Maximum recovery diluent

Manufactured by Merck Group

Sourced in Germany

Maximum recovery diluent is a laboratory product designed to provide a stable and consistent environment for the dilution of samples. It is formulated to maintain the integrity and characteristics of the analytes being tested, facilitating accurate and reliable results. The core function of this diluent is to enable the preparation of diluted samples for various analytical procedures without compromising the sample's properties.

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Maximum Recovery Diluent (MRD, (2 citations)

12 protocols using maximum recovery diluent

Fluid Uptake Ability of Powders

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Franz diffusion cell was used to determine the fluid uptake ability of powders. About 8 mg of powder was placed on a previously weighed membrane disc filter (PES, 0.45 µm, 25 mm, Pall Corporation, Port Washington, NY, USA). The membrane was lying down on the vertical receptor compartment of the Franz cell, filled with simulated wound fluid (SWF), composed of 50% fetal calf serum (Sigma Aldrich, Milan, Italy) and 50% maximum recovery diluent (Sigma Aldrich, Milan, Italy) consisting in 0.1% (w/v) peptone, a peptic digest of animal tissue, and 0.9% (w/v) sodium chloride [47 (link),48 (link)]. The SWF was thermostated at 37 °C, and during the experiment, the receptor compartment was filled with SWF to maintain a constant fluid volume in the compartment. At regular time intervals, the weight of the gel was measured after removing the membrane from the receptor until a constant weight was observed [49 (link)].
The swelling ratio of each formulation was determined as the ratio between the weight of the swollen gel (W_s) and the weight of the dried powder producing the gel (W_d), using the following Equations [50 (link)]:
All experiments were performed at least in triplicate.

Amante C., Esposito T., Del Gaudio P., Di Sarno V., Porta A., Tosco A., Russo P., Nicolais L, & Aquino R.P. (2021). A Novel Three-Polysaccharide Blend In Situ Gelling Powder for Wound Healing Applications. Pharmaceutics, 13(10), 1680.

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Microbiological Analysis of VP Meat Samples

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For microbiological analyses of the VP meat samples, a 10-g sample was weighted, transferred into a stomacher bag containing 90 mL Maximum Recovery Diluent (Sigma-Aldrich, Dublin, Ireland), homogenized on Stomacher 400 (Stomacher Lab System, United Kingdom) for 2.5 min and plated on agar media in several 1:10 serial dilutions. Parallel to this, the MRS agar was used for colony counts of lactic acid bacteria (LAB) (ISO 15214, 1998), PCA was used for total mesophilic aerobic bacteria (TMAB), and the results were presented in log CFU (colony-forming units)/g values.

Santovito E., Elisseeva S., Cruz-Romero M.C., Duffy G., Kerry J.P, & Papkovsky D.B. (2021). A Simple Sensor System for Onsite Monitoring of O2 in Vacuum-Packed Meats during the Shelf Life. Sensors (Basel, Switzerland), 21(13), 4256.

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Wound Dressing Materials Characterization

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Phosphate buffered saline (PBS) tablets, maximum recovery diluent and foetal bovine serum were obtained from Sigma-Aldrich. Hydrosorb® came from Hartmann. DuoDERM® ET, Kaltostat®, Aquacel® Extra, Aquacel® Ag and Carboflex® were purchased from ConvaTec. Mepilex®, Mepilex® Ag and Exufiber® were obtained from Mölnlycke. Carbonet^◊ and Acticoat^◊ were purchased at Smith&Nephew. All materials were used as received.

Minsart M., Van Vlierberghe S., Dubruel P, & Mignon A. (2022). Commercial wound dressings for the treatment of exuding wounds: an in-depth physico-chemical comparative study. Burns & Trauma, 10, tkac024.

