined by analyzing cellular viability and oxidative markers such as lipid peroxidation and protein carbonylation, and DNA damage was assessed by micronucleus assay. ResultsThe AS extract, as traditionally prepared, had estimated concentrations of 409.973 ± 31.107 μg/mL, 0.1041 ± 0.0246 mg GAE/mL, and 63.309 ± 3.178 mg QE/mL of total tannins, total polyphenols, and flavonoids, respectively. At concentrations of 30 and 100 μl/mL, AS decreased apoptotic events, whereas the highest concentration (300 μl/mL) increased apoptosis compared to that in the control (p < 0.05). In cells exposed to rotenone, AS treatment induced cell proliferation, reduced DNA damage (as evaluated by micronuclei), and reduced lipid and protein oxidation. ConclusionsThe data indicate the non-cytotoxic and beneficial effects of AS extract on human neural cells by reducing cellular mortality and oxidative stress in neural cells triggered by rotenone exposure.

Antibiotic-Induced Changes in Efflux Transporter Expression: A Key Factor in Pseudomonas aeruginosa Biofilm Resistance

Listed by WHO as an antibiotic-resistant priority pathogen, Pseudomonas aeruginosa (P.A.) is a serious threat in nosocomial infections. Its high antibiotic resistance is attributed to major mechanisms that can be categorized into intrinsic, acquired, and adaptive resistance. This study tests the ability of three commonly used antibiotics to inhibit new biofilm formation and eradicate mature biofilm growth, as well as investigate changes in the expression levels of selected genes coding for multidrug efflux pumps in P.A. planktonic cells and biofilms before and after treatment with antibiotics to provide a conceptual estimate of the activity of the efflux transporters that work to extrude antibiotics leading to a reduction in their effectiveness. Antimicrobial susceptibility testing was conducted with Ofloxacin (OFLX), Tobramycin (TOB), and Ceftazidime (CAZ) to determine Mean Inhibitory Concentration (MIC) and Mean Bactericidal Concentration (MBC) using microtiter plate-based biofilm assay and spectrophotometric quantification. Extraction of total RNA was performed from planktonic cultures, inhibition phase, and eradication phase P.A. biofilms. Real-time quantitative reverse transcriptase PCR was utilized to analyze the changes in expression of the mexAB, mexXY, and oprM genes. Three (3) antibiotics that have proven to show less resistance are OFLX, TOB, and CAZ when tested against overnight cultures of P.A. strain PA01. Results showed that OFLX is best for bactericidal properties, which is also supported by the viability assay data obtained from Propidium Iodide staining. Our study showed that the PAO1 strain is susceptible to OFLX for both inhibition and eradication of mature biofilms. TOB was most effective at higher concentrations in the eradication phase.

Effective Spectrophotometric Quantification of Yttrium Using Clayton Yellow Dye in Different Rare Earth Concentrates

Effective Spectrophotometric Quantification of Yttrium Using Clayton Yellow Dye in Different Rare Earth Concentrates

Standardisation of an efficient RNA isolation protocol from the seeds of Momordica charantia var. muricata (Willd.): A Comparative Evaluation

Extraction of high-quality total RNA from Momordica charantia var. muricata seeds is difficult due to the presence of polyphenolic and polysaccharide compounds which bind to nucleic acids and thus they co-precipitate and contaminate the RNA samples. The current study aimed to find out a reliable and reproducible RNA extraction method for obtaining good-quality RNA from the seeds of Momordica charantia var. muricata (Willd). The RNA was isolated from seed tissues with four different extraction protocols including the cetyltrimethylammonium bromide (CTAB) method, TRIzol (Invitrogen) method, GenElute™ total RNA purification kit (Sigma-Aldrich) and Plant RNeasy mini kit (Qiagen). The Qiagen RNeasy plant mini kit was the most efficient extraction method providing the highest RNA yield and good quality. The method successfully enhanced the yield of RNA to 923.27 ± 58.64 ng/μl and electrophoresis revealed two distinct bright 28S and 18S rRNA bands and spectrophotometric quantification detected A260/ A280 ratio of 2.04±0.01 within the desired range of 1.8-2. To verify that the RNA quality obtained was appropriate for downstream molecular applications, the isolated RNA samples were converted into cDNA and PCR amplification of the cDNA was performed using a housekeeping gene (Actin 7) and a dormancy-specific gene (DOG1). The current investigation reports on a method for isolating high-quality RNA from M. charantia var. muricata seeds for future dormancy gene expression studies.

