Comparative Large Amplitude Oscillatory Shear (LAOS) Study of Ionically and Physically Crosslinked Hydrogels.

Hydrogels are highly versatile and widely applicable materials within various scientific, technological, and food sectors. Alginate and gelatin hydrogels, along with their crafted variations, are possibly the most common ones. However, the ionic crosslinking of alginate-Ca++ is a different gelation mechanism than the physical crosslinking of gelatin. In this work, we prepare alginate-Ca++ hydrogels using individual layer gelation and experimentally evaluate LAOS rheological behavior. We apply shear-stress decomposition using the MITlaos software and obtain the elastic and viscous contributions within the nonlinear response of the individual alginate-Ca++ layer. We compare these results with the nonlinear responses of the gelatin-alginate ex situ individual layer. The strain-sweep patterns are similar, with loss modulus overshoot. The applied shear can destroy the larger-scale structural units (agglomerate/aggregates), resulting in analogous patterns. However, the critical strain points are different. Based on the shear-thickening ratio T of the LAOS analysis, it can be assumed that the common feature of ex situ preparation, i.e., gelation as individual layers, provides a matching bulk microstructure, as the hydrogels differ significantly at a molecular-binding level.

Medical applications of biopolymer nanofibers.

A wide array of biomedical applications, extending from the fabrication of implant materials to targeted drug delivery, can be attributed to polymers. The utilization of chemical monomers to form polymers, such as polypropylene, polystyrene, and polyethylene, can provide high mechanical stability to them and they can be utilized for diverse electronic or thermal applications. However, certain chemical-based synthetic polymers are toxic to humans, animals, plants, and microbial cells. Thus, biopolymers have been introduced as an alternative to make them utilizable for biomedical applications. Even though biopolymers possess beneficial biomedical applications, they are not stable in biological fluids and exhibit toxicity in certain cases. Recent advances in nanotechnology have expanded its applicational significance in various domains, especially in the evolution of biopolymers to transform them into nanoparticles for numerous biomedical applications. In particular, biopolymers are fabricated as nanofibers to enhance their biological properties and to be utilized for exclusive biomedical applications. The aim of this review is to present an overview of various biopolymer nanofibers and their distinct synthesis approaches. In addition, the medical applications of biopolymer nanofibers, including antimicrobial agents, drug delivery systems, biosensor production, tissue engineering, and implant fabrication, are also discussed.

Synthesis approach-dependent antiviral properties of silver nanoparticles and nanocomposites.

Numerous viral infections are common among humans, and some can lead to death. Even though conventional antiviral agents are beneficial in eliminating viral infections, they may lead to side effects or physiological toxicity. Silver nanoparticles and nanocomposites have been demonstrated to possess inhibitory properties against several pathogenic microbes, including archaea, bacteria, fungi, algae, and viruses. Its pronounced antimicrobial activity against various microbe-mediated diseases potentiates its use in combating viral infections. Notably, the appropriated selection of the synthesis method to fabricate silver nanoparticles is a major factor for consideration as it directly impacts antiviral efficacy, level of toxicity, scalability, and environmental sustainability. Thus, this article presents and discusses various synthesis approaches to produce silver nanoparticles and nanocomposites, providing technological insights into selecting approaches to generate antiviral silver-based nanoparticles. The antiviral mechanism of various formulations of silver nanoparticles and the evaluation of its propensity to combat specific viral infections as a potential antiviral agent are also discussed.

Bioactive components from Moringa oleifera seeds: production, functionalities and applications - a critical review.

A readily distinguishable and indigenous member of the plant kingdom in the Indian subcontinent is the 'drumstick tree', i.e. Moringa oleifera Lam. In addition to India, this drought-tolerant and rapidly evolving tree is currently extensively disseminated across the globe, including subtropical and tropical areas. The plant boasts a high nutritional, nutraceutical and therapeutic profile, mainly attributing to its significant repertoire of the biologically active components in different parts: protein, flavonoids, saponins, phenolic acids, tannin, isothiocyanate, lipids, minerals, vitamins, amongst others. M. oleifera seeds have been shown to elicit a myriad of pharmacological potential and health benefits, including: antimicrobial, anticancer, antidiabetic, antioxidant, antihypertensive, anti-inflammatory and cardioprotective properties. Additionally, the seed cakes obtained from post-extraction process are utilized for: coagulation, flocculation and sedimentation purposes, benefiting effluent management and the purification of water, mainly because of their capability in eliminating microbes and organic matter. Despite the extraordinary focus on other parts of the plant, especially the foliage, the beneficial aspects of the seeds have not been sufficiently highlighted. The health benefits of bioactive components in the seeds are promising and demonstrate enough potential to facilitate the development of functional foods. In this review, we present a critical account of the types, characteristics, production and isolation of bioactive components from M. oleifera seeds. Furthermore, we appraise the: pharmacological activities, cosmetic, biodiesel, lubricative, modern farming, nutritive and wastewater treatment applications of these functional ingredients. We infer that there is a need for further human/clinical studies and evaluation, despite their health benefits. Additionally, the safety issues need to be adequately clarified and assessed, in order to establish a conventional therapeutic profile.

