Base Hydrolysis Research Articles

Synthesis of a novel polymer and design of carboxylate-terminated hyperbranched PEI-incorporated PVDF membranes for efficient oil-in-water emulsion separation

Polyvinylidene fluoride (PVDF) membranes utilizing ultrafiltration (UF) and microfiltration (MF) configurations have widely been used in many separation processes, including oil/water emulsions. Recently, there have been tremendous research interests in designing PVDF membranes on their structural-property relations as well as high separation performances. Herein we report the synthesis of a novel carboxylate-terminated hyperbranched polyethylenimine (CHPEI) using a facile three-step one-pot strategy to enhance the characteristics of PVDF membranes for oil/water emulsion separation. The first step involved the production of one amide and one carboxylate functional group per maleic anhydride molecule on the outer surface of polyethylenimine (PEI). The second step involved producing two additional carboxylate groups via amine-based Michael addition of alkenes using aspartic acid dimethyl ester hydrochloride, followed by basic hydrolysis. CHPEI-n-PVDF membranes having various CHPEI contents are further fabricated by a phase-inversion process, obtaining as free-stable flat-sheet membranes. The synthesized polymer and membranes were thoroughly characterized by FTIR and 1H NMR, SEM, EDX, mapping, AFM, and contact angle analysis. The incorporation of CHPEI into PVDF positively enhanced the hydrophilicity, and the WCA was significantly reduced from 80.6° to 59.5°. The maximum permeation flux was obtained using CHPEI-3-PVDF membranes, which was 200 % more than that obtained using pristine PVDF. The CHPEI-2-PVDF membranes exhibited a rejection performance of >99 % for separating oil-in-water emulsions. The incorporation of CHPEI into PVDF significantly improved the antifouling characteristics of the membranes. After 5 h of operation in separating oil-in-water emulsions, the flux recoveries of CHPEI-2-PVDF and CHPEI-3-PVDF were found to be 92 % and 97 %, respectively. CHPEI has demonstrated a considerable positive influence in improving the PVDF membrane’s performance, making it an excellent candidate for oily wastewater treatment because of its high rejection performance and excellent flux recovery ratio.

Development and validation of stability indicating HPTLC method for simultaneous estimation of Dapagliflozin and Linagliptin

For the treatment of Type 2 Diabetes Mellitus, Dapagliflozin selectively inhibits the sodium-glucose co-transporter-2, while Linagliptin competitively and reversibly inhibits dipeptidyl peptidase-4 in a fixed dose combination (1:1). Dapagliflozin and Linagliptin estimation in bulk and tablet formulation was accomplished by the development and validation of a precise and accurate HPTLC method. The procedure used Toluene: Chloroform: Methanol: Triethylamine (7:2:1:0.2 v/v/v) as the mobile phase and HPTLC aluminum plates pre-coated with silica gel 60 F254 as the stationary phase used. Dapagliflozin and Linagliptin were determined to have Rf values of 0.23 and 0.40 correspondingly. At 224 nm, densitometric analysis was performed in the absorbance mode. For Dapagliflozin and Linagliptin 200–1200 ng/band were the linear ranges in which the procedure produced results. Forced degradation studies were performed on both Dapagliflozin and Linagliptin active pharmaceutical ingredients like acid hydrolysis, base hydrolysis, chemical oxidation, dry heat and photodegradation studies. Dapagliflozin and Linagliptin are susceptible to acid hydrolysis, base hydrolysis, chemical oxidation while both the chemical substances are stable to dry heat and photolytic studies.

Diketopyrrolopyrrole based molecular semiconductors with intrinsic conductivity

Intrinsically conducting diketopyrrolopyrrole (DPP) compounds functionalised with pendant quaternary ammonium groups have been synthesised. The hydroxide forms were found to be the product of base hydrolysis of the DPP amide during synthesis, likely made possible by restricted resonance. This gave mixed chemical compositions of solutions and conductive films formed by drop-casting. Chemical decomposition of the hydrolysed compounds was also observed at temperatures above 85 °C. Conductivities of approximately 3 × 10−4 S m−1 were observed in processed films of DPP ammonium hydroxides. It was also found that trifluoroacetic acid (TFA) DPP precursors gave clear electron paramagnetic resonance (EPR) signals indicative of doping and exhibited conductivities one order of magnitude larger than the hydroxides. This opens the possibility of using ammonium TFAs as moieties for doping organic semiconductors.

