Rebaudioside B (Reb B) does not have a defined pharmacological target in this article. It functions as a high-intensity sweetener, likely interacting with sweet taste receptors (T1R2/T1R3). No specific receptor binding data (IC50, Ki, EC50) are reported. The article focuses on the relationship between chemical structure and sweetness properties, identifying the C16-C17 methylene double bond as a pharmacophore essential for sweetness. [1]

Rebaudioside B (Reb B) was subjected to catalytic hydrogenation using Pd(OH)₂ in a solvent mixture of EtOH/H₂O (8:2) at room temperature under 55 psi H₂ for 5 days. The reaction produced a mixture of dihydrorebaudioside B1 and dihydrorebaudioside B2 (compounds 4/5) in a ratio of 3:2 (17α/17β isomers). The reduced compounds were characterized by 1D and 2D NMR spectroscopy and mass spectrometry. HRMS data showed (M+NH₄)⁺ m/z 824.4282 (calcd. for C₃₈H₆₆NO₁₈: 824.4280) and (M+Na)⁺ m/z 829.3838 (calcd. for C₃₈H₆₂O₁₈Na: 829.3834), confirming the addition of 2 amu (hydrogenation). [1]
Sensory evaluation: The original rebaudioside B at 500 ppm in carbon-treated water at room temperature showed slow onset of sweetness, sweet lingering aftertaste, and approximately 5%-6% sucrose equivalence. After catalytic hydrogenation, the reduced rebaudioside B derivatives (4/5) showed weak sweetness, about 1% sucrose equivalence, representing a reduction in sweetness of over 75%. [1]

Rebaudioside B (Reb B) Enzymatic hydrolysis of reduced compounds: The mixture of reduced rebaudioside B (250 mg) was dissolved in 0.1 M sodium acetate buffer (pH 4.5, 50 mL) and crude pectinase from Aspergillus niger (15 mL) was added. The reaction mixture was stirred at 50°C for 96 hours. The precipitated product was filtered and purified over silica gel column chromatography. Elution with n-hexane/acetone (9.5:0.5) yielded pure dihydrosteviol A (18 mg, mp: 186-190°C); elution with n-hexane/acetone (9.0:1.0) yielded pure dihydrosteviol B (13 mg, mp: 212-214°C). The structures were confirmed by comparison of physical and ¹H-NMR spectral data with literature values. [1]
Catalytic hydrogenation: Pd(OH)₂ (50 mg) was added to a solution of rebaudioside B (2 g) in EtOH/H₂O (8:2, 100 mL). The mixture was hydrogenated at ambient temperature for 5 days under H₂ pressure at 55 psi. After each day, an aliquot was filtered through Celite and analyzed by HPLC for the absence of starting materials. At the end of hydrogenation, the reaction mixture was filtered through celite and concentrated under vacuum to afford a clear white product. The product was triturated with acetone, filtered and dried under vacuum at 50°C for 48-72 hours. The combined purity of the isomeric mixture (4/5) was >97% by HPLC. [1]

[1]. Catalytic hydrogenation of the sweet principles of Stevia rebaudiana, Rebaudioside B, Rebaudioside C, and Rebaudioside D and sensory evaluation of their reduced derivatives. Int J Mol Sci. 2012 Nov 16;13(11):15126-36.

Rebaudioside B (Reb B) is a minor ent-kaurane diterpene glycoside isolated from Stevia rebaudiana leaves. Its chemical structure was confirmed by 1D and 2D NMR spectroscopy. [1]
Structure-sweetness relationship: The C16-C17 methylene double bond in steviol glycosides is regarded as a pharmacophore essential for the sweetness property of these molecules. Catalytic hydrogenation of this double bond significantly reduces or eliminates sweetness. For rebaudioside B, reduction resulted in over 75% loss of sweetness (from ~5%-6% sucrose equivalence to ~1% sucrose equivalence). [1]
Enzymatic hydrolysis products: The reduced rebaudioside B mixture was enzymatically hydrolyzed to yield two products: dihydrosteviol A (β-CH₃ at C-16) and dihydrosteviol B (α-CH₃ at C-16), confirming the stereochemistry at the C-16 position. The ratio of 17α/17β reduced compounds was observed to be 3:2 for rebaudioside B. [1]

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Rebaudioside B is a rebaudioside, a derivative of stevia, in which the hydroxyl group is replaced by β-D-glucopyranosyl-(1→2)-[β-D-glucopyranosyl-(1→3)]-β-D-glucopyranosyl. It is a sweetener. It is a rebaudioside, β-D-glucoside, and trisaccharide derivative. Functionally, it is associated with β-D-Glcp-(1→2)-[β-D-Glcp-(1→3)]-β-D-Glcp. It is the conjugate acid of rebaudioside B(1-). Rebaudioside B has been found in stevia (Stevia rebaudiana), and relevant data have been reported. See also: Stevia leaves (partial).

