Search

The presence of various secondary metabolites in the crude plant extracts have always been a source of potential therapeutic activities in animals and humans. The varying phytochemical composition and concentrations are instrumental in deciding and shortlisting raw materials for extractions and isolation of active compounds. Therefore, preliminary phytochemical screening of the plant extracts along with their high-performance thin layer chromatography analysis for classes of compounds become prerequisite for any type of pharmacological study. Detailed profiling of secondary metabolites like cardiac glycosides, triterpenoids and steroids which are found to be instrumental in inducing contraception in male experimental model organisms will help us further understands the mechanism of natural male contraception. In this study, crude seed extracts of Carica papaya Linn., Momordica charantia Linn. and Abrus precatorius Linn. in suitable solvents were checked for the high-performance thin layer chromatography profiling of the above-mentioned metabolites. The results obtained were recorded and analysed for repeated authentication of plant parts and standardisation of extract preparation. [ABSTRACT FROM AUTHOR]

Deferasirox is used in the treatment of chronic iron overload due to blood transfusions. Many studies have reported the high-performance liquid chromatography, high-performance thin-layer chromatography and ultraviolet spectroscopic methods for the estimation of deferasirox. The current work is intended for the development of a stability indicating method by high-performance thin layer chromatography for the estimation of deferasirox. The chromatographic development was performed on aluminium plates coated with silica gel 60 F254 using chloroform:methanol:triethylamine (9:1:0.5 v/v/v) as the mobile phase. Densitometric scanning was achieved at the absorbance maxima 248 nm. Deferasirox was subjected to stress conditions like hydrolysis under different pH conditions, acid, alkali, oxidation, thermal and photolytic stress conditions. A well-separated band was observed with an retardation factor value of 0.44±0.03. The calibration curve plotted in the concentration range 250-1250 ng/band exhibited an excellent linear relationship with the coefficient of determination value of 0.9951. The method was found to comply with all the validation parameters as per the International Council for Harmonisation guidelines Q2(R1). This stability indicating method ensures a short analysis time compared to other reported analytical methods. This validated method can be used by quality control laboratories for monitoring the stability of deferasirox. [ABSTRACT FROM AUTHOR]

Background: Plants are extremely useful to humankind as medicine, and hence the properties of medicinal plants have to be scientifically evaluated. Aim: The aim of this study is to establish pharmacognostical and phytochemical parameters for stem bark of Ximenia americana Linn. (Olacaceae). Materials and Methods: Macroscopical, microscopical evaluation, powder microscopical studies, quantitative microscopy, physicochemical evaluation, fluorescence analysis, analysis of inorganic elements, and determination of heavy metal contamination were performed as per the World Health Organization guidelines and standard methods. To evaluate various phytoconstituents, phytochemical investigations such as preliminary phytochemical screening, high-performance thin-layer chromatography, and high-performance liquid chromatography were performed. Results: Microscopical studies showed the presence of thick cortex, periderm cells, phloem parenchyma, phloem fibers, sieve elements, prismatic and granular calcium oxalate crystals, and resinous cells. The length and width of fibers, stone cells, and diameter of starch grains were measured. Physicochemical evaluation like ash values has shown total ash – 8.01% ± 0.02% w/w, acid-insoluble ash – 1.53% ± 0.02% w/w, water-soluble ash – 1.54% ± 0.01% w/w, sulfated ash – 12.02% ± 0.07% w/w, water-soluble hot extraction – 26.48 ± 0.02% w/w, cold maceration – 11.76% ± 0.04% w/w, alcohol soluble – 15.69% ± 0.03%w/w, ether soluble – 2.06% ± 0.03% w/w, and loss on drying – 4.88% ± 0.01% w/w. Conclusion: Pharmacognostical study on the stem bark of Ximenia americana may be reported as first. The results of pharmacognostical and phytochemical studies will be helpful for authentication and provide quality control parameters for the stem bark of this plant.

The current research developed a high‐performance thin layer chromatographic method based on green analytical chemistry and integrated experimental approaches of analytical quality by design for simultaneous estimation of solifenacin succinate and silodosin. The integration was planned to reduce energy and solvent usage, maximize efficacy, and minimize any negative environmental effects. Taguchi design for screening and optimization a Box–Behnken design was used. The n‐butanol:ethanol:triethylamine (6:4:0.1, v/v/v) mobile phase and ultraviolet detection at 221 nm were used to develop the method. As per International Council for Harmonization Q2(R1) specifications, the method was validated. Rf value for solifenacin succinate and silodosin was found to be 0.29 ± 0.02 and 0.58 ± 0.02 respectively. For solifenacin succinate and silodosin, the linearity was found over the concentration ranges of 500–2500 ng/band and 800–4000 ng/band. For solifenacin succinate, the limits of detection and quantification were found to be 13.00 and 39.34 ng/band, respectively, while for silodosin, they were 33.29 and 100.86 ng/band, respectively. For solifenacin succinate, the method revealed a percent recovery ranging from 98.49% to 99.50%, while for silodosin, it exhibited a range from 98.86% to 99.79%. The tools predicted greenness score to be 63 and 0.78, respectively, indicating the proposed method to be green. [ABSTRACT FROM AUTHOR]

The condition known as benign prostatic hyperplasia, which causes enlargement in the prostate gland and male pattern hair loss are both treated with a family of drugs known as 5 alpha reductase inhibitors. This study shows that Solanum nigrum L. has 5 alpha reductase inhibitory action, that is helpful in the treatment of androgenic diseases. It was carried out to screen for phytochemicals and to analyze high-performance thin layer chromatography. Using petroleum ether and methanol extract, the 5 alpha reductase inhibitory activity of the plant was compared to the renowned 5 alpha-reductase inhibitor finasteride. The inhibition of extracts to the enzyme was assessed using a biochemical approach to measure the activity of 5 alpha-reductase. Because the substrate nicotinamide adenine dinucleotide phosphate has a particular absorbance at 340 nm, the optical density value of each sample was monitored with an ultraviolet spectrophotometer. Nicotinamide adenine dinucleotide phosphate concentration rises over time in the presence of a 5 alpha-reductase inhibitor because the enzyme 5 alpha-reductase needs nicotinamide adenine dinucleotide phosphate as a substrate. Thus, this technique implies 5 alpha-reductase activity. The method was quite successful in evaluating the plant capacity to block 5 alpha-reductase. It was determined that Solanum nigrum L. petroleum ether extract 125.528±0.634 (µg/ml) and linoleic acid (chemical biomarker of the plant material) 89.672±0.669 (µg/ml) were promising candidates for future investigation into their anti-androgenic activities. [ABSTRACT FROM AUTHOR]

Dispersive liquid–liquid microextraction (DLLME) in combination with high-performance thin layer chromatography (HPTLC) is proposed as a green, and reliable technique for simultaneous determination of sodium dodecylbenzene sulfate (SDBS) and sodium dodecyl sulfate (SDS) as milk adulterations. The surfactant's models were prepared in different concentration ranges (100–400 µg ml−1). At first, in DLLME procedure, the extraction conditions of anionic surfactants from milk were optimized by using central composite design (CCD). To obtain this purpose, volume of extraction solvent, speed of centrifuge and time of extraction were considered as effective factors. The proposed method needed only 5 ml of sample, and 500 µl of chloroform as microextraction solvent. Next, milk samples were analyzed by HPTLC. Subsequently HPTLC images were digitized and non-linear curve-fitting was implemented to the digitized measuring points chromatogram for elimination of random fluctuation, efficient denoising order to provide the reliable estimations of peak area. The fitted models were assessed by coefficient of determination (R2), adjusted coefficient of determination (Adj R2), sum of squares error (SSE), standard deviation (SD) and root mean square error (RMSE) which were estimated 0.97, 0.96, 0.001, 0.032 and 0.002 respectively. As a result, detection limits (LOD), limit of quantification (LOQ) and R2 for SDS and SDBS were 27, 33 µg ml−1 and 89, 94 µg ml−1 and 0.9932, 0.9917 respectively. The repeatability (RSD%) of the method for seven analyses and the extraction recovery were found to be 2.7–5.7% and 76.4–105.6% and 71–88.9% for SDS and SDBS respectively. [ABSTRACT FROM AUTHOR]

