Curcuminoid Contents in Rhizomes of Some Zingiberaceous Plants Sold via Online Platforms: Influence of Species and Cultivation Location.

Zingiberaceous plants are versatile and find application in various fields, including food, medicine, and cosmetics. Recently, turmeric and other Zingiberaceous plants have become readily available through online platforms. However, the quality, specifically the curcuminoid content, has not been adequately assessed. In light of this issue, this study is aimed at analyzing the curcuminoid contents, including bisdemethoxycurcumin, demethoxycurcumin, and curcumin, using high-performance liquid chromatography. The analysis targets the rhizomes of Zingiber montanum (ZM), Curcuma aromatica (CA), Curcuma wanenlueanga (CW), Curcuma zedoaria (CZ), and sixteen Curcuma longa (CL) samples sold on online platforms. The influence of species and cultivation locations was evaluated, compared, and clustered. The results indicated that CL exhibited the highest curcuminoid contents, followed by CA, CZ, ZM, and CW, respectively. Curcumin was not detected in CW, while bisdemethoxycurcumin and demethoxycurcumin were absent in ZM. Cluster analysis revealed that CW was closely related to ZM, and CA was closely related to CZ, while CL was not closely related to either cluster. Among the sixteen CL samples, the most commonly found curcuminoids were curcumin, followed by bisdemethoxycurcumin and demethoxycurcumin, respectively. Three samples contained curcuminoid contents of less than 5%, failing to meet the standard level specified in the Thai Herbal Pharmacopoeia. Furthermore, ten samples had total curcuminoid contents higher than 10%, with three samples exceeding 15%. The top three samples with the highest total curcuminoid contents from different locations were as follows: Tha Yang District, Phetchaburi Province (17.02%); Phop Phra District, Tak Province (16.97%); and Pak Tho District, Ratchaburi Province (15.45%). Cluster analysis revealed that CL samples could be grouped into two major categories: low curcuminoid and high curcuminoid groups. This study offers valuable insights for consumers seeking high-quality rhizomes of Zingiberaceous plants with high curcuminoids, through online platforms. By focusing on the curcuminoid content, consumers can make informed decisions when purchasing Zingiberaceous plants online. This information not only aids in selecting superior quality rhizomes but also enhances the overall consumer experience by ensuring the potency and efficacy of the purchased products.

Maximizing Curcuminoid Extraction from Curcuma aromatica Salisb. Rhizomes via Environmentally Friendly Microwave-Assisted Extraction Technique Using Full Factorial Design.

Curcuma aromatica Salisb. contains a high content of curcuminoids, which can be utilized for cosmetic purposes. The objective of this study was to optimize the extraction conditions of C. aromatica rhizomes in castor oil to maximize curcuminoid content using a simple and environmentally friendly microwave-assisted extraction method. A 32 full factorial design was employed, with two factors-microwave power and time-varying between 600-800 W and 30-90 s, respectively. Five responses were monitored, including extraction yield, bisdemethoxycurcumin, demethoxycurcumin, curcumin, and total curcuminoid contents. The results demonstrated that increasing microwave power and time led to an increase in all five responses. The optimal condition, which simultaneously maximized extraction yield and total curcuminoid content, was achieved at a microwave power of 800 W for 90 s. This condition resulted in an extraction yield of 71.020%, bisdemethoxycurcumin content of 0.036%, demethoxycurcumin content of 0.210%, curcumin content of 0.080%, and total curcuminoid content of 0.326%. The computer program accurately predicted the results with a percentage error of less than 2%. Stability data revealed that the total curcuminoid content remained stable with a percentage remaining above 90% when stored at 4°C, 30°C ± 75%RH, and 40°C ± 75%RH for three months. In summary, this study successfully applied a full factorial design to maximize curcuminoid extraction from C. aromatica rhizomes using an environmentally friendly microwave-assisted extraction method for cosmetic purposes.

Fabrication of Orally Fast-Disintegrating Wafer Tablets Containing Cannabis Extract Using Freeze-Drying Method.