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Isolation and Identification of Oral Anaerobes

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Subgingival plaque samples were placed in 300 µL of maximum recovery diluent (Sigma) and dilutions of plaque (neat to 10⁻⁵) were spread onto agar plates. Strains were routinely grown for 2 days for aerobes and up to 14 days for anaerobes, at 38°C on Columbia Blood Agar (CBA) plates (OXOID). For isolating anaerobic strains, CBA was supplemented after autoclaving with Hemin (5 µg/ml) and Menadione (1 µg/mL) (Sigma). For liquid culture, Brain Heart Infusion (BHI) broth (OXOID) supplemented with yeast extract (5 g/L, Sigma) and cysteine hydrochloride (0.75 g/L, Sigma) and adjusted to a pH of 8.5. Hemin and Menadione were added to cooled broth after autoclaving, as above. Anaerobic strains were incubated in a MACS1000 anaerobic workstation (Don Whitley, UK) with an atmosphere of 85% N₂, 10% CO₂, 5% H₂. After observation for growth, single colonies were sub-cultured onto fresh CBA plates (Hemin and Menadione for anaerobes as above). Following further incubation at 38°C as described above, colonies on agar plates were examined for purity. Cultures were then harvested for 16S rDNA sequencing to confirm their identity and the isolates stored at −80°C in BHI with 15% glycerol.

Dewhirst F.E., Klein E.A., Bennett M.L., Croft J.M., Harris S.J, & Marshall-Jones Z.V. (2014). The Feline Oral Microbiome: A Provisional 16S rRNA gene based Taxonomy with Full-length Reference Sequences. Veterinary microbiology, 175(0), 294-303.

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Anaerobic Microbial Culture Preparation

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Sodium chlorate solutions were acquired from Sigma and stored at 4 °C. Other chemicals used in the study were ammonium chloride (VWR), potassium phosphate dibasic anhydrous, sodium phosphate monobasic monohydrate, ferrous chloride tetrahydrate, sodium selenite, sodium molybdate dihydrate, nickel chloride hexahydrate, zinc chloride, manganese chloride tetrahydrate, cobalt (II) chloride hexahydrate, copper (II) chloride dihydrate, boric acid, hydrochloric acid, EDTA disodium salt dihydrate, sodium hydroxide, lactate, acetate, l-cysteine HCl (Sigma), Bacto agar, resazurin (Difco), milk plate count agar and maximum recovery diluent (Sigma).

McCarthy W.P., Srinivas M., Danaher M., Connor C.O., Callaghan T.F., van Sinderen D., Kenny J, & Tobin J.T. (2023). Isolation and identification of chlorate-reducing Hafnia sp. from milk. Microbiology, 169(7), 001347.

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Total Viable Count Analysis of Sushi

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The analysis of total viable count (TVC) was performed according to ISO standard no. 4833-1:2013 with a slight modification related to the amount of sample used. In case of nigiri exactly 6.25 g of rice and 3.75 g of salmon from each nigiri was homogenized with 90 g of maximum recovery diluent (Sigma-Aldrich, Saint Louis, MO, USA). In case of hosomaki, a whole hosomaki piece (13.5-14.0 g) was homogenized with maximum recovery diluent in ratio of 1:9. The homogenization was carried out using Stomacher device for 2 min followed by preparation of appropriate decimal dilutions. The analysis was performed by the pour plate method, using plate count agar (Sigma-Aldrich). The plates were incubated at 30 °C for 48 h.
The microbiological analysis was performed on day 1, 3, 5, 8 and 11 of storage at 4 °C and the results were expressed as log cfu/g of sushi sample.

Kulawik P., Alvarez C., Cullen P.J., Aznar-Roca R., Mullen A.M, & Tiwari B. (2018). The effect of non-thermal plasma on the lipid oxidation and microbiological quality of sushi. Innovative Food Science and Emerging Technologies., 45.

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Antimicrobial Formulation for Wound Care

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Chlorhexidine digluconate (20%,) polyvinylpyrrolidone (PVP)–iodine complex, sodium phosphate dibasic dehydrate, citric acid, isopropyl, polysorbate 80, catalase, hydrogen peroxide, bovine serum albumin, and iodine were obtained from Sigma Aldrich (USA). Malt extract agar (MEA) and tryptic soy agar (TSA) were purchased from Oxoid (UK). Maximum recovery diluent, lecithin, and defibrinated sheep blood were obtained from Merck, Alfa Aesar, and Thermo Fisher Scientific, respectively.

