Hydrolysis Conditions Research Articles

The use of microwave radiations and propylphosphonic anhydride (T3P®) for rapid and highly yielding amide bond formation and peptide coupling

An improved, fast, and efficient procedure for amide bond formation and peptide coupling is reported leading to the expected corresponding derivatives in good to excellent isolated yields ranging from 26 to 90%. The outcome of the reaction was also investigated in relation to the influence of different parameters such as the nature of the solvent, the temperature as well as the amine/carboxylic acid couple involved or the hydrolysis conditions.

Quantitative determination of chondroitin sulfate with various molecular weights in raw materials by pre-column derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate

A simple and reliable HPLC method was developed for quantification of chondroitin sulfate (CS). The procedure is based on precolumn hydrolysis of CS to liberate galactosamine and subsequent derivatization with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate. Hydrolysis and derivatization conditions were optimized. A linear correlation coefficient of 0.9999 was calculated within the range of 10–1500 μg/mL from the standard curve. The method produces good precision and good accuracy (100.75 % recovery). An advantage over other common methods is its ability to quantify CS of all molecular weights and structures, as evidenced by the determination of CS fractions with narrow molecular weight distributions obtained through depolymerization by different methods, while enzymatic HPLC was proven to be infeasible. Extraction recoveries of CS from monosaccharide mixed samples were > 93 %. The reliability was also validated by a small difference (−1.95 % to 4.12 %) relative to enzymatic HPLC results in analysing representative CS samples of different animal origins and suppliers.

Effects of hydrolysis conditions on the morphology of cellulose II nanocrystals (CNC-II) derived from mercerized microcrystalline cellulose

The properties of cellulose nanocrystals with allomorph II (CNC-II) vary with the sources and the treatments received. In this work, the influences of hydrolysis time, temperature, and the applied acid concentration on the crystal size of CNC-II were investigated by the surface response experimental design. The results showed that temperature was the most significant factor affecting the crystal size of CNC-II during hydrolysis from mercerized cellulose. Then the morphology and colloidal properties of CNC-II were revealed by dynamic laser scattering (DLS), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), transmission electron microscopy (TEM), thermogravimetric analysis (TGA), etc. XRD results indicated that CNC-II had slightly lower crystallinity (80.89 % vs 82.7 %) and larger crystallite size (5.21 vs. 5.13 nm) than CNC-I. TEM and AFM results showed that the morphology of CNC-II were disc-like and rod-like particles, with an average diameter of 14.6 ± 4.7 nm (TEM) and a thickness of 4– 8 nm (AFM). TG and XPS revealed the reduced thermal stability was due to the introduced sulfate groups in CNC-II during hydrolysis. This investigation has addressed the features of CNC-II derived from mercerized cellulose, and it would be promising in fabricating advanced materials.

Exploring Enzymatic Hydrolysis of Urine Samples for Investigation of Drugs Associated with Drug-Facilitated Sexual Assault.

Analyzing urine is common in drug-facilitated sexual assault cases if the analysis of blood is not optimal. The efficient enzymatic pretreatment of urine is important for cleaving glucuronides and improving the detection of the parent drug. The aim was to investigate the efficiency of three β-glucuronidases on eleven glucuronides relevant to DFSA at different incubation periods and temperatures. Human drug-free urine was fortified with 11 glucuronides, hydrolyzed with either β-glucuronidase/arylsulfatase (Helix Pomatia), recombinant β-glucuronidase B-One™ or recombinant β-glucuronidase BGTurbo™ and incubated for 5, 10, 60 min, 18 h and 24 h at 20 °C/40 °C/55 °C before UHPLC-MS/MS analysis. The stability of 141 drugs and metabolites relevant to DFSA was investigated by incubating fortified urine under the same hydrolysis conditions. B-One™ showed efficient hydrolysis (>90%) of most glucuronides in 5 min at all temperatures, while BGTurbo™ showed a similar efficiency (>90%), but the optimal temperature (20-55 °C) and incubation time (5-60 min) varied among analytes. The β-glucuronidase/arylsulfatase had the lowest efficiency and required the longest incubation (24 h) at 40-55 °C. The stability of 99% of 141 drugs and metabolites was not affected by incubation at 20-55 °C for 24 h. Recombinant enzymes show promising results for the simple and efficient hydrolysis of a broad panel of glucuronides relevant for DFSA.