Introduction: The development of a novel dosage form for cannabis extract is necessary to improve drug delivery and also enhance patient convenience. Methods: Orally fast-disintegrating wafer tablets containing cannabis extract, which were prepared using the freeze drying technique, were developed in this work. The formulation consisted of several key components: cannabis extract as the active compound, Tween® 80 as a surfactant and solubilizer, gelatin and mannitol as structural components, sucralose as a sweetening agent, and sodium methylparaben and sodium propylparaben as preservatives. Results: The optimized formulation consists of the following ingredients: 5% cannabis extract, 1.25% Tween® 80, 5% gelatin, 88.34% mannitol, 0.2% sucralose, 0.19% sodium methylparaben, and 0.02% sodium propylparaben. The resulting wafer tablets exhibited the following characteristics: a porous structure, an average weight of approximately 200 mg, minimal weight variation (less than 1.4%), slightly acidic pH (pH 5.12), disintegration within 10 s, low moisture content (less than 3%), a Δ9-tetrahydrocannabinol content of approximately 2.8 mg, and a cannabidiol content of approximately 0.9 mg. Additionally, the wafer tablets rapidly dissolved in simulated saliva fluid containing sodium lauryl sulfate. Conclusion: This work succeeded in the fabrication of orally fast-disintegrating wafer tablets containing cannabis extract with desired properties.

Plant leaf mucilage/carrageenan/Eudragit® NE30D blended films: Optimization, characterization, and pharmaceutical application.

Mucilage of C. pareira leaves was utilized, being manufactured for use in pharmaceutical products. Carrageenan and Eudragit® NE30D were used to combined. Glycerin was used as a plasticizer at a concentration of 20 % w/w based on the amount of polymer used. Computer software optimized its characteristics, including tensile properties, moisture uptake, and erosion; the optimal formulation was 1.4:1.2:2.8. The percentages of optimization error ranged from 8.48 to 13.80 %. Propranolol HCl was mixed to an optimal formulation. The film layer was tight, homogeneous, and smooth, with no holes. DSC thermogram showed no interaction peaks at 101.33 °C and 170.50 °C. Propranolol HCl concentration in the film ranged from 2.18 to 2.20 mg/cm2. Propranolol HCl was quickly released from the film. The kinetic model for the release profile was first-order kinetic. Although propranolol HCl had a high-release profile, its skin permeation was limited. The permeation lag time, Jss, and Kp were 1.60-2.65 h, 0.0182-0.0338 µg/cm2/h, and 9.10-15.35 cm/h, respectively. A significant amount of propranolol HCl residue was found on the skin's surface. Glycerin appeared to influence propranolol HCl permeability. Therefore, the plant leaf mucilage/carrageenan/Eudragit® NE30D blended film can be utilized in pharmaceutical applications to control drug release from its film layer.

Porous Deproteinized Natural Rubber Film Loaded with Silver Nanoparticles for Topical Drug Delivery.

The work demonstrated the use of natural rubber for topical drug delivery. The first objective was to fabricate a porous deproteinized natural rubber film loaded with silver nanoparticles. Characterizing and assessing its formulation was the second objective. Surface pH, mechanical properties, swelling ratio, erosion, moisture vapor transmission rate, scanning electron microscopy/energy dispersive X-ray analysis, and X-ray diffraction were evaluated. In vitro studies and antibacterial activity were assessed. It was discovered that silver nanoparticles could enter the film and that their concentrations ranged between 7.25 and 21.03 µg/cm2. The pH of the film's surface was 7.00. The mechanical properties of the film with silver nanoparticle loading differed from the blank film. After adding silver nanoparticles, the film eroded faster than before, but the swelling ratio was not affected significantly. Increased time utilization had an impact on the moisture vapor transmission rate of the film. Silver nanoparticles released easily from the film while there was less permeability. The dead pig-ear skin had significant silver nanoparticle accumulation. Potent antibacterial activity was seen in the film containing silver nanoparticles. The silver nanoparticle-loaded film may be used as a wound dressing for a topical film that promotes wound healing while also protecting the area from infection.

Plaster Gel Loaded with Silver Nanoparticle-Mediated Ganoderma applanatum: from Fabrication to Evaluation.

