Spectroscopic characterization and pharmacokinetic evaluation of amorphous solid dispersions of glibenclamide for bioavailability enhancement in Wistar rats.

Oral bioavailability of glibenclamide (Glb) was appreciably improved by the formation of an amorphous solid dispersion with Poloxamer-188 (P-188). Poloxamer-188 substantially enhanced the solubility and thereby the dissolution rate of the biopharmaceutics classification system (BCS) class II drug Glb and simultaneously exhibited a better stabilizing effect of the amorphous solid dispersion prepared by the solvent evaporation method. The physical state of the dispersed Glb in the polymeric matrix was characterized by differential scanning calorimetry, X-ray diffraction, scanning electron microscope and Fourier transform infrared studies. In vitro drug release in buffer (pH 7.2) revealed that the amorphous solid dispersion at a Glb-P-188 ratio of 1:6 (SDE4) improved the dissolution of Glb by 90% within 3 h. A pharmacokinetic study of the solid dispersion formulation SDE4 in Wistar rats showed that the oral bioavailability of the drug was greatly increased as compared with the market tablet formulation, Daonil®. The formulation SDE4 resulted in an AUC0-24h ~2-fold higher. The SDE4 formulation was found to be stable during the study period of 6 months.

Monitoring and method validation of organophosphorus/organochlorine pesticide residues in vegetables and fruits by gas chromatography.

In this study, an analytical method was developed and validated for the assessment of pesticide residues in commonly consumed vegetables and fruits. Fresh samples of apple, green peas, tomatoes, and cucumbers were processed and subjected to analysis using a modified QuEChERS (quick, easy, cheap, effective, rugged, safe) extraction technique. Subsequently, quantification of pesticide residues was conducted utilizing gas chromatography (GC)-electron capture detector. Extraction and cleanup parameters were meticulously optimized, resulting in a modification of the original QuEChERS method. This modification aimed to reduce solvent consumption, making the study more environmentally friendly. The developed method was validated in terms of selectivity, specificity, linearity, precision, and accuracy by following the SANTE guidelines. Calibration curves showed good linearity (r > 0.99) within the test range. Precision was evaluated by intra- and inter-day experiments with an acceptable relative standard deviation (<20.0%). Recovery was assessed at the limit of quantification level and was observed to fall within the range of 70%-120%, with relative standard deviations below 5.45%. The validated method presented here can be applied to analyze pesticide residues in various other vegetables, fruits, and cereals. It is essential for ongoing monitoring of pesticide residues to ensure public safety.

Validation and development of RP-HPLC method for quantification of glibenclamide in rat plasma and its application to pharmacokinetic studies in wistar rats.

Herein, we report a novel, simple, specific, accurate and cost-friendly validated reverse phase-high performance liquid chromatographic (RP-HPLC) method for the quantification of second generation sulphonylurea based antidiabetic drug, glibenclamide (GLB) in rat plasma and its application to calculate pharmacokinetic parameters in wistar rats. The internal standard used was flufenamic acid. The chromatographic separation was conducted on C18 column (250 mm × 4.6 mm x 5 µm, Agilent-Zorbax, SB) using isocratic elution with mobile phase containing Acetonitrile: Water (1:1; v/v) pH adjusted to 4.0 with 0.03 % glacial acetic acid and detected by photo-diode array as detector. Calibration curves made in the rat plasma were linear in the range of 50-1200 ng/ml with r2 = 0.998. The LLOQ was 40 ng/ml. This method was effectively applied for pharmacokinetic studies of Glibenclamide following administration through oral route as solid dispersion formulation to Wistar rats. Several methods are available in the literature which can be employed for the quantification of Glibenclamide but such methods are tedious, provide lower sensitivity, less simultaneous resolution and are time-consuming. Therefore the present methods suits best for the quantification of Glibenclamide from Wistar rats.

Dissipation kinetics, residue determination and consumer risk assessment of acephate on green pea by gas chromatography-electron capture detector.