3D graphene-based adsorbents: Synthesis, proportional analysis and potential applications in oil elimination.

The suitability and efficacy of three-dimensional (3D) graphene, including its derivatives, have garnered widespread attention towards the development of novel, sustainable materials with ecological amenability. This is especially relevant towards its utilization as adsorbents of wastewater contaminants, such as heavy metals, dyes, and oil, which could be majorly attributed to its noteworthy physicochemical features, particularly elevated chemical and mechanical robustness, advanced permeability, as well as large specific surface area. In this review, we emphasize on the adsorptive elimination of oil particles from contaminated water. Specifically, we assess and collate recent literature on the conceptualization and designing stages of 3D graphene-based adsorbents (3DGBAs) towards oil adsorption, including their applications in either batch or continuous modes. In addition, we analytically evaluate the adsorption mechanism, including sorption sites, physical properties, surface chemistry of 3DGBA and interactions between the adsorbent and adsorbate involving the adsorptive removal of oil, as well as numerous effects of adsorption conditions on the adsorption performance, i.e. pH, temperature, initial concentration of oil contaminants and adsorbent dosage. Furthermore, we focus on the equilibrium isotherms and kinetic studies, in order to comprehend the oil elimination procedures. Lastly, we designate encouraging avenues and recommendations for a perpetual research thrust, and outline the associated future prospects and perspectives.

Therapeutic Applications of Metal and Metal-Oxide Nanoparticles: Dermato-Cosmetic Perspectives.

Invention of novel nanomaterials guaranteeing enhanced biomedical performance in diagnostics and therapeutics, is a perpetual initiative. In this regard, the upsurge and widespread usage of nanoparticles is a ubiquitous phenomenon, focusing predominantly on the application of submicroscopic (< 100 nm) particles. While this is facilitated attributing to their wide range of benefits, a major challenge is to create and maintain a balance, by alleviating the associated toxicity levels. In this minireview, we collate and discuss particularly recent advancements in therapeutic applications of metal and metal oxide nanoparticles in skin and cosmetic applications. On the one hand, we outline the dermatological intrusions, including applications in wound healing. On the other hand, we keep track of the recent trends in the development of cosmeceuticals via nanoparticle engrossments. The dermato-cosmetic applications of metal and metal oxide nanoparticles encompass diverse aspects, including targeted, controlled drug release, and conferring ultraviolet and antimicrobial protections to the skin. Additionally, we deliberate on the critical aspects in comprehending the advantage of rheological assessments, while characterizing the nanoparticulate systems. As an illustration, we single out psoriasis, to capture and comment on the nanodermatology-based curative standpoints. Finally, we lay a broad outlook and examine the imminent prospects.

Polysaccharide-based skin scaffolds with enhanced mechanical compatibility with native human skin.

We report a custom-made technique to synthesize process-convenient skin scaffolds by tuning the mechanical properties of hydrogels based on a few naturally occurring polysaccharides to match the rheological properties of previously established benchmarks, i.e., the ex vivo native human skins. We studied the mechanical parameters using oscillatory shear rheology. At small strain amplitudes, the intrinsic elastic modulus showed an almost linear dependence in the middle and a changing rate profile at the two ends with concentration of the principal hydrogel component variant, i.e., kappa (κ)-carrageenan. At large strain amplitudes, the hydrogels demonstrated intercycle strain-softening behavior, the onset of which was directly proportional to the κ-carrageenan concentration. We observed a concentration match for the intrinsic elastic modulus of the benchmark within this sigmoidal curve fit. Contextually, we need to explore other potent polymeric hydrogel systems to achieve mechanical affinity in terms of multiple rheological parameters derived from both strain amplitude and angular frequency sweeps. Additionally, we carried out diffusion experiments to study caffeine permeation attributes. The hydrogels show improved barrier features with increasing κ-carrageenan concentration. In terms of the penetration flux and total cumulative amount of permeated caffeine, this enhanced mechanical adherence demonstrates comparable penetration features with the commercial 3D skin model.