Traditional Asian remedies have mainly employed the macrofungus Ganoderma applanatum, which belongs to the family Ganodermataceae, as a medicinal mushroom due to its high antibacterial and antioxidant activity. Extracts of the fungus can be synthesized into nanoparticles, which are subsequently produced as plaster gels. Synthesized silver nanoparticle-mediated G. applanatum was discovered to have the greatest ability to inhibit bacterial growth in S. epidermidis. When applied to the skin, the prepared plaster gel converted from a gel to a film; thus, both gel and film generation are characteristic of its formulation. The plaster gel that was made was found to be consistent and attractive, and the yellow color had darkened. Its viscosity and pH were appropriate for the application and allowed it to remain on the skin without dripping or reacting with the skin until it dried. A shorter duration for film formation is possible. The film's tensile was slightly reduced, and it exhibited excellent thermal stability. Decomposition of the generated film occurred at a slower rate, which constrained the polymer chain's ability to move. The semi-crystalline structure was characteristic of the film. It was found that particles were distributed in the film. Rapid release from plaster gel within 4 h was seen, and this was followed by a period of a slowly declining release rate over 12 h. The accurate first-order kinetic used to estimate the release rate of the formulation. The plaster gel demonstrated greater antibacterial activity than the MIC value indicated. The in vivo evaluation was positive and showed no skin irritation. The formulation showed good stability. Therefore, this indicated that the prepared plaster gel is appropriate for topical pharmaceutical delivery and safe for skin application.

Effect of Ionic Liquid on Silver-Nanoparticle-Complexed Ganoderma applanatum and Its Topical Film Formulation.

Imidazolium-based ionic liquids have been widely utilized as versatile solvents for metal nanoparticle preparation. Silver nanoparticles and Ganoderma applanatum have displayed potent antimicrobial activities. This work aimed to study the effect of 1-butyl-3-methylimidazolium bromide-based ionic liquid on the silver-nanoparticle-complexed G. applanatum and its topical film. The ratio and conditions for preparation were optimized by the design of the experiments. The optimal ratio was silver nanoparticles: G. applanatum extract: ionic liquid at 97:1:2, and the conditions were 80 °C for 1 h. The prediction was corrected with a low percentage error. The optimized formula was loaded into a topical film made of polyvinyl alcohol and Eudragit®, and its properties were evaluated. The topical film was uniform, smooth, and compact and had other desired characteristics. The topical film was able to control the release of silver-nanoparticle-complexed G. applanatum from the matrix layer. Higuchi's model was used to fit the kinetic of the release. The skin permeability of the silver-nanoparticle-complexed G. applanatum was improved by about 1.7 times by the ionic liquid, which might increase solubility. The produced film is suitable for topical applications and may be utilized in the development of potential future therapeutic agents for the treatment of diseases.

Optimization of Process and Formulation Variables for Semha-Pinas Extract Effervescent Tablets Using the Box-Behnken Design.

Semha-Pinas (SHPN) is a Thai traditional herbal formula used as an expectorant. Its traditional dosage form is pills, which are dispersed in water before use. The development of this recipe to effervescent tablets could enhance patient convenience with shortened the time needed to disintegrate the active ingredients. This work aimed to develop SHPN extract effervescent tablets based on process and formulation optimization using the Box-Behnken design. Four levels of three independent variables, including a compressional force, a quantity of effervescent base, and a quantity of fumed silica, were screened using the one factor at a time method. Three levels of each independent variable were included in the Box-Behnken design, including 1000 - 2000 psi, 46 - 52%, and 1.67 - 3.33%, respectively. Four responses were monitored, including tablet thickness, hardness, disintegration time, and friability. In terms of design space, the results showed that the tablet hardness was not less than 5 kP, disintegration time was not more than 5 min, and friability was not more than 1% found when 2000 psi compressional force was used. The optimal parameters were a compressional force of 2000 psi, effervescent base of 50%, and fumed silica of 2.5%. This formulation had a tablet weight of 598.86 ± 0.05 mg, a diameter of 12.68 ± 0.01 mm, a thickness of 3.67 ± 0.01 mm, a hardness of 5.57 ± 0.22 kP, a disintegration time of 1.68 ± 0.04 min, and friability of 0.43 ± 0.02%. In conclusion, this work succeeded in developing SHPN extract effervescent tablets with desired properties that were easy to use. Furthermore, the time needed to disintegrate the active ingredients was decreased when compared with the traditional dosage form due to being easily dissolved in water.