The present study on "acephate persistence on green pea" was conducted in SKUAST-Kashmir. The study aimed to determine the persistence, dissipation kinetics and waiting period of acephate on green pea. Acephate was sprayed at 75% soluble powder (SP) at 560 g a.i.ha-1 at the fruiting stage followed by another application at a 10 day interval. A rapid and accurate method (quick, easy, cheap, effective, rugged and safe, QuEChERS) was used for extraction and the residue was determined by gas chromatography-electron capture detection on a CPSIL-8CB capillary column (0.25um film thickness, 0.25 mm i.d, 30 m length). At the fortification levels of 0.05, 0.1 and 0.5 mg kg-1 , the percentage recovery of acephate on green pea was found in the range of 71-107%. The initial deposit of green pea was estimated to be 0.37 mg kg-1 . At the indicated dose, the residue of acephate on green pea dissipated below the limit of quantification of 0.05 mg kg-1 after 10 days. Acephate degradation was quick in green pea, with a half-life of 4.07 days. For safe eating of green peas, a 10 day waiting period is recommended. The gas chromatography-electron capture detection technique was validated by following the SANTE standards.

Recent bioanalytical methods for the isolation of bioactive natural products from genus Codonopsis.

INTRODUCTION: Chromatography and spectroscopy are nowadays well-validated techniques allowing to isolate and purify different class of natural products from the genus Codonopsis. Several categories of phytochemicals with drug like properties have been selectively extracted, isolated, characterised by this methodology. OBJECTIVES: The present review aims to provide up-to-date and comprehensive information on the chromatography, phytochemistry and pharmacology of natural products of Codonopsis with an emphasis on the search for natural products having various biological activities and the semi-synthetic derivatives of bioactive ones and to highlight current gaps in knowledge. MATERIALS AND METHODS: A literature search was performed in the SciFinder Scholar, PubMed, Medline, and Scopus databases. RESULTS: During the period covered in this review, several classes of compounds have been reported from genus Codonopsis. Codonopsis pilosula and Codonopsis lanceolata are the most popular in the genus especially as per phytochemical and bioactive studies. Phytochemical investigation demonstrates that Codonopsis species contain mainly xanthones, flavonoids, alkaloids, polyacetylenes, phenylpropanoids, triterpenoids and polysaccharides, which contribute to numerous bioactivities. The major bioactive compounds isolated were used for semi-synthetic modification to increase the chance to discover lead compound. CONCLUSIONS: It can be concluded that genus Codonopsis has been used as traditional medicines and food materials around the world over years due to chemical constituents with diverse structural types, exhibiting extensive pharmacological activities in immune system, blood system, cardiovascular system, central nervous system, digestive system, and so forth, with almost no obvious toxicity and side effect. Therefore, Codonopsis can be used as a promising ethnopharmacological plant source.

Development and validation of gas chromatography with electron capture detection method using QuEChERS for pesticide residue determination in cucumber.

A modified quick, easy, cheap, efficient, rugged, and safe (QuEChERS) method coupled to gas chromatography with electron capture detection was developed for the simultaneous determination of selected electronegative pesticides, namely, chlorpyrifos-methyl (1), chlorpyrifos (2), quinolphos (3), profenofos (4), myclobutanil (5), ethion (6), fenpropathrin (7), and cypermethrin (8), in vegetables with high water content. The selected compounds and some of their metabolites have even been found in human body fluids. In addition, some of them are known or suspected carcinogens according to the World Health Organization. Extraction and cleanup parameters were optimized; thus, the original QuEChERS method was modified to minimize solvent usage by making the study eco-friendly. The developed method was validated for selectivity, specificity, linearity, precision, and accuracy using SANTE guidelines. Calibration curves showed good linearity (r > 0.99) within the test range. Precision was evaluated by intra- and inter-day experiments with an acceptable range of less than 20.0% of relative standard deviation. Recovery was evaluated at limit of quantification and was found to be in the range of 70-120%, with relative standard deviations lower than 4.21%. The proposed method is applicable for detection and monitoring of selected pesticides in one run not only in fruits and vegetables with high water content but also in samples containing large quantities of pigments/dyes.

Simultaneous quantification of losartan potassium and its active metabolite, EXP3174, in rabbit plasma by validated HPLC-PDA.

Herein, we report a novel, accurate and cost-effective validated analytical method for the quantification of losartan potassium and its active metabolite, EXP 3174, in rabbit plasma by reversed-phase high-performance liquid chromatography. Valsartan was used as an internal standard. The method was validated as per International Conference on Harmonization guidelines. The analytes were extracted in rabbit plasma using liquid-liquid extraction technique and analyzed at 247 nm after separation through a reverse-phase C18 column. The isocratic mobile phase used is a mixture of acetonitrile, water and glacial acetic acid in the ratio of 60:40:1 v/v/v maintained at pH 3.4. All calibration curves showed a good linear relationship (r > 0.995) within the test range. Precision was evaluated by intra- and interday tests with RSDs <1.91% and accuracy showed validated recoveries of 86.20-101.11%. Based on our results, the developed method features good quantification parameters and can serve as an effective quality control method for the standardization of drugs.

