Exosomes/EVs: BIOXOME: A NATURAL LIPOSOME MIMETIC AS DRUG DELIVERY SYSTEM

Background & Aim: Liposomes are spherical-shaped vesicles composed of one or more lipid bilayers. The ability of liposomes to encapsulate hydro- or lipophilic drugs allowed these vesicles to become a useful drug delivery system. Natural cell membranes, such as Bioxome, have newly emerged as new source of materials for molecular delivery systems. Bioxome are biocompatible and GMP-compliant liposome-like membrane that can be produced from more than 200 cell types. Bioxome self-assemble, with in-process self-loading capacity and can be loaded with a variety of therapeutic compounds. Once close to the target tissue, Bioxome naturally fuse with the cell membrane and release the inner compound. Orgenesis is interested in evaluating the potential of Bioxome as new drug delivery system for treatment of several diseases, including skin repair, local tumour or COVID19. Methods, Results & Conclusion(s): Bioxome were obtained from adipose- derived Mesenchymal Stem Cells, with a process of organic- solvent lipid extraction, followed by lyophilization and sonication assemblage. During the sonication process, Bioxome were charged or not with several cargos. Size distribution of empty Bioxome was detected by Particle Size Analyzer (NanoSight). Electron Microscopy (EM) was performed to assess Bioxome morphology. Lipid content was evaluated by electrospray ionization system. Dose response in vitro test on human lung fibroblasts treated or not with Bioxome encapsulating a specific cargo (API) against COVID19 were performed. NanoSight analysis showed that nanoparticle size in Bioxome samples ranged between 170+/-50 nm, with a concentration ranging between 109-1010+/-106 particles/mL. EM clearly showed the double phospholipid layers that composes the Bioxome. Stability study demonstrated that Bioxome are stable in size and concentration up to 90 days at +4Cdegree or even at RT. No change in size between encapsulated Bioxome with small size (~340 Da) cargo vs empty Bioxome was observed up to 30 days storage. Lipidomic analysis approach revealed that the yield of lipids and their composition are satisfactory for a therapeutic product using Bioxome. Lastly, in the in vitro model of COVID19, Bioxome encapsulating API effectively saved cells from death (20x vs untreated cells) and at lower doses of API than these of non-encapsulated cargo (0.005 microM vs 0.1 microM). Bioxome seems to be an excellent candidate for liposome mimetic tool as drug delivery system for targeting specific organs and diseases treatment.Copyright © 2023 International Society for Cell & Gene Therapy

Foaming and Other Functional Properties of Freeze-Dried Mare's Milk.

The aim of this study was to evaluate the effect of the freeze-drying process on the preservation of mare's milk. This was achieved through the characterization of the functional properties of reconstituted freeze-dried mare's milk. The chemical composition, bulk density, foam capacity, and ability to form emulsions of the atherogenic, thrombogenic, and hypercholesterolemic fatty acid index were investigated. The freeze-drying process did not change the proportion of the milk components in the dry matter. The moisture content of the freeze-dried mare's milk was 10.3 g/kg and the bulk was below 0.1 g/mL. The foaming capacity was 111.3%; hence, the foaming capacity of the milk was very poor. The oil binding capacity was 2.19 g/g of protein. The freeze-drying process improves the binding degree and retention of oil by milk proteins, but produced foam was unstable, short-lived, and lacked the ability to retain air fractions. The atherogenic index and thrombogenic index values calculated for reconstituted milk were 1.02 and 0.53, respectively. The hypercholesterolemia fatty acid index was 25.01.