Vesicle formation mechanisms: an overview.

Vesicle structures primarily embody spherical capsules composed of a single or multiple bilayers, entrapping a pool of aqueous solution in their interior. The bilayers can be synthesised by phospholipids or other amphiphiles (surfactants, block copolymers, etc.). Vesicles with broad-spectrum applications in numerous scientific disciplines, including biochemistry, biophysics, biology, and various pharmaceutical industries, have attracted widespread attention. Consequently, a multitude of protocols have been devised and proposed for their fabrication. In this review, with a motivation to derive the basic conditions for the formation of vesicles, the associated thermodynamic and kinetic aspects are comprehensively appraised. Contextually, an all-purpose overview of the underlying thermodynamics of bilayer/membrane generation and deformation, including the chemical potential of aggregates, geometric packing and the concept of elastic properties, is presented. Additionally, the current review highlights the probable, inherent mechanisms of vesicle formation under distinct modes of manufacturing. We lay focus on vesicle formation from pre-existing bilayers, as well as from bilayers, which form when lipids from an organic solvent are transferred into an aqueous medium. Furthermore, we outline the kinetic effects on vesicle formation from the lamellar phase, with and without the presence of shearing force. Wherever required, the experimental and/or theoretical outcomes, the driving forces for vesicle size selection, and various scaling laws are also reviewed, all of which facilitate an overall improved understanding of the vesicle formation mechanisms.

Graphene-Based Macromolecular Assemblies for Scavenging Heavy Metals.

The integration of graphene or graphene oxide nanosheets into three-dimensional (3D) graphene-based macromolecular assemblies (GMAs), in the form of sponges, beads, fibres, films, and crumpled nanosheets, has greatly advanced their environmental remediation applications. This is attributed to the outstanding physicochemical characteristics and superlative mechanical features of 3D GMAs, including precise and physically linked permeable networks, enormous surface area, profound porosity, and high-class sturdiness, amongst others. In this review, the recent advancements towards the exploration of 3D GMAs as an exciting new class of high-performance adsorbents, for eliminating toxic heavy metal ions from both wastewater and freshwater, are systematically summarized and discussed, from both fundamental and applied perspectives. In particular, the numerous surface modification techniques that are actively pursued to enrich the metal adsorption capacity of 3D GMAs, are comprehensively examined. Additionally, associated challenges are pointed out and tactical research strategies and improvements are proposed, with an eye on the conceivable future.

Aptamers: an emerging class of bioaffinity ligands in bioactive peptide applications.

The food and health applications of bioactive peptides have grown remarkably in the past few decades. Current elucidations have shown that bioactive peptides have unique structural arrangement of amino acids, conferring distinct functionalities, and molecular affinity characteristics. However, whereas interest in the biological potency of bioactive peptides has grown, cost-effective techniques for monitoring the structural changes in these peptides and how these changes affect the biological properties have not grown at the same rate. Due to the high binding affinity of aptamers for other biomolecules, they have a huge potential for use in tracking the structural, conformational, and compositional changes in bioactive peptides. This review provides an overview of bioactive peptides and their essential structure-activity relationship. The review further highlights on the types and methods of synthesis of aptamers before the discussion of the prospects, merits, and challenges in the use of aptamers for bioaffinity interactions with bioactive peptides.

Characterisation of aptamer-anchored poly(EDMA-co-GMA) monolith for high throughput affinity binding.