Method validation and residue determination of mineral oil Arbofine in apple and soil using gas chromatography-flame ionisation detection under temperate conditions of Kashmir, India.

This innovative study was carried out to determine the presence of the mineral oil Arbofine in apple and soil at four locations. Arbofine kills the vast majority of dormant insects and mites (mite and asphid eggs, scales and psyllids) on fruit trees (cherry, apple, plum and peach) and thus reduces the plant diseases in summer. In this study, the mineral oil was sprayed at recommended doses of 2.0 and 0.75%, and the doses were doubled to 4.0 and 1.5% in dormant and summer seasons, respectively. The soil samples were taken for observation during the dormant season, whereas both soil and apple samples were taken during the summer season after treatment for 0, 1, 3 and 5 days. The recovery study of all the 11 paraffinic constituents (n-pentane, n-hexane, n-heptane, n-octane, n-nonane, n-decane, n-undecane, n-dodecane, n-tridecane, n-tetradecane and n-pentadecane) in soil and apple samples which constitutes 60% of mineral oil in soil and apple was carried out at the fortification level of 1.0 µg/ml, which was found to be between 72.1% and 99.0%. No residue of all the 11 paraffinic compounds of Arbofine mineral oil was detected in soil and apple samples at day 0 after the recommended doses, and the recommended doses were doubled in both seasons at four locations. Therefore, mineral oil can be used on apples without any risk.

Health Risk Assessment of Pesticide Residues in Drinking Water of Upper Jhelum Region in Kashmir Valley-India by GC-MS/MS.

Globally growing demand for agricultural and farm foods has more or less become dependent on chemical pesticides to maintain the supply chain, which undoubtedly boosts agricultural production. However, pesticides not only impact the target pests but cause hazard to human health. Pesticides are ubiquitous and can be found in almost every component of the environment. They can therefore impair human and biota health when present over the threshold level. The present study assessed the concentration of commonly used pesticides for agricultural purposes but get mixed in different sources of water, as such fifteen sampling sites along the upper Jhelum basin of Kashmir valley were chosen. For the analysis, 60 water samples were obtained from different water sources. Gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) was used to determine pesticide residues in water samples. Pesticide residues from 10 of the 26 commonly used pesticides were detected in water samples. Difenoconazole had the highest concentration among the pesticides detected, with a mean concentration of 0.412 ± 0.424 µg/L ranging from 0.0 µg/L to 0.8196 µg/L. The target hazards quotient (THQ) was used to quantify the possible noncarcinogenic health risks associated with drinking pesticide-contaminated water. Only chlorpyrifos and quinalphos were detected >1 in RWS3 (1.6571), RWS4 (1.0285), RWS14 (1.2571), and RWS15 (1.2000) sample sites, implying that the drinking water poses a health risk to humans. Hence, pesticide hazards should be mitigated and rigorous monitoring is needed to reduce pesticide residues in drinking water.

Development of an efficient micropropagation system for Dioscorea bulbifera L. and phytochemical profile of regenerated plants.

BACKGROUND: The present study was designed to develop an improved in vitro regeneration system for Dioscorea bulbifera using mature nodal explants. Direct organogenesis from nodal segments was achieved by culturing the nodal explants on Murashige and Skoog medium supplemented with 3.5 mgl-1 6-benzylaminopurine (BAP). Shoot multiplication was widely affected by the kind and concentration of plant growth regulators, and the sub-culturing of in vitro regenerated shootlets on fresh medium. After incubation for 4 weeks at optimum BAP concentration, cultures were transferred to secondary medium with BAP (optimized concentration) supplemented with different auxins. RESULTS: On medium with 3.5 mgl-1 6-benzylaminopurine, maximum regeneration (87 ± 1.85%) with an average shoot number of 2.0 ± 0.08, and length of 3.5 ± 0.04 cm were observed after 4 weeks of incubation. Maximum number of shoots (3.9 ± 0.21) was observed with 3.5 mgl-1 BAP in combination with 0.75 mgl-1 indole-3-acetic acid. The best root formation was observed on ½ MS medium supplemented with 0.75 mgl-1 α-naphthalene acetic acid, with 4.7 ± 0.03 roots per shoot. The well-rooted plantlets were successfully acclimatized in with 100% survival rate. The plantlets grew well with normal growth, flowering and showed, by High performance thin layer chromatography, almost same number of phytoconstituents compared with the mother plant. CONCLUSIONS: This is the first study on comparative phytochemical analysis of wild growing and in vitro regenerated plants of D. bulbifera which validates the medicinal and nutritional properties of in vitro raised plants.

