Modified Extraction Method Research Articles

Simulation and Performance of 99Mo/99mTc Radioisotope Separation by Modified Extraction Method

Abstract Technetium-99m (99mTc) is a nuclide in nuclear medicine, commonly derived as a decay product from molybdenum-99 (99Mo). Unfortunately, Japan and Indonesia rely entirely on imports for their 99Mo/99mTc supply. This study aims to determine the distribution coefficient (Kd) for the 99Mo/99mTc production process and assess the efficiency of Methyl Ethyl Ketone (MEK) as a solvent in the extraction process. The method Initial tests using Rhenium (Re) as a substitute for 99mTc, conducted in collaboration with the Japan Atomic Energy Agency (JAEA), revealed optimal conditions: a standing time of 5 to 15 minutes, a mixing time of 3 minutes with a motor speed of 250 rpm, and a MEK volume exceeding 20 ml. These conditions yielded a Re extraction efficiency exceeding 85.14%, with a Re elution yield of up to 98.73%. These findings served as a database for experiments involving 99Mo/99mTc conducted at the National Research and Innovation Agency (BRIN). In hot experiments, the 99mTc recovery rate through MEK exceeded 100%, while the elution process achieved a recovery rate of 97.81%. Through simulation and facility enhancements in the hot cell, the 99mTc extraction recovery increased to 44.86%, and elution recovery ranged from 90.62%. From the calculation results, the distribution coefficient value database (Kd-99mTc) was 0.45. Radionuclide and radiochemistry purity of the 99mTc solution were assessed using the Gamma Spectrometer and TLC Scanner, resulting in purity of 99.90% and 99.80%, respectively. These results underscore the potential for domestic production 99mTc, reducing reliance on imports.

Identification of volatile and odor-active compounds in pea protein fractions obtained by a modified extraction method using fermentation.

This study investigates the aromatic composition of pea albumin and globulin fractions obtained through either fermentation or conventional acidification using hydrochloric acid (control) toward the isoelectric point of pea globulins. Different lactic acid bacteria were used including S. thermophilus (ST), L. plantarum (LP), and their coculture (STLP). The volatile compounds were extracted by solvent-assisted flavor evaporation technique and quantified by gas chromatography-mass spectrometry (GC-MS). Odor-active compounds (OAC) were further characterized by gas chromatography-olfactometry (GC-O). In total, 96 volatile and 36 OACs were identified by GC-MS and GC-O, respectively. The results indicated that the protein fractions obtained by conventional acidification were mainly described by green notes for the presence of different volatile compounds such as hexanal. However, the samples obtained by fermentation had a lower content of these volatile compounds. Moreover, protein fractions obtained by coculture fermentation were described by volatile compounds associated with fruity, floral, and lactic notes. PRACTICAL APPLICATION: The insights from this study on pea protein aroma could find practical use in the food industry to enhance the sensory qualities of plant-based products. By utilizing fermentation methods and specific lactic acid bacteria combinations, manufacturers may produce pea protein with reduced undesirable green notes, offering consumers food options with improved flavors. This research may contribute to the development of plant-based foods that not only provide nutritional benefits but also meet consumer preferences for a more appealing taste profile.

Improved methods for total and chloroplast protein extraction from Cajanus species for two-dimensional gel electrophoresis and mass spectrometry.