A rapid, accurate, precise, sensitive, and robust HPTLC method was developed for the simultaneous estimation of curcumin and naringin from the standard mixture and topical gel formulation. Quality by Design-based development of HPTLC method was carried out. The factor screening was performed using a seven-factor eight-run Taguchi design. The content of toluene in the mobile phase, saturation time and distance travel were selected as critical method parameters from screening studies. The Box-Behnken design was employed for systematic optimization of critical parameters and evaluating the robustness of the method. The data obtained from screening design and optimization design were analysed statistically and graphically to derive the conclusion. The optimal HPTLC separation was achieved using toluene:acetonitrile:methanol (5:3:2 v/v/v) as mobile phase with a bandwidth of 6 mm, saturation time of 25 min and distance travel up to 80 mm and scanning slit dimension of 4.00×0.30 mm. The chromatogram was scanned at 420 nm for curcumin and 286 nm for naringin. The method was validated for linearity, specificity, precision, accuracy, limit of detection, limit of quantitation and robustness according to ICH guidelines. The developed method was successfully employed for estimation of curcumin and naringin in topical gel formulation. [ABSTRACT FROM AUTHOR]

Background: Plants are extremely useful to humankind as medicine, and hence the properties of medicinal plants have to be scientifically evaluated. Aim: The aim of this study is to establish pharmacognostical and phytochemical parameters for stem bark of Ximenia americana Linn. (Olacaceae). Materials and Methods: Macroscopical, microscopical evaluation, powder microscopical studies, quantitative microscopy, physicochemical evaluation, fluorescence analysis, analysis of inorganic elements, and determination of heavy metal contamination were performed as per the World Health Organization guidelines and standard methods. To evaluate various phytoconstituents, phytochemical investigations such as preliminary phytochemical screening, high-performance thin-layer chromatography, and high-performance liquid chromatography were performed. Results: Microscopical studies showed the presence of thick cortex, periderm cells, phloem parenchyma, phloem fibers, sieve elements, prismatic and granular calcium oxalate crystals, and resinous cells. The length and width of fibers, stone cells, and diameter of starch grains were measured. Physicochemical evaluation like ash values has shown total ash - 8.01% ± 0.02% w/w, acid-insoluble ash - 1.53% ± 0.02% w/w, water-soluble ash - 1.54% ± 0.01% w/w, sulfated ash - 12.02% ± 0.07% w/w, water-soluble hot extraction - 26.48 ± 0.02% w/w, cold maceration - 11.76% ± 0.04% w/w, alcohol soluble - 15.69% ± 0.03%w/w, ether soluble - 2.06% ± 0.03% w/w, and loss on drying - 4.88% ± 0.01% w/w. Conclusion: Pharmacognostical study on the stem bark of Ximenia americana may be reported as first. The results of pharmacognostical and phytochemical studies will be helpful for authentication and provide quality control parameters for the stem bark of this plant. [ABSTRACT FROM AUTHOR]

A combination of Azilsartan Medoxomil and Cilnidipine is prescribed in the treatment of Hypertension. The present work represents an accurate and precise high-performance thin layer chromatographic method for the estimation of Azilsartan Medoxomil (AZL) and Cilnidipine (CLN) in combined tablet dosage form. Pre-coated silica gel- G60 F254 aluminum sheet (100 × 100 mm, 0.2 mm layer thickness) were used as stationary phase and Ethyl Acetate: Toluene: Glacial Acetic Acid (5: 4.9: 0.1 %v/v/v) in the mixture was used as mobile phase. The method was linear in the concentration range of 200 - 2000 ng/band for AZL and 50 -500 ng/band for CLN with a correlation coefficient (r2) of 0.996 for AZL and 0.999 for CLN. The proposed method was validated with respect to linearity, accuracy, precision, and robustness as per ICH Q2 (R1) guideline. A forced degradation study was performed to assess the stability indicating the nature of the method. All the degradant peaks were well resolved from the peak of the drug without any interference. The method was successfully applied for the analysis of AZL and CLN in combined tablet formulation. [ABSTRACT FROM AUTHOR]

A new HPTLC method and a sensitive UV spectrophotometric method were established for estimating vilazodone hydrochloride in its pharmaceutical formulations. An environment-friendly chromatographic mobile phase consisting of ethyl acetate: toluene: formic acid (5:4:1, V/V/V) along with ultraviolet densitometric detection at 240 nm using pre-coated silica gel plates was used. First derivative UV spectrophotometric determinations were performed using methanol as solvent, and amplitudes were measured for quantification purposes. Validation studies were compliant with ICH guidance. Vilazodone shows an optimum Rf value of 0.73 ± 0.02 by HPTLC method. First derivative spectrophotometric measurements were done at 230 nm (peak) and 247 nm (valley), respectively. Best method linearity was observed in the range of 100 - 600 ng band-1 and 100 - 600 ng mL-1 for the chromatographic and spectrophotometric methods, respectively. Method detection limits were 61.4 ng band-1 and 36.03 ng mL-1, for chromatography and spectrophotometry, respectively. Higher % recovery (> 99 %) of vilazodone from the in-house formulation satisfied study intent as well as instituted selectivity of both the methods. Both the newly developed methods have the potential of application in routine quantification of vilazodone in pharmaceuticals. [ABSTRACT FROM AUTHOR]

This study reports on the analysis of eleven Jarrah (Eucalyptus marginata) honeys, of which nearly half (n = 5) were re-classified as Blackbutt (E. patens) honey on the grounds of the predominant flower pollen identified by melissopalynology. Based on a comprehensive analysis of the honeys' physico- and phytochemical characteristics and antioxidant activity data, taking into account pH, electrical conductivity, refractive index and Brix values as well as moisture content, individual fructose and glucose content and derived fructose to glucose ratio alongside total phenolic content and antioxidant activity determined by the DPPH assay, no statistically significant difference was found amongst the eleven honeys classified by pollen analysis into two honey groups, ‘Jarrah’ or ‘Blackbutt’. This study therefore draws into question the value of melissopalynology as an analysis tool to authenticate Jarrah honey.

The present study focuses on the efficiency of high-performance thin layer chromatography and gas chromatography-mass spectrometry analysis in qualitative and quantitative analysis of triterpenoid from the terpene fraction of hexane extract of Simarouba glauca leaves. The total terpenoid content of the hexane extract was assessed which shows that 73.69 % of the extract content is terpenoid. The terpene fraction extracted in petroleum ether from the hexane extract of Simarouba glauca leaves was subjected to gas chromatography-mass spectrometry and high-performance thin layer chromatography analysis. Gas chromatography-mass spectrometry analysis results indicated seventeen compounds out of which only one was triterpene (Squalene an important bioactive triterpenoid). In high-performance thin layer chromatography the separation was achieved by using hexane:ethyl acetate in a 5:5 proportion as mobile phase and anisaldehyde sulphuric acid reagent was used for derivatization which imparted violet color to the triterpenoid containing bands. Detection and quantification of betulinic acid and other triterpenes was done by densitometric scanning at 525 nm. Betulinic acid produced compact spots at Rf 0.66. Linear range of betulinic acid was prepared using concentrations 1-7 µg/spot with a correlation coefficient R2 and standard deviation of 0.96004 %±19.68 % respectively. The results indicated the presence of 10 triterpenoids showing different Rf values including betulinic acid. This study confirmed the presence of betulinic acid in the terpene fraction of the hexane extract of Simarouba glauca leaves. [ABSTRACT FROM AUTHOR]