Identification and Characterization of Rotigotine Degradation Products by HPLC Coupled DAD and CAD Detectors and HRMS Through Q-Orbitrap and Electrospray Ionization

Rotigotine (RTG) is a dopamine agonist used in the treatment of Parkinson's disease. As it is susceptible to oxidation, stability studies must be carefully designed for the identification and characterization of all possible degradation products. Here, RTG degradation was evaluated according to the International Conference on Harmonization guidelines under various stress conditions, including acidic and basic hydrolysis, oxidative, metallic, photolytic, and thermal conditions. Additionally, more severe stress conditions were applied to induce RTG degradation. Significant degradation was only observed under oxidative and photolytic conditions. The samples were analyzed by high performance liquid chromatography coupled to photodiode array detectors, charged aerosol, and high-resolution mass spectrometry. Chromatographic analyses revealed the presence of eight substances related to RTG, four of which were already described and were qualified impurities (impurities B, C, K and E) and four new degradation products (DP-1 – DP-4), whose structures were characterized by high-resolution mass spectrometry through Q-Orbitrap and electrospray ionization. In the stress testing of the active pharmaceutical ingredient in solid form, significant RTG degradation was observed in the presence of the oxidative matrix. The results corroborate the literature that confirm the high susceptibility of RTG to oxidation and the importance of using different detectors to detect degradation products in forced degradation studies.

Texturally-enhanced 55S0P and 45S10P Bioactive Glass ceramic particles: Sol-gel fabrication, nano-characterization and comprehensive Bio-evaluation for applications in Bone tissue engineering

Bioactive glass ceramics (BGC) play a pivotal role in bone tissue engineering, specifically addressing challenges associated with bone repair and regeneration. In this study, we developed silica-based 55S0P and 45S10P spherical BGC nanoparticles (NPs), without and with P2O5 content, using a modified Stöber sol-gel process with base hydrolysis. The fabricated spherical NPs with pronounced textural features effectively contribute to enhanced surface roughness, a desirable parameter for improved biological activity in physiologically relevant solutions and tissue matrices. A comprehensive evaluation of the structural properties of these novel spherical BGC-NPs using diverse characterization techniques revealed distinct nano-textural features in the form of cracks and pores. The HR-TEM confirmed the nanoscale dimensions (170–190 nm) of particles with spherical surface morphology and also systematically analysed via DLS, FE-SEM studies. An augmented surface area with mesopores when phosphate added to the silica glass network was revealed by BET analysis. On the biological front, the study records the formation of an HCA layer on the surfaces of the BGC-NPs, the thickness of which increased with an increase in both phosphate content and immersion time (0, 3, 14 & 28 days) in the SBF solution. The zeta potential values attain a maximum from −22 mV to −23.6 mV for 55S0P and −21.5 mV to −26.4 mV for 45S10P BGC-NPs, and improved with increased immersion time, thereby favouring cellular adhesion for desired biological responses. In vitro, cell culture studies on MG-63 cells, after 24 and 72 h of incubation, demonstrated higher cell adhesion and proliferation, with pronounced biocompatibility. In addition, these BGC-NPs exhibited good hemocompatibility even at the highest test concentration (15 mg/ml) and demonstrated effective antibacterial activity against K. pneumoniae and E. coli. A combinatorial analysis of structural and biological properties revealed that the 45S10P BGC-NPs are more potent than 55S0P BGC-NPs for desired biological performances. In conclusion, the results suggest that the sol-gel fabricated BGC-NPs with enhanced nano-textural features are evidently suitable for applications in bone tissue engineering and regenerative medicine.

Rapid and Convenient Potentiometric Method for Determining Fluorosulfate, a Byproduct of the Fumigant and Greenhouse Gas Sulfuryl Fluoride.