Development, method validation and simultaneous quantification of eleven bioactive natural products from high-altitude medicinal plant by high-performance liquid chromatography.

Herein, a novel, rapid, reliable, simple method validation and simultaneous quantification of 11 bioactive compounds (mostly xanthones) have been described. International Conference on Harmonization guidelines were used for the analytical method validation. Good linearity, repeatability, intra- and inter-day precision, accuracy and reliability were well-illuminated in the method validation procedure. The calibration curves showed a good linear relationship (r > 0.999) within test range. Precision was evaluated by intra- and inter-day tests with relative standard deviation <2.79% and accuracy validation recovery of 74.16%-91.84%. On quantification study, the validated method described the high content of bioactive xanthone derivatives, including 1-hydroxy-3, 5-dimethoxyxanthone (7), 2-(allyloxy)-8-hydroxy-1, 6-dimethoxyxanthone (6) 1, 7, 8-trihydroxy-3-methoxyxanthone (9) and Coxanthone E (5) in Codonopsis ovata, which is advantageous given the numerous pharmacological and biological effects associated with these compounds, which mostly exhibit anti-cancerous, antioxidant, anti-inflammatory, anti-mutagenic and anti-obesity effects. The bulk abundance of these compounds can also be used for further modification to produce better lead molecules for drug discovery with low toxicity and high potency. The proposed method makes it possible to simultaneously determine all bioactive compounds in one run and can be extended to marker-based standardization of herbal formulations in medicinal and pharmaceutical industries.

Risk assessment, development and validation of a GC-ECD-based method for the quantification of cypermethrin from green pea.

The present work describes the persistence, dissipation behaviour, half-life, risk assessment and novel gas chromatography method for the residue estimation of cypermethrin in green pea by spraying cypermethrin 10EC at 50 g a.i. ha-1 at fruiting stage followed by another application at a 10 day interval. The sample extraction and cleanup was followed bya modified quick, easy, cheap, effective, rugged, and safe method, and the residues of cypermethrin were determined using a validated gas chromatography method. The initial deposits were found to be 1.21 mg kg-1 following the application of insecticide at 50 g a.i. ha-1 . Cypermethrin residues declined to below the detection limit of 0.05 mg kg-1 after 15 days at the recommended dosage. The half-life of cypermethrin was 2.66 days at 50 g a.i. ha-1 . For risk assessment studies, the waiting period of 15 days is recommended as safe for consumption for the insecticide. The GC-ECD method was validated according to the SANTE guidelines by various analytical parameters including linearity, accuracy, detection and quantification limits. The developed method is simple, selective and repeatable, and can be used for the standardization of pesticides on fruits and vegetables.

Dissipation and gas chromatographic method for the determination of profenofos residues in/on green pea and cucumber.

Herein we report a novel, accurate and cost-effective gas chromatography method for the determination of average deposits of profenofos on green pea and cucumber following good agricultural practices. Additionally the risk assessment, dissipation and waiting period for profenofos were determined. The average initial deposits (2 h after spraying) of profenofos in/on green pea and cucumber were 3.41 and 3.62 mg kg-1 respectively following two applications at a 10 day interval of profenofos 50EC formulation. Profenofos residues on both of the substrates were below the detection limit of 0.05 mg kg-1 after 20 days at the recommended dosage. For risk assessment studies, the 20th day will be safe for consumers for consumption of green peas. The gas chromatography method was validated according to the SANTE guidelines using the various analytical parameters: linearity, accuracy, detection and quantification limits. The developed method is simple, selective and repeatable and can be extended for profenofos-based standardization of pesticide formulations for green pea/cucumber and their use as pesticides.

Method validation and simultaneous quantification of eight organochlorines/organophosphates in apple by gas chromatography.

