Cassia alata L.: A Study of Antifungal Activity against Malassezia furfur, Identification of Major Compounds, and Molecular Docking to Lanosterol 14-Alpha Demethylase.

Empirically, in Indonesia, the leaves of Cassia alata L. (candle bush or ketepeng cina) have been used as a topical antifungal agent. Malassezia furfur is a natural microorganism found in the human body. It is among the factors contributing to conditions such as pityriasis versicolor, a common, benign, superficial fungal infection of the skin that is closely associated with seborrheic dermatitis and dandruff. This study aimed to explore C. alata leaves, starting from determining antifungal activity against M. furfur and the identification of major compounds in the ethyl acetate and n-hexane fractions, and then we carried out molecular docking of the major compounds in the n-hexane fraction to lanosterol 14-alpha demethylase. The method was the disc diffusion technique to test antifungal activity, LC-MS/MS for major compound identification, and homology modeling through Swiss Models for molecular docking. The fractions of ethyl acetate and n-hexane extract showed concentration-dependent antifungal activity against M. furfur. The LCMS/MS analysis revealed five major compounds in the ethyl acetate and n-hexane fractions. The molecular docking demonstrated the highest binding affinity with stearidonic acid at -7.2 kcal/mol. It can be concluded that the compounds in the n-hexane fraction have antifungal activity against M. furfur, as supported by both in vitro and in silico studies.

Antibacterial Potency of an Active Compound from Sansevieria trifasciata Prain: An Integrated In Vitro and In Silico Study.

Sansevieria trifasciata Prain holds great potential as a valuable asset in pharmaceutical development. In this study, our focus is to explore and assess the antibacterial activity of various components derived from this plant, including extracts, fractions, subfractions, and isolates, explicitly targeting two common bacteria: Escherichia coli and Streptococcus aureus. The isolated compound, identified as a derivative pyridone alkaloid (5-methyl-11-(2-oxopyridin-1(2H)-yl)undecaneperoxoicacid), demonstrates notable antibacterial effects. The extracts, fractions, subfractions, and isolates reveal significant bacterial growth reductions (p < 0.05). The minimum inhibitory concentration (MIC) values for Escherichia coli were 1.95 ppm, 3.9 ppm, 15.62 ppm, and 7.81 ppm, respectively, while the MIC values for Streptococcus aureus were 1.95 ppm, 1.95 ppm, 15.62 ppm, and 7.81 ppm, respectively. Computational analysis showed the isolates' interaction with key residues on the active site of ß-ketoacyl-ACP synthase from Escherichia coli and TyrRS from Streptococcus aureus. The findings indicate that the isolates exhibit a strong affinity for specific residues, including His333, Cys163, and Phe392 in ß-ketoacyl-ACP synthase, as well as Arg88, His117, Glu160, and Gln213 in TyrRS. Comparative energy calculations using MMPBSA demonstrate the isolates' favorable binding energy (-104,101 kJ/mol for ß-ketoacyl-ACP synthase and -81,060 kJ/mol for TyrRS) compared to ciprofloxacin. The elucidated antibacterial activity and molecular interactions of the isolates present valuable knowledge for future in vitro studies, facilitating the development of novel antibacterial agents targeting diverse bacterial strains.

Isolation, Characterization, and Evaluation of Protease Activity of Crude Bromelain of Pineapple Peel, Core, and Crown from Subang District, Indonesia.

