Lactulose

Manufactured by Merck Group

Sourced in United States, Germany, Japan, Canada, Brazil

Lactulose is a synthetic disaccharide that is used as a laxative in laboratory settings. It functions by drawing water into the intestines, which helps to soften stool and promote bowel movements.

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Lab products found in correlation

Mannitol, by Merck Group (10 mentions) Lactose, by Merck Group (6 mentions) Epilactose, by Merck Group (4 mentions) Galactose, by Merck Group (4 mentions) D galactose, by Merck Group (4 mentions) Raffinose, by Merck Group (3 mentions) Mannitol 13c6, by Merck Group (3 mentions) Lactulose 13c12, by Merck Group (3 mentions) Electrophoresis reagents, by Bio-Rad (3 mentions) D fructose, by Merck Group (3 mentions) Maltose, by Merck Group (2 mentions) Fructose, by Merck Group (2 mentions) D mannitol, by Merck Group (2 mentions) Sodium hydroxide (naoh), by Kanto Chemical (2 mentions) Lactose monohydrate, by Kanto Chemical (2 mentions) Chelating sepharose fast flow resin, by GE Healthcare (2 mentions) Sucralose, by Merck Group (2 mentions) 6 galactobiose, by Merck Group (2 mentions) Milli q synthesis a10, by Merck Group (2 mentions) D glucose, by Merck Group (2 mentions)

42 protocols using lactulose

Quantitative Analysis of Mannitol and Lactulose

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Calibrator solutions were prepared from serial dilutions of 10 mg/mL mannitol and 40 mg/mL lactulose (Sigma-Aldrich, St Louis, MO) diluted in HPLC-grade water. Internal standards contained 750 μg/mL mannitol ¹³C6 and lactulose ¹³C12 (Sigma-Aldrich).
The previously collected pooled blank urine samples (collected in the absence of administration of lactulose and mannitol) was spiked with independently prepared solutions containing serial dilutions of 10 mg/mL for assay validation and for quality control on all runs. Linear responses in the calibration equation were observed with an r > 0.999 in the range of mannitol 0.5 to 2000 μg/mL, −0.125 to 500 μg/mL for lactulose with precisions of 5% to 10% CV.

Lee G.O., Kosek P., Lima A.A., Singh R., Yori P.P., Olortegui M.P., Lamsam J.L., Oliveira D.B., Guerrant R.L, & Kosek M. (2014). Lactulose:Mannitol Diagnostic Test by HPLC and LC-MSMS Platforms: Considerations for Field Studies of Intestinal Barrier Function and Environmental Enteropathy. Journal of Pediatric Gastroenterology and Nutrition, 59(4), 544-550.

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Lactulose and Mannitol Absorption Assay

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The IP was evaluated by providing an enteral bolus of 15 mL/kg of a 5% lactulose (Sigma-Aldrich, Copenhagen, Denmark) and 5% mannitol solution (Sigma-Aldrich, Copenhagen, Denmark) exactly 3 h before euthanasia. Urine was collected at the time of euthanasia and stored at −20 °C until assayed. Concentrations of lactulose and mannitol were measured in urine by an enzymatic spectrophotometric method (Pentra 400, Irvine, CA, USA). In the presence of mannitol dehydrogenase, mannitol was oxidized by NAD into the fructose and NADH. The amount of NADH was detected by spectrophotometry at 340 nm. lactulose was also hydrolyzed into galactose and fructose. Fructose was then catalyzed into fructose-6-phosphate and to glucose-6-phosphate by phosphoglucoisomerase. Glucose-6-phosphate was dehydrogenated by adding glucose-6-phosphate-dehydrogenase in the presence of NADP to form NADPH. The concentration of NADPH is proportional to the concentration of lactulose and can be measured spectrophotometrically at 340 nm. L/M ratios quantified by enzymatic assay are provided in the Supplementary Materials.

Alinaghi M., Nguyen D.N., Sangild P.T, & Bertram H.C. (2020). Direct Implementation of Intestinal Permeability Test in NMR Metabolomics for Simultaneous Biomarker Discovery—A Feasibility Study in a Preterm Piglet Model. Metabolites, 10(1), 22.

