Anti Quorum Sensing Activity of Kayu Manis Leaves Extracts (Cinnamomun Burmannii Ness. Ex Bl.) Against Pseudomonas Aeruginosa

Quorum sensing is a communication system among bacterial cells which correlates with biofilm formation. Biofilm can protect bacteria from environment including antibiotic of which can cause higher antibiotic concentration of 100 up to 1000 times. Inhibition of quorum sensing is expected to inhibit the biofilm formation. The cinnamon bark (Cinnamomum burmanii Ness. Ex Bl.) has been known to have antibacterial and antibiofilm activities. Leaves are available abundantly which urges a research to find out the activity as anti quorum sensing against Pseudomonas aeruginosa. The succesive maceration of dried pulverized leaves produced hexane, ethyl acetate, and methanol extracts. Antibacterial activity was observed by microdillution method with MTT assay. Afterwards, the active extract was examined for anti quorum sensing activity by diffusion method in cetrimide Agar. Quorum sensing activity was shown by dark zone (opaque) growth around sample application, observed under UV light of 366 nm. TLC bioautography method was done to observe the active spots by using silica gel F254 as the stationary phase, chloroform-methanol (6:1 v/v) as the mobile phase, loading sample used was 1.25 mg and 30 min of plate contact duration. The ethyl acetate extract inhibited growth of P. aeruginosa with shown by MIC at 8 µg/µL Quorum sensing as well as growth inhibition activities were observed at loading samples 12,5 and 25 mg/wells, while at 6,25 mg the extract only exhibited quorum sensing inhibition. Presences of substances having phenolic, flavonoid, alkaloid and aldehyde/keton as functional groups were detected by TLC method of the extract but no active spot identified on bioautography

Perbandingan Daya Antiquorum Sensing Ekstrak N-heksan, Etil Asetat Dan Metanol Kulit Batang Krangean (Litsea Cubeba (Lour.) Pers.) Terhadap Pseudomonas Aeruginosa

Quorum sensing adalah suatu bentuk komunikasi bakteri yang membantu mengatur perilaku koloni bakteri. Quorum sensing merupakan mekanisme komunikasi berdasarkan ekspresi gen dan populasi bakteri yang mempengaruhi perkembangan biofilm, pompa effluks, produksi toksin, dan faktor virulen lainnya. Quorum sensing inhibitor mengurangi patogenisitas organisme, mengurangi sifat virulen organisme, dan membantu sistem imun untuk membersihkan infeksi bakteri. Quorum sensing inhibitor dapat dikombinasi dengan antibiotik untuk membersihkan patogen yang persisten. Minyak atsiri kulit batang Krangean (Litsea cubeba) diketahui dapat menghambat pertumbuhan bakteri dan pembentukan biofilm Streptococcus mutans. Karena quorum sensing berperan dalam pembentukan biofilm maka dilakukan penelitian aktivitas daya antiquorum sensing kulit batang Krangean terhadap Pseudomonas aeruginosa. Penentuan kadar hambat minimal dari ekstrak n-heksan, etil asetat, dan metanol dilakukan dengan metode mikrodilusi. Dari hasil penelitian, didapatkan hasil bahwa ekstrak etil asetat merupakan ekstrak yang paling aktif terhadap Pseudomonas aeruginosa dengan kadar hambat minimal 8µg/µl. Ekstrak aktif etil asetat kemudian diuji daya antiquorum sensing dengan metode sumuran. Hasilnya menunjukkan bahwa ekstrak aktif etil asetat dengan loading sampel 25 mg per sumuran, memiliki aktivitas antibakteri dan hambatan produksi pioverdin. Senyawa aktif diidentifikasi dengan KLT kemudian dilakukan uji bioautografi. Active compound group was unable to be determined by TLC then bioautography assay. Golongan senyawa yang terdapat di dalam ekstrak etil asetat yaitu alkaloid dan fenolik

Optimatization of Sucrose and Aspartame Composition as Sweetening Agent in Mengkudu Fruit Ethanolic Extract Effervescent-tablet Formulation

Mengkudu (Morinda citrifolia L.) has been widely used as traditional medicine. Its unpleasant smell and flavor urge a more acceptable dosage formulation. The aim of this research was to optimize the composition of sucrose and aspartame as sweetening agent in effervescent tablet formulation by using Simplex Lattice Design method. Effervescent tablets were produced by fusion method in five (5) different formulas, i.e. formula I (100% sucrose), II (suucrose-aspartame=75%:25%), III (sucrose-aspartame=50%:50%), IV (sucrose-aspartame=25%:75) dan V (100% aspartame). Effervescent granules were evaluated for mass density, flowing time, tapping index and compactibility characteristics. The effervescent tablets were tested for weight uniformity, hardness, friability, and disintegration time characteristics as well as TLC profile chromatogram. Data was analyzed by one way ANOVA, Scheffe method and Kruskall-Wallis with significance level 95%. The tablet acceptability was tested among 30 respondents. The results showed that the different composition of sucrose-aspartame influence the physical characteristics of granules and tablets effervescent produced. More sucrose content will increase the hardness, lower the friability but prolong the disintegration time. 70% respondents chose formula III as the best formula. Evaluation of SLD data recommended sucrose and aspartame in 42:58 proportion as the most optimum formula