Antioxidants can scavenge free radicals in the human body and have been associated with a protective effect on the human body against a variety of diseases. As a part of normal cellular function, free radical production occurs continuously within the body at a cellular level. Excess-free radical production from endogenous or exogenous sources may result in a variety of diseases. Vitamins (A, C, and E), carotenoids, polyphenols, and flavonoids are some examples of common natural antioxidants. Among the different antioxidants quercetin, curcumin, and ascorbic acids have been extensively studied, and numerous reports on their antioxidant activity are available in the literature. Antioxidants are regarded as a key player in the current pandemic situation, and they play a key role as a preventive and controlling tool. There are a variety of herbal formulations and immune booster therapies available, many of which claim to contain various antioxidants and are effective against covid-19. Numerous herbal formulations and immune booster therapies are available and claimed to have different antioxidants and other constituents. In the present study simple, rapid, accurate, precise, and sensitive high-performance thin-layer chromatographic analytical method was developed for simultaneous estimation of quercetin, curcumin, and ascorbic acid. The developed method can effectively employ for different polyherbal formulations. The developed method was validated according to ICH guidelines. In the present study, an aluminum plate precoated with 60F 254 silica gel was used as a stationary phase whereas chloroform: ethyl acetate: formic acid (6:6:2.5 v/v/v) was used as a mobile phase. Chromatographic detection was performed with a Camag TLC scanner at 265 nm. The linearity range for the proposed method was found to be 500–1000 ng/band for all three analytes. The antioxidant potentiality of selected antioxidants was evaluated by an optimized UV visible spectroscopic-reducing power method. The reducing power method was optimized for the concentration and volume of reagent, incubation temperature, and time. The results of the study revealed that quercetin is having the best antioxidant potential compared to curcumin and ascorbic acid. [ABSTRACT FROM AUTHOR]

Montelukast is a highly potent and selective receptor antagonist of cysteinyl leukotrienes (CysLT1) used in chronic asthma and vasoconstriction reduction and also used in allergies, swelling, hay fever, and shortness of breath. For the measurement of montelukast sodium in biological and pharmaceutical samples, there are different asynchronous and concurrent analytical methods, such as high‐performance thin layer chromatography, UV spectroscopy, capillary electrophoresis, high‐performance liquid chromatography, and liquid chromatography‐mass spectrometry, have been developed. The pathogenesis of asthma, mechanism of action, and various analytical methods for the detection of montelukast sodium and its combination of dosage forms and biological samples have been described in the present review. Ranjan et al. suggested the reverse phase high‐performance liquid chromatography approach for estimating montelukast in rabbit plasma and found the best limit of detection result of 1.0 ng/mL. Ezzeldin et al. developed a liquid chromatography with tandem mass spectrometry system for the simultaneous detection of combinations of montelukast, gliclazide, and nifedipine. The authors conclude that the studies described in the present manuscript will assist researchers in choosing a simple, rapid, and responsive method for the analysis of montelukast sodium and its combinations. [ABSTRACT FROM AUTHOR]

The present study deals with the extraction and quantification of phytochemicals, which play a key role in the pharmaceutical world. In this article, we focused on the study of plant secondary metabolites and analyzed ethanolic and methanolic extracts of bark and roots of two species of Ixora using the high performance thin layer chromatography technique. Alcoholic extracts of bark and root of both the plants revealed the presence of alkaloids, terpenoids, glycosides, phenols, saponins, sterols, tannins, etc., but both extracts were devoid of flavonoids and coumarins. Analysis of alkaloid content revealed the highest amount of total alkaloid content in the methanolic extract of roots of Ixora coccinea (146 mg/ gm) followed by the methanolic extract of bark of Ixora coccinea (138 mg/g). The lowest total alkaloid content was observed in the methanolic extract of bark of Ixora barbata (64 mg/g). Both ethanolic, as well as methanolic extracts of Ixora coccinea, showed higher amounts of alkaloid content when compared to Ixora barbata in similar solvents. As per high-performance thin layer chromatography analysis, the highest amount of camptothecin was recorded in ethanolic extract of root of Ixora coccinea (117 µg/ml) followed by ethanolic extract of bark of Ixora barbata (64.43 µg/ml). The lowest amount of camptothecin was observed in methanolic extract of bark of Ixora coccinea (12.64 µg/ml). The results obtained from the present high-performance thin layer chromatography analysis also revealed that the above-mentioned selected plants are a source of a number of other phytoconstituents. This is the first reported study on the high-performance thin layer chromatography analysis of Ixora barbata. The study includes quantification of camptothecin compound in both methanolic and ethanolic extracts of Ixora barbata for which there are no previously published reports. [ABSTRACT FROM AUTHOR]

Setaria italica (common name- foxtail, kangni) is one of the major food crops which is prominently cultivated in southern regions of India and in certain regions of Uttar Pradesh. Besides the crop's consumption as a general source of carbohydrate rich cereal, the seeds of the crop are comprised of more fiber. So, it is recommended to add in the dietary supplementation of the diabetic people across the country. In this paper, it intends to investigate the antidiabetic activity and antioxidant activity of S. italica (foxtail millet) seeds in diabetic rats. The six genotypes of foxtail millets (S. italica) namely Kangni-1, Kangni-4, Kangni-5, Kangni-6, Kangni-7 & Kangni-10 respectively were subjected to in vitro investigations via. comprehensive metabolic panel (CMP) involving blood glucose study, Kidney & Liver function test, and antioxidant study (Catalase test; Glutathione S-transferase (GST); Superoxide Dismutase (SOD); glutathione (GSH); hiobarbituric acid reactive substances (TBARS) & Glutathione peroxidase (GPx) and were performed in vivo animal investigations in Wistar rats. The STZ induced diabetic rats were fed with doses of different S. italica seed aqueous extract to evaluate its anti-hyperglycemic activity by oral administration of SISAE. Further, it was compared with Glibenclamide which acts as one of the standard oral hypoglycemic agents. From achieved outcomes, a significant fall of blood glucose level (70%) produced 300 mg SISAE/kg b.w. after 6 h of extract administration. However, no change could be produced by these doses of the SISAE in normal rats' blood glucose levels. A significant fall in glucose level along with significant glycemic control by lower HbA1c levels was observed in diabetic treated rats after 3 weeks of treatment with 300 mg of SISAE/kg b.w./day when comparing to untreated diabetic rats. Among these five genotypes of S. italica , the differences in the glycemic index were found. a significant fall could be found in blood glucose levels of Wistar rats, when every experimental rat was incorporating with the extract of different genotypes of Setaria italica L. Beauv than the rats treated with Glibenclamide in every 7 days of interval. The level of catalase, SOD, GST, GPx, GSH and TBARS showed variation while the rats were fed with the extract of S. italica in the liver test of rats. In kidney function test, the result shows that there is significant relationship between foxtail extract and kidney function of STZ induced diabetes rats. They show the change in their serum creatinine level, serum urea and serum uric acid. The result obtained from the study shows that the extract of S. italica seeds is capable for the hypolipidemic and antihyperglycemic activities, thereby, they serve as one of the good sources for herbal medicinal items. [ABSTRACT FROM AUTHOR]

Aim: Representatives of Nepetoideae Burnett subfamily are promising species of the Lamiaceae Martinov family because of accumulating such valuable groups of secondary metabolites as terpenoids and polyphenols. A high-performance thin layer chromatography (HPTLC) fingerprint method for the qualitative determination of phenolic compounds and for the quantification of rosmarinic acid (RA) content in methanol extracts of five species of this subfamily was developed for the first time. Materials and Methods: Dried aerial parts of Dracocephalum moldavica L., Ocimum americanum L., Lophanthus anisatus (Nutt.) Benth., Monarda fistulosa L., and Satureja hortensis L. collected in flowering period were macerated with methanol. The HPTLC analysis was conducted using the CAMAG analytical system (Muttenz, Switzerland). The comparative analysis of RA contents was performed by HPTLC densitometric detection at λ = 366nm. Results: Identification of polyphenols in the investigated herbs was performed by comparison of a color and Rf of the chromatographic zones with six reference standards: rutin, apigenin, luteolin, caffeic acid, chlorogenic acid, and RA. HPTLC method was also validated for the quantification of RA in the extracts of investigated herbs. RA contents decreased in such a sequence: D. moldavica (24.83 ± 0.78mg/g) > M. fistulosa (20.32 ± 0.64mg/g) > O. americanum (19.59 ± 0.61mg/g) > S. hortensis (18.77 ± 0.52mg/g) > L. anisatus (12.61 ± 0.43mg/g). Conclusion: Obtained data can facilitate the differentiation of investigated species using the chromatographic fingerprints of their phenolic compounds. Developed and validated HPTLC method provides an approach to estimate RA content as a common marker of investigated herbs.

