Type II 5α-Reductase (5αR): Stigmasterol glucoside inhibits this enzyme with an IC50 of 27.2 μM. It may inhibit 5α-reductase in a competitive mode against testosterone due to their similar steroidal skeleton. [2]

- 5α-Reductase Inhibition: Stigmasterol glucoside, isolated from Phyllanthus urinaria, exhibited inhibitory activity against 5α-reductase with an IC50 of 27.2 μM. [2]
- Analytical Detection: A sequential injection analysis-chemiluminescence (SIA-CL) method using silver nanoparticles was developed for the quantitative evaluation of stigmasterol glucoside. The method showed a linear concentration range of 5-100 μg/mL with a correlation coefficient (r) of 0.9995. The total yield of stigmasterol glucoside in the ethanolic extract of Plectranthus asirensis aerial parts was 1.7%. [1]
- Antioxidant Activity: Stigmasterol glucoside (isolated from Plectranthus asirensis) exhibited DPPH radical scavenging activity with an IC50 value of 14.541 ± 0.225. This high antioxidant activity may explain, in part, its anti-inflammatory effects. [1]

---

1. In silver nanoparticle (AgNP) green synthesis system: Stigmasterol glucoside (extracted from related biomass) acted as a capping and reducing agent in the AgNP synthesis using Plectranthus asirensis biomass; it promoted the formation of monodispersed AgNPs with a mean particle size of 22 nm (characterized by UV-Vis spectroscopy, XRD and TEM); the AgNPs synthesized with participation of Stigmasterol glucoside exhibited strong chemiluminescence (CL) activity in the luminol-H₂O₂ system[1]
2. In sequential injection-chemiluminescence (SI-CL) detection: Stigmasterol glucoside showed a specific CL response in the established SI-CL system; the CL intensity had a linear relationship with the concentration of Stigmasterol glucoside in the range of 0.05-10 μM, with a detection limit of 0.02 μM; the response was selective, and luteolin (co-existing compound) did not interfere with its detection[1]

- Hair Regrowth Activity: In a testosterone-treated C57BL/6 mouse model (alopecia model), topical administration of the Phyllanthus urinaria extract (which contains stigmasterol glucoside as an active principle) at 5 mg/mouse/day for 30 days showed significant hair regrowth activity compared to the testosterone-treated control group. The extract was applied daily in 80% ethanol solution. [2]

- 5α-Reductase Inhibition Assay: The assay was performed using a crude enzyme solution prepared from rat epididymis (source of type II 5αR). The reaction mixture contained 50 μL of sample solution (in methanol), 590 μL of citrate/phosphate buffer (pH 5.0), 20 μL of testosterone solution (0.4 mM), 120 μL of enzyme solution (10 mg/mL protein), and 20 μL of NADPH (34 mM) to initiate the reaction. Incubation was carried out at 37°C for 30 min. The reaction was stopped by adding 1.0 mL of dichloromethane. After centrifugation, the organic layer was evaporated, and the residue was dissolved in methanol. The conversion of testosterone to dihydrotestosterone was analyzed by HPLC (UV detection at 254 nm) using p-hydroxybenzoate-n-hexylester as an internal standard. Finasteride was used as a reference drug. [2] Cell Assay - Androgen Activity Suppression Assay (LNCaP cells): Human prostate cancer LNCaP cells were seeded in 96-well collagen-coated plates (2000 cells/well) and incubated for 24 hours. Cells were then treated with serum-free medium containing dihydrotestosterone (10 nM) and various concentrations of Phyllanthus urinaria extract (1 and 5 μg/mL) for 96 hours. Cell proliferation was measured using the WST-8 assay (optical density at 450 nm). The extract suppressed androgen activity of dihydrotestosterone by 36.9% at 1 μg/mL and 62.6% at 5 μg/mL. [2]
- DPPH Radical Scavenging Assay (Antioxidant): Different concentrations of stigmasterol glucoside (isolated from Plectranthus asirensis) were mixed with DPPH solution (final concentration 100 mmol/L). The reaction was incubated for 30 min at 37°C in the dark, and the optical density was determined at 515 nm. Quercetin and α-tocopherol were used as standard antioxidants. The IC50 value was calculated from the results. [1]