A fluorosulfate ion (FSO3-) is a hydrolysis product of sulfuryl fluoride (SO2F2), which is widely used to fumigate buildings, soil, construction materials, and postharvest commodities, and is a potent greenhouse gas. It is a potential marker for biological exposure to SO2F2 and for monitoring the progress of reactions used to scrub SO2F2 from fumigation vent gases. Here, we report a simple and inexpensive potentiometric method for determining FSO3- using a commercial nitrate-selective electrode and discuss its application. The method is suitable for solutions between 0.0025 mM and 660 mM FSO3- at initial pH between 5 and 9. Halide interference depends on its molar ratio to FSO3- and follows the sequence, F- < Cl- < Br- ≪ I-. Halide interference can be eliminated by adding silver sulfate. Interference by bicarbonate can be eliminated by H2SO4 pretreatment, and interference by phosphate or pyrophosphate by MgSO4 addition. Sulfate does not interfere, as it does in ion chromatography. Satisfactory method detection limits for FSO3- in spiked aqueous extracts of 11 fruits were obtained. The method accurately quantified the yield of FSO3- relative to that of F- in base hydrolysis of SO2F2. This study demonstrates that the developed method is highly selective, convenient, and sensitive and thus can be of great value in practice.

Alkaline Fading of Malachite Green in β-Cyclodextrins

The basic hydrolysis of Malachite Green (MG) in the presence of β-Cyclodextrin (β-CD) has been studied using UV-Vis spectroscopic techniques and at 20 °C. β-CD was found to catalyze the basic hydrolysis. Indeed, this basic hydrolysis is catalyzed by the interaction cyclodextrin hydroxyl group, in its deprotonated form with the carbocation in the host-guest complex. The proposed model has been successfully applied to a reaction catalyzed by CD. It considers two simultaneous pathways in the aqueous medium involving free hydroxyl ions and the substrate-CD complex. The model allows us to obtain the kinetic parameters including the bimolecular rate constant between MG and HO− in bulk water (kw = 1.47 ± 0.01 mol−1s−1), the rate constant between MG and the deprotonated hydroxyl group of β-CD inside the host-guest complex (kCD = 0.25 ± 0.03 s−1) and the binding constant of MG inside the β-CD (KS = 2500 ± 50). This behavior is like the hydrolysis of Cristal Violet (CV) in the same reaction media.

Sensitive and validated UPLC-MS/MS method for the identification and characterization of forced degradation products for felodipine

AbstractA rapid, stability indicating reverse phase liquid chromatographic method was developed for the determination of purity of Felodipine in active pharmaceutical substance form in the presence of its impurity and its degradation products. To develop the method which is also compatible to liquid chromatographic mass spectroscopic technique. The developed method is also used to determine the assay of Felodipine in bulk drug form. The drug is subjected to various stress conditions like acidic, basic, oxidation, UV light and thermal conditions. Considerable degradation was observed during base hydrolysis. Two degradation products were identified. The Waters Acquity UPLC BEH C18, 2.1 × 100 mm, 1.7 µm Column was used to achieve chromatographic separation. The gradient conditions, diluent and injection volume were optimized to achieve the acceptable resolution between impurities and its degradation products from Felodipine and to get good peak shapes. The masses were determined for main compound and its identified degradation products. Further, the characterization studies for main compound and its degradation products were performed using LCMSMS Q-TOF.

Validated chromatographic methods for determination of teriflunomide and investigation of its intrinsic stability

Two rapid, precise, and sensitive stability-indicating high performance chromatographic methods for the measurement of Teriflunomide in its degradation products’ existence were developed. These were RP-HPLC and HPTLC using UV detector. HPLC separation was accomplished utilizing Thermo BDS hypercil C18 column (250 × 4.6 mm, 5 μm) and acetonitrile: 0.03 M potassium dihydrogen phosphate: triethylamine (50:50:0.1%, by volume) as mobile phase at flow rate of 1mL/min. The separated peaks were detected at 250.0 nm. The densitometric approach was conducted utilizing HPTLC 60 F254 silica gel plates, and a developing system of benzene: ethanol: acetic acid (7.5:1:0.25, by volume) and detection was done at 250.0 nm. The developed approaches were evaluated regarding the International Conference on Harmonization (ICH) instructions. The calibration curves of both techniques were constructed with linearity ranges of (5-100) µg/mL and (2–10) µg /band, for HPLC and densitometric determination, consecutively. Teriflunomide was exposed to base and acid hydrolysis, oxidation using H2O2 and finally, thermal degradation as stated in ICH guidelines. The degradation product structures’ elucidation was achieved through LC-MS.

