Animals:** Male CD-1 mice (6 weeks old, 26-28 g) were used. They were housed under standard conditions with free access to food and water and a 12h light/dark cycle. [1]
- **Aβ Injection Model:** Aβ₁₋₄₂ was prepared by dissolving in 1.0% NH₄OH, diluting with PBS to 1 mg/mL, and incubating at 37°C for 24 hours to obtain oligomeric species. Mice were anesthetized and 5 μL of Aβ (10 μM) or vehicle (PBS) was acutely injected into the left lateral ventricle (i.c.v.) by hand. [1]
- **Drug Administration:** β-Amyrin (4 mg/kg) was suspended in 10% Tween 80 solution and administered orally (p.o.) once daily for 5 days, starting one day after the Aβ injection. Minocycline (30 mg/kg, i.p.) was used as a positive control. The control group received vehicle (10% Tween 80). [1]
- **Behavioral Tests:** Behavioral tests were conducted 5 days after Aβ injection.
- **Object Recognition Test:** Mice were habituated to an open field for 10 min. In the training phase, they were placed in the same box with two distinct objects for 10 min. After 2 h, in the test phase, one object was displaced to a novel location, and mice were allowed to explore for 5 min. Time spent exploring the displaced and non-displaced objects was measured, and a preference ratio was calculated. [1]
- **Passive Avoidance Test:** In the training trial, mice were placed in an illuminated room. When they entered the dark room, a guillotine door closed, and an electric shock (0.5 mA for 3 s) was delivered. The next day, in the test trial, mice were re-introduced to the illuminated room, and the step-through latency to enter the dark room was measured (max 300 s). [1]
- **Immunohistochemistry:** After behavioral tests, brains were fixed in 4% paraformaldehyde, cryoprotected in 30% sucrose, and frozen. Coronal sections (30 μm) were cut using a cryostat. Sections were incubated with goat anti-doublecortin (DCX, immature neuron marker) or rat anti-Ki67 (proliferation marker) antibodies, followed by biotinylated secondary antibodies and avidin-biotin-peroxidase complex. Staining was visualized with DAB. DCX- and Ki67-positive cells in the subgranular zone of the hippocampal dentate gyrus were counted. [1]

[1]. β-Amyrin Ameliorates Alzheimer's Disease-Like Aberrant Synaptic Plasticity in the Mouse Hippocampus. Biomol Ther (Seoul). 2019 Jul 30.

β-Amyrinol is a pentacyclic triterpenoid compound with a structure in which oleanane is substituted at the 3β position with a hydroxyl group, forming a double bond between the 12 and 13 positions. It is one of the most common triterpenoids in higher plants, with metabolites found in both plants and Aspergillus fungi. It is a pentacyclic triterpenoid and a secondary alcohol derived from the hydride of oleanane. β-Amyrinol has been reported in tea (Camellia sinensis), elderberry (Sambucus chinensis), and other organisms with relevant data. See also: Calendula (partial); Viburnum bark (partial); Cornflower (partial).

C30H50O

426.73

426.386

559-70-6

73145

White to off-white solid powder

1.0±0.1 g/cm3

490.7±44.0 °C at 760 mmHg

187-190°C

217.7±20.7 °C

0.0±2.8 mmHg at 25°C

1.539

11.06

1

1

0

31

790

8

C[C@@]12CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)O)C)C)[C@@H]1CC(CC2)(C)C)C

JFSHUTJDVKUMTJ-QHPUVITPSA-N

InChI=1S/C30H50O/c1-25(2)15-16-27(5)17-18-29(7)20(21(27)19-25)9-10-23-28(6)13-12-24(31)26(3,4)22(28)11-14-30(23,29)8/h9,21-24,31H,10-19H2,1-8H3/t21-,22-,23+,24-,27+,28-,29+,30+/m0/s1

(3S,4aR,6aR,6bS,8aR,12aR,14aR,14bR)-4,4,6a,6b,8a,11,11,14b-octamethyl-1,2,3,4a,5,6,7,8,9,10,12,12a,14,14a-tetradecahydropicen-3-ol

beta-Amyrin beta-Amyrenol Amyrinβ-amyrin

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

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

DMF : 10 mg/mL (~23.43 mM)

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.)