Room-temperature-storable chemiluminescence freeze-drying mixes for detection of SARS-CoV-2 neutralizing antibody

As the vaccine was successfully developed, the spread of the epidemic (COVID-19) was effectively controlled. But there are still thousands of people affected COVID-19 after being vaccinated. Neutralizing activity has become a critical method for quantifying neutralizing antibody against SARS-CoV-2. However, limited to the strict conditions of cold chain transportation, the neutralizing activity test has not been widely promoted. In this study, a room-temperature-storable chemiluminescence freeze-drying mixes for SARS-CoV-2 neutralizing antibody detection was developed to decrease the cost of lyophilization step for promoting its application in third world countries. Several freeze concentrated solutions were used to protect the antigen bioactivity. The mixes can be stored at room temperature over 12 months and still exhibited great accuracy and precision. Thus, the proposed room-temperature-storable chemiluminescence freeze-drying mixes offers a cheap and stable storage method for SARS-CoV-2 neutralizing antibody detection and shows a great potential for promoting the neutralizing activity test.

Airborne SARS-CoV- 2 detection in outdoor air

Background: The potential aerosol spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV- 2) has been suggested. While indoor air sampling for SARS-CoV- 2 has demonstrated detectable viral RNA and has been related to virus transmission, the contribution of outdoor air to the spread of the viral infection is not yet known. We aimed at developing a methodology to detect the virus in outdoor air. Method(s): T he s ampling w as p erformed u sing a C HEMVOL v olumetric impactor (Butraco) equipped with 2 stages (PM > 10 & 2.5 > PM > 10um). Filters were collected and preserved at -80 degreeC. Total RNA extraction was performed directly from the collected filters with the Phenol-Chloroform method using TRItidy GTM reagent according to the manufacturer's instructions. For total RNA purification samples were purified with the commercial kit E.Z.N.A. Total RNA Kit I. Real-Time Reverse Transcription PCR was executed to detect the N gene from the Sarbecovirus family and RdRp gene from SARS-CoV- 2 using the ViroReal Kit SARS-CoV- 2 Multiplex. A protein-rich fraction was obtained with ammonium bicarbonate buffer extraction followed by lyophilization. SARS-CoV- 2 spike protein was assessed by specific immunological detection (SARS-CoV- 2 Antigen Test Kit). Result(s): RT-PCR for N gene results, identifying Sarbecovirus family, were positive and Cq > 33, in the samples from the last week of December 2020 and the first and second weeks of January 2021, in both PM > 10 and 2.5 > PM > 10. The RdRp gene was undetectable, probably due to low virus concentration. The protein samples from the same days tested positive for the specific antigen spike protein. All results combined confirm the detection of SARS-CoV- 2 in outdoor air. Conclusion(s): Airborne SARS-CoV- 2 was detected in ambient air. These results will contribute to an early detection of SARS-Cov- 2 in ambient air, thus eventually providing the base for early alert systems allowing the implementation of preventive measures to control outbreaks.

Lipid based nanocarriers: Production techniques, concepts, and commercialization aspect

Over the past decade, compared to all other macromolecules lipid-based nanocarriers have proven to be an excellent carrier and delivery system for various pharmaceutical drugs of poor bioavailability. In addition to that, they exhibit exceptional qualities such as minimal toxicity, economical scale-up production, great biocompatibility, and high drug loading efficiency. In this study, we have discussed the various types of lipid nanoparticles, such as liposomes, nanostructured lipid carriers, solid lipid nanoparticles, and lipid polymer hybrid nanoparticles. We have also conferred in detail, the composition, shape and size, methods of preparation, advantages, and certain limitations associated with these lipid-based nanocarriers. Additionally, we have exclusively accounted for several examples of lipid-based nanomedicines that have either been approved and commercialized or are under the different phases of clinical trials. The current review overall focuses on the up-to-date research that has recently been published in view of developing lipid-based nanocarriers for various biological applications, including gene therapy, breast cancer therapy, and vaccine development.Copyright © 2022

Continuous freeze-drying of messenger RNA lipid nanoparticles enables storage at higher temperatures.