Estimation of Lexisinatide in Bulk and Tablets by RP‐UPLC Method Developed by Quality by Design Concept

AbstractThe primary goal of the proposed study is to establish and validate a low‐cost, sensitive, reliable, and trouble‐free RP‐UPLC process with improved performance for the direct estimation of Lixisenatide in pure powder and fixed‐dose tablet form. The proposed method is optimized by implementing quality by design studies. The method is optimized using quadratic model of central composite design (CCD) of randomized response surface study. Successful separation of Lixisenatide is obtained by the use of Phenyl‐XBD (1.7 µm, 2.1 mm × 100 mm) column and a mobile phase of acetonitrile: 0.1% w/v OPA (60:40) with 0.5 mL min−1 flow rate. The separated Lixisenatide and its degradants are explorated with PDA detector which has the ability to provide comprehensive spectral information, rapid data acquisition, quantitative analysis, and peak purity analysis of all compounds simultaneously. The RT of Lixisenatide is found to be 1.34 min. The lowest concentration of Lixisenatide's limit of detection and limit of quantification makes guarantees regarding the method's sensitivity. For the provided range of linear concentrations (1.25–7.5 µg mL−1), the regression coefficient is found to be 0.999. The computed average percentage recoveries of Lixisenatide in spiked solutions ranged from 98.9% to 101.2%. The % degradation of Lixisenatide in the presence of a variety of forced conditions confirms the existing method's stability. Lixisenatide is significantly photo sensitive when compared to other stressful conditions such as acid, base hydrolysis, and oxidation. The proposed UPLC approach separates Lixisenatide with high sensitivity, a short retention time, and an affordable solvent system.

LC-HRMS and NMR studies for the characterization of degradation impurities of ubrogepant along with the in silico approaches for the prediction of degradation and toxicity

Ubrogepant is the first oral calcitonin gene-related peptide (CGRP) receptor antagonist which is used for the acute treatment of migraine in adults. The present study employs liquid chromatography-high resolution mass spectrometry (LC-HRMS) and nuclear magnetic resonance spectroscopy (NMR) techniques for the identification and characterization of degradation impurities of ubrogepant. The forced degradation study of ubrogepant was performed as per the International Council for Harmonisation (ICH) Q1A and Q1B guidelines. The in silico degradation profile of ubrogepant was predicted by Zeneth. It was observed that ubrogepant was labile to acidic hydrolysis, basic hydrolysis, and oxidative degradation conditions (H2O2), although it was stable in neutral hydrolysis and photolytic (UV light and visible light) conditions. Eight degradation impurities were formed, which were separated on reversed-phase HPLC with a gradient program on an InertSustain C8 column (4.6 × 250 mm, 5 µm) using 10 mM ammonium formate (pH unadjusted) and acetonitrile as the mobile phase. The structures of all the degradation impurities were characterized using the exact masses obtained from the HRMS/MS. Further, NMR studies were conducted on two major degradation impurities (UB-4 and UB-7). A plausible mechanism was proposed to support the structures of all the degradation impurities of UBR. In silico toxicity and mutagenicity assessment were done by DEREK Nexus, SARAH Nexus, and ProTox-II.

Quality by Design-driven Analytical Method: A Quality Risk Management-Based Liquid Chromatography Method for Daclatasvir and Characterization of its Putative Degradants by LC-MS/MS

Background:: Antiviral drugs can cure more than 95 percent of people with hepatitis C, but the inaccessibility of quality affordable medicines and the lack of their uninterrupted supply poses a major challenge. Impurities in drugs have a significant impact on their quality and are one of the substantial causes of drug recalls, ultimately leading to the unavailability of the drug in the market. Hence, there is a need for a robust, quality risk management and quality by design-driven analytical method that can detect the antiviral drug, Daclatasvir dihydrochloride, in the presence of its probable impurities. Objective:: This study aimed to develop a Quality by Design-driven stability- indicating liquid chromatography method for Daclatasvir dihydrochloride and the characterization of its putative degradants by LC-MS. Method:: The fishbone diagram and quality risk assessment investigated twenty-four process parameters and concluded that three risk parameters, i.e., flow rate, buffer pH, and stationary phase type, were the critical process parameters. The critical quality attributes viz. resolution between impurity 6 and DCV and impurity 2 &amp; 3 (Rs˃1.5), the shape of the peak of DCV which is decided by the Number of Theoretical Plates (NTP˃5000), and the retention time of Daclatasvir (tR14-23 mins) were optimized using a two-level three-factor full factorial design with five center points. Results:: The optimized method is stability-indicating in its true sense as it can separate the sample with its degradants generated in basic (three), acidic (two), oxidative (H2O2: three, Azobisisobutyronitrile: one), photo (three), and dry heat (one) conditions. Degradants structures were elucidated, and degradation routes were established, using LC-MS and LC-MS/MS analyses. Conclusion:: The drug is highly susceptible to acid, base hydrolysis, and oxidation degradation conditions and poses a significant risk to the analytical method to fail in system suitability criteria. Hence, a robust and flexible chromatographic method with the capacity for continuous improvement was developed and successfully validated within the criteria of design space.