Context: Bromelain is distributed to all parts of pineapple (Ananas comusus (L.) Merr), including the peel, core, and crown, which are agricultural waste that has not been utilized properly. Aims: The purpose of this study was to determine the character and protease activity of crude bromelain from the Indonesian pineapple peel, core, and crown. The pineapple was collected from Subang district, West Java Province, Indonesia. Methods and Material: The three crude bromelains were obtained through the precipitation process with ethanol, then a protein qualitative and quantitative analysis was performed. Protease activity was determined by measuring the tyrosine produced from casein hydrolysis. The characters of crude bromelains were determined by evaluating the protease activity in various pH, temperature, and substrate concentration. Statistical Analysis Used: One-way analysis of variance was conducted to analyze the data statistically. Results: The three crude bromelains can be isolated from the peel, core, and crown of pineapple fruit with protease activity in the range of 38.32-46.78 units. Crude bromelains have an optimum temperature of 55°C for the peel and core and 35°C for the crown. All crude bromelains have an optimum pH of pH 7. The three crude bromelains have Vmax in the range of 140.85 to 285.71 units and KM in the range of 15298.59 to 18370.86 ppm. Conclusions: It concluded that the three crude bromelains have protease activity with the specific character and kinetic parameter.

Anti-Alopecia Activity of Coumarin Derivatives Isolated from Merremia peltata Leaves and Computational Study of Their Binding to Androgen Receptors Using Molecular Docking and Molecular Dynamic Simulation.

Alopecia is a condition in which hair on the scalp or other areas of the body is lost or falls out excessively. Nutritional deficiency causes blood flow to the head to decrease causing the hormone testosterone to be changed by the enzyme 5-α-reductase to dihydrotestosterone, which inhibits the growth phase and accelerates the death phase. One of the methods developed to treat alopecia is through inhibition of the 5-α-reductase enzyme, which converts testosterone to its more potent metabolite, dihydrotestosterone (DHT). Ethnomedicinally, Merremia peltata leaf is used by the people of Sulawesi as a remedy for baldness. Therefore, in this research, an in vivo study was conducted on rabbits to determine the anti-alopecia activity of M. peltata leaf compounds. The structure of the compounds isolated from the M. peltata leaf ethyl acetate fraction was determined by analysis of NMR and LC-MS data. An in silico study was then carried out using minoxidil as a comparison ligand; scopolin (1) and scopoletin (2) isolated from M. peltata leaf were identified as anti-alopecia compounds by predicting docking, simulating molecular dynamics and predicting absorption, distribution, metabolism, excretion, and toxicology (ADME-Tox). Compounds 1 and 2 had a better effect on hair growth compared to positive controls, and NMR and LC-MS analysis showed that they had comparable binding energies to receptors in the molecular docking interaction study: -4.51 and -4.65 kcal/mol, respectively, compared to -4.8 kcal/mol for minoxidil. Molecular dynamics simulation analysis with the parameters binding free energy calculated using the MM-PBSA method and complex stability based on SASA, PCA, RMSD, and RMSF showed that scopolin (1) has a good affinity for androgens receptors. The ADME-Tox prediction for scopolin (1) showed good results for the parameters of skin permeability, absorption and distribution. Therefore, scopolin (1) is a potential antagonist to androgen receptors and could be useful in the treatment of alopecia.

Estrogen receptor targeting with genistein radiolabeled Technetium-99m as radiotracer of breast cancer: Its optimization, characterization, and predicting stability constants by DFT calculation.

Objective: Genistein is an isoflavone molecule with a high affinity for estrogen receptors (ER), which could lead to the mechanism of selective estrogen receptor modulators (SERMs) in breast cancer. Genistein labeling with technetium-99m can be a new promising strategy for diagnostic breast cancer. In this research, we evaluate the physicochemical characteristics of the [99mTc]Tc-genistein complex and describe the optimal labeling method parameters. We also calculated density functional theory to study the stability constants to support complex formation analysis (DFT). Methods: The genistein was directly labeled with 99mTc, and its stability as well as its potential for usage as a radiotracer were all investigated. DFT calculations with thermodynamic cycles to determine chemical coordination models and calculate thermodynamic constants of complex more accurately. Results: The radiochemical purity of [99mTc]Tc-genistein showed a high yield of 93.25% ± 0.30% and had good physicochemical properties. The stability of the Tc(IV)-genistein complex was confirmed by DFT calculations at a value of 99.0822. Conclusions: As a result, [99mTc]Tc-genistein could be a potential radiotracer kit for SPECT imaging of breast cancer.

