Senecionine was purchased from Shanghai R&D Center for Standardization of Traditional Chinese Medicine (Shanghai, China). Senecionine purity was determined to be 92% through quantitative 1H nuclear magnetic resonance (NMR) and liquid chromatography−mass spectrometry (LC−MS) (Figures S1 and S2 , Supporting Information ). Bile acid standards were purchased from Sigma−Aldrich Co. (St. Louis, MO, USA), including CA, deoxycholic acid (DCA), chenodeoxycholic acid (CDCA), ursodeoxycholic acid (UDCA), hyodeoxycholic acid (HDCA), lithocholic acid (LCA), GCA, GDCA, GCDCA, glycoursodeoxycholic acid (GUDCA), glycolithocholic acid (GLCA), TCA, TDCA, taurochenodeoxycholic acid (TCDCA), TUDCA, taurohyodeoxycholic acid (THDCA), and taurolithocholic acid (TLCA). Acetonitrile and methanol were of HPLC grade and purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Ammonium acetate (99.0%, MS grade) was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Formic acid (99.0%, HPLC grade) and ammonia solution (25.0%, HPLC grade) were purchased from Tedia Co. (Fairfield, OH, USA). Water was purified using a Millipore Milli-Q system (Billerica, MA, USA).
Ursodeoxycholic acid
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany, Switzerland
Ursodeoxycholic acid is a chemical compound that is used as a pharmaceutical ingredient in various medical products. It is a naturally occurring bile acid that can be synthesized for use in laboratory and medical applications. The core function of ursodeoxycholic acid is to act as a bile acid and assist in the regulation of bile production and metabolism.
Automatically generated - may contain errors
Lab products found in correlation
Deoxycholic acid, by Merck Group (30 mentions)
Cholic acid, by Merck Group (28 mentions)
Chenodeoxycholic acid, by Merck Group (27 mentions)
Lithocholic acid, by Merck Group (23 mentions)
Taurochenodeoxycholic acid, by Merck Group (18 mentions)
Taurocholic acid, by Merck Group (18 mentions)
Taurodeoxycholic acid, by Merck Group (18 mentions)
Tauroursodeoxycholic acid, by Merck Group (14 mentions)
Taurolithocholic acid, by Merck Group (13 mentions)
Glycodeoxycholic acid, by Merck Group (13 mentions)
Glycocholic acid, by Merck Group (12 mentions)
Glycochenodeoxycholic acid, by Merck Group (11 mentions)
Glycoursodeoxycholic acid, by Merck Group (10 mentions)
Hyodeoxycholic acid, by Merck Group (8 mentions)
Glycolithocholic acid, by Merck Group (5 mentions)
β muricholic acid, by Merck Group (4 mentions)
Ammonium acetate, by Merck Group (4 mentions)
Milli q system, by Merck Group (3 mentions)
Formic acid, by Avantor (3 mentions)
Methanol, by Merck Group (3 mentions)
46 protocols using ursodeoxycholic acid
1
Quantitative Analysis of Senecionine and Bile Acids
2
Cecal Bile Acid Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cecal contents (5 controls and 8 from L. paracasei treated mice) from trial 3 were used to quantify primary and secondary bile acids. Bile acids analysis was performed at Bioaster (Lyon, France). Samples were prepared from 50 to 60 mg of −80 °C frozen cecal content using aqueous methanol extraction process. Bile acids extracts were analysed by LC-MS/MS on a triple quadrupole Thermo Quantum Ultra (SN: TQU00665) combined to a Dionex Ultimate 3000 HPLC system (SN: 8074045 & 8087183). Samples were separated on a C18 column (2,7 μM, 150 × 2,1 mm) from Ascentis Express using a methanol/water gradient containing 5 mM ammonium acetate and 0,012% formic acid. All bile acid standards: LithoCholic acid, ChenoDeoxyCholic acid, DeoxyCholic acid, UrsoDeoxyCholic acid, Cholic acid, GlycoDeoxyCholic acid, GlycoursoDeoxyCholic acid, GlyChenoDeoxyCholic acid, Glychocolic acid, TauroLithoCholic acid, TauroDeoxyCholic acid, TauroCholic acid, were bought from Sigma Aldrich. Results of bile salt quantification were expressed in peak area, corrected for sample weight.
