Bile acid derivative

After receiving a typical diet for a week, Sprague-Dawley rats were split into two groups at random. The NAFLD group was given a high-fat diet (60 kcal%; 18 weeks), while the control group was fed a typical diet (10 kcal%) [1]. Ten BAs, including four taurine-conjugated BAs (taurolithocholic acid (TLCA), taurodeoxycholic acid (TDCA), and tauromega-murinecholic acid), were considerably reduced in the liver of the NAFLD group. (TMC). Furthermore, the NAFLD group had reduced levels of βDCA, HDCA, 6-ketoLCA, 23-nordeoxycholic acid (NorDCA), LCA, and 3β-chenodeoxycholic acid (βCDCA) [2].

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Prematurity significantly influenced ‘rare’ BA levels [1]
The influence of prematurity on ‘rare’ BA metabolism was investigated in a group of 22 non-septic PT neonates. In summary, C-6-hydroxylated BA levels in PT neonates were below the detection limit except for TAMCA, whose median concentration was 0.1 µmol/L (IQR: 0–0.2). In total, C-6-hydroxylated BA levels were significantly higher in non-septic FT neonates than in non-septic PT neonates (Fig. 2; p < 0.01). Interestingly, TAMCA was the only ‘rare’ BA found in healthy PT neonates (Fig. 3b). Unlike in non-septic FT neonates, GMCA, TGMCA, TωMCA/TOMCA, GGMCA, and BMCA were absent in non-septic PT neonates.

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EOS FT neonates show alterations in ‘rare’ BA’ composition but not in levelss [1]
C-6-hydroxylated BA levels were determined in 20 FT neonates with diagnosed EOS. Interestingly, levels were comparable in FT neonates with EOS and in FT controls with 0.5 µmol/L (IQR: 0.3–1.3) and 0.6 µmol/L (IQR: 0.1–1.6), respectively. However, the ‘rare’ BA profile in EOS term neonates differed from that in healthy FT neonates: TωMCA/TOMCA was significantly higher in EOS (p < 0.01) and accounted for 95% of all C-6-hydroxylated BA (Fig. 3c). TAMCA was significantly lower in EOS (p < 0.01). TGMCA, GMCA, BMCA, and GGMCA, which were—in addition to TAMCA—the most abundant ‘rare’ BA in FT controls, were below the detection limit in EOS.

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EOS in PT infants was marked by significantly increased ‘rare’ BA levels caused by a rise of TωMCA/TOMCAs [1]
The influence of EOS was investigated since ‘classical’ human BA levels were found to be significantly decreased in EOS.1 C-6-hydroxylated BA levels were determined significantly higher in 13 EOS PT neonates compared to PT controls (0.6 µmol/L [IQR: 0.2–1.5] vs. 0 µmol/L [IQR: 0.0–0.2]; p < 0.01). As in EOS FT neonates, the BA profile in EOS PT neonates included a significantly higher amount of TωMCA/TOMCA (p < 0.01) than did that of PT controls. Overall, TOMCA accounted for 72% of all ‘rare’ BA, followed by GBMCA (17%), TAMCA (8%), and BMCA (3%; Fig. 3d). TAMCA values—the only C-6-hydroxylated BA in PT controls—were significantly lower in EOS (p < 0.01).

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TOMCA/TωMCA levels are independently associated with EOSs [1]
Last, we evaluated the autonomous potential of TOMCA/TωMCA as a biomarker in EOS. EOS biomarker are listed in Table 3. Multivariate regression analysis revealed that TOMCA levels were independent of EOS biomarkers CRP, PCT, IL-6, and bilirubin (r = 0.75, p = 0.85).

Study design and patient characteristics [1]
Prematurity was diagnosed when neonates were born at <37 weeks’ gestation. EOS was confirmed by complete white blood cell count with a positive blood culture and CRP concentrations above 5 mg/L. All neonates with primary liver or biliary-tract disorders or asphyxia were excluded and EOS was excluded in controls. Fasting blood sampling in non-septic and EOS neonates was performed during routine screening for phenylketonuria or routine measurements, respectively. Blood sampling was within 48 h after starting antibiotics. Neonates not fed within 2 h were considered ‘fasted’. Overnight fasting blood sample collection was not possible since newborn infants are fed at least every 4 h. Finally, serum samples were stored at −80°C until assays. Analyses including concentrations of CRP, IL-6, PCT, and bilirubin were measured by standard laboratory methods. In addition, C-6 hydroxylated BA levels were measured by HPLC-HRMS, including unconjugated and taurine (T)- and glycine (G)-conjugated species (Table 1). Individual BA were separated by HPLC using a reversed-phase C18 column and a kinetex pentafluorophenyl column. Quantification and characterization was achieved using a mass spectrometer Q Exactive™ MS/MS and a high-performance quadrupole precursor selection with high-resolution and accurate-mass (HR/AM) Orbitrap™ detection according to the method published by Amplatz 2017.14