The recent advances in pigeon pea genomics, including high-quality whole genome and chloroplast genome sequence information helped develop improved varieties. However, a comprehensive Cajanus proteome, including the organelle proteome, is yet to be fully mapped. The spatial delineation of pigeon pea proteins at sub-cellular levels and inter-organelle communication could offer valuable insights into its defense mechanism against various stresses. However, the major bottleneck in the proteomic study is the lack of a suitable method of protein extraction and sample preparation compatible with two-dimensional gel electrophoresis (2D-PAGE), liquid chromatography-mass spectrometry (LCMS), or matrix-assisted laser desorption ionization-time of flight (MALDi-ToF). Our study introduces two efficient methods, one for isolating total proteins and another for organelle (chloroplast) proteins from various Cajanus spp. For total protein extraction, we have optimized a protocol using phenol in combination with a reducing agent (DTT) and protease inhibitor cocktail, also washing (6-7 times) with ice-cold acetone after overnight protein precipitation of total proteins. Our modified extraction method using phenol for total leaf protein yielded approximately 2-fold more proteins than the previously reported protocols from C. cajan (3.18 ± 0.11 mg/gm) and C. scarabaeoides (2.06 ± 0.08 mg/gm). We have also optimized a protocol for plastid protein extraction, which yielded 1.33 ± 0.25 mg/10 gm plastid proteins from C. cajan and 0.88 ± 0.19 mg/10 gm plastid proteins from C. scarabaeoides. The 2D-PAGE analysis revealed 678 ± 08 reproducible total protein spots from C. cajan and 597 ± 22 protein spots from C. scarabaeoides. Similarly, we found 566 ± 10 and 486 ± 14 reproducible chloroplast protein spots in C. cajan and C. scarabaeoides, respectively. We confirmed the plastid protein fractions through immunoblot analysis using antibodies against LHCb1/LHCⅡ type Ⅰ protein. We found both methods suitable for 2D-PAGE and mass spectrometry (MS). This is the first report on developing protocols for total and chloroplastic protein extraction of Cajanus spp. suitable for advanced proteomics research.

Determination of Multiclass Pharmaceutical Residues in Milk Using Modified QuEChERS and Liquid-Chromatography-Hybrid Linear Ion Trap/Orbitrap Mass Spectrometry: Comparison of Clean-Up Approaches and Validation Studies.

A multi-residue method was developed to identify and quantify pharmaceutical drug residues in full-fat milk, using a modified QuEChERS extraction procedure and sonication combined with Ultra-High-Performance Liquid Chromatography-High-Resolution Orbitrap Mass Spectrometry (UHPLC-HR-Orbitrap-MS). Sample preparation involves three different QuEChERS extraction procedures and sorbents for the purification step. The optimized modified extraction method, combined with the clean-up approaches using C18 and the EMR-Lipid sorbent, has been validated in terms of linearity, recovery, precision, LOD and LOQ, matrix effects (ME) and expanded uncertainty. The optimized method showed a linearity >0.9903, recoveries within the range 65.1-120.1%, precision (expressed as %RSD) <17.5%, medium (<39.9%) to low (<16.7%) matrix effects and acceptable expanded uncertainty (<33.1%). Finally, the proposed method was applied to representative real samples of milk (by local markets), revealing the existence of one pharmaceutical drug (imidocarb) in one sample.

The Relationship of Chemical Compounds and Crystal Development in Porang Tuber (Amorphophallus muelleri Blume)

Chemical compounds are presumed to interact with each other and potentially affect the crystal development of porang tubers. This study focused on glucomannan and calcium oxalate (CaOx) as chemical compounds found in porang. Crystal development was analyzed by microscopic observation. The tuber tissue was used for microscopic slides and was harvested two weeks before the plants lost their leaves, when the plants lost their leaves, and two weeks after the plants lost their leaves. The CaOx and glucomannan content was measured using a modified extraction method. The increasing of CaOx content tends to increase the number of CaOx crystals. The analysis showed that the crystal density could affect the increasing of tuber weight possibility. There was a development process in raphide crystals that showed by different sizes at three harvest times, which the others were not. The results also indicated that glucomannan might influence the crystal density and the CaOx content. It can be concluded that glucomannan can potentially induce CaOx synthesis and crystal count increase. Keywords: Caox, Crystal, Development, Glucomannan, Interact, Porang.

Optimization of InnoXtract™ extraction and purification system for DNA extraction from skeletal samples.