Determination of Phenylethanolamine A Residues in Milk by Solid Phase Extraction-Liquid Chromatography Tandem Mass Spectrometry

A method was established for the determination of phenethanolamine A residue in milk by solid phase extraction, liquid chromatography-tandem mass spectrometry. The samples were extracted with ammonium acetate buffer solution under the condition of enzymatic hydrolysis and purified by MCX solid phase extraction column. The eluent was blown dry with nitrogen at 45℃ and dissolved with 1.0 mL 0.1% formic acid solution. The samples were isolated by Poroshell 120EC-C18, 100 mm ×3.0 mm (i.d.), 2.7 μm column, and then determined by mass spectrometry in multiple reaction monitoring mode, and quantified by internal standard method. The results showed that phenethanolamine A had good linear relationship in the concentration range of 0.5-50.0 μg/L, the correlation coefficient r=0.9984, the limit of detection was 0.17 μg/kg, and the limit of quantitation was 0.57 μg/kg. The average recovery was 98.3-108.7% and the precision was 2.4-3.7% at different supplemental levels. The established method can purify the samples effectively, improve the detection efficiency, and is suitable for the detection of phenethanolamine A residue in milk.

Effects of Hydrolysis Condition and Detection Method on the Monosaccharide Composition Analysis of Polysaccharides from Natural Sources

Monosaccharide composition analysis is essential to the structural characterization and research into the biological activity of polysaccharides. In this study, a systematic comparison was performed among commonly used monosaccharide composition analysis methods, including colorimetric and chromatographic methods. These were tested on 16 aldoses, ketoses, alditols, amino sugars, and uronic acids. Furthermore, the effect of hydrolysis methods was also investigated. The results showed that the phenol sulfuric acid method is greatly affected by the type of monosaccharide that is used as the reference substance. The determination of uronic acid using sulfuric acid carbazole is less affected by neutral sugars than that method using m-hydroxybiphenyl. The high-performance thin-layer chromatography (HPTLC) method can simultaneously analyze multiple samples and accurately determine the type of uronic acid. High-performance liquid chromatography (HPLC) can provide a good qualitative and quantitative analysis of aldose, amino sugars, and uronic acids, while gas chromatography–mass spectrometry (GC-MS) can detect aldose, ketose, and alditols. Fructose was detected in a large amount in inulin and Codonopsis pilosula after one-step hydrolysis, while it was totally destroyed in two-step hydrolysis. The release of galacturonic acid significantly increased after two-step hydrolysis in pectin and Lycium barbarum, which indicated that one-step hydrolysis is not enough for acidic polysaccharides. The results of this study are beneficial for selecting appropriate hydrolysis and analysis methods in order to accurately analyze the monosaccharide compositions of natural polysaccharides.

Itaconate and citrate releasing polymer attenuates foreign body response in biofabricated cardiac patches

Application of cardiac patches to the heart surface can be undertaken to provide support and facilitate regeneration of the damaged cardiac tissue following ischemic injury. Biomaterial composition is an important consideration in the design of cardiac patch materials as it governs host response to ultimately prevent the undesirable fibrotic response. Here, we investigate a novel patch material, poly (itaconate-co-citrate-co-octanediol) (PICO), in the context of cardiac implantation. Citric acid (CA) and itaconic acid (ITA), the molecular components of PICO, provided a level of protection for cardiac cells during ischemic reperfusion injury in vitro. Biofabricated PICO patches were shown to degrade in accelerated and hydrolytic conditions, with CA and ITA being released upon degradation. Furthermore, the host response to PICO patches after implantation on rat epicardium in vivo was explored and compared to two biocompatible cardiac patch materials, poly (octamethylene (anhydride) citrate) (POMaC) and poly (ethylene glycol) diacrylate (PEGDA). PICO patches resulted in less macrophage infiltration and lower foreign body giant cell reaction compared to the other materials, with corresponding reduction in smooth muscle actin-positive vessel infiltration into the implant region. Overall, this work demonstrates that PICO patches release CA and ITA upon degradation, both of which demonstrate cardioprotective effects on cardiac cells after ischemic injury, and that PICO patches generate a reduced inflammatory response upon implantation to the heart compared to other materials, signifying promise for use in cardiac patch applications.

