Citrate acid

Manufactured by Merck Group

Sourced in United States

Citric acid is a weak organic acid found in citrus fruits. It is commonly used in laboratory settings as a pH adjuster and buffer to maintain specific pH levels in various experiments and processes.

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5 protocols using citrate acid

Synthesis of Radiolabeled Peptide Conjugates

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Protected N^α-Fmoc-amino acid derivatives, coupling reagents, and Rink amide 4-methylbenzhydrylamine (MBHA) resin were purchased from Calbiochem-Novabiochem (Laufelfingen, Switzerland). The Fmoc-21-amino-4,7,10,13,16,19-hexaoxaheneicosanoic acid (Fmoc-Ahoh-OH) and Fmoc-8-amino-3,6-dioxaoctanoic acid (Fmoc-AdOO-OH) were purchased from Neosystem (Strasbourg, France). DOTA(OtBu)₃-OH (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate tert-butyl ester) was purchased from CheMatech (Dijon, France). Citrate acid, sodium citrate, and sodium chloride were obtained from Sigma-Aldrich Corp. (St. Louis, MO, USA). All other chemicals were commercially available by Sigma-Aldrich, Fluka (Bucks, Switzerland) or LabScan (Stillorgan, Dublin, Ireland) and were used as received unless otherwise stated. Preparative HPLCs were carried out on a LC8 Shimadzu HPLC system (Shimadzu Corporation, Kyoto, Japan) equipped with a UV Lambda-Max Model 481 detector. UV-Vis measurements were carried out on Thermo Fisher Scientific Inc. (Wilmington, Delaware USA) Nanodrop 2000c spectrophotometer equipped with a 1.0-cm quartz cuvette (Hellma). ¹⁷⁷LuCl₃ and ¹¹¹InCl₃ were obtained from IDB (Petten, The Netherlands) and Mallinckrodt Radiopharmaceuticals Italia (Segrate, Italy), respectively.

Accardo A., Galli F., Mansi R., Del Pozzo L., Aurilio M., Morisco A., Ringhieri P., Signore A., Morelli G, & Aloj L. (2016). Pre-clinical evaluation of eight DOTA coupled gastrin-releasing peptide receptor (GRP-R) ligands for in vivo targeting of receptor-expressing tumors. EJNMMI Research, 6, 17.

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Skeletal Muscle Enzyme Activities

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Skeletal muscle samples (soleus and mixed gastrocnemius) were homogenized in the buffer containing 5 mM HEPES, 1 mM EGTA, 0.1% Triton X-100, 1 mM DTT and 10 N KOH. COX activity was measured as a time-course increase in absorbance at 550 nm due to oxidation of its substrate 1 mM reduced cytochrome C from horse (Sigma Aldrich, St. Louis, MO) according to the equation: reduced Cyt C+O₂→oxidized Cyt C+H₂O. CS activity was determined as the increase of absorbance at 412 nm according to reactions: acetylCoA (0.3 mM)+oxalacetic acid (0.5 mM)+H₂O→citrate acid+CoASH+DTNB (0.1 M)→CoAS+H+ +C₆O₄S²⁻ (all reagents were from Sigma Aldrich, St. Louis, MO). Subsequently, enzyme activities were normalized to protein concentration in the samples (measured with BCA assay) and the values were used to calculate COX and CS activities.

Wojewoda M., Kmiecik K., Ventura-Clapier R., Fortin D., Onopiuk M., Jakubczyk J., Sitek B., Fedorowicz A., Majerczak J., Kaminski K., Chlopicki S, & Zoladz J.A. (2014). Running Performance at High Running Velocities Is Impaired but V′O2max and Peripheral Endothelial Function Are Preserved in IL-6−/− Mice. PLoS ONE, 9(2), e88333.