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Resistant Starch Compounds and Lactulose

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The two RSs, HAMS (Hylon VII) and modified HAMS (HAMSA-Crispfilm) were obtained from Ingredion, USA. Hylon VII is composed of 70% amylose and 30% amylopectin. It is estimated that Hylon VII contains 50 g of RS per 100 g [11 (link)]. Crispfilm (CF) has a Hylon VII backbone and has undergone a further esterification process to form a starch acetate. The degree of acetylation of CF is less than 2.5%, the limit imposed by U.S. Food and Drug Administration (FDA) food regulation and CF has Generally Recognized as Safe (GRAS) status. Lactulose was obtained from Sigma-Aldrich, Australia.

Gopalsamy G., Mortimer E., Greenfield P., Bird A.R., Young G.P, & Christophersen C.T. (2019). Resistant Starch Is Actively Fermented by Infant Faecal Microbiota and Increases Microbial Diversity. Nutrients, 11(6), 1345.

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Cloning and Characterizing Carbohydrate-Active Enzymes

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The restriction endonucleases NheI and EcoRI, T4 DNA ligase, and EX taq polymerase were obtained from Takara (Kyoto, Japan). The expression vector pET28a (+) was obtained from Novagen (Darmstadt, Germany). Plasmid DNA was purified by using DNA mini-preparation kits (Qiagen, Hilden, Germany). Escherichia coli BL21 (DE3) (New England Biolabs, Hertfordshire, UK) was used for protein expression. C. saccharolyticus DSM 8903 cell was purchased from DSMZ (Braunschweig, Germany). Standard sugars (lactose, epilactose, and lactulose) for high-performance liquid chromatography (HPLC) were purchased from Sigma-Aldrich (MO, USA).

Park A.R., Kim J.S., Jang S.W., Park Y.G., Koo B.S, & Lee H.C. (2017). Rational modification of substrate binding site by structure-based engineering of a cellobiose 2-epimerase in Caldicellulosiruptor saccharolyticus. Microbial Cell Factories, 16, 224.

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Intestinal Permeability Assessment via L/M Test

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Intestinal permeability was assessed using the L/M test preoperatively (the morning of the surgery) and on the 3^th and 10^th day postoperatively [1 (link),11 (link)]. After an overnight fast, all subjects were given the oral test solution containing 10 g of lactulose (Sigma-Aldrich, Tokyo, Japan) and 5 g of mannitol (Sigma-Aldrich) in 60 ml of physiological saline. For the next 6 h, the subjects were rested and no food or water was allowed. Complete 6 h urine collections were taken and a further 10-mL urine sample was frozen at −20°C until analysis. Urinary lactulose and mannitol concentrations were measured by gas–liquid chromatography.

Liu Z., Li C., Huang M., Tong C., Zhang X., Wang L., Peng H., Lan P., Zhang P., Huang N., Peng J., Wu X., Luo Y., Qin H., Kang L, & Wang J. (2015). Positive regulatory effects of perioperative probiotic treatment on postoperative liver complications after colorectal liver metastases surgery: a double-center and double-blind randomized clinical trial. BMC Gastroenterology, 15, 34.

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Enzymatic Synthesis of Epilactose

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Epilactose was enzymatically synthesized using the cellobiose 2-epimerase enzyme from Caldicellulosiruptor saccharolyticus produced by Saccharomyces cerevisiae [17 (link)]. After enzymatic production, a mixture containing 87% of epilactose was obtained by a two-step purification methodology using β-galactosidase and yeast treatment [11 (link)]. Lactulose and raffinose (analytical grade) were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Cardoso B.B., Amorim C., Franco-Duarte R., Alves J.I., Barbosa S.G., Silvério S.C, & Rodrigues L.R. (2024). Epilactose as a Promising Butyrate-Promoter Prebiotic via Microbiota Modulation. Life, 14(5), 643.

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Lactulose and Mannitol Assay Validation

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Calibrator solutions prepared via serial dilutions of 10 mg/mL mannitol and lactulose (Sigma-Aldrich) diluted in HPLC-grade water. Internal standards contained 0.01 mg/mL mannitol ¹³C6 and lactulose ¹³C12 (Sigma-Aldrich). Pooled blank urine (urine collected before the administration of lactulose and mannitol) was spiked with independently prepared solutions containing serial dilutions of 10 mg/mL for assay validation and for quality control on all runs. Linear responses in the calibration equation were observed with an r > 0.999 in the range of 10 to 4000 μg/mL for both lactulose and mannitol with 95% confidence intervals of 2%. A second transition was monitored for each analyte to confirm identity and required to be within 20% of the relative peak area of the first transition used for quantification.