Messenger RNA (mRNA) lipid nanoparticles (LNPs) have emerged at the forefront during the COVID-19 vaccination campaign. Despite their tremendous success, mRNA vaccines currently require storage at deep freeze temperatures which complicates their storage and distribution, and ultimately leads to lower accessibility to low- and middle-income countries. To elaborate on this challenge, we investigated freeze-drying as a method to enable storage of mRNA LNPs at room- and even higher temperatures. More specifically, we explored a novel continuous freeze-drying technique based on spin-freezing, which has several advantages compared to classical batch freeze-drying including a much shorter drying time and improved process and product quality controlling. Here, we give insight into the variables that play a role during freeze-drying by evaluating the impact of the buffer and mRNA LNP formulation (ionizable lipid to mRNA weight ratio) on properties such as size, morphology and mRNA encapsulation. We found that a sufficiently high ionizable lipid to mRNA weight ratio was necessary to prevent leakage of mRNA during freeze-drying and that phosphate and Tris, but not PBS, were appropriate buffers for lyophilization of mRNA LNPs. We also studied the stability of optimally lyophilized mRNA LNPs at 4 °C, 22 °C, and 37 °C and found that transfection properties of lyophilized mRNA LNPs were maintained during at least 12 weeks. To our knowledge, this is the first study that demonstrates that optimally lyophilized mRNA LNPs can be safely stored at higher temperatures for months without losing their transfection properties.

Design of solid powder particles for a needle-free injection

Background: In 2020, the COVID19 pandemic has shown the medical need for vaccinations. The conventional method of vaccine application is intravascular injection of a liquid solution. However, this method is associated with some disadvantages, such as a high risk of infection. Purpose(s): The approach of a needle-free ballistic administration accelerates solid powder particles to a sufficient speed so that they are able to penetrate into the skin and address target Langerhans cells. For this purpose, the particles require certain characteristics (Weissmueller et al., 2017). The main criteria for a successful application is the particle size as well as the density (Maa et al., 2004). Method(s): One potential production process is freeze-drying out of a solution with a subsequent milling step (abbreviated to FD). Another modified approach is spray-freeze drying (abbreviated to SFD). Dried powders are treated afterwards by ultrasonic microsieving (6000 vibrations per seconds for ten minutes) in order to segregate a useable fraction (38 mum to 75 mum). Tap density was determined according to the protocol by Ph.eur. guidelines. Helium pycnometry determines the true density. The magnitude of density is described by the quotient of tap density rho tap and pycometric density rho He-pycnometer. Result(s): Estimated density of examined samples containing trehalose and mannitol could not exceed 50%. Conclusion(s): The described techniques reveal a quite porous structure of the product. This structure might not be sufficient for particles to successfully penetrate into the skin. These powder particles might burst upon the surface. However the dimension of the speed has to be considered as well as it plays a crucial role as well.

Freeze-Drying of a Capsid Virus-like Particle-Based Platform Allows Stable Storage of Vaccines at Ambient Temperature.

The requirement of an undisrupted cold chain during vaccine distribution is a major economic and logistical challenge limiting global vaccine access. Modular, nanoparticle-based platforms are expected to play an increasingly important role in the development of the next-generation vaccines. However, as with most vaccines, they are dependent on the cold chain in order to maintain stability and efficacy. Therefore, there is a pressing need to develop thermostable formulations that can be stored at ambient temperature for extended periods without the loss of vaccine efficacy. Here, we investigate the compatibility of the Tag/Catcher AP205 capsid virus-like particle (cVLP) vaccine platform with the freeze-drying process. Tag/Catcher cVLPs can be freeze-dried under diverse buffer and excipient conditions while maintaining their original biophysical properties. Additionally, we show that for two model cVLP vaccines, including a clinically tested SARS-CoV-2 vaccine, freeze-drying results in a product that once reconstituted retains the structural integrity and immunogenicity of the original material, even following storage under accelerated heat stress conditions. Furthermore, the freeze-dried SARS-CoV-2 cVLP vaccine is stable for up to 6 months at ambient temperature. Our study offers a potential solution to overcome the current limitations associated with the cold chain and may help minimize the need for low-temperature storage.

Spray freeze dried niclosamide nanocrystals embedded dry powder for high dose pulmonary delivery.