Immobilisation of aptameric ligands on solid stationary supports for effective binding of target molecules requires understanding of the relationship between aptamer-polymer interactions and the conditions governing the mass transfer of the binding process. Herein, key process parameters affecting the molecular anchoring of a thrombin-binding aptamer (TBA) onto polymethacrylate monolith pore surface, and the binding characteristics of the resulting macroporous aptasensor were investigated. Molecular dynamics (MD) simulations of the TBA-thrombin binding indicated enhanced Guanine 4 (G4) structural stability of TBA upon interaction with thrombin in an ionic environment. Fourier-transform infrared spectroscopy and thermogravimetric analyses were used to characterise the available functional groups and thermo-molecular stability of the immobilised polymer generated with Schiff-base activation and immobilisation scheme. The initial degradation temperature of the polymethacrylate stationary support increased with each step of the Schiff-base process: poly(Ethylene glycol Dimethacrylate-co-Glycidyl methacrylate) or poly(EDMA-co-GMA) [196.0 °C (±1.8)]; poly(EDMA-co-GMA)-Ethylenediamine [235.9 °C (±6.1)]; poly(EDMA-co-GMA)-Ethylenediamine-Glutaraldehyde [255.4 °C (±2.7)]; and aptamer-modified monolith [273.7 °C (±2.5)]. These initial temperature increments reflected in the associated endothermic energies were determined with differential scanning calorimetry. The aptameric ligand density obtained after immobilisation was 480 pmol/µL. Increase in pH and ionic concentration affected the surface charge distribution and the binding characteristics of the aptamer-modified disk-monoliths, resulting in the optimum binding pH and ionic concentration of 8.0 and 5 mM Mg2+, respectively. These results are critical in understanding and setting parametric constraints indispensable to develop and enhance the performance of aptasensors.

Structure-informed separation of bioactive peptides.

The application of proteomic and peptidomic technologies for food-derived bioactive peptides is an emerging field in food sciences. These technologies include the use of separation tools coupled to a high-resolution spectrometric and bioinformatic tools for prediction, identification, sequencing, and characterization of peptides. To a large extent, one-dimensional separation technologies have been extensively used as a continuous tool under different optimized conditions for the identification and analysis of food peptides. However, most one-dimensional separation technologies are fraught with significant bottlenecks such as insufficient sensitivity and specificity limits for complex samples. To address this limitation, separation systems based on orthogonal, multidimensional principles, which allow for the coupling of more than one analytical separation tool with different operational principles, provide a higher separation power than one-dimensional separation tools. This review describes the structure-informed separation and purification of protein hydrolyzates to obtain peptides with desirable bioactivities. PRACTICAL APPLICATIONS: Application of bioactive peptides in the formulation of functional foods, nutraceuticals, and therapeutic agents have increasingly gained scholarly and industrial attention. The bioactive peptides exist originally in protein sources and are only active after hydrolysis of the parent protein. Currently, several tools can be configured in one-dimensional or multidimensional systems for the separation and purification of protein hydrolyzates. The separations are informed by the structural properties such as the molecular weight, charge, hydrophobicity or hydrophilicity, and the solubility of peptides. This review provides a concise discussion on the commonly used analytical tools, their configurations, advantages and challenges in peptide separation. Emphasis is placed on how the structural properties of peptides assist in the separation and purification processes and the concomitant effect of the separation on peptide bioactivity.

Structure-informed detection and quantification of peptides in food and biological fluids.

Peptides with biological properties, that is, bioactive peptides, are a class of biomolecules whose health-promoting properties are increasingly being exploited in food and health products. However, research on targeted techniques for the detection and quantification of these peptides is still in its infancy. Such information is needed in order to enhance the biological and chemometric characterization of peptides and their subsequent application in the functional food and pharmaceutical industries. In this review, the role of classic techniques such as electrophoretic, chromatographic, and peptide mass spectrometry in the structure-informed detection and quantitation of bioactive peptides are discussed. Prospects for the use of aptamers in the characterization of bioactive peptides are also discussed. PRACTICAL APPLICATIONS: Although bioactive peptides have huge potential applications in the functional foods and health area, there are limited techniques in enhancing throughput detection, quantification, and characterization of these peptides. This review discusses state-of-the-art techniques relevant in complementing bioactive detection and profiling irrespective of the small number of amino acid units. Insights into challenges, possible remedies and prevailing areas requiring thorough research in the extant literature for food chemists and biotechnologists are also presented.

Nonlinear viscoelastic properties of native male human skin and in vitro 3D reconstructed skin models under LAOS stress.