[1]. Neonatal sepsis leads to early rise of rare serum bile acid tauro-omega-muricholic acid (TOMCA).Pediatr Res. 2018 Jul;84(1):66-70.
[2]. Turnover of bile acids in liver, serum and caecal content by high-fat diet feeding affects hepatic steatosis in rats. Biochim Biophys Acta Mol Cell Biol Lipids. 2019 Oct;1864(10):1293-1304

Background: We investigated 'rare' bile acids (BA) as potential markers in septic neonates. Methods: 'Rare' (C-6 hydroxylated BA) and 'classical' BA were determined in 102 neonates using high-performance liquid chromatography-high-resolution mass spectrometry (HPLC-HRMS). Four groups according to maturity (full term, FT vs. preterm, PT) and septic status (early-onset neonatal sepsis, EOS vs. CTR; non-septic controls) were formed: FT-CTR; (n = 47), PT-CTR (n = 22), FT-EOS (n = 20), PT-EOS (n = 13). Results: Firstly, FT-CTR had a significant higher amount of 'rare' BA than PT (FT-CTR: 0.5 µmol/L, IQR: 0.3-1.3 vs. PT-CTR: 0.01 µmol/L, IQR 0.01-0.2; p < 0.01). The most common 'rare' BA in FT-CTR were tauro-γ- (TGMCA) and tauro-α-muricholic acid (TAMCA). Secondly, in EOS, absolute 'rare' BA levels were comparable in both gestational age groups (FT-EOS: 0.6 µmol/L, IQR: 0.1-1.6 and PT-EOS: 0.6 µmol/L, IQR: 0.2-1.5). Therefore, EOS had significantly higher median 'rare' BA values than non-septic PT neonates (p < 0.01). In PT and term neonates, the relative amount of tauro-ω-muricholic acid (TOMCA) within the 'rare' BA pool was significantly higher in EOS than in controls (FT-CTR vs. "FT-EOS and PT-CTR vs. PT-EOS; p < 0.01). It was hence the predominant 'rare' BA in EOS. Conclusion: TOMCA is an independent factor associated with EOS. It has diagnostic potential.[1]

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Background: Bile acids (BAs) participate in lipid absorption and serve as metabolic regulatory factors in gut-liver communication. To date, there are no studies on the systemic patterns of BAs in the serum, liver, and gut in the same non-alcoholic fatty liver disease (NAFLD) model. Methods: A targeted metabolomics approach and 16S rRNA sequencing were used to identify the profile of BAs and connection between BAs and microbiota. The role and mechanism of altered BAs on hepatic steatosis were investigated. Findings: In the liver, the composition of taurocholic acid (TCA) was increased, but taurohyodeoxycholic acid (THDCA) and ursodeoxycholic acid (UDCA) were decreased. In the gut, the deconjugated form of TCA (cholic acid (CA)) was increased, while the deconjugated forms of THDCA (α-hyodeoxycholic acid (HDCA)) and ω-muricholic acid (ωMCA) were decreased. In the serum, the composition of TCA was increased, while both HDCA and THDCA were decreased. THDCA induced the gene expression of apolipoprotein, bile secretion-related proteins, and cytochrome P450 family but suppressed inflammatory response genes expression in steatotic hepatocytes by RNAseq analysis. THDCA ameliorated neutral lipid accumulation and improved insulin sensitivity in primary rat hepatocytes. The decreased HDCA level correlated with the level of Bacteroidetes, while the level of CA correlated with the levels of Firmicutes and Verrucomicrobia but correlated inversely with Bacteroidetes. Conclusion: BAs profiles in the serum, liver and caecal content were altered in a rat NAFLD model, which may affect hepatic lipid accumulation and correlate with gut dysbiosis. [2]

C26H44NNAO7S

537.684838294983

537.273

2456348-84-6

146047145

White to off-white solid powder

4

7

7

36

897

11

C[C@H](CCC(=O)NCCS(=O)(=O)[O-])[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2[C@H]([C@@H]([C@H]4[C@@]3(CC[C@H](C4)O)C)O)O)C.[Na+]

NYXROOLWUZIWRB-JTNLMKNRSA-M

InChI=1S/C26H45NO7S.Na/c1-15(4-7-21(29)27-12-13-35(32,33)34)17-5-6-18-22-19(9-11-25(17,18)2)26(3)10-8-16(28)14-20(26)23(30)24(22)31;/h15-20,22-24,28,30-31H,4-14H2,1-3H3,(H,27,29)(H,32,33,34);/q;+1/p-1/t15-,16-,17-,18+,19+,20+,22+,23-,24-,25-,26-;/m1./s1

sodium;2-[[(4R)-4-[(3R,5R,6R,7R,8S,9S,10R,13R,14S,17R)-3,6,7-trihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]ethanesulfonate

Tauro-ω muricholic acid sodium; T-ω-MCA; TOMCA; 2456348-84-6; Tauro-omega-muricholic acid sodium; Tauro-; O-muricholic acid (sodium); Tauro-?-muricholic Acid Sodium Salt; Sodium 2-((R)-4-((3R,5R,6R,7R,8S,9S,10R,13R,14S,17R)-3,6,7-trihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)pentanamido)ethanesulfonate; Tauro-?-muricholic Acid (sodium salt); Tauro-; Tauro ω muricholic acid sodium

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)