Based on the drug repositioning strategy, niclosamide (NCL) has shown potential applications for treating COVID-19. However, the development of new formulations for effective NCL delivery is still challenging. Herein, NCL-embedded dry powder for inhalation (NeDPI) was fabricated by a novel spray freeze drying technology. The addition of Tween-80 together with 1,2-Distearoyl-sn-glycero-3-phosphocholine showed the synergistic effects on improving both the dispersibility of primary NCL nanocrystals suspended in the feed liquid and the spherical structure integrity of the spray freeze dried (SFD) microparticle. The SFD microparticle size, morphology, crystal properties, flowability and aerosol performance were systematically investigated by regulating the feed liquid composition and freezing temperature. The addition of leucine as the aerosol enhancer promoted the microparticle sphericity with greatly improved flowability. The optimal sample (SF- 80D-N20L2D2T1) showed the highest fine particle fraction of ∼47.83%, equivalently over 3.8 mg NCL that could reach the deep lung when inhaling 10 mg dry powders.

Design and lyophilization of lipid nanoparticles for mRNA vaccine and its robust immune response in mice and nonhuman primates.

mRNA and lipid nanoparticles have emerged as powerful systems for the preparation of vaccines against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. The emergence of novel variants or the necessity of cold chain logistics for approved mRNA vaccines undermines the investigation of next-generation systems that could preserve both potency and stability. However, the correlation between lipid nanoparticle composition and activity is not fully explored. Here, we screened a panel of ionizable lipids in vivo and identified lead lipid nanoparticles with a branched-tail lipid structure. Buffer optimization allowed the determination of lyophilization conditions, where lipid nanoparticle-encapsulated mRNA encoding SARS-CoV-2 spike protein could induce robust immunogenicity in mice after 1 month of storage at 5°C and 25°C. Intramuscularly injected lipid nanoparticles distributed in conventional dendritic cells in mouse lymph nodes induced balanced T helper (Th) 1/Th2 responses against SARS-CoV-2 spike protein. In nonhuman primates, two doses of 10 or 100 µg of mRNA induced higher spike-specific binding geometric mean titers than those from a panel of SARS-CoV-2-convalescent human sera. Immunized sera broadly inhibited the viral entry receptor angiotensin-converting enzyme 2 (ACE2) from binding to the spike protein in all six strains tested, including variants of concern. These results could provide useful information for designing next-generation mRNA vaccines.

mRNA-Lipid Nanoparticle COVID-19 Vaccines: Structure and Stability

A drawback of the current mRNA-lipid nanoparticle (LNP) COVID-19 vaccines is that they have to be stored at (ultra)low temperatures (2). Understanding the root cause of the instability of these vaccines may help to rationally improve mRNALNP product stability and thereby ease the temperature conditions for storage. In this presentation we discuss proposed structures of mRNALNPs, factors that impact mRNA-LNP stability and strategies to optimize mRNA-LNP product stability. Analysis of mRNA-LNP structures reveals that mRNA, the ionizable cationic lipid and water are present in the LNP core. The neutral helper lipids are mainly positioned in the outer, encapsulating, wall. mRNA hydrolysis is an important driver for mRNA-LNP instability. It is currently a matter of debate whether water in the LNP core can freely interact with the mRNA and to what extent the degradation prone sites of mRNA are protected through a coat of ionizable cationic lipids. To improve the stability of mRNALNP vaccines, optimization of the mRNA nucleotide composition should be prioritized. Secondly, a better understanding of the milieu the mRNA is exposed to in the core of LNPs may help to rationalize adjustments to the LNP structure to preserve mRNA integrity. Moreover, drying techniques, such as lyophilization, are promising options still to be explored. As vaccines turn out to be the major weapon against the COVID-19 viral attack, the urge to develop more stable formulations is still growing and alternative, not-mRNA based products, may come to the forefront in situations where the (ultra) cold chain cannot be guaranteed. (Table Presented).

