High-performance Thin-layer Chromatography Technique Research Articles

Simultaneous Determination of Caffeine and Paracetamol in Commercial Formulations Using Greener Normal-Phase and Reversed-Phase HPTLC Methods: A Contrast of Validation Parameters.

There has been no assessment of the greenness of the described analytical techniques for the simultaneous determination (SMD) of caffeine and paracetamol. As a result, in comparison to the greener normal-phase high-performance thin-layer chromatography (HPTLC) technique, this research was conducted to develop a rapid, sensitive, and greener reversed-phase HPTLC approach for the SMD of caffeine and paracetamol in commercial formulations. The greenness of both techniques was calculated using the AGREE method. For the SMD of caffeine and paracetamol, the greener normal-phase and reversed-phase HPTLC methods were linear in the 50–500 ng/band and 25–800 ng/band ranges, respectively. For the SMD of caffeine and paracetamol, the greener reversed-phase HPTLC approach was more sensitive, accurate, precise, and robust than the greener normal-phase HPTLC technique. For the SMD of caffeine paracetamol in commercial PANEXT and SAFEXT tablets, the greener reversed-phase HPTLC technique was superior to the greener normal-phase HPTLC approach. The AGREE scores for the greener normal-phase and reversed-phase HPTLC approaches were estimated as 0.81 and 0.83, respectively, indicated excellent greenness profiles for both analytical approaches. The greener reversed-phase HPTLC approach is judged superior to the greener normal-phase HPTLC approach based on numerous validation parameters and pharmaceutical assays.

Phytochemical and Comparative Assessment of Bark and Root Extracts of Ixora barbata Roxb. ex. Sm. and Ixora coccinea L. Using High-Performance Thin Layer Chromatography Assay

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 highperformance 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.

Qualitative and quantitative estimation of gallic acid in flower galls of Crataeva religiosa through FT-IR and HPTLC techniques

In the present study, FT- IR (FourierTransform Infrared spectroscopy) analysis of galled flower was conducted to qualitatively identify gallic acid while comparative HPTLC (High-Performance ThinLayer Chromatography) studies of normal and galled flower extracts confirmed the gallic acid quantitatively. The quantitative estimation of gallic acid was carried out on silica gel 60 F24HPTLC plates (stationary phase). The plates were developed in a solvent system as prepared with Toluene, Acetone and Glacial acetic acid at a ratio of 3:1:2, in a CAMAG glass twin trough chamber of 20X10 cm, at a temperature of 2 20C. Detection was carried out densitometrically at 220 nm. The FTIR analysis was done with a spectrophotometer system, which was used to identify and analyze the characteristic peaks and their respective functional groups. HPTLC analysis revealed that gallic acid content increased almost two folds in gall tissues as compared to normal tissues. C. religiosa flower galls can be utilized medicinally due to enhanced gallic acid.This is the first report of the occurrence of gallic acid in flower galls of C.religiosa.

Rapid, Highly-Sensitive and Ecologically Greener Reversed-Phase/Normal-Phase HPTLC Technique with Univariate Calibration for the Determination of Trans-Resveratrol

The rapid, highly-sensitive and ecologically greener reversed-phase (RP)/normal-phase (NP) high-performance thin-layer chromatography (HPTLC) densitometric technique has been developed and validated for the determination of trans-resveratrol (TRV). The reversed-phase HPTLC-based analysis of TRV was performed using ethanol–water (65:35, v v−1) combination as the greener mobile phase, while, the normal-phase HPTLC-based estimation of TRV was performed using chloroform–methanol (85:15, v v−1) combination as the routine mobile phase. The TRV detection was carried out at 302 nm for RP/NP densitometric assay. The linearity was recorded as 10–1200 and 30–400 ng band−1 for RP and NP HPTLC techniques, respectively. The RP densitometric assay was observed as highly-sensitive, accurate, precise and robust for TRV detection in comparison with the NP densitometric assay. The contents of TRV in commercial formulation were recorded as 101.21% utilizing the RP densitometric assay, while, the contents of TRV in commercial formulation were found to be 91.64% utilizing the NP densitometric assay. The greener profile of RP/NP technique was obtained using the analytical GREEnness (AGREE) approach. The AGREE scales for RP and NP densitometric assays were estimated 0.75 and 0.48, respectively. The recorded AGREE scale for the RP densitometric assay indicated that this technique was highly green/the ecologically greener compared to the NP densitometric assay. After successful optimization of analytical conditions, validation parameters, AGREE scale and chromatography performance, the RP densitometric assay with univariate calibration was found to be better than the NP densitometric assay for the analysis of TRV.