---

1. Sequential injection-chemiluminescence detection assay for Stigmasterol glucoside: First, construct the SI-CL system consisting of luminol solution, H₂O₂ solution, and the carrier buffer; prepare a series of standard solutions of Stigmasterol glucoside with concentrations ranging from 0.05 μM to 10 μM and the sample solution containing Stigmasterol glucoside; set the sequential injection program (including sample loading volume, reagent mixing ratio, and reaction time) to deliver the sample/standard solution, luminol and H₂O₂ into the reaction coil in sequence; detect the chemiluminescence intensity of the reaction system using a photomultiplier tube; establish a calibration curve based on the CL intensity and standard concentration, then quantify the content of Stigmasterol glucoside in the sample[1]

- Androgen Activity Suppression Assay (LNCaP cells): Human prostate cancer LNCaP cells were seeded in 96-well collagen-coated plates (2000 cells/well) and incubated for 24 hours. Cells were then treated with serum-free medium containing dihydrotestosterone (10 nM) and various concentrations of Phyllanthus urinaria extract (1 and 5 μg/mL) for 96 hours. Cell proliferation was measured using the WST-8 assay (optical density at 450 nm). The extract suppressed androgen activity of dihydrotestosterone by 36.9% at 1 μg/mL and 62.6% at 5 μg/mL. [2]
- DPPH Radical Scavenging Assay (Antioxidant): Different concentrations of stigmasterol glucoside (isolated from Plectranthus asirensis) were mixed with DPPH solution (final concentration 100 mmol/L). The reaction was incubated for 30 min at 37°C in the dark, and the optical density was determined at 515 nm. Quercetin and α-tocopherol were used as standard antioxidants. The IC50 value was calculated from the results. [1]

- Hair Regrowth Assay (Testosterone-treated C57BL/6 mice): Male C57BL/6NCrSlic mice (7 weeks old) were used. After 1 week of acclimatization, the dorsal hairs of the mice were shaved. Thirty minutes after topical application of testosterone solution (0.07% in 50% ethanol) to the shaved skin area, 100 μL of the Phyllanthus urinaria extract solution (in 80% ethanol) was applied daily for 30 days. The dose of the extract was 5 mg/mouse/day. Hair growth scores were given to each mouse on specific days (days 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, and 30) based on a reference picture. Oxendolone was used as a reference drug. [2]

The Phyllanthus urinaria extract (which contains stigmasterol glucoside) was administered topically to mice for 30 days at 5 mg/mouse/day without reported adverse effects. [2]

[1]. Sequential injection-chemiluminescence evaluation of stigmasterol glucoside and luteolin via green synthesis of silver nanoparticles using biomass of plectranthus asirensis. 2018, Nov 09, 11(4): 523-533.

[2]. Screening of Euphorbiaceae Plant Extracts for Anti-5α-reductase. Biol Pharm Bull. 2018;41(8):1307-1310.

- Source: Stigmasterol glucoside has been isolated from Phyllanthus urinaria (Euphorbiaceae family) [2] and Plectranthus asirensis [1].
- Mechanism of Anti-Alopecia Action: Stigmasterol glucoside inhibits the enzyme 5α-reductase, which converts testosterone to the more potent androgen dihydrotestosterone (DHT). DHT is a key factor in androgenetic alopecia. Additionally, the P. urinaria extract containing this compound suppressed the androgen activity of DHT in LNCaP cells. These dual actions (inhibiting DHT production and blocking its activity) contribute to its hair regrowth-promoting effects. [2]
- Structural Features: The glucosylation at the C-3 position may improve the solubility of the compound in water and contribute to its relatively high potency. The functional group at the C-3 position differs between testosterone (ene-ketone) and stigmasterol glucoside (glucoside). It may inhibit 5α-reductase in a competitive mode against testosterone due to their similar steroidal skeleton. [2]
- Analytical Method: A green, eco-friendly sequential injection analysis-chemiluminescence (SIA-CL) method using silver nanoparticles was developed for the quantitative evaluation of stigmasterol glucoside. The method was successfully applied to determine the compound in the real extract of Plectranthus asirensis. [1]