Anti-Alopecia Activity of Alkaloids Group from Noni Fruit against Dihydrotestosterone-Induced Male Rabbits and Its Molecular Mechanism: In Vivo and In Silico Studies.

Androgenic alopecia (AA) is a condition that most commonly affects adult men and is caused by an increase in the hormone dihydrotestosterone (DHT) in the hair follicles. Anti-alopecia drugs should be discovered for hair follicles to enter the anagen growth phase. Therefore, this study evaluated the hair growth-promoting activity of Noni fruit's water, ethyl acetate, n-hexane fractions, and sub-fractions from the active fraction in the alopecia male white rabbit model. The Matias method was modified by inducing rabbits using DHT for 17 days, followed by topical application of Noni fruit solution for 21 days. Meanwhile, hair growth was evaluated by histological observation of the follicular density and the anagen/telogen (A/T) ratio in skin tissue. In the first stage, five groups of male white rabbits were studied to obtain the active fraction; DHT+Minoxidil as standard, DHT+vehicle (NaCMC 1%), DHT+FW, DHT+FEA, and DHT+FH. The FEA as the active fraction was followed by open-column chromatography separation (DCM:Methanol) with a gradient of 10% to produce sub-fractions. In the second stage, the six main sub-fraction groups of male rabbits studied were DHT+FEA-1 to DHT+FEA-6. The follicular density of groups FEA-3 was 78.00 ± 1.52 compared with 31.55 ± 1.64 and 80.12 ± 1.02 in the Vehicle and Minoxidil groups. Additionally, group FEA-3 showed large numbers of anagen follicles with an A/T ratio of 1.64/1 compared to the vehicle group of 1/1.50 and 1.39/1 for Minoxidil control. Group FEA-3 was identified by LC-MS/MS-QTOF, followed by molecular docking to the androgen receptor (PDB: 4K7A), causing alopecia. The results showed that three alkaloid compounds with skeleton piperazine and piperidine, namely (compounds 2 (−4.99 Kcal/mol), 3 (−4.60 Kcal/mol), and 4 (−4.57 Kcal/mol)) had a binding affinity similar to Minoxidil, with also has alkaloid skeleton piperidine−pyrimidine (−4.83 Kcal/mol). The dynamic behavior showed the stability of all androgen receptor compounds with good RMSD, SMSF, and SASA values after being studied with 100 ns molecular dynamics (MD) simulations. This study produced a common thread in discovering a class of alkaloid compounds as inhibitors of androgen receptors that cause alopecia.

Herbal Therapy for the Treatment of Seborrhea Dermatitis.

Seborrhea dermatitis is a skin disorder that usually appears on parts of the body that have high density of sebaceous glands, such as the face, chest, and scalp. Clinical manifestations that generally appear as scaly skin and erythema. Seborrhea dermatitis is also known as one of the causes of alopecia. Treatments that can be used for seborrhea dermatitis are antifungal, anti-inflammatory, keratolytic, and coal tar. There are concerns about poor adherence, resistance, and some side effects of drugs that have been used in the treatment of seborrhea dermatitis. Concerns regarding these issues increase the urgency for the development of new therapeutic agents in the treatment of seborrhea dermatitis. Research on medicinal plants has enormous potential to produce compounds with new structures and bioactivity. This review discusses clinical and in vitro studies related to the activity of several medicinal plants that have potential as a treatment for seborrhea dermatitis, as well as the compounds that play a role in these activities. Literature searches were carried out on the PubMed, Taylor & Francis, and SpringerLink databases using Boolean Operators to get 25 articles that match the keywords used. Of the 25 articles, six were clinical trials, while 19 were in vitro studies of Malassezia. Several plants have potential as promising therapeutic agents for the treatment of seborrhea dermatitis by inhibiting the growth of Malassezia, decreasing sebum secretion, and decreasing symptoms associated with seborrhea dermatitis such as itching, pain or burning sensation, and redness.