BACKGROUND: The increasing and extensive use of pesticides worldwide has resulted in a significant loss of non-target populations particularly humans by direct or indirect exposures. Also, various methods have been used for the estimation of pesticide residues in fruits and vegetables from recent past which are either tedious, time consuming or expensive. Therefore, the present study was performed to determine the pesticide residues from apple by simple and novel validated gas chromatography. RESULTS: A novel, accurate, ecofriendly and cost-effective gas chromatography method was developed for simultaneous quantification of eight pesticides, namely chlorpyrifos-methyl (1), chlorpyrifos (2), quinolphos (3), profenofos (4), myclobutnil (5), ethion (6), fenpropathrin (7) and cypermethrin (8). The developed method was validated as per the SANTE guidelines. All calibration curves showed a good linear relationship (r > 0.99) within the test range. Precision was evaluated by intra- and inter-day tests with relative standard deviations (RSDs) < 2.0%, recovery in between 70% and 120% with RSDs < 2.00%. CONCLUSION: The results demonstrate that the concentration of pesticides 1 to 8 were found below the detectable limit. Method validation parameters like linearity, precision, accuracy, specificity, robustness, detection and quantification limits were found within the acceptable range. The proposed method makes it possible to determine simultaneously pesticides 1-8 in one run which can be extended for residue-based standardization of pesticides from apple and other fruits and vegetables. © 2019 Society of Chemical Industry.

Risk assessment, dissipation behavior and persistence of quinalphos in/on green pea by gas chromatography with electron capture detector.

Chemical investigation was carried out to examine the risk assessment, dissipation behavior, persistence, and half-life period of quinalphos in/on green pea fruit by spraying quinalphos at fruiting stage followed by another application after 10-day interval. The samples were extracted by using the quick, easy, cheap, effective, rugged, and safe method, and the residues of quinalphos were analyzed by gas chromatography with electron capture detector. Herein, we report a novel, accurate, and cost-effective gas chromatography method for the determination of average deposits of quinalphos in/on green pea. The initial deposits and half-life of quinalphos were found to be 1.20 mg/kg and 2.77 days, respectively, following the application of insecticide. Residues of quinalphos reached below detection limit of 0.05 mg/kg after 10 days at recommended dosage. For risk assessment studies, the tenth day will be safe for consumers for consumption of green pea. The developed method is simple, selective, and repeatable, and it can be extended for quinalphos-based standardization of herbal formulations containing green pea and its use in pesticide industries.

Quantification, dissipation behavior and risk assessment of ethion in green pea by gas chromatography-electron capture detector.

Residue investigation was carried out to scrutinize the persistence, dissipation behavior, half-life, and risk assessment of ethion on green pea fruit by spraying ethion at the fruiting stage followed by another application at 10 day intervals. The samples were extracted by using a quick, easy, low-cost, effective, rugged, and safe method, and the residues of ethion were analyzed by gas chromatography with electron capture detection. Here we report a novel, accurate, and cost-effective gas chromatography method for the determination of average deposits of ethion on green pea. The initial deposits were found to be 4.65 mg/kg following the application of insecticide. Residues of ethion reached below the detection limit of 0.10 mg/kg after 25 days at recommended dosage. The half-life of ethion was found to be 4.62 days. For risk assessment studies, the 25th day will be safe for consumers for the consumption of green peas. The developed method is simple, sensitive, selective, and repeatable and can be extended for ethion-based standardization of herbal formulations containing green pea and its use in pesticide industries.

Biologically active xanthones from Codonopsis ovata.

Five new xanthones, named coxanthones A-E (1-5), together with 21 known secondary metabolites (6-26) that include seven xanthones, five flavonoids, two steroids and seven triterpenoids were isolated from the chemically unexplored whole plant Codonopsis ovata. The structures of new metabolites were elucidated by HRMS, interpretation of NMR spectra and other spectroscopic techniques. The absolute configuration of the stereogenic centre of coxanthone B (2) was determined by electronic circular dichroism (ECD) spectroscopy. This is the first report of xanthones from the genus Codonopsis. All isolated metabolites were evaluated for cytotoxic activity by SRB assay against six human cancer cell lines A549 (lung), PC-3 (prostate), HCT-116 (colon), MCF-7 (breast), SF-295 (CNS), and MDAMB-435 (melanoma). Among the new compounds, coxanthone B (2) exhibited significant inhibitory activity against SF-295 and MDAMB-435 with IC50 values of 7.0 and 15.0 µM, respectively. Coxanthone A (1) displayed cytotoxicity against A549 cell line at IC50 value of 22.5 µM. Cytotoxic activity of 1-hydroxy-3,5-dimethoxyxanthone (7), swertiperenine (9) and 1,7,8-trihydroxy-3-methoxyxanthone (10) are reported here first time that exhibited the IC50 values of 3.0, 5.0 and 21.0 µM against A549, MDAMB-435, and A549 cell lines, respectively. Kaempferol (13) showed most potent cytotoxic activity with an IC50 values in the 1.0-2.3 µM range against all tested cancer cell lines.