Antitumor Effects of Delta (9)-Tetrahydrocannabinol and Cannabinol on Cholangiocarcinoma Cells and Xenograft Mouse Models.

Cholangiocarcinoma (CCA) is a very aggressive tumor. The development of a new therapeutic drug for CCA is required. This study aims to evaluate the antitumor effect of ∆9-tetrahydrocannabinol (THC), the major psychoactive component of marijuana (Cannabis sativa), and cannabinol (CBN), a minor, low-psychoactive cannabinoid, on CCA cells and xenograft mice. THC and CBN were isolated, and their identities were confirmed by comparing 1H- and 13C-NMR spectra and mass spectra with a database. Cell proliferation, cell migration, and cell apoptosis assays were performed in HuCCT1 human CCA cells treated with THC or CBN. The phosphorylation of signaling molecules in HuCCT1 cells was detected. To determine the effects of THC and CBN in an animal model, HuCCT1 cells were inoculated subcutaneously into nude mice. After the tumors reached an appropriate size, the mice were treated with THC or CBN for 21 days. Tumor volumes were monitored and calculated. The 1H- and 13C-NMR data of THC and CBN were almost identical to those reported in the literature. THC and CBN significantly inhibited cell proliferation and migration and induced apoptosis in HuCCT1 cells. The phosphorylation of AKT, GSK-3α/ß, and ERK1/2 decreased in HuCCT1 cells treated with THC or CBN. CCA xenograft mice treated with THC showed significantly slower tumor progression and smaller tumor volumes than control mice. THC and CBN induced apoptosis in CCA by inhibiting the AKT and MAPK pathways. These findings provide a strong rationale for THC and CBN as therapeutic options for CCA.

Fabrication and Characterization of Buccal Film Loaded Self-emulsifying Drug Delivery System containing Lysiphyllum strychnifolium Stem Extracts.

Lysiphyllum strychnifolium has long been used as a popular herbal medicinal plant for treating fever and alcohol intoxication. This study aimed to prepare buccal film for L. strychnifolium stem extracts. These extracts were less soluble in water and were therefore loaded in self-emulsifying systems before being mixed into the film. Astilbin was selected as a chemical marker in L. strychnifolium stem extracts. Firstly, the L. strychnifolium stem extracts were entrapped in the self-emulsifying systems which were designed and optimized based on 32 factorial design. The optimal formulation was 0.60 g of surfactant-co-surfactant mixture (Tween® 80 and polyethylene glycol 400 in the ratio of 7.5:1) and 0.40 g of caprylic/capric triglyceride. Secondly, the optimal self-emulsifying system was loaded in the polymeric film which consisted of polyvinyl alcohol blended with poloxamer 407 using glycerin as a plasticizer. The properties of the prepared buccal film were unchanged, and the film showed an amorphous state, indicating all ingredients might be completely dissolved in the film. The buccal film could be placed in direct contact with the mouth without oral mucosal irritation, and showed a smooth and homogeneous surface with a rough and compact cross-sectional morphology. Astilbin content in the buccal film was 61.39 ± 11.45 µg/cm2. Astilbin was released from the buccal film while the permeation rate was low. The release mechanism was both swelling and diffusion, and followed anomalous or non-Fickian transfer. The permeability coefficient of the cumulative amount of astilbin permeated from buccal film was 1.0192 ± 0.1395 ×10-3 cm/h. Thus, the buccal film can be prepared by using a self-emulsifying system for herbal applications and shows potential as a safe and convenient form of oral drug administration.

Design and optimization of process parameters of polyvinyl alcohol-graft-lactic acid films for transdermal drug delivery.