Copper Soaps Formation in Verdigris-Linseed Oil Painting Mixtures: A Multispectroscopic Characterization.

Copper acetate (also known as verdigris) is a bimetallic Cu(II) greenish pigment widely used in oil paintings. Since ancient times, this pigment has been known as a degradation-prone compound, especially when combined with lipidic binders. However, the degradation mechanism and the involved species have not yet been disclosed. In this article, we study verdigris interactions with linseed oil in painting mock-ups, stressing out the formation of copper-based complexes and proposing reaction routes. Such complex systems are studied by applying a complementary multispectroscopic approach: a combination of continuous-wave (CW) electron paramagnetic resonance (EPR), electron spin echo envelope modulation (ESEEM), and Raman and attenuated total reflection (ATR) Fourier transform infrared (FTIR) spectroscopies. Based on the experimental evidence, we propose the following mechanism: the neutral copper acetate shifts to a basic verdigris-promoting triglyceride hydrolysis, aided by the coordination of Cu(II) cations toward the carboxylic functions. The increased amount of free fatty acids in the mixtures triggers the formation of monomeric Cu(II) complexes. Afterward, the oil polymerization reaction occurs, and secondary oxidation species, containing OH groups in the alkyl chain, act as further ligands for copper nuclei. This is the first time, to the best of our knowledge, that a comprehensive view is proposed.

Complex compound of trinitrotriamminecobalt(III): in theoretical studies

[Co(NH3)3(NO2)3] is an octahedral complex compound that can have several isomers. The complex compound has magnetic properties. Its stability has been explained. It can be easily synthesized and is known as a bioinorganic synthesis reagent, oxidant compound and base hydrolysis.

A newly developed and validated environmentally friendly RP-HPLC stability indicating method for the COVID-19 antiviral Molnupiravir: Application to degradation kinetics structure suggestion using LC-MS and the effect on viable cells of the major degradation products

The SARS-CoV-2 coronavirus pandemic also referred to as COVID-19 is a severe respiratory illness. Molnupiravir (Moln) is a frequently prescribed antiviral medication used in the treatment regimen for mild to moderate cases of COVID-19. Since medications might experience chemical deterioration over time it is crucial to create a stability-indicating technique to evaluate the drug's stability. This study presents the development and validation of a novel stability-indicating approach employing reversed-phase high-performance liquid chromatography. The purpose of this method is to accurately quantify Moln in the presence of its degradation products. The technique was determined to be linear throughout the concentration range of 10–200 μg/mL. It exhibited precision accuracy and specificity. Moln was shown to be susceptible to oxidative acidic and basic hydrolysis. The proposed structure of the degradants was determined using LC-MS. The degradation kinetics were examined revealing that both acidic and oxidative hydrolysis conditions adhere to first-order kinetics. The sulforhodamine B (SRB) test was used to assess the impact of these degradants on Human skin fibroblast cell. The method was assessed using four green metric scales; National environmental metric index (NEMI) Analytical-ECO scale Green Analytical Procedure Index (GAPI) and Analytical GREEnness Metric Approach (AGREE); and it was determined to be ecologically friendly.

Analytical quality by design aided stability indicating a robust ultra‐performance liquid chromatographic technique for the quantification of sonidegib and its organic impurities in bulk drug substance

AbstractA simple quality by design aided stability indicating method was developed for quantification of sonidegib (SONI) and its process related impurities using on ultra‐performance liquid chromatography in bulk drug substance. AutoChrom and Design‐Expert software were used to predict physicochemical properties, draw Ionization graphs, and generate analytical target profile. SONI was subjected to forced degradation conditions, such as oxidative, acid hydrolysis, base hydrolysis, hydrolytic, thermal, and photolytic hydrolysis. All degradation products and process contaminants were separated using an Acquity Ethylene Bridged Hybrid C18 column in gradient elution mode with a mobile phase containing 0.02 M ammonium acetate buffer and acetonitrile: methanol (80:20 v/v). The predicted physicochemical properties are accurate and they facilitated for selection of robust conditions in development of chromatographic method with minimal trials. The developed method can be used for quantification of drug and its process related impurities in bulk drugs.