Development of Improved Spectrophotometric Assays for Biocatalytic Silyl Ether Hydrolysis.

Reported herein is the development of assays for the spectrophotometric quantification of biocatalytic silicon-oxygen bond hydrolysis. Central to these assays are a series of chromogenic substrates that release highly absorbing phenoxy anions upon cleavage of the sessile bond. These substrates were tested with silicatein, an enzyme from a marine sponge that is known to catalyse the hydrolysis and condensation of silyl ethers. It was found that, of the substrates tested, tert-butyldimethyl(2-methyl-4-nitrophenoxy)silane provided the best assay performance, as evidenced by the highest ratio of enzyme catalysed reaction rate compared with the background (uncatalysed) reaction. These substrates were also found to be suitable for detailed enzyme kinetics measurements, as demonstrated by their use to determine the Michaelis-Menten kinetic parameters for silicatein.

OxyHbMeter-a novel bedside medical device for monitoring cell-free hemoglobin in the cerebrospinal fluid-proof of principle.

Delayed cerebral ischemia (DCI) occurs in up to one third of patients suffering from aneurysmal subarachnoid hemorrhage (aSAH). Untreated, it leads to secondary cerebral infarctions and is frequently associated with death or severe disability. After aneurysm rupture, erythrocytes in the subarachnoid space lyse and liberate free hemoglobin (Hb), a key driver for the development of DCI. Hemoglobin in the cerebrospinal fluid (CSF-Hb) can be analyzed through a two-step procedure of centrifugation to exclude intact erythrocytes and subsequent spectrophotometric quantification. This analysis can only be done in specialized laboratories but not at the bedside in the intensive care unit. This limits the number of tests done, increases the variability of the results and restricts accuracy. Bedside measurements of CSF-Hb as a biomarker with a point of care diagnostic test system would allow for a continuous monitoring for the risk of DCI in the individual patient. In this study, a microfluidic chip was explored that allows to continuously separate blood particles from CSF or plasma based on acoustophoresis. An in vitro test bench was developed to test in-line measurements with the developed microfluidic chip and a spectrometer. The proof of principle for a continuous particle separation device has been established with diluted blood and CSF samples from animals and aSAH patients, respectively. Processing 1 mL of blood in our microfluidic device was achieved within around 70 min demonstrating only minor deviations from the gold standard centrifugation (7% average error of patient samples), while saving several hours of processing time and additionally the reduction of deviations in the results due to manual labor.

Spectrophotometric Determination of Hydroxycinnamic Acids in Herbal Drugs. Part 2. Determination of Hydroxycinnamic Acids in the Presence of Polyphenolic Compounds (Review)

INTRODUCTION. Polyphenolic compounds of various classes have pronounced absorption bands overlapping in almost the same ultraviolet region. This superposition of bands complicates the quantification of total hydroxycinnamic acids in herbal drugs by direct spectrophotometry without additional sample preparation.AIM. This study aimed to establish the possibility of quantifying hydroxycinnamic acids in herbal drugs by spectrophotometry in the presence of other polyphenolic compounds.DISCUSSION. This review analyses publications on analytical procedures for the spectrophotometric quantification of hydroxycinnamic acids that are based on the preliminary separation or chemical modification of polyphenolic compounds (combining the use of spectrophotometry, chromatography, extraction, Arnow’s reagent, etc.). The described sample preparation can help to resolve absorption bands in the spectra of polyphenolic compounds. The results of assessing the characteristic parameters of these analytical procedures (selectivity, sensitivity, labour intensity, etc.) can serve as a basis for searching novel approaches to the quantitative determination of hydroxycinnamic acids in plants.CONCLUSIONS. To determine the content of hydroxycinnamic acids in mixtures of polyphenolic compounds, analysts may use special sample preparation (combining spectrophotometry with chromatography and extraction) or chemical modification of the initial compounds. The acquired spectra could benefit from specific mathematical processing (derivative photometry and Vierordt’s method). However, even with the abovementioned approaches, it may not always be possible to increase the selectivity of an analytical procedure. Therefore, there is a need to further improve existing analytical procedures for the quantification of hydroxycinnamic acids and develop new ones.