The InnoXtract™ extraction and purification system is a purification method designed for DNA extraction from low-template samples, specifically rootless hair shafts. Its ability to successfully capture highly fragmented DNA suggests its suitability for use with other challenging sample types, including skeletal remains. However, the lysis and digestion parameters required modifications to successfully optimize the method for this sample type. A two-part digestion was developed utilizing a homebrew digestion buffer (0.5M EDTA, 0.05% Tween 20, and 100mM NaCl) and a supplemental lysis with the Hair Digestion Buffer included in the InnoXtract™ kit. Additionally, the magnetic bead volume was modified to improve DNA recovery from these challenging samples. With the altered protocol, the quality and quantity of DNA recovered from InnoXtract™ extracts were comparable to another commercial skeletal extraction method (PrepFiler™ BTA). This modified extraction method successfully purified sufficient amounts of quality DNA from a variety of skeletal samples to produce complete STR profiles. Successful STR typing from surface decomposition, burned, cremated, buried, and embalmed remains indicates the potential of this new method for challenging human identification and missing-person cases.

Karst spring recession and classification: efficient, automated methods for both fast- and slow-flow components

Abstract. Analysis of karst spring recession hydrographs is essential for determining hydraulic parameters, geometric characteristics, and transfer mechanisms that describe the dynamic nature of karst aquifer systems. The extraction and separation of different fast- and slow-flow components constituting a karst spring recession hydrograph typically involve manual and subjective procedures. This subjectivity introduces a bias that exists, while manual procedures can introduce errors into the derived parameters representing the system. To provide an alternative recession extraction procedure that is automated, fully objective, and easy to apply, we modified traditional streamflow extraction methods to identify components relevant for karst spring recession analysis. Mangin's karst-specific recession analysis model was fitted to individual extracted recession segments to determine matrix and conduit recession parameters. We introduced different parameter optimization approaches into Mangin's model to increase the degree of freedom, thereby allowing for more parameter interaction. The modified recession extraction and parameter optimization approaches were tested on three karst springs under different climate conditions. Our results showed that the modified extraction methods are capable of distinguishing different recession components and derived parameters that reasonably represent the analyzed karst systems. We recorded an average Kling–Gupta efficiency KGE &gt; 0.85 among all recession events simulated by the recession parameters derived from all combinations of recession extraction methods and parameter optimization approaches. While there are variabilities among parameters estimated by different combinations of extraction methods, optimization approaches, and seasons, we found much higher variability among individual recession events. We provided suggestions to reduce the uncertainty among individual recession events and raised questions about how to improve confidence in the system's attributes derived from recession parameters.

The extraction of Peste Des Petits Ruminants Virus RNA from paraffin-embedded tissues using a modified extraction method

Peste des petits ruminants (PPR) which is caused by small ruminant morbillivirus (PPRV) has an important economic impact on small ruminant farming due to high mortality rates, weight loss and restrictions on the export of small ruminants products. Molecular assays are commonly used in the diagnosis of the disease. Extraction of RNA from formalin-fixed paraffin-embedded (FFPE) tissues is challenging because of the RNA is often degraded by formalin fixation process. Although commercial kits have been developed for extraction of nucleic acids from FFPE tissues, they are expensive than other extraction kits. In this study, a modified extraction method was evaluated for detection of PPRV from FFPE tissues. A total of 20 FFPE tissue samples including 15 PPRV positive and 5 PPRV negative FFPE tissue samples were used. Two years ago, these selected FFPE tissue samples were analysed by nucleoprotein gene based one step real time RT-PCR method before they were fixed with formalin and embedded in paraffin. FFPE tissue samples were extracted using modified extraction method and were tested by fusion (F) gene based one step RT-PCR. PPRV specific RNA was detected in 12 FFPE tissue samples whereas 3 positive samples were found negative by one-step RT-PCR. Furthermore, 5 negative FFPE tissue samples were also found negative. Three false negative results were from samples with high real-time RT-PCR cycle threshold. Therefore, false negative results could be related with lower viral loads which might be lower than detection limit of the one-step RT-PCR. The results of the study show that modified extraction method could be used for RNA extraction from FFPE tissues which had been stored for 2 years.

