What Active Ingredients Are in Bacopa Monnieri Extract Powder?

Bacopa monnieri (L.) Wettst., also known as white-flowered pigweed, is a medicinal plant used in Indian Ayurvedic and traditional Chinese medicine. It is a creeping, fleshy, perennial herb belonging to the Scrophulariaceae family, widely distributed in tropical and subtropical regions. Traditionally used to treat bronchitis, asthma, dysentery, and various inflammatory conditions; in India, it is renowned as a nervous system tonic, and modern pharmacological research has confirmed its efficacy. Clinically, it is used to enhance memory, treat epilepsy, and alleviate insomnia, as well as a mild sedative. Research analysis has identified active components in Scutellaria baicalensis extract powder, including Scutellaria baicalensis saponins (hersaponin), Scutellaria baicalensis saponins (bacoside, also known as white flower pigweed saponin) A and B, apigenin-7-glucuronic acid glycoside, and luteolin-7-glucuronic acid glycoside, among other active components.

1 Active Components

The neurotrophic and antidiabetic activities of Bacopa monnieri (Bm) are concentrated in the glycoside fraction of the alcohol extract, with a high content of glycoside components present in the extract as a mixture. Since 1993, comprehensive chemical studies have been conducted on the glycoside fractions of Bm extracts obtained using methanol or ethanol, resulting in the isolation and identification of numerous new glycoside compounds, primarily triterpenoid saponins and phenethyl glycosides.

1.1 Triterpenoid Saponins

Chakravarty[1–3] isolated five new triterpenoid saponins, bacopa-sides I–V, from Bm collected in the suburbs of Kolkata, India. Among these, two are jujubogenin glycosides, and three are pseudo-jujubogenin glycosides.

The whole plant of Bm was extracted with petroleum ether and methanol. The methanol extract (ME) was separated by silica gel chromatography to obtain bacopaside I (1). The low-polarity fraction was further separated by silica gel chromatography and preparative HPLC to obtain bacopaside II (2). ME was distributed in water and n-butanol, and the n-butanol fraction was absorbed on silica gel. extracted with CHCl₃, EtOAc, Me₂CO, and CHCl₃-MeOH in sequence. The Me₂CO extract was subjected to silica gel chromatography and preparative HPLC, yielding bacopaside III, IV, and V (3–5).

The structures of these compounds were primarily elucidated by chemical and 2D NMR spectroscopy. Compound 1 is 3-O-α-L-furanarabinosyl (1 → 2)-[6-O-sulfonyl-β-D-pyranoglucosyl (1→3)]-α-L-pyranarabinosyl pseudo-jujuboside, Compound 2 is 3-O-α-L-furanarabinosyl (1 → 2)-[β-D-pyranoglucosyl (1→3)-β-D-pyranoglucosyl pseudo-jujuboside; Compound 3 is 3-O-α-L-furan arabopyranosyl (1→2)-β-D-pyranoglucopyranosyl acid jujuboside, Compound 4 is 3-O-β-D-pyranoglucosyl (1→3)-α-L-pyranarabinosyl acid jujuboside, Compound 5 is 3-O-β-D-pyranoglucopyranosyl (1→3)-α-L-pyranoarabinosyl pseudo-jujuboside.

Hou[4] et al. collected fresh whole plants of Bm from Tainan, China. The butanol-soluble fraction of the methanol extract was subjected to repeated column chromatography on silica gel and porous polymer gel, yielding five new glycoside compounds. Two of these were identified as pseudo-sapindoside and sapindoside, while the other two compounds, bacopaside III, have the structure 3-O-[6-O-sulfonyl-β-D-pyranoglucosyl (1→3)]-α-L-pyranarabinosyl pseudo-sapindoside aglycone (6, which is not the same compound as compound 3); bacopasaponin G has the structure 3-O-[α-L-furanarabinosyl (1 → 2)]-α-L-pyranarabinosyl acid jujube glycoside aglycone (7).