Spectrophotometric Determination of Hydroxycinnamic Acids in Herbal Drugs. Part 1. Direct Spectrophotometry (Review)

INTRODUCTION. Current scientific research aims to discover new plant sources of hydroxycinnamic acids for the development of novel medicinal products. Most commonly, the quantitative determination of hydroxycinnamic acids in herbal drugs involves testing procedures based on spectrophotometry and its modifications.AIM. This study aimed to establish the possibility of the quantification of hydroxycinnamic acids in herbal drugs by direct spectrophotometry.DISCUSSION. This review presents organised information on the use of direct spectrophotometry to quantify hydroxycinnamic acids in 70 plant species growing or cultivated in the Russian Federation. The review covers publications in Russian for the period from 2007 to 2023. According to the reviewed publications, direct spectrophotometry is an easy-to-use and highly reproducible method that does not require additional reagents or labour-intensive steps. Direct spectrophotometry may result in an overestimation of the hydroxycinnamic acid content due to overlapping absorption bands of polyphenolic compounds with different structures.CONCLUSIONS. Direct spectrophotometry can be considered as a method for measuring the total content of phenolic compounds (as well as hydroxycinnamic acids) in herbal drugs. Different approaches to direct spectrophotometric quantification of hydroxycinnamic acids in herbal drugs by can serve as a methodological basis for developing analytical procedures specific to the chemical compositions of biologically active compounds in herbal drugs and their extracts.

The binding of auranofin at DNA/RNA nucleobases: A DFT assessment

A variety of therapies are based on cytotoxicity induced via hampering the DNA and/or RNA homeostasis. The metallodrugs’ forerunner, cisplatin, is exemplificative of the tremendous biological effects of nucleobase targeting, and a large body of researches to discover novel metallodrugs. In this paper, DFT approaches were employed to investigate the structure, stability and electronic properties of the complexes formed by the [Et3PAu]+ metal fragment generated by auranofin -or its derivatives- and the DNA/RNA nucleobases. Therefore, TDDFT calculations were performed to assess the viability of the spectrophotometric quantification of the [Et3PAu]+-nucleobase adducts.

Spectrophotometric quantification of amitriptyline hydrochloride in pharmaceutical tablets: method development and validation

The development and validation of two uncomplicated, selective, and non-extraction-based visible spectrophotometric methods was carried out for the quantification of a tricyclic antidepressant amitriptyline hydrochloride as pharmaceutical tablets as well as in its pure form. The methods rely on the formation of ion pair complexes between amitriptyline base and two acidic dyes, bromothymol blue and bromocresol purple, which were subsequently measured at 410 and 400 nm, respectively. The developed methods included numerous reaction variables that were examined and improved. Beer's law is applicable under optimal conditions for the bromothymol blue and bromocresol purple methods, respectively, over concentrations from 0.5 to 12.5 μg mL−1 and from 0.5 to 10.0 μg mL−1 amitriptyline hydrochloride. The computed values of molar absorptivity were 1.79 × 104 and 2.32 × 104 L.mol−1.cm−1 for bromothymol blue and bromocresol purple methods, respectively, with Sandell’s sensitivity values of 0.0155 and 0.0119 μg cm−2. Both detection limits and quantification limits were computed and determined to be 0.55 and 1.67 μg mL−1 for bromothymol blue method, and 0.49 and 1.49 μg mL−1 for bromocresol purple method, respectively. The developed methods were successfully applied to estimate the concentration of amitriptyline hydrochloride in bulk form and commercial pills. Therefore, these methods demonstrate promising potential for testing this drug in labs for quality control.Graphical abstractSpectrophotometric quantification of amitriptyline hydrochloride in pharmaceutical tablets: method development and validation

Spectrophotometric Quantification of Trace Elements and Toxic Metals in Raw Milk Samples of Peshawar City, Pakistan

The present study focused on the physicochemical characterization and spectrophotometric determination ofthe concentrations of trace elements (Zn, Cu, Fe and Mn) and toxic heavy metals (Cd and Pb) in raw milk samplescollected from various commercial sites of Peshawar city, Pakistan. Evaluation of the nutritional values as well as thepossible health concerns associated with the consumption of contaminated milk shown the average percentagecomposition of protein, fats, solid non-fats, lactose, salts and added water in the milk sample as 2.78, 4.81, 6.41, 3.05,0.47 and 31.1 %, respectively. The values of pH, conductivity, density and freezing point were observed as 6.59, 3.51mS/cm, 1.04 g/mL and -0.3674 ºC at 26 ºC. The physicochemical characteristics such as percentage protein, fat,lactose, salts, water, solid not fat, total solids, pH, conductivity and density were evaluated and found in the safe limitsof the WHO/FAO. The average concentration of trace elements such as zinc and copper were found to be 1.4035 mg/Land 0.2588 mg/L, respectively. The concentration of Zn and Cu was found within permissible limits suggested byWHO. However, the average concentration of Fe, Mn, Cd and Pb were found higher than the permissible limits ofWHO. The observed concentrations were: Fe = 1.5553, Mn = 0.4354, Cd = 0.1865 and Pb = 1.1162 mg/L. Thepresence of the non-essential and hazardous Cd and Pb in milk samples even in lower concentrations may cause severehealth problems since these are the most noxious pollutants due to their non-biodegradable and bio-accumulativenature. Over-dose of Fe and Mn also leads to several health issues. A thorough and more precise investigation of rawmilk consumption in Peshawar is highly recommended to ascertain the actual reasons and sources in the larger publicinterests.