This work discusses the first set of rheometric measurements carried out on commercially accessible juvenile and aged skin models under large amplitude oscillatory shear deformations. The results were compared with those of native male whole human and dermis-only foreskin specimens, catering to a few ages from 0.5 to 68 years, including specimens from a 23-year-old male abdomen. At large strains, strain thinning was more pronounced for the dermis of the young skins and for their whole skin counterparts. An inverse qualitative tendency was observed for the adult skins and the skin models. This can be explained by the high dermal collagen compactness associated with an incomplete epidermal proliferation. The qualitative Lissajous plots as well as the quantitative dimensionless indices analyzed using the MITlaos software indicated predominant nonlinear intracycle elastic strain stiffening and viscous shear thinning for all the native specimens at the maximum deformation. For the full thickness models, we have evidence of structure collapse and yielding under similar conditions. The whole skin specimen from the 68-year-old male showed smaller age-dependent nonlinear elastic contributions than the dermis, which we relate to the epidermal degeneration taking place during aging. Regardless of the age group, the models manifested more pronounced intercycle and intracycle elastic nonlinearities, and their magnitudes were significantly larger. The nonlinear elastic trends will serve as advanced standards for understanding and delineating the mechanical limits of destructive and non-destructive deformations of such unique biomaterials.

Binding Characterization of Aptamer-Drug Layered Microformulations and In Vitro Release Assessment.

Efficient delivery of adequate active ingredients to targeted malignant cells is critical, attributing to recurrent biophysical and biochemical challenges associated with conventional pharmaceutical delivery systems. These challenges include drug leakage, low targeting capability, high systemic cytotoxicity, and poor pharmacokinetics and pharmacodynamics. Targeted delivery system is a promising development to deliver sufficient amounts of drug molecules to target cells in a controlled release pattern mode. Aptameric ligands possess unique affinity targeting capabilities which can be exploited in the design of high pay-load drug formulations to navigate active molecules to the malignant sites. This study focuses on the development of a copolymeric and multifunctional drug-loaded aptamer-conjugated poly(lactide-co-glycolic acid)-polyethylenimine (PLGA-PEI) (DPAP) delivery system, via a layer-by-layer synthesis method, using a water-in-oil-in-water double emulsion approach. The binding characteristics, targeting capability, biophysical properties, encapsulation efficiency, and drug release profile of the DPAP system were investigated under varying conditions of ionic strength, polymer composition and molecular weight (MW), and degree of PEGylation of the synthetic core. Experimental results showed increased drug release rate with increasing buffer ionic strength. DPAP particulate system obtained the highest drug release of 50% at day 9 at 1 M NaCl ionic strength. DPAP formulation, using PLGA 65:35 and PEI MW of ∼800 Da, demonstrated an encapsulation efficiency of 78.93%, and a loading capacity of 0.1605 mg bovine serum albumin per mg PLGA. DPAP (PLGA 65:35, PEI MW∼25 kDa) formulation showed a high release rate with a biphasic release profile. Experimental data depicted a lower targeting power and reduced drug release rate for the PEGylated DPAP formulations. The outcomes from the present study lay the foundation to optimize the performance of DPAP system as an effective synthetic drug carrier for targeted delivery.

Cardiovascular therapies utilizing targeted delivery of nanomedicines and aptamers.

Cardiovascular ailments are the foremost trigger of death in the world today, including myocardial infarction and ischemic heart diseases. To date, extraordinary measures have been prescribed, from the perspectives of both conventional medical therapies and surgeries, to enforce cardiac cell regeneration post cardiac traumas, albeit with limited long-term success. The prospects of successful heart transplants are also grim, considering exorbitant costs and unavailability of suitable donors in most cases. From the perspective of cardiac revascularization, use of nanoparticles and nanoparticle mediated targeted drug delivery have garnered substantial attention, attributing to both active and passive heart targeting, with enhanced target specificity and sensitivity. This review focuses on this aspect, while outlining the progress in targeted delivery of nanomedicines in the prognosis and subsequent therapy of cardiovascular disorders, and recapitulating the benefits and intrinsic challenges associated with the incorporation of nanoparticles. This article categorically provides an overview of nanoparticle-mediated targeted delivery systems and their implications in handling cardiovascular diseases, including their intrinsic benefits and encountered procedural trials and challenges. Additionally, the solicitations of aptamers in targeted drug delivery with identical objectives, are presented. This includes a detailed appraisal on various aptamer-navigated nanoparticle targeted delivery platforms in the diagnosis and treatment of cardiovascular maladies. Despite a few impending challenges, subject to additional investigations, both nanoparticles as well as aptamers show a high degree of promise, and pose as the next generation of drug delivery vehicles, in targeted cardiovascular therapy.