Teaching a graduate aourse in pharmaceutical excipients: personal experience and perspectives

Excipients are critically important in converting active pharmaceutical ingredients (API) into drug products that have optimal stability, bioavailability, manufacturability, duration of action, and therapeutic benefits. They will play even greater roles in the future to enable drug targeting, delivery of biotech products and vaccines, gene therapy, continuous manufacturing, 3D printing, and so forth. This commentary describes the author’s experience in teaching a graduate course on excipients at St. John’s University to train students on optimal selection and appropriate use of excipients in formulating dosage forms and development of drug delivery systems. The course is offered in 15 two-hour sessions over a semester, and the course materials are divided into 13 modules on chemistry of different classes of polymeric and non-polymeric excipients and their application in dosage form development, including the use as solubilizing agents, lyophilizing agents, cryoprotectants, buffers, biodegradable materials, and carriers for amorphous solid dispersions and 3D printing. The development of coprocessed excipients, the need for new excipients, and the regulatory aspects of excipients are also covered. The course includes presentations by guest speakers from the industry, and the students also watch virtual presentations from experts that are publicly available from the internet. It is a popular course at St. John’s University taken by all graduate students in the pharmaceutics program. It is recommended that such courses are introduced in other pharmacy schools and academic institutions. The course may be adapted to meet specific needs of different academic programs. Professional associations, such as AAPS and CRS, industry groups like IPEC, and the pharmaceutical industry may be able to help in introducing such courses by providing lecture materials and guest lecturers.

Dexamethasone and Nutraceutical Therapy Can Reduce the Myalgia Due to COVID-19 - a Systemic Review of the Active Substances that Can Reduce the Expression of Interlukin-6.

Background: Myalgia reflects generalized inflammation and cytokine response and can be the onset symptom of 36% of patients with COVID-19. Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF- α) levels in plasma and upper respiratory secretions directly correlate with the magnitude of viral replication, fever, and respiratory and systemic symptoms, including musculoskeletal clinical manifestations. Objective: The aim of our work is to report literature scientific investigation clinical protocol to reduce the immunomodulation and inflammatory response nutraceutical therapy associated with dexamethasone and how can reduce the expression of Interlukina-6(IL-6) and myalgia due to COVID-19. Methods: We searched in Pubmed and Cochrane the nautriceutical drugs to treat the immune modulation of organism to COVID-19. We put these keywords: immune inflammation, desease descriptions, epidemiology COVID-19; immunomodulations; IL-6; Rheumatic Symptoms; Joint; Musculoskeletal Disorders; dexamethasone; Polydatin; Zinc; Melatonin; N- Acetyl Cysteine; Colostrum; L- Glutamine; Vitamin D3. Results: We found 61 papers. All the authors analyze them. After the Analyze we suggest the use of response nutraceutical therapy associated with dexamethasone can reduce the expression of Interlukina-6(IL-6) and myalgia due to COVID-19. Conclusion: According the scientific literature nutraceutical therapy associated with dexamethasone can reduce the expression of Interlukina-6(IL-6) and myalgia due to COVID-19.

Clathrate Hydrates of Organic Solvents as Auxiliary Intermediates in Pharmaceutical Research and Development: Improving Dissolution Behaviour of a New Anti-Tuberculosis Drug, Perchlozon.

There is an urgent need for new drugs to overcome the challenge of the ever-growing drug resistance towards tuberculosis. A new, highly efficient anti-tuberculosis drug, Perchlozone (thioureidoiminomethylpyridinium perchlorate, Pz), is only available in an oral dosage form, though injectable forms and inhalation solutions could be better alternatives, offering higher bioavailability. To produce such forms, nano- and micro-particles of APIs would need to be prepared as dispersions with carriers. We use this case study to illustrate the principles of selecting solvents and excipients when preparing such formulations. We justify the choice of water-THF (19.1 wt % THF) as solvent and mannitol as carrier to prepare formulations of Pz-a poorly soluble compound-that are suitable for injection or inhalation. The formulations could be prepared by conventional freeze-drying in vials, making the proposed method suitable for industrial scaling. A similar strategy for selecting the organic solvent and the excipient can be applied to other compounds with low water solubility.