Active Antialopecia Chemical Identification of Merremia peltata Leaves and Computational Study toward Androgen Receptor Using Molecular Docking and Molecular Dynamic Simulation.

Alopecia is a health condition in which the hair loses its function in some or all of the body. Alopecia occurs due to various genetic, environmental, and nutritional factors. One of the methods developed to treat alopecia is through inhibition of the enzyme 5-α-reductase, which converts testosterone into its more potent metabolite, dihydrotestosterone (DHT). In ethnomedicine, the leaves of Merremia peltata are used by the people of Sulawesi as a remedy for baldness. Therefore, in this study, an in vivo study was conducted on rabbits to investigate the antialopecia activity of the ethanolic extract of M. peltata leaves. The purified M. peltata leaf extract was fractionated using vacuum liquid chromatography with several solvents to produce fractions F1-F5. Each fraction was then retested in vivo in rabbits, and its content was then analyzed by LC-MS. An in silico study was then carried out using minoxidil as a comparison ligand; 17 compounds derived from M. peltata leaves were identified as antialopecia compounds through prediction of molecular interactions and molecular dynamics simulation and prediction of absorption, distribution, metabolism, excretion, and toxicology (ADME-Tox). The assay results showed that fractions F2 and F3 had a better effect on hair growth compared to the positive control, and the test compound obtained from the LC-MS analysis, bufotalinin, had a strong binding energy to the receptor in the molecular docking interaction study: -5.99 kcal/mol compared to -4.8 kcal/mol for minoxidil. Molecular dynamics simulation analysis with complex stability parameters based on solvent-accessible surface area (SASA), principal component analysis (PCA), root mean square deviation (RMSD), and root mean square fluctuation (RMSF) showed that bufotalinin has good affinity for androgen receptors. ADME-Tox prediction for bufotalinin showed good results for the parameters of skin permeability, absorption, and distribution. Therefore, bufotalinin, a steroid compound, is a potential androgen receptor antagonist and could be useful in the treatment of alopecia.

Unrevealing the Potential of Sansevieria trifasciata Prain Fraction for the Treatment of Androgenetic Alopecia by Inhibiting Androgen Receptors Based on LC-MS/MS Analysis, and In-Silico Studies.

Androgenetic Alopecia (AGA) occurs due to over-response to androgens causing severe hair loss on the scalp, and requires the development of new and efficient drugs to treat this condition. This study explores and identifies secondary metabolites from Sansevieriatrifasciata Prain using the LC-MS/MS and in-silico method. The inhibitory activity of bioactive compounds from S. trifasciata Prain against androgen receptors (PDB ID: 4K7A) was evaluated molecularly using docking and dynamics studies by comparing their binding energies, interactions, and stability with minoxidil. The results of the LC-MS/MS analysis identified Methyl pyrophaeophorbide A (1), Oliveramine (2), (2S)-3', 4'-Methylenedioxy-5, 7-dimethoxyflavane (3), 1-Acetyl-ß-carboline (4), Digiprolactone (5), Trichosanic acid (6) and Methyl gallate (7) from the leaves subfraction of this plant. Three alkaloid compounds (compounds 1, 3, and 4), and one flavonoid (compound 2), had lower docking scores of -7.0, -5.8, -5.2, and -6.3 kcal/mol, respectively. The prediction of binding energy using the MM-PBSA approach ensured that the potency of the four compounds was better than minoxidil, with energies of -66.13, -59.36, -40.39, and -40.25 kJ/mol for compounds 1, 3, 2, and 4, respectively. The dynamics simulation shows the stability of compound 1 based on the trajectory analysis for the 100 ns simulation. This research succeeded in identifying the compound and assessing the anti-alopecia activity of Sansevieria trifasciata Prain. Seven compounds were identified as new compounds never reported in Sansevieria trifasciata Prain. Four compounds were predicted to have better anti-alopecia activity than minoxidil in inhibiting androgen receptors through an in silico approach.