Synthesis of α-santonin derived acetyl santonous acid triazole derivatives and their bioevaluation for T and B-cell proliferation.

A new series of α-santonin derived acetyl santonous acid 1,2,3-triazole derivatives were synthesised using Huisgen 1,3-dipolar cyclo-addition reaction (click chemistry approach) and evaluated for their in vitro inhibition activity on concanavalin A (ConA) induced T cell proliferation and lipopolysaccharide (LPS) induced B cell proliferation. Among the synthesised series, compounds 2-10 and 19 exhibited significant inhibition against ConA and LPS stimulated T-cell and B-cell proliferation in a dose dependent manner. More significantly compounds 4, 9-10 and 19 exhibited potent inhibition activity with remarkably lower cytotoxicity on the mitogen-induced T cell and B cell proliferation at 1 µM concentration. The compound 6 displayed potent immunosuppressive effects with ∼89% against LPS induced B-cell and ∼83% against ConA stimulated T-cell proliferation at 100 µM concentration without cytotoxicity. Compound 10 was more selective against B cell proliferation and exhibited 81% and 69% suppression at 100 and 1 µM concentration respectively. The present study led to the identification of several santonin analogs with reduced cytotoxicity and strong inhibition activity against the cell proliferation induced by the mitogens.

Isolation, cytotoxic evaluation, and simultaneous quantification of eight bioactive secondary metabolites from Cicer microphyllum by high-performance thin-layer chromatography.

Chemical investigation of Cicer microphyllum resulted in the isolation and characterization of eight natural products viz. Stigmasterol, Oleanolic acid-3-acetate, Oleanolic acid, Biochanin A, Genistein, Pratensein, Chrysoeriol, and Luteolin. Herein, we report a novel, accurate, and cost-effective high-performance thin-layer chromatography method for the simultaneous quantification of the isolated natural products on silica-gel 60F254 plates using the solvent system n-hexane/ethyl acetate/formic acid (9.0:6.5:0.8, v/v/v). Natural products were quantified after postchromatographic derivatization with ceric ammonium sulfate. The method was validated as per the International Conference on Harmonization guidelines. All calibration curves showed a good linear relationship (r > 0.9943) within the test range. Precision was assessed by intra- and interday tests with relative standard deviations <1.82%, accuracy validation recovery 98.38-99.57% with relative standard deviations <1.00%. On quantification, Pratensein was a major constituent (0.921%). The screening for cytotoxic activity using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay resulted into identification of Luteolin as potent molecule with IC50 3.5 and 25.6 µg/mL against murine melanoma and human epidermoid carcinoma cell lines, respectively.

Simultaneous quantification of eight bioactive secondary metabolites from Codonopsis ovata by validated high performance thin layer chromatography and their antioxidant profile.

Chemical investigation of Codonopsis ovata resulted in the isolation and identification of ß-sitosterol-3-O-glycoside, luteolin, apigenin, gentiacaulein, swertiaperenine, ß-sitosterol, taraxeryl-3-acetate, and 3ß-acetoxyoleanane-12-one. A rapid, precise, sensitive and validated HPTLC method for simultaneous quantification of these natural products (NPs) was developed on silica-gel 60F254 plate using ternary solvent system, n-hexane:ethyl acetate:formic acid (10.5:3.5:0.43, v/v/v). Markers were quantified after post chromatographic derivatization with cerric ammonium sulfate reagent. The method was validated for accuracy, precision, LOD, LOQ and all calibration curves showed a good linear relationship (r>0.9924) within test range. Precision was evaluated by intra- and inter-day tests with RSDs <2.59%, accuracy validation recovery 92.43-99.50% with RSDs <1.00%. Apigenin was found major component (natural abundance: 1.103%) and ß-sitosterol the least (0.0263%). The NPs displayed antioxidant activity with luteolin exhibiting maximum effect at 1µg/mL concentration (75.9% for DPPH and 43.7% for ABTS) and others at 10 and 25µg/mL, suggesting thereby their apparent potential use for the prevention of free radical induced diseases or as an additive element to food and pharmaceutical industry.