ŞAHİNER A., HALAT E, & ALĞIN YAPAR E. (2019). Comparison of bactericidal and fungicidal efficacy of antiseptic formulations according to EN 13727 and EN 13624 standards. Turkish Journal of Medical Sciences, 49(5), 1564-1567.

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Quantification of Biocontrol Agent Viability

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For both biocontrol products, viable populations were determined by quantifying colony-forming units (CFUs) using the viable plate count technique. The total populations were estimated with a Neubauer haemocytometer under a microscope. Cell viability was confirmed by culturing B. subtilis on NA and G. catenulatum on MEA at 25 °C for 48 h. The non-viable population was estimated as the difference between the total counts made with the haemocytometer and the cultured viable populations. The two products were serially diluted with the maximum recovery diluent (Merck Life Science UK Limited, Gillingham, Dorset, U.K.). For statistical analyses all the data for CFUs was log transformed. After dilution, 500 μL of the BCA suspension was transferred into a 1.5 mL Eppendorf, the total cell concentration in the 500 μL volume was used. This showed that in this volume the B. subtilis cells/propagules present was 7.96 log₁₀ CFUs, and of these 6.96 log₁₀ CFUs were viable. The total conidial concentration in the 500 μL volume of the G. catenulatum contained 8.64 log₁₀ units, of which 5.7 log₁₀ CFUs were viable in the formulation.

Tut G., Magan N., Brain P, & Xu X. (2021). Molecular Assay Development to Monitor the Kinetics of Viable Populations of Two Biocontrol Agents, Bacillus subtilis QST 713 and Gliocladium catenulatum J1446, in the Phyllosphere of Lettuce Leaves. Biology, 10(3), 224.

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Isolation and Characterization of Liquorilactobacillus nagelii

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Liquorilactobacillus nagelii AGA58 was isolated from fermented turnip juice (Shalgam) produced in Adana, Turkey. A 10 mL of the Shalgam sample was diluted with 90mL of Maximum Recovery Diluent (Merck, GmbH, Darmstadt, Germany) in Schott bottle and vortexed for 1 minute with highspeed vortex (MS-3 Basic, IKA-Werke GmbH, Staufen, Germany). A 100µL of sample from serial dilutions were spread on MRS agar (Merck) followed by incubation at 30 for 48h anaerobically. The isolate later named as AGA58 was picked and subjected to colony purification twice. Gram staining and catalase tests were performed to pure isolate of L. nagelii AGA58, respectively. The cryovial stocks of AGA58 was prepared using MRS broth (Merck) with 25% glycerol and were stored at -80 .

Yetiman A.E, & Ortakci F. (2023). Genomic, probiotic, and metabolic potentials of Liquorilactobacillus nagelii AGA58, a novel bacteriocinogenic motile strain isolated from lactic acid-fermented shalgam. Journal of bioscience and bioengineering, 135(1).

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Microbial Analysis of Meat Samples

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For the microbiological analyses, 10 g of meat sample was homogenized with 90 ml of Maximum Recovery Diluent (Merck, 112535) in a stomacher (MAYO, HG-400, Australia). Appropriate dilutions were plated to get total mesophilic aerobic bacteria (TMAB) counts as previously described by Duran and Kahve (2020) (link) in detail. Baird Parker Agar (BPA, Sigma Aldrich) was used for Staphylococcus aureus. The selected colonies were subjected to a coagulase assay and verified. In the Staphylococcus aureus analysis, the incubation temperature was 37 ± 2 °C for 48 h. After incubation, the black colonies with a clear zone were identified to be Staphylococcus aureus (ISO 6888-1, 1999) . Additionally, Pseudomonas spp. count (ISO 13720, 2010) were determined using Pseudomonas Selective Agar Base (CFC agar, Merck) which was incubated at 30ºC ± 2 °C for 72 h. The blue-violet colonies surrounded by red-violet-colored zones were counted. The bacterial counts were given in cfu/g.

Kahve H.İ., & Duran A. (2020). KİTOSAN KAPLAMANIN SOĞUTULARAK SAKLANAN ETLERİN MİKROBİYOLOJİK VE OKSİDATİF ÖZELLİKLERİNE ETKİSİNİN BELİRLENMESİ. GIDA / THE JOURNAL OF FOOD, 45(6), 1154-1162.

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