New Hair Growth Cream Formulation with Cocoa Pod Peel (Theobroma cacao L.).

Our previous study verified that the waste skin of cocoa (Theobroma cacao L) fruit or waste cocoa pod husks had the efficacy to overcome hair loss or alopecia. This study aims to determine the formula and activity of hair cream of cocoa pod peel water fraction, which is effective in stimulating hair growth. Activity testing uses the modified Tanaka method. The results showed that the cocoa husk wastewater fraction could be formulated into hair cream, but there were changes in viscosity and pH after the freeze-thaw test, but still within the allowed limit. The hair cream water fraction gel stimulated hair growth activity based on the hair length data with a significant difference in concentration of the preparation. The best activity in hair cream preparation was at 12.5% concentration. In addition, there were no signs of irritation to the rabbit's skin where hair cream preparation was applied. The results of this study indicated that cocoa fruit peel cream can be used for antialopecia treatments.

Efficacy, Safety, and Drug-Drug Interactions for Insomnia Therapy in COVID-19 Patients.

Coronavirus disease-19 (COVID-19) is a systemic viral infection. COVID-19 patients show diverse clinical presentations ranging from asymptomatic, mild symptoms to severe symptoms characterized by severe respiratory distress. Sleep disorders or insomnia is one of the psychiatric problems that arise during the COVID-19 pandemic. The term used to define this particular insomnia is coronasomnia or COVID-19 insomnia. Data show that the prevalence of this problem is increasing, especially in the confirmed COVID-19 patient group. Anti-insomnia drugs such as hypnotics, sedatives, and anxiolytics are the easiest option. As with drugs generally, anti-insomnia drugs are associated with various safety issues, especially in people with COVID-19. Therefore, their use may be hazardous. The literature review aims to make health practitioners aware of the anti-insomnia drugs that have the best efficacy and safety issues that are clinically relevant from the use of anti-insomnia drugs and the interactions of anti-insomnia drugs with various drugs used in the treatment of COVID-19. The articles were explored on PubMed and Cochrane Library, whereas the drug-drug interactions between the anti-insomnia and COVID-19 drugs were searched on Drugs.com Interaction Checker and Lexiomp-interact. Overall anti-insomnia drugs have efficacy in improving sleep parameters. Orexin receptor antagonist drugs have good efficacy in increasing WASO, LPS, and SE with an acceptable safety profile. Meanwhile, the combination of zolpidem, lorazepam, and diphenhydramine improved TST parameters better than other drugs. Side effects such as drowsiness and dizziness were among the most commonly reported effects. Therefore, attention and monitoring of the use of anti-insomnia drugs in COVID-19 patients need to be carried out by considering the side effects and interactions that are very risky.

Molecular docking studies and ADME-Tox prediction of phytocompounds from Merremia peltata as a potential anti-alopecia treatment.

Alopecia is a condition in which some or all of the hair from the scalp is lost. One recent preventative measure is the inhibition of the enzyme 5-α-reductase. Inhibition of the enzyme 5-α-reductase converts circulating testosterone to its more potent metabolite, dihydrotestosterone. Ethnobotically, Merremia peltata is used as a baldness medicine by utilising compounds contained within the leaves. This research aimed to test activity of 18 known compounds contained within M. peltata) as anti-alopecia. Activity was based on their interaction with the androgen receptor (PDB code 4K7a) using molecular docking and ADME-Tox prediction. The stages of research performed were: preparation of androgen protein structure databases; preparation and optimization of three-dimensional structures of compounds using ChemDraw 8.0; molecular docking to the androgen receptor protein using Autodock 1.5.6.; and ADME-Tox prediction using the pkCSM tool. The following test compounds had strong bond energies (ΔG): compound 16 (olean-12-en-3beta-ol, cinnamate)-7.71 kcal/mol, compound 17 (alpha-amyrine)-6.34 kcal/mol, and Finasteride-6.03 kcal/mol. Interestingly, the ΔG of compound 16 (olean-12-en-3beta-ol, cinnamate) is better than of minoxidil (-4.8 kcal/mol) and also to gold-standard treatment compound, finasteride. ADME-Tox prediction for compound 16 showed favorable results in several metrics such as skin permeability, absorption, and distribution. Compound 16 (olean-12-en-3beta-ol, cinnamate) is therefore a potential androgen receptor antagonist and may be beneficial in the treatment of alopecia.