Bio-Ethanol Production from Wheat Straw Using Yeast Isolates

In the first hand, the cost of fossil fuel is increasing alarmingly. On the second hand, combustion of fossil fuels contributes for global warming. Therefore, it need to strength the production of renewable energies. The aim of this study was to produce bioethanol from wheat straw using yeast isolates. The isolates were isolated from decomposed soil, termite soil and rotten wood samples using yeast extract peptone dextrose media (YPD) and characterized chemically and morphologically. The wheat straws were powdered and hydrolyzed with dilute sulfuric acids. After neutralization, it was used to produce ethanol. Response surface methodology was employed to optimize the ethanol production process from wheat straws. The isolates were grown optimally at a temperature of 30oC, pH nearly 5, and sugar concentration 70 to 120 g/L. Among hydrolysis conditions, lower acid concentration (less than 1.5%) and temperature of 60oC resulted higher reducing sugars. The optimization study showed that the highest bio-ethanol concentration of 6.8g/l was observed by SWX under the optimum conditions of with 1% H2S04, 60oC temperature and 52.5-minute time hydrolysis at 30°C for 48 hour incubation time. Wheat straws could be good candidate for ethanol production.

Effects of soybean endogenous enzyme hydrolysis on the quality of soymilk after blanching

This study examined the relationship between the endogenous enzymatic hydrolysis conditions and the quality and flavor of soymilk derived from blanched soybeans. Results represented that following blanching, the lipoxygenase activity of soybeans decreased by 88.50% and the endogenous protease activity greatly improved compared to unblanched. During the subsequent endogenous enzymatic hydrolysis, the stability factor and sensory score dropped as the amount of okara rose, although the content of peptides and amino acids nitrogen increased. At an enzymolysis temperature of 50 °C, the soymilk had the highest sensory score. After 60 min of enzymolysis, the beany odor was weakened maximumly; the levels of polypeptide and amino acid nitrogen increased by 45.00% and 43.33%, respectively. Enzymatic hydrolysis at 50 °C for 60 min without okara generated the finest soymilk. Blanching diminished the primary components of the beany odor of raw soymilk. Enzymolysis of blanched pretreatment soymilk further dropped the content of beany odor components and exhibited a richer variety and concentration of volatile components. Following the hydrolysis of endogenous enzymes generated after blanching, the overall quality of soymilk was enhanced.

Study on the hydrolytic degradation behaviour of bictegravir by LC-PDA-Q/TOF-MS/MS NMR and in silico toxicity assessment

Forced degradation studies provide rapid access to degradation products (DPs), where structural characterization and assessment of their potential toxicity are vital for pharmaceutical safety and regulatory compliance. As per the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) Q1A R(2) guidelines, a forced degradation study of Bictegravir (BIC), a USFDA-approved drug for HIV wild type, in hydrolytic conditions (acid, base, and neutral) revealed the formation of six DPs in RP-HPLC (Reverse Phase- High-Performance Liquid Chromatography) gradient elution program using a C18 (4.6 × 250 mm, 5 µm) column. DP-1, 2, and 3 were characterized using liquid chromatography-tandem mass spectrometry (LC-MS/MS), whereas DP-4, 5, and 6 posed difficulties in characterization due to their isomeric nature. Using characteristic NOEs (Nuclear Overhauser Effect) from 2D ROESY NMR (Nuclear Magnetic Resonance) studies, we have elucidated diastereomeric DP-4/5 and isomeric DP-6/BIC configurational structures. Furthermore, in silico toxicity studies for the six degradation products were predicted for toxicity endpoints by employing DEREK, SARAH, and Pro Tox-II application tools. The DP-1 (methanamine) and DP-3 (carboxylic acid) resulting from acid-catalyzed hydrolysis, were predicted to have potential carcinogenic and mutagenic properties. These findings contribute significantly to our understanding of BIC's stability and safety profile in pharmaceutical development and underscore the rigorous characterization of stereoisomers by NMR that were further utilized for toxicity prediction.

A novel stability indicating mass compatible reversed‐phase high‐performance liquid chromatography for the determination of Diroximel fumarate and its related impurities

AbstractDiroximel fumarate (DRF) is a novel oral fumarate used in the treatment of multiple sclerosis. Comprehending DRF's stability behavior in different degradation conditions is crucial and has been studied under photolysis, oxidative, hydrolysis, and thermal conditions. Nucleosil C18 column (250 × 4.6 mm, 5 μm) was employed to achieve drug separation, with a mobile phase comprising of water and acetonitrile (65:35, v/v). The flow rate was 1 mL/min and detection was performed at 210 nm using a photodiode array detector. The fully validated reversed phase‐high‐performance liquid chromatography stability method as per the International Council for Harmonization Q2 R1 demonstrated excellent selectivity, accuracy, and precision with good sensitivity. During the analysis, two prominent degradant product peaks of DRF were identified across all stress conditions. The usage of liquid chromatography‐mass spectrometry compatible solvents in the mobile phase permits further characterization of DRF and its degradants peak. Significant levels of degradation were found in alkaline and acidic hydroxide, peroxide, photolytic, and thermal conditions with hydrolysis being postulated as a possible mechanism of degradation. The fully validated stability‐indicating analytical method can be used for routine quality control and stability testing of DRF, requiring storage in a protected primary container, as it is sensitive to light and moisture.