Linear viscoelastic and microstructural properties of native male human skin and in vitro 3D reconstructed skin models.

The study reports first ever account of measurements of linear viscoelastic moduli under small amplitude oscillatory shear deformations, for commercially available juvenile and aged in vitro 3D reconstructed skin models. The results were compared with those of native male whole human and dermis-only foreskin samples, catering to a wide age group from 0.5 to 68 years, including samples from a 23-year-old male abdomen. In the strain sweep tests, the dermis of the juvenile/young age group assumed a higher intrinsic elastic modulus than the whole skin. A reverse qualitative trend was noted for the adult/aged age group. Confirmed by the histological examination of the stained cross-sections, this is attributed to the nascent epidermal differentiation and the high fiber density of dermal collagen. The oscillation frequency sweeps exposed a greater dependence of the elasticity on the frequency for the native male dermis foreskin samples as compared to the whole skins, irrespective of age. This is anticipated since the extremely structured epidermis confers higher resistance to the whole skins towards intracycle deformations compared to the dermis, thereby storing smaller elastic energy. The 3D skin models examined in this work exhibited a broader linear viscoelastic region, a larger viscoelasticity, and much higher dynamic moduli, compared to the native skin. The rheological trends are a significant addition to the literature and may be used as a reference for the design of next generation of scaffolds.

Bioprocessing of Functional Ingredients from Flaxseed.

Flaxseeds (Linum usitatissimum L.) are oilseeds endowed with nutritional constituents such as lignans, lipids, proteins, fibre, carbohydrates, and micronutrients. Owing to their established high nutritional profile, flaxseeds have gained an established reputation as a dietary source of high value functional ingredients. Through the application of varied bioprocessing techniques, these essential constituents in flaxseeds can be made bioavailable for different applications such as nutraceuticals, cosmetics, and food industry. However, despite their food and health applications, flaxseeds contain high levels of phytotoxic compounds such as linatine, phytic acids, protease inhibitors, and cyanogenic glycosides. Epidemiological studies have shown that the consumption of these compounds can lead to poor bioavailability of essential nutrients and/or health complications. As such, these components must be removed or inactivated to physiologically undetectable limits to render flaxseeds safe for consumption. Herein, critical description of the types, characteristics, and bioprocessing of functional ingredients in flaxseed is presented.

Parametric Investigation of Batch Adsorption of Proteins onto Polymeric Particles.

BACKGROUND: Effective bimolecular adsorption of proteins onto solid matrices is characterized by in-depth understanding of the biophysical features essential to optimize the adsorption performance. RESULTS: The adsorption of bovine serum albumin (BSA) onto anion-exchange Q-sepharose solid particulate support was investigated in batch adsorption experiments. Adsorption kinetics and isotherms were developed as a function of key industrially relevant parameters such as polymer loading, stirring speed, buffer pH, protein concentration and the state of protein dispersion (solid/aqueous) in order to optimize binding performance and adsorption capacity. Experimental results showed that the first order rate constant is higher at higher stirring speed, higher polymer loading, and under alkaline conditions, with a corresponding increase in equilibrium adsorption capacity. Increasing the stirring speed and using aqueous dispersion protein system increased the adsorption rate, but the maximum protein adsorption was unaffected. Regardless of the stirring speed, the adsorption capacity of the polymer was 2.8 mg/ml. However, doubling the polymer loading increased the adsorption capacity to 9.4 mg/ml. CONCLUSIONS: The result demonstrates that there exists a minimum amount of polymer loading required to achieve maximum protein adsorption capacity under specific process conditions.

A Proposal for Six Sigma Integration for Large-Scale Production of Penicillin G and Subsequent Conversion to 6-APA.

Six Sigma methodology has been successfully applied to daily operations by several leading global private firms including GE and Motorola, to leverage their net profits. Comparatively, limited studies have been conducted to find out whether this highly successful methodology can be applied to research and development (R&D). In the current study, we have reviewed and proposed a process for a probable integration of Six Sigma methodology to large-scale production of Penicillin G and its subsequent conversion to 6-aminopenicillanic acid (6-APA). It is anticipated that the important aspects of quality control and quality assurance will highly benefit from the integration of Six Sigma methodology in mass production of Penicillin G and/or its conversion to 6-APA.