Improved extraction and recrystallization of high-quality aleuritic acid from natural resins using a modified technique

ABSTRACT The current extraction procedures for aleuritic acid from natural resins (seedlacs) are time-consuming and provide a variable yield of 10–20% with a purity of 70–94%. Furthermore, the perfume industries do not consider this low purity level to be a viable starting material for commercial purposes, since it necessitates further purification by multistage crystallization, resulting in 40–50% aleuritic acid losses. The current study developed a modified extraction method that combines alkaline hydrolysis with a salting-out mechanism and a polarity-manipulated solvent system suited for purification via a single-step recrystallization procedure. The extraction method consistently produces 17–18 grams of aleuritic acid per 100 grams of fresh resins, with a purity of > 99% and a melting point of 100 0C. Furthermore, the developed recrystallization technique resulted in a recovered yield of 85–95%, unlike the traditional recrystallization techniques, which results in yield losses of 40–50%. The purity of aleuritic acid was determined using HPLC equipped with an RI detector, and the melting points and crystals were characterized using Differential Scanning Calorimetry and Field Emission Scanning Electron Microscope. The new approach was less expensive, more ecologically friendly, and provides standard-quality aleuritic acid while using fewer chemicals.

Novel Extraction Method for Combined Lipid and Metal Speciation From Caenorhabditis elegans With Focus on Iron Redox Status and Lipid Profiling

Parkinson´s disease progression is linked to iron redox status homeostasis via reactive oxygen species (ROS) formation, and lipids are the primary targets of ROS. The determination of iron redox status in vivo is challenging and requires specific extraction methods, which are so far tedious and very time-consuming. We demonstrated a novel, faster, and less laborious extraction method using the chelator ethylene glycol l-bis(β-aminoethyl ether)-N,N,N′,N′-tetra acetic acid (EGTA) as a stabilizing agent and synthetic quartz beads for homogenization under an argon atmosphere. Additionally, we combined the metal extraction with a well-established lipid extraction protocol using methyl-tert-butyl ether (MTBE) to avoid the problems of lipid precipitation in frozen samples and to determine lipid profiles and metal species from the same batch. The nonextractable matrix, such as the debris, is removed by centrifugation and digested to determine the total metal content of the sample as well. Lipid profiling using RP-LC–MS demonstrated high accordance of the modified extraction method to the reference method, and the organic solvent does not affect the iron redox status equilibrium. Furthermore, rigorous testing demonstrated the stability of the iron redox status equilibrium during the extraction process, secured by complexation, inert atmosphere, fast preparation, and immediately deep frozen extracts.

Large-Scale Multi-Omics Genome-Wide Association Studies (Mo-GWAS): Guidelines for Sample Preparation and Normalization.

Both gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) are widely used metabolomics approaches to detect and quantify hundreds of thousands of metabolite features. However, the application of these techniques to a large number of samples is subject to more complex interactions, particularly for genome-wide association studies (GWAS). This protocol describes an optimized metabolic workflow, which combines an efficient and fast sample preparation with the analysis of a large number of samples for legume crop species. This slightly modified extraction method was initially developed for the analysis of plant and animal tissues and is based on extraction in methyl tert-butyl ether: methanol solvent to allow the capture of polar and lipid metabolites. In addition, we provide a step-by-step guide for reducing analytical variations, which are essential for the high-throughput evaluation of metabolic variance in GWAS.

Large-Scale Multi-Omics Genome-Wide Association Studies (Mo-GWAS): Guidelines for Sample Preparation and Normalization

Both gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) are widely used metabolomics approaches to detect and quantify hundreds of thousands of metabolite features. However, the application of these techniques to a large number of samples is subject to more complex interactions, particularly for genome-wide association studies (GWAS). This protocol describes an optimized metabolic workflow, which combines an efficient and fast sample preparation with the analysis of a large number of samples for legume crop species. This slightly modified extraction method was initially developed for the analysis of plant and animal tissues and is based on extraction in methyl tert-butyl ether: methanol solvent to allow the capture of polar and lipid metabolites. In addition, we provide a step-by-step guide for reducing analytical variations, which are essential for the high-throughput evaluation of metabolic variance in GWAS.

Validation of High-Sensitivity Severe Acute Respiratory Syndrome Coronavirus 2 Testing for Stool-Toward the New Normal for Fecal Microbiota Transplantation.