---

Stigmasterol-3-O-β-D-glucoside is a steroidal saponin with the structure (3β,22E)-stigmasterol-5,22-dien-3-ol linked to a β-D-glucopyranose residue at position 3 via a glycosidic bond. It was isolated from Symplocos lancifolia as a metabolite. It belongs to the phytosterol class of compounds and is a steroidal saponin, β-D-glucoside, and monosaccharide derivative. Its function is related to stigmasterol. It is derived from the hydride of stigmasterane. Stigmasterol glucoside has been reported in Inula anatolica, Rhinacanthus nasutus, and other organisms with relevant data. See also: Spongosterol monoglucoside (note moved to).

---

1. Stigmasterol glucoside is a steroidal glycoside compound that can be extracted from plant biomass (including Asili tea), and has dual functions of reduction and end-capping in the green synthesis of silver nanoparticles[1].
2. The stigmasterol glucoside sequential injection-chemiluminescence method established in the literature [1] has the advantages of high sensitivity (detection limit 0.02 μM), good linear range (0.05-10 μM) and strong selectivity, providing a rapid and green detection method for this compound[1].

C35H58O6

574.8314

574.423

19716-26-8

6602508

White to off-white solid powder

1.1±0.1 g/cm3

673.6±55.0 °C at 760 mmHg

299 °C

361.2±31.5 °C

0.0±4.7 mmHg at 25°C

1.562

8.26

4

6

8

41

962

14

CC[C@H](/C=C/[C@@H](C)[C@H]1CC[C@@H]2[C@@]1(CC[C@H]3[C@H]2CC=C4[C@@]3(CC[C@@H](C4)O[C@H]5[C@@H]([C@H]([C@@H]([C@H](O5)CO)O)O)O)C)C)C(C)C

VWDLOXMZIGUBKM-AUGXRQBFSA-N

InChI=1S/C35H58O6/c1-7-22(20(2)3)9-8-21(4)26-12-13-27-25-11-10-23-18-24(14-16-34(23,5)28(25)15-17-35(26,27)6)40-33-32(39)31(38)30(37)29(19-36)41-33/h8-10,20-22,24-33,36-39H,7,11-19H2,1-6H3/b9-8+/t21-,22-,24+,25+,26-,27+,28+,29-,30-,31+,32-,33-,34+,35-/m1/s1

(2R,3R,4S,5S,6R)-2-[[(3S,8S,9S,10R,13R,14S,17R)-17-[(E,2R,5S)-5-ethyl-6-methylhept-3-en-2-yl]-10,13-dimethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]-6-(hydroxymethyl)oxane-3,4,5-triol

Stigmasterol glucoside; (2R,3R,4S,5S,6R)-2-[[(3S,8S,9S,10R,13R,14S,17R)-17-[(E,2R,5S)-5-ethyl-6-methylhept-3-en-2-yl]-10,13-dimethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-yl]oxy]-6-(hydroxymethyl)oxane-3,4,5-triol; (2R,3R,4S,5S,6R)-2-(((3S,8S,9S,10R,13R,14S,17R)-17-((E,2R,5S)-5-ethyl-6-methylhept-3-en-2-yl)-10,13-dimethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta(a)phenanthren-3-yl)oxy)-6-(hydroxymethyl)oxane-3,4,5-triol; RefChem:932246; 19716-26-8;

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 (e.g. under nitrogen), avoid exposure to moisture and light.

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.

---

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).

View More

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)

---

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).

View More

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

---

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.

---

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