Stability‐indicating reversed‐phase/normal‐phase high‐performance thin‐layer chromatography technique for the determination of arbidol: Green analytical chemistry viewpoint

AbstractThe sensitive and rapid stability‐indicating green reversed‐phase (RP)/routine normal‐phase (NP) high‐performance thin‐layer chromatography (HPTLC) technique has been established for the analysis of arbidol (ADL) from green analytical chemistry viewpoint. The green reversed‐phase analysis of ADL was carried out using RP‐18 silica gel 60 F254S plates. The mobile phase for the green reversed‐phase technique was ethanol/water/ammonia (80:10:10, vol/vol/vol). The routine normal‐phase analysis of ADL was performed using NP‐18 silica gel 60 F254S plates. The mobile phase for the routine normal‐phase technique was hexane/acetone/ammonia (50:45:5, vol/vol/vol). The detection of ADL was performed at 317 nm for both of the techniques. The linearity ranges for the green reversed‐phase and routine normal‐phase techniques were recorded as 40–800 ng/band and 100–600 ng/band, respectively. The proposed reversed‐phase HPTLC technique was also found to be stability‐indicating for the quantification of ADL in the presence of impurities/degradation products. The AGREE scores for green reversed‐phase HPTLC and routine normal‐phase HPTLC techniques were obtained as 0.86 and 0.49, respectively, suggesting the excellent greenness for reversed‐phase HPTLC technique compared with routine normal‐phase HPTLC technique. Overall, the green reversed‐phase HPTLC technique was found to be superior over routine normal‐phase HPTLC technique for the detection of ADL.

A rapid and highly sensitive stability-indicating high-performance thin-layer chromatography technique for the determination of tedizolid phosphate with a classical univariate calibration

A rapid and highly sensitive stability-indicating high-performance thin-layer chromatography technique for the determination of tedizolid phosphate with a classical univariate calibration

Simultaneous Estimation of Cinnamaldehyde and Eugenol in Essential Oils and Traditional and Ultrasound-Assisted Extracts of Different Species of Cinnamon Using a Sustainable/Green HPTLC Technique.