INTRODUCTION:Mounting evidence demonstrates potential for fecal–oral transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The US Food and Drug Administration now requires SARS-CoV-2 testing of potential feces donors before the use of stool manufactured for fecal microbiota transplantation. We sought to develop and validate a high-sensitivity SARS-CoV-2 reverse transcriptase polymerase chain reaction (RT-PCR) procedure for testing stool specimens.METHODS:A modified extraction method was used with an RT-PCR assay adapted from the Centers for Disease Control and Prevention PCR protocol for respiratory specimens. Contrived specimens were created using pre-COVID-19 banked stool specimens and spiking in known concentrations of SARS-CoV-2-specific nucleic acid. The highest transcript concentration at which 2/2 or 1/2 SARS-CoV-2 targets were detected in 9/10 replicates was defined as the dual-target limit and single-target limit of detection, respectively. The clinical performance of the assay was evaluated with stool samples collected from 17 nasopharyngeal swab RT-PCR-positive patients and 14 nasopharyngeal RT-PCR-negative patients.RESULTS:The dual-target and single-target limit of detection were 56 copies/μL and 3 copies/μL, respectively. SARS-CoV-2 was detected at concentrations as low as 0.6 copies/μL. Clinical stool samples from known COVID-19-positive patients demonstrated the detection of SARS-CoV-2 in stool up to 29 days from symptom onset with a high agreement with nasopharyngeal swab tests (kappa statistic of 0.95, P value < 0.001).DISCUSSION:The described RT-PCR test is a sensitive and flexible approach for the detection of SARS-CoV-2 in stool specimens. We propose an integrated screening approach that incorporates this stool test to support continuation of fecal microbiota transplantation programs.

Comparison of Two Different Methods for the Extraction of Outer Membrane Vesicles from the Bordetella pertussis as a Vaccine Candidate.

Despite the availability of a vaccine, pertussis is still a worldwide health problem. Outer membrane vesicles (OMVs) in gram-negative bacteria can stimulate the immune system due to several outer membrane proteins and are very good candidates in vaccine development. OMVs obtained from Bordetella pertussis contain several antigens, which are considered immunogenic, and could make them a potential candidate for vaccine production. The current study aimed to compare the current OMV extraction method (with ultracentrifuge) and a modified extraction method (without ultracentrifuge) and to evaluate the physicochemical properties as well as the expression of their main virulence factors. Vaccinal strain BP134 grown on Bordet Gengo agar were inoculated in Modified Stainer-Scholte medium for mass cultivation. OMVs were prepared using two different methods. They were then stained and examined with a transmission electron microscope. Protein contents were measured by the Bradford method, and then the protein profile was evaluated by SDS-PAGE. The presence of immunogenic antigens was detected by Western blotting. The size and shape of the OMVs obtained from the modified method without the use of ultracentrifuge were similar to the current method and had a size between 40 and 200 nm. The total protein yields of the OMV isolated using the current and modified methods were 800 and 600 µg/ml, respectively. Evaluating the protein profile of extracted OMVs showed the presence of different proteins. Finally, the presence of PTX, PRN, and FHA was observed in OMVs extracted from both methods. Comparison of the two OMV extraction methods showed that the obtained vesicles have a suitable and similar shape and size as well as the expression of three important pathogenic factors as immunogens. Despite the relatively low reduction in protein yield as the modified method does not require ultracentrifuge, this extraction method can be used as a suitable alternative for extracting the outer membrane vesicles from B. pertussis, especially in developing countries. It should be noted that further experiments including immunogenicity determination of OMVs obtained as vaccine candidates in animal models are required.