3
Quantitative Analysis of Bile Acids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pure standards of all the studied BAs, namely cholic acid (CA), chenodeoxycholic acid (CDCA), deoxycholic acid (DCA), ursodeoxycholic acid (UDCA), lithocholic acid (LCA), and their respective glycine and taurine conjugated, were purchased from Sigma-Aldrich (ST. Louis, MO, USA). All the studied BAs were identified and quantified by high-pressure liquid chromatography-electrospray-tandem mass spectrometry (HPLC-ES-MS/MS) using a previously validated method [21 (link)] suitable for BAs determination in plasma or serum after appropriate clean-up of pre-analytical procedures. Liquid chromatography analysis was performed using an Alliance HPLC system model 2695 from Waters combined with a triple quadruple mass spectrometer QUATTRO-LC (Micromass; Waters) using an electrospray interface. BAs were separated by elution gradient mode with a mobile phase composed of a mixture ammonium acetate buffer 15 mM, pH 8.0 (Solvent A) and acetonitrile: methanol = 75:25 v/v (Solvent B). All the chromatograms were acquired in electrospray negative ionization with the mass spectrometer operating in multiple reactions monitoring mode. Up to 15 BAs where identified and quantified in plasma with a limit of quantification suitable for their accurate analysis even in patients with low BAs concentration.
4
Metabolite Identification in Biological Samples
5
Bile Acid Analysis in Bronchoalveolar Lavage Fluid
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BALF samples were treated with equal volumes of Sputolysin® (Calbiochem, Darmstadt, Germany) and processed as described previously [16 (link),26 (link)]. Each BALF sample was analysed blinded to patient data for the presence of 12 principal bile acids compared to purified reference standards. Cholic acid (CA), chenodeoxyCholic acid (CDCA), deoxyCholic acid (DCA), lithoCholic acid (LCA), ursodeoxyCholic acid (UDCA), glycodeoxyCholic acid (GDCA), taurochenodeoxyCholic acid, (TCDCA), taurodeoxyCholic acid (TDCA), tauroCholic acid (TCA), glycoCholic acid (GCA), taurolithoCholic acid (TLCA) were purchased from Sigma-Aldrich (Buchs, Switzerland) and tauroursodeoxyCholic acid (TUDCA) was purchased from Calbiochem. All chemicals used were HPLC grade.
6
Colonic Bile Acid Extraction and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Colonic bile acid extraction was performed as previously described with proper modification [20 (link)]. Briefly, colonic content (20 mg) was homogenized in 400 μL methanol and centrifuged at 12,000× g and 4 °C for 15 min. The supernatant was collected and evaporated in a centrifuge (Eppendorf, Hamburg, Germany) to remove the solvent. The obtained solute was resolved in 400 μL methanol and centrifuged under the same conditions before use. Bile acid standards (Sigma-Aldrich, St. Louis, MO, USA), including deoxycholic acid (DCA), ursodeoxycholic acid (UDCA), tauroursodeoxycholic acid (TUDCA), hyodeoxycholic (HDCA), lithocholic acid (LCA), β-muricholic acid (β-MCA), taurocholic acid (TCA), and cholic acid (CA), were used as internal standards. A UPLC-MS system (Thermo Fisher Scientific, Waltham, MA, USA) was used to analyze the prepared samples. The instrumental settings were set and data processing was performed as previously described [21 (link)].
7
Cloning and Expression of Bacterial Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
Escherichia coli TOP 10 (F−mcrA Δ(mrr‐hsdRMS‐mcrBC) ϕ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara‐leu)7697 galE15 galK16 rpsL(StrR) endA1 λ−) and BL21(DE3) (F−ompT gal dcm lon hsdSB(rB−mB−) λ(DE3 [lacI lacUV5‐T7p07 ind1 sam7 nin5]) [malB+]K‐12(λS)) were purchased from Invitrogen (Darmstadt, Germany). Tryptone, yeast extract, cholic acid, ursodeoxycholic acid, and nicotinamide adenine dinucleotide cofactors (NAD(P)+/H) were from Sigma–Aldrich (St. Louis, US). Ursodeoxycholic acid was from Tokio Chemical Industry (TCI) (Tokyo, Japan). Restriction enzymes and Phusion Q‐5 DNA polymerase mastermix were from New England Biolabs (NEB, Ipswich, US). All other reagents were of analytical grade and are commercially available.