The basic idea of the Chromatographic process is that the distinct compounds have different properties like solubility, adsorption, ion exchange, and affinity that can be regulated through various separation mechanisms causing the analytes to travel through the stationary phase at different speeds, resulting in their separation from a complex mixture. The Chromatographic techniques have been explored as multidimensional, which has been growing and evolving due to continuous advancements in technology allowing it to meet the upcoming challenges and analytical demands in various scientific scenarios including academics and industries. Phytochemistry is one of the oldest core areas of research in science where one of the major challenges in drug discovery from natural sources is the identification and isolation of closely related active molecules within these complex to the observed biological activities therefore there is utmost need to better understand their intrinsic complexity and exploit their vast commercial potential. Nowadays the hyphenated techniques with one component as the chromatographic technique have been widely explored as inevitable analytical tools for early detection and identification of bioactive compounds and driving forces in the evolution and discoveries in phytochemistry from crude plant extracts that boosted mainly due to rapid technological advancements in instrumentation. It’s also significant as an intellectual method for phytotherapeutic quality control and regulation. In the present review, we have discussed the advances and insight of chromatographic techniques which can be explored for comprehensive chemical profiles of herbal medicine preparations or extracts in phytochemistry. The review provides a systematic update of the recent advancement and published approaches to methodology in phytochemistry.

In this study, a high-performance thin layer chromatography (HPTLC) method by two step gradient elution with two mobile phases was developed for the simultaneous analysis of seven constituents in Ophiopogonis Radix. The chromatography was performed on silica gel 60 F254 plate with dichloromethane-methanol-ethyl acetate-water (70:25:12:3, v/v/v/v) and dichloromethane-methanol (300:1, v/v) as the mobile phase for two step gradient elution. Then, the HPTLC profiles were observed after derivatization with 10% sulfuric acid in ethanol solution. The obtained HPTLC images were further analyzed by chemometric approaches and the samples could be clustered based on regions and/or growth years, which were two important factors affecting the constituents in Ophiopogonis Radix. Furthermore, five compounds including ophiopogonin D, ophiopojaponin C, ophiopogonin D’, ophiopogonin C’ and methylophiopogonanone B were screened as potential lipase inhibitors from Ophiopogonis Radix by the HPTLC-bioautographic method. The binding modes and interactions between the five compounds and lipase were further explored by molecular docking analysis. The developed HPTLC method could be used for quality control of Ophiopogonis Radix and screening of the potential lipase inhibitors.

The principle motive behind this work is the densitometric high-performance thin-layer chromatography (HPTLC) method has been developed and validated for the simultaneous estimation of Olmesartan medoxomil and Chlorthalidone in three different brands of the tablet dosage form. The method involves silica gel 60 F254 high-performance thin-layer chromatography and densitometric detection at 238 nm using Toluene: Ethyl acetate: Methanol: Glacial acetic acid (5:4.7:0.3:0.1 % v/v/v/v) as a mobile phase. The calibration curve was linear over a range of 100-1200 ng/band for Olmesartan medoxomil and 62.5-750 ng/band for Chlorthalidone. Precision provided relative standard deviations (RSD) lower than 2 % of both analytes. Percent recoveries were performed and found to be 99.53-99.90 % for Olmesartan medoxomil and 98.18-99.49 % for Chlorthalidone. Validation of the method was carried out as per International Conference on Harmonization (ICH) guideline Q2 (R1). The HPTLC densitometric driven method is highly suitable for analysis of Olmesartan medoxomil and Chlorthalidone in dosage form without any interference. [ABSTRACT FROM AUTHOR]

A novel ecofriendly, cost and time saving high‐performance thin‐layer chromatographic method was developed and validated for simultaneous determination of metoclopramide, ergotamine, caffeine, and paracetamol in bulk and pharmaceutical formulation. The separation was carried out on silica gel plates, using ethyl acetate:ethanol:ammonia (9:1:0.1, v/v/v) as a developing system. Ultraviolet detection was carried out at 272 nm. The resulting retention times were 0.15, 0.36, 0.49, and 0.74 min for metoclopramide, ergotamine, caffeine, and paracetamol, respectively. The greenness profile assessment was achieved to the proposed method to evaluate its greenness characters to the environment with acceptable results. Validation parameters were checked according to International Conference of Harmonization guidelines to achieve the international requirements for quality control analysis of the proposed drugs. [ABSTRACT FROM AUTHOR]

Aim: Representatives of Nepetoideae Burnett subfamily are promising species of the Lamiaceae Martinov family because of accumulating such valuable groups of secondary metabolites as terpenoids and polyphenols. A high-performance thin layer chromatography (HPTLC) fingerprint method for the qualitative determination of phenolic compounds and for the quantification of rosmarinic acid (RA) content in methanol extracts of five species of this subfamily was developed for the first time. Materials and Methods: Dried aerial parts of Dracocephalum moldavica L., Ocimum americanum L., Lophanthus anisatus (Nutt.) Benth., Monarda fistulosa L., and Satureja hortensis L. collected in flowering period were macerated with methanol. The HPTLC analysis was conducted using the CAMAG analytical system (Muttenz, Switzerland). The comparative analysis of RA contents was performed by HPTLC densitometric detection at λ = 366 nm. Results: Identification of polyphenols in the investigated herbs was performed by comparison of a color and Rf of the chromatographic zones with six reference standards: rutin, apigenin, luteolin, caffeic acid, chlorogenic acid, and RA. HPTLC method was also validated for the quantification of RA in the extracts of investigated herbs. RA contents decreased in such a sequence: D. moldavica (24.83 ± 0.78 mg/g) > M. fistulosa (20.32 ± 0.64 mg/g) > O. americanum (19.59 ± 0.61 mg/g) > S. hortensis (18.77 ± 0.52 mg/g) > L. anisatus (12.61 ± 0.43 mg/g). Conclusion: Obtained data can facilitate the differentiation of investigated species using the chromatographic fingerprints of their phenolic compounds. Developed and validated HPTLC method provides an approach to estimate RA content as a common marker of investigated herbs. [ABSTRACT FROM AUTHOR]

This study optimised a high-performance thin-layer chromatography (HPTLC) method that could facilitate the visualisation and quantification of antioxidative phytochemical compounds in Caulerpa lentillifera. This method involved the use of 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and ferric chloride (FeCl3) as derivatising agents. It was found that the incubation period for DPPH• was consistent with 60 minutes in the dark, while the plate that was derivatised with FeCl3 needed to be neutralised using NH4OH and then dried at 100°C for 5 minutes. The antioxidant activity of the seaweed extracts can now be analysed using Videoscan and quantified as gallic acid equivalents (GAE). [ABSTRACT FROM AUTHOR]