Potential anti-alopecia constituents from Theobroma cacao: An in silico study.

Tinea capitis is local alopecia caused by a dermatophyte infection of the scalp. Trichophyton rubrum produces the squalene epoxidase enzyme, which has a crucial role in prolonged dermatophyte infection, as well as in synthesizing fatty acids in this dermatophyte group. This study analyzes Trichophyton cacao compounds as anti-alopecia by inhibiting the squalene epoxidase enzyme formation, in silico. The structure of T. cacao compounds was prepared using the MolView Web application. The compound docked to squalene epoxidase using AutoDock Vina in PyRx 0.8, followed by PyMOL for visualization, and the Proteins Plus program to analyze the complexity. The binding affinity value of catechin, epicatechin (-8.0 kcal/mol), and anthocyanin (-7.8 kcal/mol) compounds was higher than the positive control (terbinafine, -6.7 kcal/mol). Pre-ADMET demonstrated that catechin and epicatechin had moderate Human Intestinal Absorption (66.71%), but anthocyanin was very good (100%). Caco-2 parameters for catechin and epicatechin were relatively low (<4 nm s - 1), while anthocyanin, theobromine, and terbinafine were within 4-70 nm s - 1. Plasma protein binding shows catechin, epicatechin, and anthocyanin diffuse through the plasma membrane and interact with plasma proteins. The toxicity results for all compounds are mutagenic, and only terbinafine is carcinogenic. Based on the Lipinski's "Rule of Five," compounds from T. Cacao can be given orally. Catechin and epicatechin compounds have the potential to act as anti-alopecia. These two compounds can diffuse and interact with plasma proteins so they are directly on the target when given orally.

Antibacterial Activity Test of Extracts and Fractions of Cassava Leaves (Manihot esculenta Crantz) against Clinical Isolates of Staphylococcus epidermidis and Propionibacterium acnes Causing Acne.

This study is aimed at determining antibacterial activity from ethanol extracts and the most active fraction of cassava leaves against clinical isolates of Staphylococcus epidermidis and Propionibacterium acnes. Research carried out by the experimental method involved determination of plants, extraction with maceration method, fractionation with liquid-liquid extraction, antibacterial activity testing of extracts and fractions by agar diffusion method, determination of most active fraction from the extract, and minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) testing of most active fraction by microdilution method. The results showed that ethanol extracts of cassava leaves had antibacterial activity against both bacteria with the most active fraction indicated by ethyl acetate. MIC values of ethyl acetate fraction against S. epidermidis were in the concentration range of 2.5%-5.0% (w/v) and against P. acnes were in the concentration range of 1.25%-2.5% (w/v). The MBC value of ethyl acetate fraction against S. epidermidis was at a concentration of 5% (w/v), while P. acnes was at a concentration of 2.5% (w/v). From the results of this study, it can be concluded that the ethanol extract of cassava leaves (Manihot esculenta Crantz) has antibacterial activity against clinical isolates of Staphylococcus epidermidis as well as on Propionibacterium acnes. The fraction with the best activity from the ethanol extract of cassava leaves to the two test bacteria was shown by ethyl acetate fraction. It is suggested that cassava leaves are possible to be developed into standardized antiacne herbal.