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Melanoma Cell Culture and Treatments

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The human melanoma MNT-1 cell line was cultured as previously described with minor modifications [36] (link). Briefly, the cells were maintained at 37°C in MEM (Gibco, Carlsbad, CA, USA) with 20% fetal bovine serum (Gibco). Wortmannin (Cell signaling, Danvers, MA, USA), YM201636 (Cell signaling), 740 Y-P (R&D systems, Minneapolis, MN, USA) and Cyclohexamide (Sigma, St. Louis, MO, USA) were purchased and dissolved in DMSO to produce stock solutions. Sucrose, trehalose, cellobiose, maltose, lactose and lactulose (all from Sigma) were purchased and dissolved in culture medium and sterilized by filtration with a 0.45-μm pore size filter before use. Designed siRNA mixtures (Genolution, Seoul, Korea) and Lipofectamine RNAimax (Invitrogen, CA, USA) were purchased and treated as the manufacturer describes.

Bin B.H., Bhin J., Yang S.H., Choi D.H., Park K., Shin D.W., Lee A.Y., Hwang D., Cho E.G, & Lee T.R. (2014). Hyperosmotic Stress Reduces Melanin Production by Altering Melanosome Formation. PLoS ONE, 9(8), e105965.

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Electrodialysis-Assisted Extraction and Purification

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The high-performance
liquid chromatography (HPLC)-grade sugars such as lactose, lactulose,
galactose, glucose, and fructose were obtained from Sigma-Aldrich
(Ottawa, Ontario, Canada), whereas, the high-purity (purity ≥95%)
chemicals and reagents of analytical grade were procured from different
suppliers. The potassium hydroxide (KOH), potassium sulfate (K₂SO₄), hydrochloric acid (HCl), and phenolphthalein
(C₂₀H₁₄O₄) were purchased from Fisher
Chemical (Fair Lawn, NJ), Sigma-Aldrich Co. (St. Louis, MO), Fisher
Chemical (Geel, Belgium), and MAT Laboratory Inc. (Laboratoire MAT
Inc., Quebec, Canada), respectively. The food-grade whey powder (lactose,
75%; total proteins, 12%; ash, 7%; and moisture content, <5%) was
obtained from Agropur Co-operative (St-Hubert, Quebec, Canada). The
cation-exchange membrane (CMI 7000S) and anion-exchange membrane (AMI
7001S) were bought from Membrane International Inc. (Ringwood, NJ)
and were directly used in the EA reactor without any pretreatment.

Karim A, & Aider M. (2020). Sustainable Valorization of Whey by Electroactivation Technology for In Situ Isomerization of Lactose into Lactulose: Comparison between Electroactivation and Chemical Processes at Equivalent Solution Alkalinity. ACS Omega, 5(14), 8380-8392.

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Enzymatic Synthesis of Galactooligosaccharides

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Unless otherwise stated, all materials and reagents were of analytical grade or higher supplied by Sinopharm Chemical Reagent (Shanghai, China). Isopropyl β-d-1-thiogalactopyranoside (IPTG) and ampicillin sodium were supplied by Sangon Biological Engineering Technology and Services (Shanghai, China). Electrophoresis reagents were obtained from Bio-Rad (Hercules, CA, USA). Escherichia coli BL21 (DE3) and DH5α were obtained from Promega (Madison, WI). The Ni²⁺-chelating Sepharose Fast Flow resin was purchased from GE Healthcare (Uppsala, Sweden). Epilactose and lactulose were of the highest grade and were obtained from Sigma (St Louis, MO, USA). Cheese whey powder was purchased from Apple Foods Tech (Shanghai, China).

Chen Q., Xiao Y., Shakhnovich E.I., Zhang W, & Mu W. (2019). Semi-rational design and molecular dynamics simulations study of the thermostability enhancement of cellobiose 2-epimerases. International journal of biological macromolecules, 154, 1356-1365.

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