Background: Dillenia pentagyna Roxb. (Dilleniaceae) is an endangered medicinal plant, commonly known as "agai" and distributed randomly in the Terai belt of North East states of India. D. Pentagyna extract has been studied for therapeutic possessions. Objective: The proximate parameters and high-performance thin layer chromatography (HPTLC) analysis in the same plant of D. pentagyna of four-time harvest stage during the maturation of fruits were studied and also the antihypoglycemic activity of hydroalcoholic extract of D. pentagyna fruit. Materials and Methods: The proximate compositions were determined by measuring the amount of water removed from the food AOAC method and the chemical fingerprinting was carried out by HPTLC method. An acute oral toxicity study was carried out in healthy male Wister rats (110 g). The dose was finally made to 200 and 400 mg/kg body weight for oral administration after the LD50 estimation. Results: The most phenolic concentration was continuously decrease with during the maturation of fruits. After 21 days of treatment, level of blood glucose, as well as albumin, protein, creatinine, urea, and uric acid were significantly decreased when compared with the diabetic control. Hydroalcoholic (80% alcohol to 20% DW) extracts will be subjected to further extensive studies to isolate and identify their active constituents which are useful for against antidiabetic. Conclusion: D. pentagyna fruit could act as a source of functional compounds for the control of diabetes mellitus. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • First report on pentacyclic triterpenoids in Indian Swertia from Western Himalayas. • A validated and simultaneous high-performance thin-layer chromatography was performed. • Microwave, heat reflux and Soxhlet extractions were carried out for the triterpenoids. • Microwave coupled with aqueous ethanol was the most efficient method for extraction. • S. paniculata showed promise to substitute S. chirata in triterpenoid content. Abstract Swertia chirata (Family: Gentianaceae) is a potent and valuable medicinal herb used in various herbal formulations in Indian system of medicine. Different species of Swertia are being used as constituents in various herbal formulations. Present study was undertaken for the screening of an efficient extraction technique and for the development of a rapid and simultaneous quantification of four pentacyclic triterpenoids in different populations of Swertia species by a high-performance thin-layer chromatography (HPTLC)-densitometric method. This method involved the separation of four triterpenoids with a single mobile phase composition consisting of n-hexane, ethyl acetate, toluene (14.2, 4, 1.8 v/v). The retention factor (R f) values for oleanolic acid (OA), ursolic acid (UA), betulinic acid (BA), and lupeol (Lup) were 0.25, 0.50, 0.58 and 0.74 respectively. In this study, microwave assisted extraction (MAE) using aqueous ethanol proved to be the most efficient method for extraction for all the studied triterpenoids when compared with the other conventional methods. The present HPTLC method was found to be precise, fast, and cost effective and can be used for the quality control of any Swertia samples with regards to its pentacyclic triterpenoids content. Since, S. chirata is a critically endangered species, the present study depicts the utility of other species of Swertia viz. S. paniculata and S. angustifolia as promising substitutes for S. chirata in herbal preparations. [ABSTRACT FROM AUTHOR]

Detoxified Nux-vomica (detoxified Strychnos nux-vomica Linn.) is a very popular crude herb used in Unani and Ayurvedic pathy after detoxification for many years. The first person to identify Nux-vomica with the Indian drug kuchla was Haji Zainuddin. Detoxified Nux-vomica has significant effects on various disorders like rheumatic arthritis, inflammation, and gout (Anonyms, 2007; Bentley and Trimen, 1983). It has proved the best remedy for paralysis, nervous disorders, nervous debility, tremor, diarrhoea, and mental emotions (Nadkarni, 2000). The present study is designed to develop the qualitative as well as quantitative parameters of crudely detoxified Nux-vomica seeds and their comparative study by Thin layer chromatography (TLC) and High-performance thin layer chromatography (HPTLC). Organoleptic characters, ash values, extractive values, phytochemical analysis, fluorescence analysis, TLC and HPTLC profiles, etc. were the parameters used for the standardisation of the test drug. Total ash values, water, alcohol, ether-soluble extractive values, and volatile oil percentages were found to be 1.925%, 6.54%, 3.25%, 3.20%, and 2.00%, respectively. The TLC profile of Detoxified Nux-vomica shows three spots for aqueous extract and five spots for hydroalcoholic extract in Toluene: Ethyl Acetate: Triethylamine (7:2:1) and four spots in Methanol: Ethyl Acetate: Triethylamine (0.5:8.5:1) with exposure in Dragendorff's Reagent. The percentage yield of strychnine in the hydro-alcoholic and aqueous extracts of Detoxified Nux-vomica is 7.48 and 2.10, while it is 6.17 and 1.85 for brucine, respectively. The present study used secondary data for the identification and quantification of active constituents in detoxified Nux vomica seeds. [ABSTRACT FROM AUTHOR]

Context: A high-performance thin layer chromatography (HPTLC) is an analytical technique, which can be used for the determination of constituents or marker components in various parts of the plants. Earlier studies have estimated phytoconstituents from the stem and other aerial plant parts of Cissus quadrangularis Linn. Estimation of hydrocarbons can also be successfully done using HPTLC technique using suitable derivatization. Aims: To develop and validate a simple and rapid method for the estimation of long chain aliphatic hydrocarbons from the leaves of C. quadrangularis using HPTLC technique. Methods: Precoated silica gel 60 F254 plates were used as stationary phase. The mobile phase used was hexane (100 %). The detection of spots was carried out using berberine sulphate as detecting reagent. Results: The method was validated in terms of linearity, sensitivity, accuracy, and precision. Linearity range was found to be 2-10 µg/mL, limit of detection 0.127 µg/mL, and limit of quantification 0.384 µg/mL. Conclusions: A novel, simple, accurate, precise and sensitive HPTLC method has been developed and validated for the estimation of long chain aliphatic hydrocarbons obtained from the leaves of C. quadrangularis Linn.

Aim: Detection of aflatoxin B1 in Livestock compound Feed and feed ingredients by high-performance thin layer chromatography (HPTLC). Materials and Methods: Chromatography was performed on HPTLC silica gel 60 F 254, aluminum sheets by CAMAG automatic TLC sampler 4, with mobile phase condition chloroform:acetone:water (28:4:0.06). Extraction of aflatoxin B1 from samples was done as per AOAC method and screening and quantification done by HPTLC Scanner 4 under wavelength 366 nm. Results: A total of 97 livestock feed (48) and feed ingredients (49) samples received from different livestock farms and farmers were analyzed for aflatoxin B1of which 29 samples were contaminated, constituting 30%. Out of 48 livestock compound feed samples, aflatoxin B1 could be detected in 16 samples representing 33%, whereas in livestock feed ingredients out of 49 samples, 13 found positive for aflatoxin B1 representing 24.5%. Conclusion: HPTLC assures good recovery, precision, and linearity in the quantitative determination of aflatoxin B1 extracted from Livestock compound feed and feed ingredients. As more number of feed and feed ingredients are contaminated with aflatoxin B1 which causes deleterious effects in both animal and human beings, so there is a need for identifying the source of contamination, executing control measures, enabling better risk assessment techniques, and providing economic benefits.

Aim: Wheatgrass (WG) is the shoot of Triticum aestivum Linn. belongs to the family Gramineae, and possess high chlorophyll content and essential vitamins, minerals, vital enzymes, amino acids, dietary fibers etc., It has been shown to possess anti-cancer, anti-ulcer, antioxidant, and anti-arthritic activity due to the presence of biologically active compounds, and minerals. Therefore, in the present study, high-performance thin layer chromatography (HPTLC), and high-performance liquid chromatography (HPLC) methods for qualitative and quantitative analysis have been proposed, which will help in quality evaluation of wheat grass extract. Materials and Methods: Samples for analysis were prepared in methanol and water simply by sonication. These were applied on pre-coated silica plate and chromatograms were developed using toluene: Ethyl acetate: Formic acid. HPLC analysis was done on Waters HPLC system using water, methanol, and acetonitrile as mobile phase. Merck C18 column has been used. Results: HPTLC finger printing of alcoholic extracts of WG was carried out and found 10–11 spots at different wavelengths 254, 366, and 435 nm. HPLC fingerprinting produced 22 peaks at 256 nm. Quantitative HPTLC analysis was done to determine the gallic acid content, and was found to be 0.077% w/w in aqueous extract. By HPLC, the content of gallic acid and rutin was found to be 0.07%, and 0.04% w/w in aqueous extract of WG. Conclusion: The developed HPLC and HPTLC fingerprinting method can be used for the quality control, and standardization of WG and its extracts used as nutritional supplement.