A wide range of analytical techniques are reported for the determination of cinnamaldehyde (CCHO) and eugenol (EOH) in plant extracts and herbal formulations either alone or in combination. Nevertheless, sustainable/green analytical techniques for the estimation of CCHO and EOH either alone or in combination are scarce in the literature. Accordingly, the present research was carried out to establish a rapid, highly sensitive, and sustainable high-performance thin-layer chromatography (HPTLC) technique for the simultaneous estimation of CCHO and EOH in the traditional and ultrasound-assisted methanolic extracts of Cinnamomum zeylanicum, C. burmannii, and C. cassia and their essential oils. The simultaneous estimation of CCHO and EOH was performed through NP-18 silica gel 60 F254S HPTLC plates. The cyclohexane/ethyl acetate (90:10, v v−1) solvent system was optimized as the mobile phase for the simultaneous estimation of CCHO and EOH. The greenness score of the HPTLC technique was predicted using AGREE software. The entire analysis was carried out at a detection wavelength of 296 nm for CCHO and EOH. The sustainable HPTLC technique was observed as linear in the range 10–2000 ng band−1 for CCHO and EOH. The proposed technique was found to be highly sensitive, rapid, accurate, precise, and robust for the simultaneous estimation of CCHO and EOH. The content of CCHO in traditional methanolic extracts of C. zeylanicum, C. burmannii, and C. cassia was found to be 96.36, 118.49, and 114.18 mg g−1, respectively. However, the content of CCHO in ultrasound-assisted methanolic extracts of C. zeylanicum, C. burmannii, and C. cassia was found to be 111.57, 134.39, and 129.07 mg g−1, respectively. The content of CCHO in essential oils of C. zeylanicum, C. burmannii, and C. cassia was found to be 191.20, 214.24, and 202.09 mg g−1, respectively. The content of EOH in traditional methanolic extracts of C. zeylanicum, C. burmannii, and C. cassia was found to be 73.38, 165.41, and 109.10 mg g−1, respectively. However, the content of EOH in ultrasound-assisted methanolic extracts of C. zeylanicum, C. burmannii, and C. cassia was found to be 87.20, 218.09, and 121.85 mg g−1, respectively. The content of EOH in essential oils of C. zeylanicum, C. burmannii, and C. cassia was found to be 61.26, 79.21, and 69.02 mg g−1, respectively. The amounts of CCHO and EOH were found to be significantly higher in ultrasound-assisted extracts of all species compared to its traditional extraction and hence ultrasound extraction has been proposed as a superior technique for the extraction of CCHO and EOH. The AGREE analytical score of the present analytical technique was predicted as 0.75, suggesting excellent greenness profile of the proposed analytical technique. Based on all these observations and results, the proposed sustainable HPTLC technique can be successfully used for the simultaneous estimation of CCHO and EOH in different plant extracts and herbal products.

Combining Normal/Reversed-Phase HPTLC with Univariate Calibration for the Piperine Quantification with Traditional and Ultrasound-Assisted Extracts of Various Food Spices of Piper nigrum L. under Green Analytical Chemistry Viewpoint.

Due to unavailability of sustainable analytical techniques for the quantitation of piperine (PPN) in food and pharmaceutical samples, there was a need to develop a rapid and sensitive sustainable analytical technique for the quantitation of PPN. Therefore, the current research presents a fast and highly sensitive normal/reversed-phase high-performance thin-layer chromatography (HPTLC) technique with classical univariate calibration for the quantitation of PPN in various food spices of black pepper with traditional (TE) and ultrasound-assisted extracts (UBE) of various food spices of Piper nigrum L. under green analytical chemistry viewpoint. The amount of PPN in TE of four different spices of black pepper—namely BPMH, BPLU, BPSH, and BPPA—was found to be 309.53, 304.97, 282.82, and 232.73 mg g−1, respectively using a sustainable normal-phase HPTLC technique. However, the amount of PPN in UBE of BPMH, BPLU, BPSH, and BPPA was recorded as 318.52, 314.60, 292.41, and 241.82 mg g−1, respectively using a sustainable normal phase HPTLC technique. The greenness of normal/reversed-phase HPTLC technique was predicted using AGREE metric approach. The eco-scale was found to be 0.90, suggested excellent greenness of normal/reversed-phase technique. UBE of PPN was also found to be superior over TE of PPN. Overall, the results of this research suggested that the proposed normal/reversed-phase densitometry technique could be effectively used for the quantitation of PPN in food and pharmaceutical samples.

Quantitative determination of canagliflozin in human plasma samples using a validated HPTLC method and its application to a pharmacokinetic study in rats.