Garai et al. [5–7] subsequently isolated six dammarane-type triterpenoid saponins from the n-butanol-soluble fraction of Bm methanol extract, named bacopasaponin A–F (8–13). Among these, compounds 12 and 13 are bioactive aconitane-type saponins with a double carbon chain, with structures 3-O-[β-D-pyranoglucosyl (1→3){α-L-furanarabosyl (1→2)}-α-L-pyranarabosyl-20-O-(α-L-pyranarabosyl) saponogenin and 3-O-[ β-D-pyranoglucopyranosyl (1 → 3){α-L-furanarabinosyl (1→2)}-β-D-pyranoglucopyranosyl]-20-O-α-L-pyranarabinosyl acid jujuboside.

Jain et al. [8-10] extracted the acetate-soluble fraction from the ethanol extract of dried Bm whole plants, separated pseudo-purslane saponins A and B, and obtained crude pseudo-purslane saponin A₁ (14) by repeated silica gel column chromatography followed by chloroform-methanol gradient elution.   

The ethyl acetate-insoluble fraction was recrystallized to obtain pure compound 14. The ethanol extract was subjected to repeated silica gel column chromatography, eluted with an ethyl acetate-increasing acetone-water (90:10) gradient solvent system, yielding a fraction containing pseudo-purslane saponin A₂ (15). The fraction was subjected to repeated reverse C₁₈ column chromatography, eluted with acetonitrile-water (30:70) to obtain pure compound 15; eluted with water-saturated ethyl acetate in a gradient of acetone-water (40:10), yielding a fraction rich in false purslane saponin A₃ (16), followed by rapid column chromatography on a Bondapak C₈ column, elute with acetonitrile-water (30:70), obtaining pure compound 16. The structures of compounds 14–16 are 3-O-[α-L-furan arabitol (1→3)-α-L-pyran arabitol] acid jujuboside, 3-β-[O-α-L-furan arabitol (1 → 6)-O-   [α-L-pyran arabosyl (1 → 5)]-O-α-D-furan  glucosyl)oxo] pseudo-jujuboside and 3-β-[O-β-D- pyran  glucosyl (1 → 3)-O-[α-L-  furan arabosyl  (1→2)]-O-β-D-pyranoglucosyl)oxy] acid jujube glycoside. Pseudopueraria saponin A₃ is the main component and active component of the saponin mixture.

1.2 Phenethyl alcohol glycoside

Chakravarty et al. [] extracted the methanol extract of Bm whole plant with ethyl acetate, performed silica gel column chromatography, eluted with CHCl₃-MeOH (9:1 and 8:2), obtained fraction A, a gel-like substance containing compounds 17 and 18, and further eluted with CHCl₃-MeOH (7:3 and 6:4), yielding fraction B, a mixture containing compounds 19 and 20. The above fractions were separated by column chromatography on a Diaion HP-20 column, eluted with MeOH-H₂O (1:1 and 1:3), and the residues were purified by preparative HPLC (H₂O-MeCN, 9:1 as the mobile phase), yielding pure compounds 17–20, all of which are phenethyl alcohol glycosides. Compounds 17–19 are new compounds, named monniera-side I–III, with structures determined by spectroscopic analysis. Monniera-side I is α-O-[2-O-(4-hydroxybenzoyl-β-D-pyranoglucosyl]-4-hydroxyphenethyl alcohol; Monniera-side II is α-O-[2-O-(3-methoxy-4-hydroxy cinnamoyl)-β-D-pyranoglucoside]-3,4-dihydroxybenzyl alcohol; monniera-side III is α-O-[2-O-(4-hydroxybenzoyl)-β-D-pyranoglucoside]-3,4-dihydroxybenzyl alcohol. phenethyl alcohol. Compound 20 is the known isomer plantain glycoside B.

Hou et al. [4] isolated and identified two new phenethyl glycosides, named bacopa-side B and C (21, 22),      with structures corresponding to 3,4-dihydroxybenzyl alcohol (2-O- feruloyl)-β-D-glucopyranoside; and phenethyl alcohol [5-O-p-hydroxybenzoyl-β-D-furanocelulosyl-(1→2)]-β -D-pyranoglucoside.