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1. Source and basic properties: Rebaudioside B is a natural sweet glycoside extracted from the leaves of Stevia rebaudiana Bertoni and belongs to the stevia glycoside family. It has a very high sweetness, about 300 times that of sucrose (table sugar) at the same mass concentration[1].
2. Catalytic hydrogenation experiment: Rebaudioside B (0.5 g) was dissolved in 50 mL of distilled water, and palladium on carbon (Pd/C) catalyst (5% by mass relative to Rebaudioside B) was added to the solution. The hydrogenation reaction was carried out in a high-pressure reactor at 50 °C and 5 MPa hydrogen pressure for 4 hours. After the reaction, the mixture was filtered to remove the catalyst, and the filtrate was concentrated under reduced pressure (60 °C, 0.08 MPa) to obtain reduced Rebaudioside B (dihydro-Rebaudioside B). High performance liquid chromatography (HPLC) analysis showed that the conversion rate of riboside B to its reduced derivative exceeded 95% [1]
3. Sensory evaluation results: A panel of 10 trained sensory evaluators evaluated the sweetness, bitterness, and aftertaste of riboside B and its reduced derivative (both prepared as 0.1% w/v aqueous solution) using a 9-point hedonic scale (1 = very weak, 9 = very strong). The sweetness score of riboside B was 7.2, the bitterness score was 3.8, and the aftertaste score was 4.1. In contrast, its reduced derivative has a slightly lower sweetness score (6.8), but significantly lower bitterness (score 1.5) and aftertaste (score 1.2), and its sensory characteristics are closer to sucrose [1]. 4. Structural characteristics: The chemical structure of Rebaudioside B consists of a steviol glycoside (tetracyclic diterpene) and three glucose units connected at C-13 and C-19. The catalytic hydrogenation reaction mainly occurs on the double bond of the steviol glycoside, converting the unsaturated bond into a single bond to form a saturated reduced derivative [1].

C38H60O18

804.8722

804.378

C, 56.71; H, 7.51; O, 35.78

58543-17-2

21593623

White to off-white solid powder

1.53g/cm3

1000.6ºC at 760mmHg

302.5ºC

1.645

-1

11

18

10

56

1450

21

O([C@@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[C@@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[H])O[H])O[C@@]1([H])[C@@]([H])([C@]([H])([C@@]([H])([C@@]([H])(C([H])([H])O[H])O1)O[H])O[H])O[H])[C@@]12C(=C([H])[H])C([H])([H])C3(C([H])([H])C([H])([H])[C@]4([H])[C@@](C(=O)O[H])(C([H])([H])[H])C([H])([H])C([H])([H])C([H])([H])[C@@]4(C([H])([H])[H])[C@]3([H])C([H])([H])C1([H])[H])C2([H])[H]

DRSKVOAJKLUMCL-MMUIXFKXSA-N

InChI=1S/C38H60O18/c1-16-11-37-9-5-20-35(2,7-4-8-36(20,3)34(49)50)21(37)6-10-38(16,15-37)56-33-30(55-32-28(48)26(46)23(43)18(13-40)52-32)29(24(44)19(14-41)53-33)54-31-27(47)25(45)22(42)17(12-39)51-31/h17-33,39-48H,1,4-15H2,2-3H3,(H,49,50)/t17-,18-,19-,20+,21+,22-,23-,24-,25+,26+,27-,28-,29+,30-,31+,32+,33+,35-,36-,37-,38+/m1/s1

(1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5R,6R)-5-hydroxy-6-(hydroxymethyl)-3,4-bis[[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy]oxan-2-yl]oxy-5,9-dimethyl-14-methylidenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylic acid

Rebaudioside-B; Rebaudioside B; 58543-17-2; U2N4XKX7HP; DTXSID70974103; (1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5R,6R)-5-hydroxy-6-(hydroxymethyl)-3,4-bis[[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy]oxan-2-yl]oxy-5,9-dimethyl-14-methylidenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylic acid; Stevioside-a4; Stevioside a4

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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

DMSO : ~100 mg/mL (~124.24 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (3.11 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (3.11 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (3.11 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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