Spectrophotometric Quantification of Atomoxetine Hydrochloride Based on Nucleophilic Substitution Reaction with 1,2-Naphthoquinone-4-Sulfonic Acid Sodium Salt (NQS).

A simple, sensitive, selective, and cost-effective colorimetric method has been established for the quantitative estimation of atomoxetine hydrochloride in bulk and formulation.A simple, sensitive, selective and cost effective colorimetric method has been entrenched for the quantitative estimation of Atomoxetine hydrochloride in bulk and formulation. It was established based on the visible reaction between atomoxetine hydrochloride and 1,2-naphthoquinone-4-sulfonic acid sodium salt in a basic medium (potassium hydroxide). The resulting orange colored chromogen exhibited an absorption maximum at 474 nm. Based on the optimization studies, distilled water as the solvent, 1% w/v potassium hydroxide (2 mL), and 0.3% w/v 1,2-naphthoquinone-4-sulfonic acid sodium salt (2 mL) were used in the method. The developed method was validated per the International Council for Harmonization (ICH) guidelines. The linearity was found at a concentration of 10-50 μg/mL. The method showed a good correlation between the concentration of atomoxetine hydrochloride and its absorbance. The correlation coefficient (r2) of 0.999 evidenced the same. The limits of detection and quantification were 0.20 and 0.606 μg/mL, respectively, for atomoxetine hydrochloride. The accuracy and precision of the method were also evaluated and the results obtained were within the acceptance criteria (relative standard deviation % < 2.00). The percentage assay of atomoxetine hydrochloride proved to be 101.52, which is in accordance with its label claim. The developed method is non-complex and can be effectively employed in the analytical practices of atomoxetine hydrochloride in pharmaceutical dosage forms.

Subcritical water extraction of red cabbage regarding cholinesterase and tyrosinase inhibitory activity

Pressurized high-temperature processing of water where polarity can be regulated has been an advanced extraction technology considering sustainable management and efficient use of natural resources. In this study, subcritical water extraction of red cabbage and its enzyme inhibitory potential were investigated using response surface methodology. Spectrophotometric quantification was utilized for total phenol, flavonoid, and anthocyanin contents, while dominant anthocyanin molecules were identified with UPLC-DAD, ESI-MS/MS. The 200 bar-120°C-30 min-30 solvent:solid ratio extract was determined with the highest total anthocyanin content (2.33 mg Cya3GluE/g), where cyanidin-3-sophoroside-5-glucoside, cyanidin-3-(sinapoyl)-sophoroside-5-glucoside, and cyanidin-3-(sinapoyl)(sinapoyl)-sophoroside-5-glucoside were identified as the dominant anthocyanins (1.15 mg Cya3GluE/g). IC50 values for acetylcholinesterase, butyrylcholinesterase, and tyrosinase inhibitory activities were found to be 0.058 mg/ml, 0.215 mg/ml, and 0.86 mg/ml, respectively (tenfold superior to solvent extract). Despite the well-known high-temperature lability of phenolic compounds, subcritical water proved to be useful as a more selective system for extracting red cabbage bioactives with enzyme inhibitory activity when appropriate process conditions could be determined.

Evaluation of organo-vermiculites as sorbent phases for solid-phase extraction of ibuprofen from water.

The study of ibuprofen (IBU) preconcentration was carried out making use of a homemade column for solid-phase extraction (SPE), using vermiculite (VT) or organo-vermiculites (OVTs) as sorbent phases. Aqueous samples (50.0 mL) percolated the column and IBU was sorbed onto the VT or OVT and then desorbed using acetonitrile. Employing this SPE system and OVT, calibration curves were generated for IBU, by spectrophotometric quantification using the α-naphthylamine method. R2 values higher than 0.9950 and LOD between 12 and 18 μg L-1 were observed, for real enrichment factors of 21 and 31, by using OVTs. The analytical protocol was applied to three water samples, which were spiked with IBU solutions to evaluate the precision and accuracy of the method. Recoveries between 77 and 110% at three different IBU concentrations and RSD lower than 18% were observed, even by using the spectrophotometric method. The protocol developed in this study demonstrated that the OVT was appropriate to work as a preconcentration phase for IBU determination in water samples.