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Isoliquiritigenin and TGF-β1 in vitro

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Isoliquiritigenin (ISL; C₁₅H₁₂O₄; CAS number: 961-29-5; purity ≥ 98%; Sigma-Aldrich, St. Louis, MO, USA) is a yellow powder insoluble in water with a molecular weight of 256.25 kDa. The powder was dissolved in dimethyl sulphoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA) to prepare a stock solution of 100 mM and stored at −20 °C until use. The carrier solvent (0.1% DMSO) was added to the control group. Recombinant human transforming growth factor-β (TGF-β1; CAS number: 100–21; purity ≥ 98%; Peprotech, Rocky Hill, NJ, USA) derived from HEK293 cells is a 25.0 kDa protein. The powder was dissolved in citrate acid (Sigma-Aldrich, St. Louis, MO, USA) containing 0.1% BSA to prepare a stock solution of 100 mg/mL and stored at −20 °C until use. The carrier solvent (citrate acid + 0.1% BSA) was added to the control group.

Chen H.Y., Chiang Y.F., Huang J.S., Huang T.C., Shih Y.H., Wang K.L., Ali M., Hong Y.H., Shieh T.M, & Hsia S.M. (2021). Isoliquiritigenin Reverses Epithelial-Mesenchymal Transition Through Modulation of the TGF-β/Smad Signaling Pathway in Endometrial Cancer. Cancers, 13(6), 1236.

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Fabrication of Stretchable Bioresorbable Electronic Patch

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The Mo serpentine structure was fabricated using a laser cutter (LPKF ProtoLaser R, LPKF Laser & Electronics). The POCO pre‐polymer was synthesized according to procedures based on a previous report.^[²⁷^] Briefly, 1,8‐ontandiol (Sigma–Aldrich), citrate acid (Sigma–Aldrich), and octanol (Acros Organics) were mixed and melted at 165 °C, followed by reaction at 140 °C for 3 h. The mixture was then dissolved in ethanol and precipitated in DI water for purification. The precipitate was collected and lyophilized for 24 h to obtain the final product. To fabricate the POCO patch, about 1 mL of the 40% POCO (w/w in ethanol) prepolymer solution was added to a glass slide (7.5 × 5 cm²) and left at room temperature overnight for solvent evaporation. Then, the slide was placed in an oven set to 80 °C for 4 days to cure the POCO patch. To fabricate the BCEP, the Mo serpentine structure was placed atop a viscous POCO sol after solvent evaporation and cured at 80 °C for 4 days.

Ryu H., Wang X., Xie Z., Kim J., Liu Y., Bai W., Song Z., Song J.W., Zhao Z., Kim J., Yang Q., Xie J.J., Keate R., Wang H., Huang Y., Efimov I.R., Ameer G.A, & Rogers J.A. (2023). Materials and Design Approaches for a Fully Bioresorbable, Electrically Conductive and Mechanically Compliant Cardiac Patch Technology. Advanced Science, 10(27), 2303429.

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HPLC Analysis of Organic Acids

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Chromatographic analysis of PR and fPR was carried out using an HPLC system (1260 infinity, Agilent Technologies, Santa Clara, CA, USA) equipped with a UV detector, a column oven, and an autosampler. Sulfuric acid at a concentration of 0.008 N was used as the mobile phase. Separation of the samples was achieved on an Eclipse XDB-C18 column (5 µm, 250 mm × 4.6 mm, Agilent Technologies) at 25 °C with the mobile phase flow rate maintained at 0.6 mL/min. Detection of the ingredients was carried out at 210 nm using malic acid, lactic acid, and citrate acid (Sigma-Aldrich, Saint Louis, MO, USA) as standards for calibration (Figure S1).

Choi Y., Bose S., Shin N.R., Song E.J., Nam Y.D, & Kim H. (2020). Lactate-Fortified Puerariae Radix Fermented by Bifidobacterium breve Improved Diet-Induced Metabolic Dysregulation via Alteration of Gut Microbial Communities. Nutrients, 12(2), 276.

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