The objective was to apply a simplex lattice design to determine the properties of polyvinyl alcohol-graft-lactic acid (PVA-g-LA) with different values for two independent variables: curing time (X1) and LA ratio (X2). Each independent variable was varied among three levels: -1, 0, and +1. Three coded levels were 120 min, 150 min, and 180 min for X1 and 2.5 g, 5 g, and 7.5 g for X2. Dependent variables of swelling behavior in various swelling media and thermal analysis parameters were monitored. The optimal formulation was selected based on the desirability value. The prediction was accurate, showing a low value of percent error. The morphology of the selected formulation with the highest desirability value showed a compact and dense film. Propranolol hydrochloride used as a model drug, was loaded into PVA-g-LA film. The propranolol hydrochloride content was 4.19 ± 1.05 mg/cm2. The cumulative release and permeation of drug were 61.94 ± 8.03% and 59.96 ± 6.61%, respectively. Thus, response surface methodology can be used as a tool to predict or optimize the process parameters for PVA-g-LA transdermal films in an accurate manner. PVA-g-LA could control the release and permeation of drug from the film layer.

Simplex lattice design for optimization of the mass ratio of Curcuma longa L., Curcuma zedoaria (Christm.) Roscoe and Curcuma aromatica Salisb. to maximize curcuminoids content and antioxidant activity.

The aim of this work was to optimize the mass ratio of three Curcuma plants' rhizomes to obtain the highest curcuminoid content and antioxidant activity using the simplex lattice design. The selected Curcuma plants were C. longa, C. zedoaria and C. aromatica. The simplex lattice design was applied in the work. The individual curcuminoids (curcumin, demethoxycurcumin and bis-demethoxycurcumin) and total curcuminoid content were determined using high-performance liquid chromatography. Curcuma longa alone provided the highest content of bis-demethoxycurcumin and demethoxycurcumin. A mixture of C. longa and C. aromatica in the mass ratio of 72:28 % provided the highest curcumin content. The results showed that C. longa alone exhibited the highest antioxidant activity.

Investigation of the Interaction of Herbal Ingredients Contained in Triphala Recipe Using Simplex Lattice Design: Chemical Analysis Point of View.

The aim of this work was to investigate the interaction of herbal ingredients contained in Triphala recipe (Terminalia chebula, Terminalia bellirica, and Phyllanthus emblica in equal proportion) using simplex lattice design. This work focused on chemical analysis of four phenolic compounds including gallic acid, corilagin, chebulagic acid, and chebulinic acid by validated high-performance liquid chromatography. The effect of the extraction technique (decoction vs. infusion) and gamma irradiation was also examined. The combination index was used as a tool for determination of interaction of the ingredients contained in the herbal recipe. Results showed that the extraction technique and gamma irradiation slightly altered the content of some phenolic compounds as well as the combination index. The positive interaction seems to be found at the equal proportion of the three plants. This work scientifically supported the suitable formula of the Triphala recipe in the traditional use.

Validated Reversed-Phase Ion-Interaction High-Performance Liquid Chromatography for Quantitation of Nitrate Content of Clausena anisata (Willd.) Hook. f. ex Benth. Leaves.

This work sought to validate the reversed-phase ion-interaction high-performance liquid chromatography for quantifying the nitrate content in the extract and raw material of Clausena anisata (Willd.) Hook. f. ex Benth. leaves. Three extraction methods (i.e., decoction, infusion, and ultrasound-assisted extraction) were investigated and compared. Furthermore, the effect of the solid-to-solvent ratio and defatting was also evaluated. The validation result showed that the high-performance liquid chromatographic method had a linear response (R 2 = 0.9999) in the range of 1-50 µg/mL. The limit of detection and limit of quantitation were 0.25 µg/mL and 0.75 µg/mL, respectively. In addition, the method was specific, precise, and accurate. So the validated method was suitable for determination of the nitrate content in C. anisata leaves. Infusion of a nondefatted sample using a solid-to-solvent ratio of 1 : 10 gave the highest nitrate content in the raw material, 0.251 ± 0.003%. In case of a defatted sample, decoction provided the highest nitrate content, 0.309 ± 0.001%. Increasing the solid-to-solvent ratio and defatting had a huge effect on the nitrate content of C. anisata leaves extracted from decoction. To the best of our knowledge, this is the first report that used the reversed-phase ion-interaction high-performance liquid chromatography for quantifying the nitrate content in C. anisata leaves. Furthermore, the authors suggested that nitrate could be used as a standard marker for quality control of C. anisata leaves' extract and raw material.

Optimal condition of cannabis maceration to obtain the high cannabidiol and Δ9-tetrahydrocannabinol content.