Response surface methodology optimisation enhancing lactic acid production from Prosopis africana pods by Rhizopus oryzae

Microbial fermentation of lignocellulosic biomass into lactic acid (LA) has received considerable attention because it ensures the valorisation of wastes and reduces dependence on fossil sources. First, the proximate and phytochemical compositions of Prosopis africana pods (PAP) were determined. Previous biologically-pretreated pods of Prosopis africana were then saccharified using dilute acid hydrolysis following a full factorial design. The factors optimized were acid type (HCl and H2SO4), acid concentration (1 %, 3 % and 5 %), solid loading ratio (5 %, 10 % and 20 %) and reaction time (15, 30 and 60 minutes). Several Rhizopus oryzae isolates were screened for LA production and the most prolific was molecularly identified. The factors affecting LA yield from PAP hydrolysate were screened using a half-factorial design. The significant factors were then optimised using Box-Behnken Design of Response Surface Methodology (RSM). The proximate analysis showed high levels of protein, lipid, ash and carbohydrates. The phytochemical analysis of PAP revealed the presence of flavonoids, alkaloids, saponins, phenols, tannins, glycosides and terpenoids. The hydrolysis conditions of 3 % HCl, 20 % solid loading and 15 minutes hydrolysis time yielded the highest reducing sugars of 42.5 g/L. The most promising isolate, identified as R. oryzae strain AK-22, produced 19.7 g/L after RSM optimization, a 38.1 % increase over yields from non-optimised conditions. These findings are on the biotechnological production of LA from the pods of Prosopis africana, an abundant yet under-utilised tree crop.

3-isocyanatopropyltriethoxysilane block-graft engineering tailoring carbon fiber surface to manipulate interface properties of carbon fiber/polyamide 6 composites

Introducing the isocyanate group on carbon fiber (CF) to improve its surface activity is an effective method for CF surface modification. However, its application in industry is limited by the fact that the modification process is harmful to the environment and the isocyanate is prone to fail. In this paper, caprolactam (CPL) was used to block the isocyanate bond of 3-isocyanopropyltriethoxysilane (IPTS). By adjusting the hydrolysis conditions and concentration of the modified coupling agent solution, the CPL-blocked IPTS was grafted onto the surface-oxidized CF, and the CF-reinforced polyamide 6 (PA6) composites were prepared. The results showed that the interlaminar shear strength (ILSS) of the composites reached 52.53 MPa after treating CF with 1.0 wt% of coupling agent solution, which was 23.83% higher than that of the desized CF/PA6 composites. During the composite molding process, the high temperature unblocked the IPTS, and the released isocyanate reacted with the terminal amino group of PA6 by covalent grafting to establish a high-strength composite interface, which was the main reason for the improvement of ILSS. This study shows that this method is simple and environmentally friendly, and can effectively improve the interface properties of CF/PA6 composites.

Production and characterization of amaranth amyloid fibrils from food protein waste

Plant proteins self-assembly into amyloid fibrils is a promising modification to be introduced in emerging food and materials applications. Although the valorization of protein-rich side streams in amaranth oil production is completely missing, we show that double salt extraction provides the recovery of globulin fraction of amaranth seed from food waste with high purity and complete protein solubility at acidic pH. We demonstrate in silico the self-assembly propensity with a high density of amyloidogenic regions of the 11S seed storage globulin of Amaranthus hypochondriacus. We further show that the recovered amaranth proteins can self-assemble into amyloid fibrils under heat-induced acidic hydrolysis. The amyloid nature of the fibrils was studied by thiazole orange (TO) fluorescence, SDS-PAGE, FTIR, CD, TEM, and AFM. We systematically investigated how the temperature of fibrillization affected protein unfolding, peptide release, as well as its effect on tuning the formation, growth, and polymorphism of amaranth amyloid fibrils. The β-sheet-rich structured proteins produced from the amaranth food waste exhibited mesoscopic polymorphism of the rigid amyloid fibrils with the manifestation of left-handed ribbons of varying polymorphism. We show that favourable conditions for extensive hydrolysis of the amaranth proteins promote the growth of well-ordered thick twisted ribbons with small-pitch and extraordinary rigidity and we interpret these finding based on structural mesoscopic models of amyloid fibrils. These results provide additional insight into the formation of plant amyloid fibrils as functional food and biocompatible materials, and introduce a pathway to convert amaranth food protein waste into amyloid nanofibrils.

