SCREENING OF ANTIOXIDANT ACTIVITY AND TOTAL PHENOLIC CONTENT IN RAPHANUS SATIVUS POD

Objective: The aim of this study was to evaluate the antioxidant activity and total phenolic content of Raphanus sativus L. Var. caudatus Alef pod extracts. Methods: In this study, the extract of R. sativus pod was prepared in three different types of solvent. The antioxidant activitiy of R. sativus extract was determined using a spectrophotometric technique, based on a free radical diphenyl-picryhydrazyl scavenging assay (DPPH assay) and a ferric reducing antioxidant power assay (FRAP assay). Total phenolic content was also observed using the spectrophotometric technique. Results: The result showed that the antioxidant activity which was expressed by IC50 values varied from 1,365 to 4,371 mg/ml and 312 to 6,478 mg/ml, based on DPPH assay and FRAP assay, respectively. Total phenolic content was also evaluated and calculated as gallic acid equivalents which ranged from 0.26 to 34.60 mg gallic acid per 100 g fresh sample. Conclusion: It was suggested that hexane extract of R. sativus pod contained the highest amount of phenolic compounds in comparison with those of dichloromethane part and ethanol part. The result from FRAP assay was positively correlated to total phenolic content which the highest antioxidant value belongs to the hexane extract of R. sativus pod. It was concluded that R. sativus pod contained phenolic compounds which showed mild antioxidant activity

Development and Validation of Oseltamivir Carboxylate

Objective: To develop a simple HPLC analysis for the quantification of oseltamivir carboxylate (OC) and  oseltamivir phosphate (OP) in human plasma.  Method: Chromatographic separation was carried out using Hypersil BDS cyano 5 μm column (length 250 mm, inner diameter 4.6 mm) with UV detection at 230 nm. The gradient system composed of 50 mM ammonium acetate: acetonitrile (95:5) for 4 min, followed by 50 mM ammonium acetate: acetonitrile (70:30) for 7 min, and finally 50 mM ammonium acetate: acetonitrile (95:5) for 4 min.  Results: Coefficients of determination for the quantification of OC and OP were 0.9994 and 0.9991, respectively, with retention times of 4.35 and 11.04 min, respectively. The method limit of quantification for both compounds was 1.5  μg/ml. Coefficients of variation for the between- and within-day precision of OC were 2.15 - 7.19% and 1.97 - 6.63%, respectively, and those of OP were 2.41 - 8.52% and 1.90 - 8.57%, respectively. Conclusion: A simple and reliable HPLC quantification method of oseltamivir carboxylate and oseltamivir phosphate in human plasma was successfully validated. Keywords: oseltamivir carboxylate, oseltamivir phosphate, HPLC, method validatio

DEVELOPMENT AND VALIDATION OF LC-MS METHOD FOR QUANTITATIVE ANALYSIS OF A TRADITIONAL THAI ANTIHYPERTENSIVE HERBAL RECIPE

Objective: To develop a new validated LC-MS method in which can be used for identification and quantification of the active pharmaceutical compounds in a traditional Thai antihypertensive herbal recipe (TTAH). Methods:  First, a quadrupole-ion trap MS was used to identify the active pharmaceutical compounds to facilitate a set of markers in the TTAH using multiple reactions monitoring (MRM) method. Coupling to LC in gradients mode, the chromatogram of the TTAH was established. Second, the method validation was conducted to observe several parameters, such as, carryover, linearity, a limit of detection, a limit of quantification, precision, accuracy, robustness, specificity, and system suitability. Results: Piperine, imperatorin, and pinostrobin were identified using the quadrupole-ion trap MS by comparison the mass spectrum of the TTAH samples and reference standards. The parent and product ions were optimized using MRM method. Good chromatographic peaks were achieved along with a simple and fast analysis. All parameters were validated and found that the method reported in this article can be used to quantify the amount of piperine, imperatorin, and pinostrobin in the TTAH. Conclusion: Piperine, imperatorin, and pinostrobin were selected as the markers in LC-MS analysis of the TTAH. This new validated LC-MS method is readily to use in a quality assurance of the TTAH

Antioxidant Activity and Lipid-Lowering Effect of Essential Oils Extracted from Ocimum sanctum L. Leaves in Rats Fed with a High Cholesterol Diet

It has been reported that Ocimum sanctum L. (OS) leaves decrease serum lipid profile in normal and diabetic animals. No experimental evidences support the anti-hyperlipidemic and antioxidative actions against hypercholesterolemia. Moreover the identity of the specific chemical ingredients in OS leaves responsible for these pharmacological effects are unknown. Since OS leaves are rich in essential oil (EO). Therefore the present study was conducted to investigate the anti-hyperlipidemic and antioxidative activities of EO extracted from OS leaves in rats fed with high cholesterol (HC) diet. EO was extracted by the hydrodistillation method and the chemical constituents were then identified by Gas Chromatography-Mass Spectrometry. The experiment was performed in Male Wistar rats fed with 2.5 g%(w/w) of cholesterol diet for seven weeks. During the last 3 weeks, rats were daily fed with EO. The results showed that phenyl propanoid compounds including eugenol and methyl eugenol were the major constituents of EO. EO suppressed the high serum lipid profile and atherogenic index as well as serum lactate dehydrogenase and creatine kinase MB subunit without significant effect on high serum levels of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase in rats fed with HC diet. In addition, EO was found to decrease the high levels of thiobarbituric acid reactive substances (TBARS), glutathione peroxidase (GPx) and superoxide dismutase (SOD) without impacting catalase (CAT) in the cardiac tissue while in the liver, it decreased high level of TBARS without significantly effecting GPx, SOD and CAT. Histopathological results confirmed that EO preserved the myocardial tissue. It can be concluded that EO extracted from OS leaves has lipid-lowering and antioxidative effects that protect the heart against hypercholesterolemia. Eugenol that is contained in EO likely contribute to these pharmacological effects

Lipid-Lowering and Antioxidative Activities of Aqueous Extracts of Ocimum sanctum L. Leaves in Rats Fed with a High-Cholesterol Diet

The present study was conducted to investigate the lipid-lowering and antioxidative activities of Ocimum sanctum L. (OS) leaf extracts in liver and heart of rats fed with high-cholesterol (HC) diet for seven weeks. The results shows that OS suppressed the high levels of serum lipid profile and hepatic lipid content without significant effects on fecal lipid excretion. Fecal bile acids excretion was increased in HC rats treated with OS. The high serum levels of TBARS as well as AST, ALT, AP, LDH, CK-MB significantly decreased in HC rats treated with OS. OS suppressed the high level of TABARS and raised the low activities of GPx and CAT without any impact on SOD in the liver. As for the cardiac tissues, OS lowered the high level of TABARS, and raised the activities of GPx, CAT, and SOD. Histopathological results show that OS preserved the liver and myocardial tissues. It can be concluded that OS leaf extracts decreased hepatic and serum lipid profile, and provided the liver and cardiac tissues with protection from hypercholesterolemia. The lipid-lowering effect is probably due to the rise of bile acids synthesis using cholesterol as precursor, and antioxidative activity to protect liver from hypercholesterolemia

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 to 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 disintegrating wafer tablets containing cannabis extract with desired properties