Background: Momordica charantia Linn. (Cucurbitaceae) fruits are well known for their beneficial effects in diabetes that are often attributed to its bioactive component charantin. Objective: The aim of the present study is to develop and optimize an efficient protocol for the extraction of charantin from M. charantia fruits. Materials and Methods: Response surface methodology (RSM) was used for the optimization of ultrasound-assisted extraction (UAE) conditions. RSM was based on a three-level, three-variable Box-Behnken design (BBD), and the studied variables included solid to solvent ratio, extraction temperature, and extraction time. Results: The optimal conditions predicted by the BBD were: UAE with methanol: Water (80:20, v/v) at 46°C for 120 min with solid to solvent ratio of 1:26 w/v, under which the yield of charantin was 3.18 mg/g. Confirmation trials under slightly adjusted conditions yielded 3.12 ± 0.14 mg/g of charantin on dry weight basis of fruits. The result of UAE was also compared with Soxhlet extraction method and UAE was found 2.74-fold more efficient than the Soxhlet extraction for extracting charantin. Conclusions:A facile UAE protocol for a high extraction yield of charantin was developed and validated.

Achyranthes coynei is a rare, endemic perennial shrub reported from Karnataka and Maharashtra states of India. The plant is used to treat various disorders by folk healers and was proven to have antimicrobial and antioxidant properties. The present study was undertaken to evaluate microscopic and macroscopic characters of A. coynei stem, along with its physicochemical parameters. ProgRes ® CapturePro and Microsoft Excel were used for statistical analysis. Perennial, shrubby nature and woody stem were the distinguishing morphological characters observed. Transverse section (TS) illustrated quadrangular outline of the stem and showed the presence of two types of trichomes on the thick-walled epidermis. TS also showed number of rosette calcium oxalates crystals; prismatic and microsphenoid crystals; conjoint, collateral open secondary vascular bundles; and two amphixylic medullary bundles in the pith. Ash and extractive values, micro and macro elements and nutritive factors were estimated in the present study. The presence of alkaloids, saponins and triterpenoids were observed in preliminary phytochemical screening. High-performance thin layer chromatographic analysis yielded different bands and also indicated the presence of oleanolic acid. The studied parameters for A. coynei stem will be useful for identification and authentication of the plant material.

Objective: Estimation of terizidone in pharmaceutical dosage form by using high-performance thin-layer chromatography (HPTLC) CAMAG system. Materials and Methodology: The separation was achieved using precoated silica gel 60 F254 aluminum plates with mobile phase toluene: n-Butanol 9:1 mL % v/v at wavelength 268 nm. Results: The linearity range was found between 50 and 300 ng/mL-band having correlation coefficient 0.997. The recovery study of method showed the recovery between 99.3% and 101.6%. The limit of detection (LOD) and limit of quantification (LOQ) values for method were 0.981 and 2.973, respectively. The R.S.D. of precision study was <2. The method parameters were validated as per the ICH guidelines. The forced degradation studies revealed that the drug found stable in acidic, thermal, and photodegradation. Conclusion: The liquid chromatography-mass spectrometry study of degraded products gave the idea about the fragmentation patterns of degraded products. The developed method can be used for routine analysis of terizidone and its formulations. [ABSTRACT FROM AUTHOR]

Abstract: The combination of sofosbuvir and daclatasvir or sofosbuvir and ledipasvir is now widely used as an ideal treatment for hepatitis C virus infection. For this purpose, a simple, sensitive, accurate, economic, and precise high‐performance thin‐layer chromatography was developed and validated for the determination of sofosbuvir, daclatasvir, and ledipasvir in their pure form as well as their different pharmaceutical products. The method used Merck high‐performance thin‐layer chromatography aluminum plates precoated with silica gel 60 F254 as a stationary phase and mobile phase consisting of methylene chloride/methanol/ethyl acetate/ammonia (25%) (6:1:4:1, v/v/v/v). This system was found to give compact symmetric peaks of sofosbuvir, daclatasvir, and ledipasvir with retardation factors of 0.27 ± 0.01, 0.50 ± 0.007, and 0.68 ± 0.008, respectively. The densitometric scanner was set at 275 nm using a deuterium lamp. The calibration curves were linear over the range of 100–3000 ng/spot for sofosbuvir, and daclatasvir, and range of 50–3000 ng/spot for ledipasvir. The detection limits were 22.5, 31.90, and 15.80 for sofosbuvir, daclatasvir, and ledipasvir. The quantitation limits were 67.50, 95.60, and 47.50 for sofosbuvir, daclatasvir, and ledipasvir. The proposed method was validated according to International Conference on Harmonization (ICH) guidelines and the results were acceptable. [ABSTRACT FROM AUTHOR]

A simple, accurate and precise high-performance thin layer chromatography (HPTLC) method has been developed for the simultaneous determination of allopurinol (AP), a xanthine oxidase inhibitor and its active metabolite oxypurinol (OP) in human plasma and AP in tablet dosage form. The proposed method was developed and validated on aluminium plates pre-coated with silica gel 60G F254 as the stationary phase using methanol: chloroform: ammonia (2.0:7.9: 0.1, v/v/v) as the developing solvent system. Densitometric measurements were made at 206 nm and the retention factors (Rf) obtained were 0.38 ± 0.01 and 0.65 ± 0.01 for AP and OP respectively. System suitability testing parameters were evaluated to determine the performance of the chromatographic method. The regression plots were linear (r2 > 0.9993) in the concentration range of 100-700 ng/band for both the analytes. The limit of detection and limit of quantitation of the method were 19.56 and 59.29 ng/band for AP and 19.01 and 57.59 ng/band for OP respectively for plasma samples. For spiked plasma samples, protein precipitation with formic acid in acetonitrile afforded mean recovery of 84.67 % and 86.21 % for AP and OP, respectively. The method was validated for linearity, specificity, accuracy and precision, repeatability, stability and robustness. [ABSTRACT FROM AUTHOR]

Lipid transmethylation methods described in the literature are not always evaluated with care so to insure that the methods are effective, especially on food matrix or biological samples containing polyunsaturated fatty acid (PUFA). The aim of the present study was to select a method suitable for all lipid species rich in long chain n-3 PUFA. Three published methods were adapted and applied on individual lipid classes. Lipid (trans)methylation efficiency was characterized in terms of reaction yield and gas chromatography (GC) analysis. The acid-catalyzed method was unable to convert triglycerides and sterol esters, while the method using an incubation at a moderate temperature was ineffective on phospholipids and sterol esters. On the whole only the method using sodium methoxide and sulfuric acid was effective on lipid classes taken individually or in a complex medium. This study highlighted the use of an appropriate (trans)methylation method for insuring an accurate fatty acid composition. [ABSTRACT FROM AUTHOR]

Background: Ageratum conyzoides L. is a traditionally used herb for various ethnoveterinary purposes. There are no earlier reports on simultaneous high-performance thin layer chromatography (HPTLC) quantification of bioactive markers. Objective: To develop a sensitive, robust, and replicable HPTLC method for simultaneous estimation of precocene I (PI) and precocene II (PII) in A. conyzoides L. collected from the Western Himalaya region of India, with an aim to understand the level of chemotypic differences arising in the intraspecific population of this ethnobotanically important plant. Materials and Methods: A sensitive HPTLC method was developed to resolve PI and PII using toluene: ethyl acetate (9.8:0.2 v/v) as mobile phase. The method was validated for selectivity, specificity, linearity, and precision as per International Conference on Harmonization guidelines. Results: Good linearity was achieved over a five-point concentration range with a correlation coefficient of 0.986 and 0.988 for PI and PII, respectively. The PI content varies in the range of 0.0016% (NAC-77) to 0.0834% (NAC-82), whereas PII was reported to be present in the range of 0.016% (NAC-85) to 0.143% (NAC-91) on a dry weight basis. A principal component analysis biplot of samples based on the content of PI and PII identified four elite chemotypes, namely, NAC-81, NAC-82, NAC-85, and NAC-91. Conclusion: The study identifies superior germplasms for commercial prospection and develops a validated method that can be used for the quality control of herbal drug/formulation using A. conyzoides as an ingredient. [ABSTRACT FROM AUTHOR]