An easy and simple method for the immobilization of dyes through click reactions: activated alkyne, copper not needed.

The copper-free azide-alkyne click reaction has shown to be a successful alternative to immobilize covalently a fluorescente compound onto poly(-l-lactic) acid (PLLA) surfaces. Proceded by basic hydrolysis and amidation reaction, typical surface characterization techniques have validated each functionaliztion step and the success of the conjugation. This method offers a catalyst-free option for various surface conjugations, extremely demanded in biomedical and biosensory fields.

STABILITY INDICATING UPLC METHOD FOR ESTIMATION OF BENAZEPRIL AND HYDROCHLOROTHIAZIDE IN BULK AND COMBINED DOSAGE FORM

Objective: The main objective was to develop stability indicating UPLC technique for simultaneous estimation of Benazepril and Hydrochlorothiazide in bulk and formulation. Methods: 0.1% Triethylamine phosphate: Methanol (25:75v/v) was used as the mobile phase. Benazepril linearity was found to be 4-20 µg/ml and Hydrochlorothiazide linearity was found to be 5-25 g/ml. The detection wavelength was 236 nm, and the retention period of Benazepril was 3.4 min and Hydrochlorothiazide was 5.4 min with a flow rate of 1.0 ml/min. According to the ICH guidlines, the proposed method was validated and stress studies revealed that the drugs are prone to alkali and peroxide stress conditions. Results: The calibration curve was plotted, and the regression equations for Benazepril were y = 2,01,491.67x+60,532.30 with a correlation coefficient (r2) of 0.9997 and Hydrochlorothiazide were y = 64,635.86x-74,607.10 with a correlation coefficient (r2) of 0.9994. According to the accuracy research, the percent recovery of Benazepril is 99.09-100.69 % and that of Hydrochlorothiazide is 98.27-101.88%, both of which are within the ICH recommendations. Benazepril has a limit of detection of 0.08 g/ml-0.24 g/ml and Hydrochlorothiazide has a limit of quantitation of 0.03 g/ml-0.10 g/ml. The procedure was found to be straightforward, linear, rapid, exact, repeatable, and robust. It was determined that the % RSD was within ICH norms. Stress degradation tests showed the drug's vulnerability to oxidative, thermal, photolytic, acid, basic, and neutral hydrolysis stress conditions. Under the circumstances of alkali and peroxide stress, it was discovered that the drug degraded most quickly. Conclusion: The developed chromatographic technique under consideration was suitable for the accurate, precise, and quick simultaneous measurement of hydrochlorothiazide and benazepril in both their bulk and medicinal dose forms.

Alcohol synthesis based on the SN2 reactions of alkyl halides with the squarate dianion.

A convenient method has been developed for transforming alkyl halides into the corresponding alcohols via an SN2 reaction. Treatment of an alkyl halide with the squarate dianion at high temperature produces mono-alkyl squarate, and a one-pot basic hydrolysis of the intermediate affords the alcohol in good yield.

The effect of amaranth, a pseudo-cereal, on the activity of L. acidophilus probiotic bacteria and its antioxidant activity in the gastrointestinal digestion process

Pseudo-cereals are an excellent source of nutrients, rich in carbohydrates, dietary fiber, protein, lipids, phytochemicals, and minerals such as magnesium, zinc, copper, sodium, potassium, and calcium. The positive effects of gluten-free pseudo-cereals on the digestive system are an alternative to natural cereals. Pseudo-cereals have prebiotic properties and strengthen digestion by positively affecting the development of probiotic bacteria, especially Lactobacillus. Therefore, the effect of amaranth, a pseudo-cereal, on the activity of L. acidophilus probiotic bacteria, which helps digestion, was determined. First, solvent, acidic, and basic hydrolysis extractions of amaranth in eight different solvent media were performed, and total phenolic content and antioxidant activity values were determined. The total phenolic content values in the gastrointestinal digestion process were investigated by applying three different consumption methods, milling, boiling, and drying, to amaranth grains. L. acidophilus probiotic bacteria were activated with milled, dried, and boiled amaranth, and the increase in viability was examined. While the viability of L. acidophilus activated with milled and dried amaranth increased by 9.47% and 7.46%, respectively, the viability of bacteria activated with boiled amaranth almost did not increase (0.60%).