Canagliflozin (CNZ) is the first sodium-glucose co-transporter-2 inhibitor approved for treatment of type 2 diabetes mellitus. In the proposed work, a sensitive, rapid and validated high-performance thin-layer chromatography (HPTLC) method was established for the estimation of CNZ in human plasma for the first time. HPTLC analysis of CNZ and internal standard (sildenafil) was performed on glass coated silica gel 60 F254 HPTLC plates using a binary mixture of chloroform-methanol 9:1 (%, v/v) as the mobile phase. Densitometric detection was done at 295 nm. Retardation factor values were obtained as 0.22 and 0.52 for the CNZ and the IS, respectively. The linearity range of CNZ was obtained as 200-3,200 ng/ml. A simple protein precipitation method was used for the extraction of analyte from plasma using methanol. The proposed HPTLC technique was validated for linearity, accuracy, precision and robustness. The proposed HPTLC technique was successfully utilized for the assessment of pharmacokinetic profile of CNZ in rats after oral administration. After oral administration, the peak plasma concentration of CNZ was obtained as 1458.01 ng/ml in 2 h. The proposed HPTLC method could be applied to the study of the pharmacokinetic profile of pharmaceutical formulations containing CNZ.

Simple and Accurate HPTLC-Densitometric Method for Quantification of Delafloxacin (A Novel Fluoroquinolone Antibiotic) in Plasma Samples: Application to Pharmacokinetic Study in Rats.

Delafloxacin (DLX) is a recently-approved fluoroquinolone antibiotic, which is recommended for the treatment of “acute bacterial skin and skin structure infections”. A thorough literature survey revealed only a single published method for the estimation of DLX using UPLC-MS/MS technique in biological samples. There is no high-performance thin-layer chromatography (HPTLC) method has been reported for the estimation of DLX in dosage forms and/or biological samples. Therefore, a selective, sensitive, rapid and validated HPTLC-densitometry technique has been used for the estimation of DLX in human plasma for the first time. HPTLC quantification of DLX and internal standard (IS; gatifloxacin) was carried out on glass coated silica gel 60 F254 HPTLC plates using the ternary mixture of ethyl acetate:methanol:ammonia solution 5:4:2 (%, v/v/v) as the mobile phase. Densitometric detection was done at 344 nm. The Rf values were recorded as 0.43 and 0.27 for the DLX and the IS, respectively. The linearity range of DLX was obtained as 16–400 ng/band. A simple protein precipitation method was used for the extraction of analyte from plasma using methanol. The proposed HPTLC technique was validated for “linearity, accuracy, precision, and robustness”. The proposed HPTLC technique was successfully utilized for the assessment of pharmacokinetic profile of DLX in rats after oral administration. After oral administration, the peak plasma concentration of DLX was obtained as 194.19 ng/ml in 1 h. The proposed HPTLC method could be applied in study of pharmacokinetic profile and therapeutic drug monitoring of DLX in clinical practice.

Chromatographic investigation of phytoconstituents in milk yam (Ipomoea digitata L.) tubers

Background: Milk yam commonly called Ksheervidhari is a perennial climber; its tubers are an integral crude drug in different Ayurvedic formulations as well as folkloric medicine. It is used as antidiabetic, restorative, carminative, expectorant, galactagogue, stomachic, and appetizer. Objective: The present study is done with the aim of identifying the maturity stage of milk yam tuber containing maximum concentration of umbelliferone – a coumarin present in the tubers. Materials and Methods: Umbelliferone present in immature tubers (6 months after planting), overmature tubers (36 months after planting), and tubers at optimum maturity (21 months after planting) was assessed using high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) analysis. Results: HPLC and HPTLC techniques revealed the presence of umbelliferone and it was quantified to be higher in over mature tubers (0.44 and 0.42 μg/g, respectively), followed by optimally mature (0.31 and 0.22 μg/g, respectively) and immature tubers (0.20 and

Validated Stability Indicating HPTLC and UV-Spectrophotometric Techniques for the Determination of Avanafil