The contents of ganoderic acids in mycellium of different Ganoderma species (Ganodermataceae) obtained by different methods of cultivation

The effect of different cultivation methods on the content of ganoderic acids of 7 species, 10 strains of the Ganoderma genus (Ganoderma applanatum 1899; Ganoderma сarnosum 2502; Ganoderma lucidum 1904; Ganoderma resinaceum 2477, 2503; Ganoderma sinense 2516; Ganoderma tsugae 1848, 2024, 2566, Ganoderma oregonense 2560) genus fungi from the IBK Mushroom Culture Collection M.G. Kholodny Institute of Botany of National Academy of Sciences of Ukraine was investigated. It has been shown that the submerged cultivation method is more efficient for the accumulation of ganoderic acids for five strains. In the mycelium of the strain G. sinense 2516 was the highest content of ganoderic acids – 25.2 ± 1.5 mg / g. The productivity (yeld) of ganoderic acids synthesis is much higher with using the submerged culture cultivation method for mycelium of all used species and strains since the use of this method provides the accumulation of much more biomass in comparison with the static liquid cultivation method. The highest yield amount of ganoderic acids was in the mycelium of the G. tsugae 2024 and G. tsugae 2566 species, namely: 0.35 ± 0.019 and 0.36 ± 0.028 g / l. It was proved that the modified extraction method significantly reduces the extraction time of ganoderic acids. Extraction time is reduced from 14 to 2 days. For the G. sinense 2516 and G. tsugae 2024 strains was determined content of the ganoderic acids and their yield in dynamics of grows in the submerged culture on 6, 8, 10, 12, 14, 16, 18 and 20 day of cultivation. The highest amount of the ganoderic acids content was accumulated by the mycelium of the strain G. sinense 2516 – it was 26.4 ± 1.5 mg / g on the 14th day of cultivation. The highest yield of the ganoderic acids was in G. sinense 2516 on 14th day, and G. tsugae 2024 mycelium on the 16th day of cultivation with the next numbers 0.6 ± 0.031, 0.62 ± 0.033 and 0.62 ± 0.027 g/l.

Retrospective phylogenetic analyses of formalin-fixed paraffin-embedded samples from the 2011 Rift Valley fever outbreak in South Africa, through sequencing of targeted regions

The last major Rift Valley fever outbreak in South Africa was between 2008 and 2011. Viruses isolated between 2008 and 2010 were phylogenetically assigned to Lineage C, Lineage K and the novel lineage H. The 2011 outbreaks occurred primarily in the Eastern, Western and Northern Cape provinces, with no sequence data or phylogenetic relationship published. Samples from these outbreaks were submitted to the Faculty of Veterinary Sciences, University of Pretoria, for immunohistochemical confirmation of Rift Valley fever phlebovirus presence. These samples were formalin-fixed and paraffin-embedded (FFPE) and stored at the Pathology section for several years. This study describes a modified extraction method used to obtain RNA from the FFPE samples, as well as the primer combinations used to phylogenetically classify them as belonging to the novel lineage H.

An innovative continuous sample drop flow microextraction for GC–MS determination of pesticides in grape juice and water samples

In this work, for the first time, low-density organic solvents were employed in continuous sample drop flow microextraction (CSDF-ME) by changing the design of extraction vessel. To examine the effectiveness of the modified extraction method, different organophosphorus pesticides (OPPs) were extracted using proposed method followed by detection by gas chromatography–mass spectrometry (GC–MS). In fact, unlike conventional CSDF approach, in this study, we applied halogen-free organic solvents that are lighter than water for analytes extractions. These solvents are not only less toxic but also more environmentally friendly compared to the halogenated solvents. A conical open-end vial fixed in a small container filled with double-distilled water was used as the extraction vessel. A small amount of extraction solvent was placed at the bottom of the vial and the sample solution was pumped into the extraction vessel through a needle located inside the vial using a peristaltic pump. The sample droplets were continuously passed through the extraction solvent (hexane; 13 μL) to extract the analytes. Once the extraction was completed, the extraction solvent left at the top of aqueous phase was collected and injected into the GC–MS for further analysis. Under optimized conditions, the repeatability of the proposed method was found to be in range of 3.9–5.8 % for samples with low concentration of 5.0 μg L−1. The LOD (limit of detection) and LOQ (limit of quantification) of the method were determined in the ranges of 0.02−0.30 and 0.07–1.0 μg L−1, respectively. The EF (enrichment factor) was found to be between 510 and 960 for 8.0 mL sample solutions. The proposed method allows an extraction recovery ranging between 25.5 % and 48.0 %.