+ Expand
Escherichia coli TOP 10 (F−mcrA Δ(mrr‐hsdRMS‐mcrBC) ϕ80lacZΔM15 ΔlacX74 nupG recA1 araD139 Δ(ara‐leu)7697 galE15 galK16 rpsL(StrR) endA1 λ−) and BL21(DE3) (F−ompT gal dcm lon hsdSB(rB−mB−) λ(DE3 [lacI lacUV5‐T7p07 ind1 sam7 nin5]) [malB+]K‐12(λS)) were purchased from Invitrogen (Darmstadt, Germany). Tryptone, yeast extract, cholic acid, ursodeoxycholic acid, and nicotinamide adenine dinucleotide cofactors (NAD(P)+/H) were from Sigma–Aldrich (St. Louis, US). Ursodeoxycholic acid was from Tokio Chemical Industry (TCI) (Tokyo, Japan). Restriction enzymes and Phusion Q‐5 DNA polymerase mastermix were from New England Biolabs (NEB, Ipswich, US). All other reagents were of analytical grade and are commercially available.
8
Synthesis and Characterization of RR-NHEt Peptide
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The C-terminal modified RR peptide (Arg-Arg ethylamide, RR-NHEt; 357.4 Da) was synthesized by ANYGEN (Gwangju, Republic of Korea). Acetonitrile (ACN), antibiotic antimycotic solution, a cathepsin B activity assay kit, N,N-diisopropylethylamine (DIPEA), anhydrous N,N-dimethylformamide (DMF), Dulbecco’s Modified Eagle’s Medium (DMEM), diethyl ether, N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride (EDC), N-hydroxysuccinimide (NHS), methanol, phosphate-buffered saline (PBS), trifluoroacetic acid (TFA), and ursodeoxycholic acid (UDCA) were purchased from Sigma-Aldrich (Milwaukee, WI, USA). Fetal bovine serum (FBS) was obtained from Gibco (Waltham, MA, USA). An Ez Cytox kit was obtained from DoGenBio (Seoul, Republic of Korea). Neutral buffered formalin (10%) was purchased from HuBenTech (Damyang, Republic of Korea). A Pierce BCA protein assay kit was obtained from Thermo Fisher Scientific (Waltham, MA, USA). Chloroform was purchased from DAEJUNG (Siheung, Republic of Korea).
9
Bilirubin Quantification Using Spectrophotometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All chemicals were of analytical grade. PH, ursodeoxycholic acid, bilirubin, and ethanol
were obtained from Sigma Aldrich, USA; bilirubin assay kit from Randox Laboratories, UK
and sample tubes containing gel and clot activator (Y330984 IMPROVACUTER) from Guangzhou
Improve Medical Instruments Co., Ltd., China.
were obtained from Sigma Aldrich, USA; bilirubin assay kit from Randox Laboratories, UK
and sample tubes containing gel and clot activator (Y330984 IMPROVACUTER) from Guangzhou
Improve Medical Instruments Co., Ltd., China.
10
Quantification of Bile Acids and Cortisone
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cholic acid (CA), glycoCholic acid (GCA), tauroCholic acid (TCA), chenodeoxyCholic acid (CDCA), glycochenodeoxyCholic acid (GCDCA), taurochenodeoxyCholic acid (TCDCA), ursodeoxyCholic acid (UDCA), glycoursodeoxyCholic acid (GUDCA), tauroursodeoxyCholic acid (TUDCA), deoxyCholic acid (DCA), glycodeoxyCholic acid (GDCA), taurodeoxyCholic acid (TDCA), taurolithoCholic acid (TLCA), and cortisone acetate were purchased from Sigma-Aldrich (St. Louis, MO, USA). α-MuriCholic acid (α-MCA) and tauromuriCholic acid (TMCA) were purchased from Steraloids Inc (Newport, RI, USA). All other reagents and solvents were of high performance liquid chromatography (HPLC) grade. Deionized water was purified using a Milli-Q system (Millipore, Bedford, MA, USA). S. baicalensis Georgi (Hebei Province), G. uralensis Fisch (Inner Mongolia of China), P. lactiflora Pall (Anhui Province), and Z. jujuba Mill (Henan Province) were authenticated by Dr. Ehu Liu (State Key Laboratory of Natural Medicine, China Pharmaceutical University, China).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!