A novel reversed phase high performance liquid chromatography method to accurately determine low concentrations of curcumin in rat plasma

Due to its lipophilic nature, curcumin levels in plasma are very low after oral administration, and therefore hard to detect. A number of chromatographic methods, including LC/MS have been developed. Although the LC/MS method is sensitive, the matrix effect can be difficult to handle. Furthermore, LC/MS equipment is relatively expensive compared to the conventional RP-HPLC. Therefore, the aim of this study was to develop a sensitive and reliable method for the determination of curcumin concentration in plasma using an RP-HPLC. Curcuma longa extract was used which contains three curcuminoids. The method started by selection of mobile phase to optimally separate the curcuminoids. The best mobile phase composition was used to analyze the plasma samples. Rat plasma was spiked with curcumin and processed for protein precipitation followed by liquid-liquid extraction of curcuminoids and an internal standard, emodin. Chromatogaphic separation of curcuminoids and emodin was achieved using a Knauer C-18 column (250 x 4.6 mm; particle size: 5µm) and a gradient program of mobile phase of three solvents, methanol -acetonitrile-1% acetic acid. A gradient elution was applied with increasing the ratio of the volume percentages of acetonitrile to acetic acid from 50/45 to 53/42 during 5 minute and thereafter elution was isocratic for 15 minute. The methanol concentration was kept constant at 5 vol-% during the whole run. The method was validated according to the FDA guidelines.The method was selective, with an excellent resolution (Rs value > 2.5). The peak shape of both curcumin and emodin were symmetric with a tailing factor of 0.9-1.1. The method linearity (correlation coefficient of 0.999) was demonstrated at 6 to 200 ng/mL. The intra and inter-day precision was 5.90-8.50% and 5.37-11.26%, respectively; the intra- and inter-day accuracy was 92.47-103.61% and 96.17-105.70%, respectively. In conclusion, the RP-HPLC method meets the validation requirements as described in the FDA guidelines and is applicable to accurately quantify curcumin concentrations as low as 6 ng/mL in rat plasma samples

A novel reversed phase high performance liquid chromatography method to accurately determine low concentrations of curcumin in rat plasma

Due to its lipophilic nature, curcumin levels in plasma are very low after oral administration, and therefore hard to detect. A number of chromatographic methods, including LC/MS have been developed. Although the LC/MS method is sensitive, the matrix effect can be difficult to handle. Furthermore, LC/MS equipment is relatively expensive compared to the conventional RP-HPLC. Therefore, the aim of this study was to develop a sensitive and reliable method for the determination of curcumin concentration in plasma using an RP-HPLC. Curcuma longa extract was used which contains three curcuminoids. The method started by selection of mobile phase to optimally separate the curcuminoids. The best mobile phase composition was used to analyze the plasma samples. Rat plasma was spiked with curcumin and processed for protein precipitation followed by liquid-liquid extraction of curcuminoids and an internal standard, emodin. Chromatogaphic separation of curcuminoids and emodin was achieved using a Knauer C-18 column (250 x 4.6 mm; particle size: 5µm) and a gradient program of mobile phase of three solvents, methanol -acetonitrile-1% acetic acid. A gradient elution was applied with increasing the ratio of the volume percentages of acetonitrile to acetic acid from 50/45 to 53/42 during 5 minute and thereafter elution was isocratic for 15 minute. The methanol concentration was kept constant at 5 vol-% during the whole run. The method was validated according to the FDA guidelines.The method was selective, with an excellent resolution (Rs value > 2.5). The peak shape of both curcumin and emodin were symmetric with a tailing factor of 0.9-1.1. The method linearity (correlation coefficient of 0.999) was demonstrated at 6 to 200 ng/mL. The intra and inter-day precision was 5.90-8.50% and 5.37-11.26%, respectively; the intra- and inter-day accuracy was 92.47-103.61% and 96.17-105.70%, respectively. In conclusion, the RP-HPLC method meets the validation requirements as described in the FDA guidelines and is applicable to accurately quantify curcumin concentrations as low as 6 ng/mL in rat plasma samples