Extraction of phytosterols and tocopherols from rapeseed oil waste by supercritical CO2 plus co-solvent: A comparison with conventional solvent extraction

In the present study, modified extraction methods using supercritical CO2 were investigated in order to obtain high-added value compounds from rapeseed oil deodorizer distillate and comparisons were done with modified Soxhlet extraction (solvent extraction + silica). For supercritical fluid extraction (SFE), the optimal extraction parameters were temperature of 40 °C, pressure of 350 bar (for phytosterols), 400 bar (for tocopherol), 5 wt% ethanol as co-solvent, and saponification pretreatment. The optimized SFE procedure led to the recovery of three main phytosterols (50 wt % β-sitosterol, 23.91 wt % Brassicasterol, and 36.25 wt % Campesterol) and only α-tocopherol. Moreover, there was no synergistic effect with saponification pretreatment + co-solvent and the efficiency and concentration of target compounds were less than supercritical CO2 + co-solvent. Also, comparative Data showed that the efficiency of phytosterols and tocopherols was approximately three times higher (p < 0.05) in SFE relative to modified Soxhlet extraction. Furthermore, the use of ethanol (5 wt %) as co-solvent, improved phytosterols and tocopherol efficiency and purity. The SFE technique offers various advantages over the modified Soxhlet extraction technique, including increasing the solubility of tocopherols and sterols by using CO2+ co-solvent, minimized usage of toxic organic solvents and increased purity of extracted products.

Response of human periosteal cells to degradation products of zinc and its alloy.

Zinc (Zn) and its alloys are proposed as promising resorbable materials for osteosynthesis implants. Detailed studies should be undertaken to clarify their properties in terms of degradability, biocompatibility and osteoinductivity. Degradation products of Zn alloys might affect directly adjacent cellular and tissue responses. Periosteal stem cells are responsible for participating in intramembranous ossification during fracture healing. The present study aims at examining possible effects emanating from Zn or Zn-4Ag (wt%) alloy degradation products on cell viability and osteogenic differentiation of a human immortalized cranial periosteal cell line (TAg cells). Therefore, a modified extraction method was used to investigate the degradation behavior of Zn and Zn-4Ag alloys under cell culture conditions. Compared with pure Zn, Zn-4Ag alloy showed almost fourfold higher degradation rates under cell culture conditions, while the associated degradation products had no adverse effects on cell viability. Osteogenic induction of TAg cells revealed that high concentration extracts significantly reduced calcium deposition of TAg cells, while low concentration extracts enhanced calcium deposition, indicating a dose-dependent effect of Zn ions. Our results give evidence that the observed cytotoxicity effects were determined by the released degradation products of Zn and Zn-4Ag alloys, rather than by degradation rates calculated by weight loss. Extracellular Zn ion concentration was found to modulate osteogenic differentiation of TAg cells. These findings provide significant implications and guidance for the development of Zn-based alloys with an optimized degradation behavior for Zn-based osteosynthesis implants.

Optimization of extraction and quality assessment based on physicochemical properties of Carrageenan from red algae (Kappaphycus alvarezii) origin of South Sulawesi Indonesia

One of the compounds that can be developed as raw material for drugs or excipients is carrageenan which can be obtained from the red algae Kappaphycus alvarezii. This species of red algae was chosen as the object of research because in addition to its high carrageenan content, this species is easily found in the waters of South Sulawesi Indonesia. This research was started with collecting, washing and processing the red algae, then carrageenan extraction was carried out using the modified extraction method. The extracted products were separated, purified and dried to obtain carrageenan raw materials. Then, the products were analyzed physically and chemically, and also using spectrophotometry and chromatography to ensure that carrageenan obtained meets the quality standards as excipient raw materials in pharmaceutical preparations. The results obtained were that Kappaphycus alvarezii red algae raw material fulfil the quality of the Indonesian National Standard (SNI) and carrageenan produced meet the quality standards of carrageenan according to Food Agriculture Organization (FAO), Food Chemicals Codex (FCC) and European Economic Community (EEC).