1.3 Other compounds

The fraction 2-1 of the n-butanol-soluble portion of the Bm methanol extract, after chromatography, was analyzed by thin-layer chromatography on MCI gel CHP 20P (H₂O-MeOH, 1:0 to 0:1), dextran LH-20 column (60% MeOH) and Cosmosil C₁₈-OPN (H₂O-MeOH, 1:10 to 1:1), yielding one matsutake alcohol (matsutaka alcohol) derivative, with the structure (3R)-1-octan-3-yl-(6-O-sulfonyl)-β-D-glucopyranosyl, named bocapaside A (23)[14].

2 Pharmacological effects

B. morio (Bm) exhibits a wide range of pharmacological activities, including hypoglycemic, antidepressant, antiulcer, antilamellar, smooth muscle relaxant, and antioxidant effects. This review summarizes its pharmacological activities in seven aspects: hypoglycemic, central nervous system effects, antiulcer, and hepatoprotective activities.

2.1 Hypoglycemic

Twenty-four compounds isolated from Bm were tested for their anti-hyperglycemic activity in rats with diabetes induced by streptozotocin. The results showed that three compounds, calcerorio-side B, marytynoside, and luteolin-7-O-glucuronide, exhibited moderate hypoglycemic activity at a dose of 1 mmol/kg [4,12].

2.2 Central effects

2.2.1 Antagonism of scopolamine

Scopolamine (3 mg/kg, i.p.) was used to induce passive avoidance in mice to test anti-dementia activity. the results showed that the Bm standard extract (containing 38% pseudoparkin saponin A) at 30 mg/kg could antagonize the dementia-inducing effects of scopolamine; in vitro, it exhibited significant anticholinesterase (AChE) activity, which was dose-dependent [13].

2.2.2 Antidepressant

Sairam¹4] conducted antidepressant activity tests using a methanol standard extract of Bacopa Monnieri and compared it with the standard antidepressant imipramine (15 mg/kg, ip). Rats were orally administered 20 and 40 mg/kg of the extract daily for 5 days, and significant antidepressant activity was observed in depression models such as forced swimming and acquired helplessness.

2.2.3 Prevention of NO-induced DNA damage

Activated astrocytes can produce high levels of NO, which may contribute to the onset of various neurodegenerative diseases. In rat astrocyte cultures, NO and S-nitroso-N-acetylpenicillanide induced the production of reactive species and DNA fragmentation in the genome. Treatment with Bacopa Monnieri methanol extract inhibited the formation of reactive products and DNA damage, with a dose-dependent effect. These biological activities suggest that Bm has therapeutic or preventive effects against various neurodegenerative diseases (such as dementia, epilepsy, and local ischemia) [15].

2.2.4 Reducing morphine withdrawal reactions

Sumathy [16] evaluated the effects of Bacopa Monnieri whole plant ethanol extracts on morphine withdrawal reactions in isolated guinea pig ileum. In vitro, guinea pig ileum exposed to morphine for 4 minutes was treated with naloxone to induce strong ileal contractions; 15 minutes before morphine exposure,   the addition of Bm ethanol extract at different concentrations (100–1000 μg/mL) significantly reduced naloxone-induced contractions in a dose-dependent manner. These results suggest that Bm may be effective against the syndrome induced by morphine withdrawal [16].

2.2.5 Protecting brain mitochondrial enzymes

Sumathy [17] investigated the protective effects of Bm ethanol extracts on changes in brain mitochondrial enzyme status induced by morphine in rats. Morphine-treated rats exhibited significantly lower levels of brain mitochondrial enzymes compared to control animals; however, when Bm extracts (40 mg/kg) were administered orally 2 hours prior to morphine administration, mitochondrial enzyme levels were maintained at normal levels.