The Production of Bioactive Peptides by Optimization of Enzymatic Hydrolysis Process of Protein from Tilapia Fish Skin Waste (Oreochromis niloticus, Linnaeus 1758) using Alcalase 2.4.L

Aims: This study aimed at developing bioactive peptides by optimization of the enzymatic hydrolysis process of protein from tilapia fish skin waste (Oreochromis niloticus, Linnaeus 1758) using alcalase 2.4.L. Background: Natural bioactive peptides are considered to have low toxicity and therapeutic properties as antioxidants. Objective: The conditions of protein hydrolysis obtained from tilapia fish skin waste (Oreochromis niloticus, Linnaeus 1758) were optimized using alcalase 2.4.l Methods: In this study, the hydrolysis of protein obtained from tilapia fish skin waste (TFSW) was optimized using alcalase 2.4.L by central composite design (CCD). Degree of hydrolysis (DH), radical scavenging activities (DPPH), and ferric-reducing antioxidant power (FRAP) were used as dependent variables, whereas temperature, pH, and proportion of enzyme to the substrate (PE%) as independent variables. Results: The optimum degree of hydrolysis DH%, DPPH, and FRAP were achieved at a temperature of 58.4 ℃, a pH of 8.7, except for DPPH, which was achieved at a pH of 7.0. Conclusion: The present work demonstrated that TFSW could be used as a source to produce bioactive peptides with significant antioxidant activities under specific conditions of enzymatic hydrolysis.

Optimization of the Enzymatic Hydrolysis of Yellowfin tuna, (Thunnus albacares) Viscera by Response Surface Methodology

Tuna is abundant in the Philippines, but the by-products during processing in various forms may be a source of waste and environmental pollution. To minimize these wastes, they are utilized directly or fermented to serve as food for humans and animals and as a functional food ingredient since they are rich in proteins and polyunsaturated lipids. For this purpose, they are often converted into protein hydrolysates using proteolytic enzymes. This study optimized the hydrolysis of the yellowfin tuna (Thunnus albacares) viscera (YFTV) using the enzyme neutrase to produce hydrolysates with a high degree of hydrolysis (DH) and foaming capacity (FC). Using the central composite design of the Response Surface Methodology (RSM), the YFTV protein hydrolysis at varying concentrations (0.5-1.5%, w/v) and hydrolysis time (60 to 180 min) was carried out. From the RSM-generated model, the optimum conditions to obtain the highest DH was 179.50 min hydrolysis time and 1.5% enzyme concentration, and for the highest FC, 176.58 hydrolysis time and 1.5% enzyme concentration. The predicted optimum values using the generated linear and quadratic equations were 17.26% DH and 1.60% FC. The lack of a fit test for both responses yielded an insignificant value (p > 0.05) for the model, suggesting that the regression coefficient was sufficient for estimating both responses under any group of variables. The optimized protein hydrolysis conditions of YFTV using Neutrase could be applied in food production systems, especially downstream processing. Furthermore, the utilization of tuna viscera as protein hydrolysates could potentially contribute to the waste management of these processing by-products.

Tolerance of engineered Rhodosporidium toruloides to sorghum hydrolysates during batch and fed-batch lipid production

BackgroundOleaginous yeasts are a promising candidate for the sustainable conversion of lignocellulosic feedstocks into fuels and chemicals, but their growth on these substrates can be inhibited as a result of upstream pretreatment and enzymatic hydrolysis conditions. Previous studies indicate a high citrate buffer concentration during hydrolysis inhibits downstream cell growth and ethanol fermentation in Saccharomyces cerevisiae. In this study, an engineered Rhodosporidium toruloides strain with enhanced lipid accumulation was grown on sorghum hydrolysate with high and low citrate buffer concentrations.ResultsBoth hydrolysis conditions resulted in similar sugar recovery rates and concentrations. No significant differences in cell growth, sugar utilization rates, or lipid production rates were observed between the two citrate buffer conditions during batch fermentation of R. toruloides. Under fed-batch growth on low-citrate hydrolysate a lipid titer of 16.7 g/L was obtained.ConclusionsCitrate buffer was not found to inhibit growth or lipid production in this engineered R. toruloides strain, nor did reducing the citrate buffer concentration negatively affect sugar yields in the hydrolysate. As this process is scaled-up, $131 per ton of hydrothermally pretreated biomass can be saved by use of the lower citrate buffer concentration during enzymatic hydrolysis.Graphical