Background: Aśvagandhādyariṣṭa is a polyherbal formulation that is available commercially as an over the counter drug. There are three famous brands that are available in the market. However, there are no comparative reports on the physicochemical, chromatographic, and biological profiles of Aśvagandhādyariṣṭa manufactured by these famous companies. Aims: The present study deals with the physicochemical standardization, high performance thin layer chromatography (HPTLC) profiling, and biological evaluation of Aśvagandhādyariṣṭa. Materials and Methods: Aśvagandhādyariṣṭa manufactured by three leading companies were purchased from Jalandhar, Punjab. The physicochemical standardization of the samples was carried out in accordance with the Ayurvedic Pharmacopoeia of India (API). Authentified Eisenia foetida were procured from Ujjwal Ujala Vermiculture Group, Amritsar. The anthelmintic activity, 1,1-diphenyl-2-picrylhydrazyl scavenging, and hydrogen peroxide scavenging ability of Aśvagandhādyariṣṭa was determined. Statistical Analysis Used: The data of anthelmintic activity were expressed as mean ± standard error of the mean of six earthworms in each group. The statistical analysis was carried out using one-way analysis of variance, followed by Dunnet t-test. The difference in values at P < 0.05 was considered to be statistically significant. Results: Most of the physicochemical standardization parameters mentioned as per the API were found to be within limit. HPTLC profiling showed the presence of withanolide D in commercial samples. Out of three commercial brands, ASA-DAB was the most active as compared to the ASA-BDN and ASA-AVP at the concentration of 200 mg/ml for anthelmintic activity against E. foetida. ASA-DAB showed the best antioxidant activity in both the in vitro assay at the concentration of 100 μg/ml. Conclusions: The ability of this formulation to scavenge free radicals supports its medical claim of antistress formulation. The anthelmintic potential of this formulation helps us conclude that it can also be considered as a general tonic because it provides relief from helminths.

Introduction: Gamma amino butyric acid (GABA) is an important ubiquitous four carbon nonprotein amino acid with an amino group attached to gamma carbon instead of beta carbon. It exists in different organisms including bacteria, plants, and animals and plays a crucial role in humans by regulating neuronal excitability throughout the nervous system. It is directly responsible for the regulation of muscle tone and also effective in lowering stress, blood pressure, and hypertension. Aim and Objective: The aim of the study was to develop the fingerprint profile of selected medicinally and economically important plants having central nervous system (CNS) activity and to determine the quantity of GABA in the selected plants grown under natural conditions without any added stress. Materials and Methods: The high-performance thin layer chromatography analysis was performed on precoated silica gel plate 60 F -254 plate (20 cm × 10 cm) in the form of bands with width 8 mm using Hamilton syringe (100 μl) using n-butanol, acetic acid, and water in the proportion 5:2:2 as mobile phase in a CAMAG chamber which was previously saturated for 30 min. CAMAG TLC scanner 3 was used for the densitometric scanning at 550 nm. Specific marker compounds were used for the quantification. Results and Conclusion: Among the screened medicinal plants, Zingiber officinale and Solanum torvum were found to have GABA. The percentage of GABA present in Z. officinale and S. torvum were found to be 0.0114% and 0.0119%, respectively. The present work confirmed that among the selected CNS active medicinal plants, only two plants contain GABA. We found a negative correlation with plant having CNS activity and accumulation of GABA. The GABA shunt is a conserved pathway in eukaryotes and prokaryotes but, although the role of GABA as a neurotransmitter in mammals is clearly established, its role in plants is still vague.

Background: Coleus forskohlii is a well-known industrially important medicinal plant, for its high forskolin content. Objective: A simple, selective, and sensitive high-performance thin layer chromatography (HPTLC) method was developed and validated for simultaneous quantification of forskolin and iso-forskolin in C. forskohlii germplasm collected from the Eastern Ghats, India. Materials and Methods: Chromatographic separation of the targeted marker(s) was obtained on precoated silica plates using toluene: ethyl acetate: methanol (90:30:0.5, v/v/v) as the mobile phase. Results: Densitometric quantification of forskolin and iso-forskolin was carried out at 545 nm. Forskolin and iso-forskolin were identified by comparing the ultraviolet spectra of standard and sample track at Rfof 0.64 ± 0.02 and 0.36 ± 0.01, after derivatization with anisaldehyde sulfuric acid reagent. The linearity of both the analytes was obtained in the range of 300–1200 ng/spot with the regression coefficient (R2) of 0.991 and 0.986. Recovery of analyte (s) at three levels, namely, 100, 150, and 200 ng/spot was found to be 100.46% ± 0.29%, 99.64% ± 0.33%, 100.02% ± 0.76% and 99.76% ± 0.62%, 99.56% ± 0.35%, 100.02% ± 0.22%, respectively, for forskolin and iso-forskolin. The content of forskolin and iso-forskolin varies from 0.046% to 0.187% and 0.002% to 0.077%, respectively (dry weight basis), the maximum content of both the markers was found in NBC-31, from Thakurwada, Maharashtra. Conclusion: The developed HPTLC method was linear, accurate, and reliable as per the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The study aids in the identification of elite chemotype for commercial prospection of industrially viable medicinal crop. [ABSTRACT FROM AUTHOR]

Background: Sansevieria roxburghiana Schult. and Schult. f. (Asparagaceae) grows in India, Indonesia, Sri Lanka, and tropical Africa. Even though the plant has been traditionally used for the treatment of many ailments, the antioxidant and antiproliferative activities of S. roxburghiana methanol extract and its fractions have not yet been explored. Materials and Methods: Quantitative estimation of phenols and different antioxidant assays were performed using standard methods. Anti-proliferative effect of the extract and fractions were evaluated in HCT-116, HeLa, MCF-7, HepG2, and A-549 cancer cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and sulforhodamine B (SRB) assay methods. High-performance liquid chromatography (HPLC) and high-performance thin layer chromatography (HPTLC) fingerprint profiling were carried out for extract and different fractions. Results: Significant antioxidant and anti-proliferate activity were detected in ethyl acetate fraction. Ethyl acetate fraction showed prominent scavenging activity in 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, and nitric oxide antioxidant assays with an concentration yielding 50% inhibition (IC50) 15.33 ± 1.45, 45.3 ± 1.93 and 48.43 ± 0.46 µg/ml, respectively. Cytotoxicity of ethyl acetate fraction was the highest among other fractions against HCT-116, HeLa, and MCF-7cancer cell lines with IC50 values 16.55 ± 1.28, 12.38 ± 1.36, and 8.03 ± 1.9 µg/ml, respectively, by MTT assay and 15.57 ± 0.70, 13.19 ± 0.49, and 10.34 ± 0.9 µg/ml, respectively, by SRB assay. The presence of gallic acid in the ethyl acetate fraction of S. roxburghiana rhizomes was confirmed by HPLC and HPTLC analysis. Conclusion: Results suggested that ethyl acetate fraction exhibited effective antioxidant and antiproliferative activities. The phenolic compounds identified in ethyl acetate fraction could be responsible for the activities. [ABSTRACT FROM AUTHOR]