Dual targeting powder formulation of antiviral agent for customizable nasal and lung deposition profile through single intranasal administration.

Unpredictable outbreaks due to respiratory viral infections emphasize the need for new drug delivery strategies to the entire respiratory tract. As viral attack is not limited to a specific anatomic region, antiviral therapy that targets both the upper and lower respiratory tract would be most effective. This study aimed to formulate tamibarotene, a retinoid derivative previously reported to display broad-spectrum antiviral activity against influenza and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), as a novel dual particle size powder formulation that targets both the nasal cavity and the lung by a single route of intranasal administration. Spray freeze drying (SFD) and spray drying (SD) techniques were employed to prepare tamibarotene powder formulations, and cyclodextrin was used as the sole excipient to enhance drug solubility. With the employment of appropriate atomizing nozzles, particles of size above 10 µm and below 5 µm could be produced for nasal and lung deposition, respectively. The aerosol performance of the powder was evaluated using Next Generation Impactor (NGI) coupled with a glass expansion chamber and the powder was dispersed with a nasal powder device. By blending powder of two different particle sizes, a single powder formulation with dual aerosol deposition characteristic in both the nasal and pulmonary regions was produced. The aerosol deposition fractions in the nasal cavity and pulmonary region could be modulated by varying the powder mixing ratio. All dry powder formulations exhibited spherical structures, amorphous characteristics and improved dissolution profile as compared to the unformulated tamibarotene. Overall, a novel dual targeting powder formulation of tamibarotene exhibiting customizable aerosol deposition profile was developed. This exceptional formulation strategy can be adopted to deliver other antimicrobial agents to the upper and lower airways for the prevention and treatment of human respiratory infections.

Room-temperature-storable chemiluminescence freeze-drying mixes for detection of SARS-CoV-2 neutralizing antibody

As the vaccine was successfully developed, the spread of the epidemic (COVID-19) was effectively controlled. But there are still thousands of people affected COVID-19 after being vaccinated. Neutralizing activity has become a critical method for quantifying neutralizing antibody against SARS-CoV-2. However, limited to the strict conditions of cold chain transportation, the neutralizing activity test has not been widely promoted. In this study, a room-temperature-storable chemiluminescence freeze-drying mixes for SARS-CoV-2 neutralizing antibody detection was developed to decrease the cost of lyophilization step for promoting its application in third world countries. Several freeze concentrated solutions were used to protect the antigen bioactivity. The mixes can be stored at room temperature over 12 months and still exhibited great accuracy and precision. Thus, the proposed room-temperature-storable chemiluminescence freeze-drying mixes offers a cheap and stable storage method for SARS-CoV-2 neutralizing antibody detection and shows a great potential for promoting the neutralizing activity test.

Retention of hemostatic and immunological properties of frozen plasma and COVID-19 convalescent apheresis fresh-frozen plasma produced and freeze-dried in Canada.

BACKGROUND: Randomized clinical trial data show that early plasma transfusion may save lives among trauma patients. Supplying plasma in remote environments is logistically challenging. Freeze-dried plasma (FDP) offers a possible solution. STUDY DESIGN AND METHODS: A Terumo BCT plasma freeze-drying system was evaluated. We compared pooled frozen plasma (FP) units with derived Terumo BCT FDP (TFDP) units and pooled COVID-19 convalescent apheresis fresh-frozen plasma (CC-AFFP) with derived CC-TFDP units. Parameters measured were: coagulation factors (F) II; V; VII; VIII; IX; XI; XIII; fibrinogen; Proteins C (PC) and S (PS); antithrombin (AT); α2 -antiplasmin (α2 AP); ADAMTS13; von Willebrand Factor (vWF); thrombin-antithrombin (TAT); D-dimer; activated complement factors 3 (C3a) and 5 (C5a); pH; osmolality; prothrombin time (PT); and activated partial thromboplastin time (aPTT). Antibodies to SARS-CoV-2 in CC-AFFP and CC-TFDP units were compared by plaque reduction assays and viral protein immunoassays. RESULTS: Most parameters were unchanged in TFDP versus FP or differed ≤15%. Mean aPTT, PT, C3a, and pH were elevated 5.9%, 6.9%, 64%, and 0.28 units, respectively, versus FP. CC-TFDP showed no loss of SARS-CoV-2 neutralization titer versus CC-AFFP and no mean signal loss in most pools by viral protein immunoassays. CONCLUSION: Changes in protein activities or clotting times arising from freeze-drying were <15%. Although C3a levels in TFDP were elevated, they were less than literature values for transfusable plasma. SARS-CoV-2-neutralizing antibody titers and viral protein binding levels were largely unaffected by freeze-drying. In vitro characteristics of TFDP or CC-TFDP were comparable to their originating plasma, making future clinical studies appropriate.