Crizotinib inhibits the metabolism of tramadol by non-competitive suppressing the activities of CYP2D1 and CYP3A2.

To investigate the interaction between tramadol and representative tyrosine kinase inhibitors, and to study the inhibition mode of drug-interaction. Liver microsomal catalyzing assay was developed. Sprague-Dawley rats were administrated tramadol with or without selected tyrosine kinase inhibitors. Samples were prepared and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used for analysis. Besides, liver, kidney, and small intestine were collected and morphology was examined by hematoxyline-eosin (H&E) staining. Meanwhile, liver microsomes were prepared and carbon monoxide differential ultraviolet radiation (UV) spectrophotometric quantification was performed. Among the screened inhibitors, crizotinib takes the highest potency in suppressing the metabolism of tramadol in rat/human liver microsome, following non-competitive inhibitory mechanism. In vivo, when crizotinib was co-administered, the AUC value of tramadol increased compared with the control group. Besides, no obvious pathological changes were observed, including cell morphology, size, arrangement, nuclear morphology with the levels of alanine transaminase (ALT) and aspartate transaminase (AST) increased after multiple administration of crizotinib. Meanwhile, the activities of CYP2D1 and CYP3A2 as well as the total cytochrome P450 abundance were found to be decreased in rat liver of combinational group. Crizotinib can inhibit the metabolism of tramadol. Therefore, this recipe should be vigilant to prevent adverse reactions.

E.ANGUSTFOLİUM XAMMALINDA FLAVONOİDLƏRİN SPEKTROFOTOMETRİYA İLƏ MİQDARI TƏYİNİ VƏ ÜSULUN VALİDASİYASI

The purpose of the work is to determine the amount of flavonoids in the grass E. angustfoli-um using spectrophotometry and assess the validity of the method. The raw material of the species E. angustifolium used for research purposes was collected in the flowering phase of the plant from the Guba region of the Republic of Azerbaijan in July 2022. The validation parameters of the method for spectrophotometric quantification of flavonoids in the E. angustifolium herb were assessed in accordance with the requirements of international documents. The amount of flavonoids in the E.angustifolium herb was determined using a spectrophoto-metric method and it was found that the amount of flavonoids was ± 2.77%. A validation assessment of the spectrophotometric method for the quantitative determination of flavonoids in E.angustifolium raw materials was carried out, the reliability interval of the method accuracy indicator was 0.56; linearity index in the concentration range of 0.01-0.05 mg/ml (R2=0.999); coefficient of variation of the accuracy indicator - 0.26% on the first day, 0.31% on the second day; The limit of determination of the method was found to be 0.02 mg/ml, and the limit of determination of quantity was found to be 0.006 mg/ml. The conducted studies show that the herb E.angustifolium is quite rich in flavonoids, which lays the foundation for expanding the local medicinal plant raw material base in the future.

The effectiveness of the tissue engineering in the obtaining of the biological materials from the extracellular matrix

Background: The present work describes the possibility of manufacturing biomaterials from the extracellular matrix for the treatment of the skin wounds. Biomedical collagen-based materials are clinically effective. Collagen is the most abundant and major component of the skin. Porcine collagen is almost similar to the human collagen, it is not immunogenic when used for the therapeutic purposes. Biomaterials can be obtained from the decellularized dermis, being a matrix rich in the collagen and glycoproteins. Material and methods: 3 parallel groups of biomaterials were established and the average value was calculated. To ensure the effectiveness of the decellularization process, the decellularized porcine dermis was compared with the intact sample using qualitative and quantitative criteria. Results: Histologically, the decellularized tissues revealed the presence of fewer cells. As a result, were removed approximately 80.5% of the genetic material from porcine dermal structures, demonstrated by the spectrophotometric quantification of deoxyribonucleic acid. In vitro graft degradation study in 0.01 M phosphate buffer pH 7.4 combined with collagenase, demonstrated a significant (p &lt; 0.05) loss of collagen sponge mass by 100% over one hour in the group II compared to the decellularized dermis in group I which decreased in the weight by 91.3% during 35 hours. Conclusions: Acellular biomaterials are immunologically inert, have hydrophilic and biodegradable properties, thus they can play a key role in the wound care, exerting the transfer of the bioactive molecules and drugs directly into the wound.