The aim of this work was to optimize a maceration condition of cannabis (Cannabis sativa L.). A circumscribed central composite experimental design was applied in this work. Temperature and time were varied from 40-80 °C and 30-90 min, respectively. The three responses (i.e., extraction yield, cannabidiol content, and Δ9- tetrahydrocannabinol content) were predicted by computer software. The yield was high when cannabis was macerated using ethanol at high temperature and long duration time. While cannabidiol and Δ9- tetrahydrocannabinol content was high when macerating at a low heating temperature and short duration time. The optimal condition provided the simultaneous high of cannabidiol and Δ9- tetrahydrocannabinol content was 40 °C for 30 min. The prediction was accurate due to low percent error. This optimal condition could be used as a guide for maceration of cannabis to obtain the extract containing a high content of cannabidiol and Δ9- tetrahydrocannabinol.

Convection combined microwave drying affect quality of volatile oil compositions and quantity of curcuminoids of turmeric raw material

Abstract The aim of the work was to estimate the quality of some compositions in turmeric volatile oil and quantity of individual and total curcuminoids in turmeric powder dried under different conditions. Effect of convection time (0-30 min) and microwave time (20-60 min) on quality of volatile oil compositions and quantity of curcuminoids were investigated using computer software. Quality of volatile oil obtained from the hydrodistillation of dried turmeric was analyzed by gas chromatography-mass spectrometry. The quantity of individual curcuminoids (bisdemethoxycurcumin, demethoxycurcumin, and curcumin) and total curcuminoids were analyzed by high performance liquid chromatography. Ten volatile compounds in turmeric volatile oil were used to estimate the variation of their quality. Results showed that ar-turmerone, turmerone, and curlone were the three major compounds found in turmeric volatile oil. The quality of the ten volatile compounds varied depending on convection time and microwave time. The three principal curcuminoids were found in turmeric dried at long convection time and medium microwave time. However, curcumin was also found in high amount in turmeric dried at short convection time and long microwave time. Total curcuminoids in dried turmeric were equal to or greater than 5% (w/w) as stated in the Thai Herbal Pharmacopoeia. It was found that almost all of the drying procedure achieved the standard of the Thai Herbal Pharmacopoeia except at short convection time and short microwave time. In summary, convection combined with microwave drying affected the quality of volatile oil compositions and quantity of curcuminoids of turmeric raw material.

Effect of Temperature and Duration Time of Maceration on Nitrate Content of Vernonia cinerea (L.) Less.: Circumscribed Central Composite Design and Method Validation.

Vernonia cinerea (L.) Less. is a herbal plant in Family Asteraceae. It can be used as a smoking cessation aid due to the fact that it contains nitrate which can induce tongue numbness and cause less favor of cigarette smell and taste. The aim of this work was to investigate the effect of maceration temperature and time on the nitrate content of V. cinerea. A circumscribed central composite experimental design was applied in the work. Two factors (temperature and duration time) were investigated and two responses (yield of the extraction and nitrate content) were monitored. The high performance liquid chromatography using for quantitative analysis of nitrate content was validated. The HPLC response was linear (R2 = 1.000) in range of 10-100 µg/mL. The HPLC method was specific, precise, and accurate. The maceration temperature and time were varied from 40 to 100°C and 10 to 60 min, respectively. Results showed that maceration at high temperature and long duration time gave the simultaneous high yield of the extraction and nitrate content. The prediction by the computer software, Design-Expert, was stable, reliable, and accurate. The optimal condition that provided simultaneous high yield of the extraction and nitrate content was achieved when extraction was at 99.5°C for 56.4 min. V. cinerea extracted using the optimal condition gave the yield of the extraction and nitrate content of 15.6% and 0.610%, respectively. In conclusion, maceration temperature and duration time had a positive effect on yield of the extraction and nitrate content of V. cinerea. Furthermore, the optimal condition in this work could be used as a guide for extraction of V. cinerea to obtain the high yield of the extraction as well as nitrate content.

Optimization of Caesalpinia sappan L. heartwood extraction procedure to obtain the highest content of brazilin and greatest antibacterial activity.