Objective: Antioxidant potential has protective effects in diabetic neuropathy (DN); hence, the present study was designed with an objective to quantify quercetin from shade-dried leaves of Allium cepa Lam. and to study its effects on streptozotocin (STZ)-induced chronic DN. Materials and Methods: The shade-dried leaves of A. cepa Lam. were extracted with methanol and then fractionated using ethyl acetate (ACEA). The quantification of quercetin in ACEA was evaluated by high-performance thin layer chromatography (HPTLC). The STZ (40 mg/kg) was administered to Sprague-Dawley rats (180-250 g) maintained at normal housing conditions. The STZ was administered once a day for 3 consecutive days. The elevation in blood glucose was monitored for 3 weeks periodically using flavin adenine dinucleotide-glucose dehydrogenase method by Contour TS glucometer. Rats showing blood glucose above 250 mg/dl were selected for the study. Animals were divided into eight groups. ACEA (25, 50, and 100 mg/kg), quercetin (40 mg/kg), metformin (120 mg/kg), and gabapentin (100 mg/kg) were given orally once a day for 2 weeks. The blood glucose level was again measured at the end of treatment to assess DN. Thermal hyperalgesia, cold allodynia, motor incoordination, and neurotoxicity were studied initially and at the end of 2-week treatment. Biochemical parameters were also evaluated after 2-week drug treatment. Results: The quercetin present in ACEA was 4.82% by HPTLC. All the ACEA treatment reduces blood glucose level at the end of the 2-week study and shows a significant neuroprotective effect in STZ-induced DN in the above experimental models. Conclusion: The quercetin present in ACEA proved protective effect in STZ-induced DN. [ABSTRACT FROM AUTHOR]

A simple, accurate, and precise high-performance thin layer chromatographic (HPTLC) method has been developed and validated for the simultaneous quantification of antihypertensive drugs, amlodipine (AML), hydrochlorothiazide (HCTZ), lisinopril (LIS), and valsartan (VAL) in their pharmaceutical formulations and human plasma. Separation of the drugs was performed on aluminum-backed layer of silica gel 60 F254using a mixture of methanol–dichloromethane–glacial acetic acid (9.0:1.0:0.1, v/v/v) as the mobile phase. Densitometric determination of the separated spots was done at 215 nm. The retention factors (Rf) obtained under the optimized conditions were 0.56, 0.75, 0.29, and 0.67 for AML, HCTZ, LIS, and VAL, respectively. Linearity of the method was established in the range of 200–1,500 ng/band for AML, 300–1,500 ng/band for HCTZ, 400–2,000 ng/band for LIS, and 1,000–7,000 ng/band for VAL. The limit of detection/limit of quantitation of the method found were 54.21/164.28, 77.27/234.15, 83.45/252.87, and 156.48/474.19 ng/band for AML, HCTZ, LIS, and VAL, respectively. To determine the drugs in spiked plasma samples, solid phase extraction was performed, which provided highly consistent and quantitative recovery for all four drugs. The method was satisfactorily applied for the analysis of different tablet formulations and proved to be specific and accurate for the quality control of these drugs. [ABSTRACT FROM AUTHOR]

An integrated screening approach was developed to assess the potential of the Botryococcus genus for the production of lipids for biofuel application (hydrocarbons and triacylglycerols). The strategy developed in this study permitted to rigorously measure mandatory parameters for the determination of strain performance – i.e. growth rate and oil content. For that purpose, mini-photobioreactors run in parallel were used together with an exhaustive lipid class analysis. The quantitative productivity measurements indicated that the ten screened strains presented very different patterns in the conditions of PBR culture, firstly run in batch, and secondly in continuous mode. Indeed, with the applied setup, only four strains presented biomass productivities close to that previously measured for the reference strain 807/1 (between 74 and 307 mg L −1 day −1 of biomass for the tested strains). HC productivity obtained under continuous light (∼14 mg L −1 day −1 ) was close to that measured for the strain 807/1 (∼20 mg L −1 day −1 ), confirming B. braunii tested strains potential for biofuel application. The analysis of the lipid chemodiversity revealed moreover that this genus should be further investigated for its polar lipids, as it suggests potential pharmaceutical applications such as production of antitumorals or immunosuppressors. [ABSTRACT FROM AUTHOR]

Non-aqueous capillary electrophoresis (NACE) method was optimised for simultaneous determination of major bioactive curcuminoids (CCD) viz., curcumin (CC), desmethoxycurcumin (DC) and bisdesmethoxycurcumin (BC), and some of the degradation products viz., vanillin (VN), vanillic acid (VA), ferulic acid (FA) and 4-hydroxybenzaldehyde (HB) in turmeric milk and herbal products. Separation was achieved using non-aqueous background electrolyte (NA-BGE), consisting of a mixture of sodium tetraborate, sodium hydroxide, methanol and 1-propanol in a single run by capillary electrophoresis (CE). In the present study, a novel ultrasonication-assisted phase separation method (US-PS) was used for extracting the analytes in turmeric milk and the extract was directly injected into the capillary without any pretreatment. High-performance thin layer chromatographic (HPTLC) method for the separation of the analytes was also evaluated on pre-coated silica gel plate. Two different high-performance liquid chromatographic (HPLC) conditions were reported in the literature for the separation of above-mentioned target compounds. The proposed method NACE is simple, fast, convenient and economical. [ABSTRACT FROM AUTHOR]

Background: Woodfordia fruticosa, a plant of Indian origin, is extensively used in folk medicine for the treatment of various ailments. Objective: The aim of the present study was to standardize the flowers of W. fruticosa, Kurz (Lythraceae), an important plant of Indian origin and explore the chemical constituents contributing to its anti-ulcer activity. Materials and Methods: High-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) profiling of the three samples of W. fruticosa flowers purchased from three different markets was done using ellagic acid as the biomarker. Two doses of the aqueous extract of the W. fruticosa (AEWF) flowers were evaluated for anti-ulcer activity by ethanol-induced ulcer model in Wistar albino rats. Omeprazole was used as the positive control. The parameters used for the assessment of the anti-ulcer potential were total titratable acidity (TTA), ulcer index, and percentage protection. Results: The HPTLC and HPLC studies confirmed the presence of ellagic acid in all the three drug samples. The AEWF showed significant reduction in terms of TTA at both doses of 100 mg/kg and 200 mg/kg. The gastroprotection indicated by a lower ulcer index and higher percentage protection was significant for 200 mg/kg dose of AEWF, better than the protection afforded by omeprazole (10 mg/kg). Conclusion: The chromatographic profiling and the anti-ulcer studies served as an efficient tool in the characterization of ellagic acid as an important biomarker for the flowers of W. fruticosa and a probable contributor to the gastroprotective capacity of the drug. The bioactivity studies further supported the traditional use of W. fruticosa in the treatment of ulcers. [ABSTRACT FROM AUTHOR]

Background: Libidibia ferrea (Mart. ex Tul.) L.P. Queiroz (Fabaceae) is a tree which is native to Brazil, widely known as "Jucá," where its herbal derivatives are used in folk medicine with several therapeutic properties. The constituents, which have already been described in the fruit, are mainly hydrolysable tannins (gallic acid [GA] and ellagic acid [EA]). Objective: The aim of this study was to investigate the phenolic variability in the fruit of L. ferrea by ultraviolet/visible (UV/VIS) and chromatographic methods (high-performance liquid chromatography [HPLC]/high-performance thin layer chromatography [HPTLC]). Materials and Methods: Several samples were collected from different regions of Brazil and the qualitative (fingerprints by HPTLC and HPLC) and quantitative analysis (UV/VIS and HPLC) of polyphenols were performed. Results: The HPTLC and HPLC profiles allowed separation and identification of both major analytical markers: EA and GA. The chemical profiles were similar in a number of spots or peaks for the samples, but some differences could be observed in the intensity or area of the analytical markers for HPTLC or HPLC, respectively. Regarding the quantitative analysis, the polyphenolic content by UV/VIS ranged from 13.99 to 37.86 g% expressed as GA or from 10.75 to 29.09 g% expressed as EA. The contents of EA and GA by liquid chromatography-reversed phase (LC-RP) method ranged from 0.57 to 2.68 g% and from 0.54 to 3.23 g%, respectively. Conclusion: The chemical profiles obtained by HPTLC or HPLC, as well as the quantitative analysis by spectrophotometry or LC-RP method, were suitable for discrimination of each herbal sample and can be used as tools for the comparative analysis of the fruits from L. ferrea. [ABSTRACT FROM AUTHOR]