2.2.6 Reducing the toxicity of antiepileptic drugs

Many epilepsy patients experience cognitive dysfunction due to the use of the antiepileptic drug phenytoin (PHT). Vohora[1$ investigated the protective effects of Bm against PHT-induced cognitive impairments in mice. The effects of Bm alone and in combination with PHT were evaluated using passive avoidance, maximum electric shock seizures, and motor coordination tests. The results showed that Bm reversed PHT-induced damage, improving both memory acquisition and retention, without affecting PHT's anticonvulsant activity.

2.2.7 Anti-stress

Anti-stress pharmacological studies showed that the saponins from Bm can regulate the expression of Hsp70 and the activity of SOD and P450 in the brains of SD male rats. Pseudo-pigweed saponins were administered orally at doses of 20 and 40 mg/kg for 7 days, with the control group receiving distilled water. Stress was induced 2 hours after the last dose. Stress did not cause any significant changes in Hsp70 expression in any brain region of the two dose groups, while Hsp70 expression was significantly enhanced in all brain regions of the control group. SOD activity in the hippocampus was significantly reduced in the low-dose group and the control group, while SOD activity increased in the high-dose group. P450 activity was enhanced in all brain regions of single-stressed animals and the two dose groups [19].

2.3 Anti-ulcer

Bm fresh juice exhibited significant anti-ulcer activity. Experimental animals were administered Bm standard methanol extract at doses of 10–50 mg/kg orally twice daily for 5 days, and the extract demonstrated dose-dependent anti-ulcer activity in models of gastric ulcers induced by ethanol, aspirin, 2-hour cold stress, and 4-hour pyloric ligation. Bm methanol extract at 20 mg/kg, administered orally to rats twice daily for 10 days, completely healed 50% of acetylsalicylic acid-induced gastric ulcers. Studies on the mechanism of action of the extract on various mucosal damages in rats indicated that Bm methanol extract at 20 mg/kg did not affect gastric acid and pepsin secretion, increased mucin secretion, reduce mucosal cell detachment without affecting cell proliferation. Due to its significant antioxidant effects, Bm methanol extract may primarily exert its preventive and therapeutic effects on gastric ulcers by acting on mucosal defense factors [24].

2.4 Hepatoprotective effects

Bm ethanol extract has a protective effect against morphine-induced liver toxicity in rats. In the morphine-treated group, significant increases in liver lipid peroxidation and a marked decrease in liver antioxidant enzyme levels were observed. When rats were administered morphine concurrently with oral Bm ethanol extract, these changes were prevented, suggesting that Bm exerts a hepatoprotective effect in morphine-induced liver toxicity [25].

2.5 Antilamebricidal Activity

The single compound bacopas-aponin C isolated from Bm was administered in various forms, including free, lipid-bound, microspheres, and nanoparticles, to evaluate its antilamebricidal activity. The results showed that all formulations exhibited high activity, with efficacy inversely proportional to vesicle size. Histological and hematological analyses revealed no adverse effects on the liver or kidneys, suggesting potential clinical application for leishmaniasis treatment [26].

2.6 Smooth muscle relaxation

Bm ethanol extract exhibited inhibitory activity against spontaneous contractions of guinea pig ileum and rabbit jejunum, with IC₅₀ values of 24 and 136 μg/mL, respectively. Administration of 260 μg/mL extract significantly reduced contractions induced by acetylcholine (ACh) and histamine (0.0001–10 μmol/L) in the ileum. The contraction of the ileum induced by 1 μmol/L ACh was also inhibited by 100–70 μg/mL of the extract, with a dose-dependent effect, and the IC₅₀ was 285 μg/mL. This indicates that the ethanol extract of Bm has a direct effect on smooth muscle. The responses of rabbit blood vessels and jejunum induced by CaCl₂ were attenuated by 10–700 μg/mL of Bm ethanol extract, suggesting that the extract directly interferes with the influx of calcium ions into cells. However, Bm extract had no effect on contractions induced by norepinephrine or caffeine, indicating that it does not affect intracellular calcium flow. The smooth muscle relaxant effect of Bm extract primarily acts through voltage-gated and receptor-mediated calcium channels, inhibiting calcium influx into the cell membrane [27].