Determination of the Major Component of Water Fraction of Katuk (Sauropus androgynous (L.) Merr.) Leaves by Liquid Chromatography-Mass Spectrometry.

BACKGROUND: The katuk leaf (Sauropus androgynous (L.) Merr.) is one of the plants that are used to overcome baldness by the people of Kampung Mak Kemas, Malaysia. It is suspected that secondary metabolites contained in katuk leaves play a key role in stimulating hair growth. AIMS AND OBJECTIVES: The aim of this study was to identify the optimum method to obtain one of the chemical compounds in the water fraction and to identify the hypothesized chemical isolates in the water fraction katuk leave's ethanol extract. MATERIALS AND METHODS: The methods used in this study included the collection and determination of the katuk plant, the processing of the katuk, phytochemical filtrating, extracting with ethanol 96%, and fractionation using the liquid-liquid extraction method with n-hexane, ethyl acetate, and water solvents The water fraction of katuk leaves was analyzed by its components by thin-layer chromatography using the stationary phase of silica gel 60 F254, developer of n-butanol:acetic acid:water (4:1:5), and detection under ultraviolet (UV) light at a wavelength of 366 and 254nm, as well as with vanillin-sulfuric acid reagent. To isolate the compounds from water fraction of katuk leaves, it was then eluted with a vacuum column chromatography by eluent with a level polarity that would get 11 subfractions. Each subfraction was checked by two-dimensional thin-layer chromatography to see subfraction purity characterized by the appearance of a spot on the chromatogram plate. The isolate was analyzed using spot test, ultraviolet-visible spectrophotometer, infrared spectrophotometer, and liquid chromatography-mass spectrometry. RESULTS: The isolate was an alkaloid compound with a molecular mass of 406.3131 m/z with the molecular formula C21H39N6O2 as S, S-5, 5'-amino-4,4'-dihexyl-propyldihydropyrazol-3, 3-one. CONCLUSION: One of the chemical compounds contained in the water fraction of the ethanol extract of the katuk leaf was an alkaloid group.

Identification of compounds in the essential oil of nutmeg seeds (Myristica fragrans Houtt.) that inhibit locomotor activity in mice.

The present study was designed to evaluate the inhibitory effect of nutmeg (Myristica fragrans Houtt.) seed essential oil on the locomotor activity of mice in a wheel cage. Active compounds in the essential oil were identified by off-line solid phase extraction (SPE-C18) and GC/MS analysis. The essential oil was administered by inhalation at doses of 0.1, 0.3, and 0.5 mL/cage. The results showed that inhalation of nutmeg seed essential oil at a dose of 0.5 mL/cage decreased locomotion by 68.62%; and inhalation of 0.1 and 0.3 mL/cage inhibited locomotion by 62.81% and 65.33%, respectively. Generally, larger doses and longer administrations of nutmeg seed essential oil exhibited greater locomotor inhibition. Subsequently, the plasma concentrations of essential oil compounds were measured. The most concentrated compound in the plasma was myristicin. Half an hour after the addition of 1 mL/cage of nutmeg seed oil, the plasma concentration of myristicin was 3.7 µg/mL; one and two hours after the addition, the blood levels of myristicin were 5.2 µg/mL and 7.1 µg/mL, respectively. Other essential oil compounds identified in plasma were safrole (two-hour inhalation: 1.28 µg/mL), 4-terpineol (half-hour inhalation: 1.49 µg/mL, one-hour inhalation: 2.95 µg/mL, two-hour inhalation: 6.28 µg/mL) and fatty esters. The concentrations of the essential oil compounds in the blood plasma were relatively low (µg/mL or ppm). In conclusion, the volatile compounds of nutmeg seed essential oil identified in the blood plasma may correlate with the locomotor-inhibiting properties of the oil when administered by inhalation.