Investigation of the Freezing Phenomenon in Vials Using an Infrared Camera.

The freezing phenomenon has a dramatic impact on the quality of freeze-dried products. Several freezing models applied to solutions in vials have been proposed to predict the resulting product morphology and describe heat transfer mechanisms. However, there is a lack of detailed experimental observations of the freezing phenomenon in vials in the literature. Thus, the present work offers new experimental observations of the freezing phenomenon in vials by infrared (IR) thermography. IR imaging allowed each vial's whole axial temperature profile to be collected during freezing, providing significant insights into the process. Spontaneous nucleation and vacuum-induced surface freezing (VISF), as a controlled nucleation technique, are investigated. Batches having vials in direct contact with the shelf (exchanging heat mainly through conduction) as well as suspended (exchanging heat mainly through natural convection and radiation) were tested. The study used three solutions: sucrose 5%, mannitol 5%, and dextran 10%. SEM images coupled with an automated image segmentation technique were also performed to examine possible correlations between the freezing observations and the resulting pore size distributions. IR thermography was found to be a promising tool for experimentally predicting the resulting product morphology in-line.

Sea Buckthorn Leaf Powders: The Impact of Cultivar and Drying Mode on Antioxidant, Phytochemical, and Chromatic Profile of Valuable Resource.

Sea buckthorn (Hippophae rhamnoides L. (HR)) leaf powders are the underutilized, promising resource of valuable compounds. Genotype and processing methods are key factors in the preparation of homogenous, stable, and quantified ingredients. The aim of this study was to evaluate the phenolic, triterpenic, antioxidant profiles, carotenoid and chlorophyll content, and chromatic characteristics of convection-dried and freeze-dried HR leaf powders obtained from ten different female cultivars, namely 'Avgustinka', 'Botaniceskaja Liubitelskaja', 'Botaniceskaja', 'Hibrid Percika', 'Julia', 'Nivelena', 'Otradnaja', 'Podarok Sadu', 'Trofimovskaja', and 'Vorobjovskaja'. The chromatic characteristics were determined using the CIELAB scale. The phytochemical profiles were determined using HPLC-PDA (high performance liquid chromatography with photodiode array detector) analysis; spectrophotometric assays and antioxidant activities were investigated using ABTS (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) and FRAP (ferric ion reducing antioxidant power) assays. The sea buckthorn leaf powders had a yellowish-green appearance. The drying mode had a significant impact on the total antioxidant activity, chlorophyll content, and chromatic characteristics of the samples; the freeze-dried samples were superior in antioxidant activity, chlorophyll, carotenoid content, and chromatic profile, compared to convection-dried leaf powder samples. The determined triterpenic and phenolic profiles strongly depend on the cultivar, and the drying technique had no impact on qualitative and quantitative composition. Catechin, epigallocatechin, procyanidin B3, ursolic acid, α-amyrin, and ß-sitosterol could be used as quantitative markers in the phenolic and triterpenic profiles. The cultivars 'Avgustinka', 'Nivelena', and 'Botaniceskaja' were superior to other tested cultivars, with the phytochemical composition and antioxidant activity.