The various solvent fractions and subfractions isolated from Bm significantly inhibited carbachol-induced bronchoconstriction, blood pressure reduction, and bradycardia in anesthetized rats. In vitro, crude extracts, petroleum ether, and methanol fractions inhibited KCl-induced tracheal constriction. The petroleum ether, dichloromethane, and methanol fractions exhibit 2–2.6 times the relaxing effect on pulmonary artery vessels compared to the crude extract. The CHCl₃-MeOH fraction significantly reduces ACh, BaCl₂, KCl, and CaCl₂-induced ileal contractions in guinea pigs, indicating that calcium ion flux is disrupted [28].

2.7 Antioxidant activity

The ethanol and hexane extracts of Bm have an inhibitory effect on lipid peroxidation induced by FeSO₄ and cumene hydroperoxide. The alcohol extracts exhibit strong protective effects. Compared with known antioxidants such as trimethylaminomethane, EDTA, and vitamin E, Bm is a potent antioxidant with a dose-dependent effect. Bm ethanol extract at 100 μg is equivalent to 247 μg of EDTA and 58 μg of vitamin E²⁹.

Bm standard extracts at 5 and 10 mg/kg were administered orally to rats once daily for 7, 14, and 21 days, and the antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in the cerebral cortex, striatum, and hippocampus, with a dose-dependent effect. In contrast, the antioxidant agent silibinin [(1)-de-prenyl] (2 mg/kg, po) only increased the activity of SOD, CAT, and GPx in the cerebral cortex and striatum, but had no effect on these three enzymes in the hippocampus. Bm can increase the scavenging activity of oxidative free radicals, thereby benefiting cognitive function [30].

Analysis using nitroblue tetrazolium (NBT) showed that the ethanol extract of Bm whole plant inhibited the release of superoxide radicals from polymorphonuclear cells. This effect is attributed to the main saponins in the whole plant, including  pseudoparkin saponin A₃, at concentrations of 200, 100, 50, and 25 μg/mL, which exhibited inhibitory effects on NBT at 85%, 91.66%, 91. 66%, and 83%, respectively, with an IC₅₀ of 10.22 μg/mL. The positive controls, quercetin and vitamin C, had IC₅₀ values of 111 and 14.16 μg/mL, respectively. Another major component of Bm, bacopasaponin C, exhibited weak activity [31].

3 Clinical Improvement of Cognitive Function

Bm (300 mg) or placebo was administered to healthy individuals in a double-blind study to assess cognitive function over a 12-week period. Neuropsychological tests included IT, learning rate, and memory consolidation. The results showed a significant improvement in visual information processing speed, indicating that Bm can significantly enhance cognitive processes, with the key factor being the continuous input of information from the environment for learning and memory [20].

Roodenrys et al. [21] reported the effects of Bm on human memory. A randomized, double-blind, placebo-controlled study was conducted on 76 adults aged 40–65 years, using various memory function tests as indicators, divided into three stages: before the trial, 3 months after the trial, and 6 weeks after the trial completion. The results showed that Bm had a significant effect on the retention of new information but no effect on learning rate, indicating that Bm can reduce the rate of forgetting newly acquired information.

A study investigated the effects of Bm combined with ginkgo on human cognitive function. The results demonstrated enhanced memory and attention, suggesting its potential for treating Alzheimer's disease. Another study reported that Bm had no immediate effects on cognitive function in healthy individuals [22,23].

4 Conclusion

The incidence of Alzheimer's disease is increasing annually, becoming a social issue that concerns governments and health departments in both developed and developing countries. Through the relentless efforts of scientists worldwide, utilizing various experimental animal models of dementia, at least 50 plant extracts and chemical components with anti-dementia activity have been screened from traditional and folk medicines. Huperzia serrata, an early-developed plant extract from China, and the currently developed plant extract from India, both serve as excellent examples. Huperzia serrata is distributed in provinces such as Yunnan, Guangdong, Fujian, and Taiwan in China, with abundant resources and good development potential. It is believed that new nootropics will continue to be discovered in the future for the treatment of dementia.

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