OBJECTIVE: The objective of the work was to optimize the extraction conditions of Caesalpinia sappan L. heartwood in order to maximize the brazilin content and antibacterial activity of the extract. METHODS: Two independent factors were studied: extraction temperature (45-95 °C) and extraction time (30-60 min). In addition, five dependent factors were monitored, including extraction yield, brazilin content, and clear zones against Staphylococcus aureus TISTR 1466, Staphylococcus epidermidis TISTR 518 and Propionibacterium acnes DMST 14961. The brazilin content was quantified by high-performance liquid chromatography and antibacterial activity was determined by disk diffusion assay. RESULTS: The high temperature provided high total extract yield as well as brazilin content, while extraction time had little effect on yield or brazilin content. Extraction time had a positive effect, while extraction temperature had little effect on clear zone against S. aureus. The largest clear zone against S. epidermidis was achieved at low extraction temperature and long extraction time. Conversely, short extraction time and high extraction temperature provided the largest clear zone against P. acnes. The optimal conditions providing the highest brazilin content was an extraction temperature and extraction time of 95 °C and 30 min, respectively. The same optimal conditions also provided the simultaneous greatest antibacterial activity against the three bacteria. Modeled optimal conditions were validated be conducting extraction using these values. Yield and antibacterial activity of the resulting extract demonstrated that the model had a low percentage error. CONCLUSION: The optimal condition will be used as a standard condition for extraction of C. sappan heartwood to maximize brazilin content and antibacterial activity.

An optimization approach of dynamic maceration of Centella asiatica to obtain the highest content of four centelloids by response surface methodology

ABSTRACT Centella asiatica (L.) Urb., Apiaceae, is commonly used as food, food supplement, and medicine. Development of the extraction process to obtain the high extent of the active compound is necessary. So, the response surface methodology was used in this work to optimize the dynamic maceration of C. asiatica to obtain the highest content of the four centelloids including asiatic acid, madecassic acid, asiaticoside, and madecassoside. Two factors: extraction temperature and extraction time, were studied. The content of four centelloids was observed. After the extraction of C. asiatica using ethanol, the content of four centelloids was analyzed using validated high performance liquid chromatography. The optimization result showed that madecassoside and asiaticoside had a similar pattern of the contour plots and response surfaces. These two centelloids were highly extracted at a high extraction time and high extraction temperature. The other two centelloids had the same pattern, they had a high content at a high temperature and time as well as at a low temperature and time. The simultaneous highest content of four centelloids was achieved when extracted at 60 °C for 120 min. The optimal condition could be used as standard condition for extraction of C. asiatica to provide the highest content of four centelloids.

Validated high performance liquid chromatography for simultaneous determination of stability of madecassoside and asiaticoside in film forming polymeric dispersions

ABSTRACT The objective of the work was to validate the high performance liquid chromatography for simultaneous determination of stability of madecassoside and asiaticoside in Centella asiatica (L.) Urb., Apiaceae, extract-loaded film forming polymeric dispersions. High performance liquid chromatography method was validated in five topics: linearity and range, limit of detection and limit of quantitation, specificity, precision, and accuracy. Results showed the method had a good linearity (R2 > 0.9990) in the range of 5-150 µg/ml and specific. The limit of detection and limit of quantitation of madecassoside were 81 and 245 ng/ml and asiaticoside were 21 and 64 ng/ml, respectively. The percent relative standard deviation of intraday and interday precision were less than 1 and 3%, respectively. The accuracy presented as percent recovery was 101.54-103.29% for madecassoside and 100.39-102.58% for asiaticoside. This validated high performance liquid chromatography method was used to determine the stability of the formulation containing Centella asiatica extract. Centella asiatica extract-loaded film forming polymeric dispersions used Eudragit® RS 30D and Eudragit® RL 30D as film former, glycerin as plasticizer, and absolute ethanol as solvent and penetration enhancer. Three formulations with different ratio of Eudragit® RS 30D and Eudragit® RL 30D were prepared and stored for 90 days at 4 ºC, 25 ºC, and 40 ºC. Stability results showed that almost all of the formulations were unstable at 25 ºC and 40 ºC. Except, two of three formulations were stable at 4 ºC. However, the formulation was further developed to improve the stability of madecassoside and asiaticoside in the formulation.