Optimization, Scale-Up, and Economic Analysis of the Ethanol Production Process Using Sargassum horneri

Recently, the extensive spread of some algae along coastlines has surged into unmanageable thick decomposition layers. This study aimed to demonstrate the use of Sargassum horneri as a biomass resource for ethanol production through the continuous hydrolysis, enzymatic saccharification, and fermentation process. Sugars from S. horneri were obtained using a combination of thermal acid hydrolysis and enzymatic saccharification. The optimal conditions for thermal acid hydrolysis involved a 10% (w/v) S. horneri slurry treated with 100 mM H2SO4 at 121 °C for 60 min; enzymatic saccharification using 16 U/mL Cellic CTec2 further boosted the monosaccharide concentration to 23.53 g/L. Fermentation experiments were conducted with mannitol-adapted Saccharomyces cerevisiae BY4741 using S. horneri hydrolysate. Enhanced ethanol production was observed in the hydrolysate, particularly with mannitol-adapted S. cerevisiae BY4741, which yielded 10.06 g/L ethanol. Non-adapted S. cerevisiae produced 8.12 g/L ethanol, as it primarily utilized glucose and not mannitol. Regarding ethanol fermentation using 5 L- and 500 L-scale fermenters, the ethanol concentrations reached 10.56 g/L and 7.88 g/L with yields of 0.51 and 0.45, respectively, at 48 h. This study confirmed the economic viability of ethanol production using waste seaweed with optimized pretreatment conditions and the adaptive evolution of S. cerevisiae to mannitol.

Optimization of ultrasound-assisted enzymatic hydrolysis of channel catfish (Ictalurus punctatus) frames yield high-calcium and high-amino acid hydrolysates

ABSTRACT In this study, a high-calcium and high-amino acid fish frame hydrolyzate (FFH) was prepared from channel catfish (Ictalurus punetaus) through ultrasound-assisted enzymatic hydrolysis (UAEH). A 4-factor and 3-level Box-Behnken design of response surface methodology was employed to optimize the hydrolysis conditions, where the independent variables were sonication power (W), sonication time (min), enzyme activity (U) and water-substrate ratio (v/w). The degree of hydrolysis (DH) and soluble calcium content (SCC) were used as response factors. The optimal hydrolysis conditions were found to be sonication power (X1) = 492.91 W, sonication time (X2) = 15.01 min, enzyme activity (X3) = 6677.88 U, and water-to-substrate ratio (X4) = 3.56 v/w. Under these conditions, UAEH effectively disintegrated fish frame collagen and converted bound calcium to soluble calcium. Nevertheless, this study investigated the pattern of collagen transformation into short peptides and free amino acids throughout the degradation process. These findings highlight the potential of the FFH from channel catfish to serve as a foundation for creating highly nutritious food additive. The outcomes of this study further enhance the food industry’s ability to capitalize on the nutritional benefits offered by fish frame hydrolyzates. This expansion of health-focused product options and endorsement of the sustainable use of fish by-products harmonizes seamlessly with the industry’s evolving goals of waste reduction and value optimization.

Stability Indicating HPTLC Method Development and Validation for Acebrophylline in Bulk and its Tablet Dosage Form

A simple, precise, accurate stability-indicating method was developed and validated for Acebrophylline in bulk and tablet dosage form. The chromatographic separation was achieved by using Methanol: Acetonitrile: Acetone: Formic acid (4:4:2:0.1 v/v/v/v) as mobile phase and UV detection at 272nm. The retention factor for Acebrophylline was found to be 0.74±0.10. The developed method was validated with respect to linearity, accuracy, precision, limit of detection, limit of quantitation, and robustness as per ICH guidelines. Results were found to be linear in the concentration range of 400-2400ng band -1 with a correlation coefficient of 0.9921. The % assay was found to be 102.4±5. The drug was subjected to stress conditions of hydrolysis (acid, base), oxidation, photolysis, and thermal degradation. The developed method can be used for the quantification of drug in the dosage form, bulk drug as well as for routine analysis in quality control laboratories.