Aims: To develop methods with complete validation according to ICH guidelines and to be applied for the determination of avanafil in pure form and in pharmaceutical formulation in the presence of its degradation products.
 Study Design: High performance thin layer chromatography (HPTLC) and different spectrophotometric methods (dual wavelength, first derivative, first derivative of ratio spectra and ratio difference are developed for simultaneous determination of avanafil in laboratory-prepared mixtures of avanafil with its degradation products and in pharmaceutical formulation.
 Place and Duration of Study: Sample: Department of Analytical Chemistry, Faculty of Pharmacy (Girls), Al-Azhar University, Cairo, Egypt, between May 2019 and September 2019.
 Methodology: Two techniques have been developed for the determination of avanafil in the presence of its degradation products. The first was HPTLC where separation was performed on silica gel 60 F254 plates, with chloroform: toluene: methanol: conc. ammonia (6:5:3:0.1, by volume) as a developing system and UV detection at 230 nm. The second one was UV- spectrophotometry which included dual wavelength between 267 and 292 nm, first derivative determination of the drug at 261 nm, first derivative of ratio of peak amplitudes at 275.6, 305.4 and 329 nm and the ratio difference with the amplitude difference between (266 and 250 nm).
 Results: HPTLC method was applied over the concentration range of 0.5-5. μg/spot, while spectrophotometric methods were linear over the concentration range 5-50 μg/mL for avanafil.
 Conclusion: Novel, simple and accurate method for the determination of avanafil in laboratory-prepared mixtures of avanafil with its degradation products and in pharmaceutical formulation.

Secondary Metabolites in Arctium lappa L.: Variation Among Plant Parts and Phenological Stages

Arctium lappa L. (Asteraceae) occupies a prominent position in the ethnotherapeutics practices. The herb is known to contain a number of bioactive metabolites, which have immense therapeutic value. The objective of our present investigation was to estimate the variation of metabolites in different parts and phonological stages of A. lappa. This investigation has been done using a high-performance thin-layer chromatography technique. The present analysis reveals that the root (vegetative stages) holds the significantly high content of triterpenoids (betulinic acid, oleanolic acid, and lupeol), in comparison to the reproductive stage.

Separation of 9-Fluorenylmethyloxycarbonyl Amino Acid Derivatives in Micellar Systems of High-Performance Thin-Layer Chromatography and Pressurized Planar Electrochromatography

The problems with separation of amino acid mixtures in reversed-phase mode are the result of their hydrophilic nature. The derivatisation of the amino group of mentioned above solutes leads to their solution. For this purpose, 9-fluorenylmethoxycarbonyl chloroformate (f-moc-Cl) as the derivatisation reagent is often used. In our study, the separation of some f-moc- amino acid derivatives (alanine, phenylalanine, leucine, methionine, proline and tryptophan) with the use of micellar systems of reversed-phase high-performance thin-layer chromatography (HPTLC) and pressurized planar electrochromatography (PPEC) is investigated. The effect of surfactant concentration, its type (anionic, cationic and non-ionic) and mobile phase buffer pH on the discussed above solute migration distances are presented. Our work reveals that the increase of sodium dodecylsulphate concentration in the mobile phase has a different effect on solute retention in HPTLC and PPEC. Moreover, it also affects the order of solutes in both techniques. In PPEC, in contrast to the HPTLC technique, the mobile phase pH affects solute retention. The type of surfactant in the mobile phase also impacts solute retention and migration distances. A mobile phase containing SDS improves system efficiency in both techniques. Herein, such an effect is presented for the first time.

Determination of Amlodipine Besilate and Azilsartan Medoxomil by UHPLC, HPTLC and Spectrophotometric Techniques

Aims: To develop methods with complete validation according to ICH guidelines and to be applied for the determination of both drugs in laboratory prepared mixtures and in synthetic tablets.
 Study Design: Ultra high performance liquid chromatography (UHPLC), High performance thin layer chromatography (HPTLC) and visible spectrophotometric methods are developed for determination of amlodipine besilate and azilsartan medoxomil in laboratory-prepared mixtures and in synthetic tablets.
 Methodology: Two techniques have been developed for the simultaneous determination of amlodipine besilate and azilsartan medoxomil in pure form and synthetic tablets. The first was UHPLC in which separation was achieved on a C18 column using 0.1% o-phosphoric acid - acetonitrile - methanol (60:10:30, by volume) as mobile phase with detection at 243nm. The second was HPTLC where separation was performed on silica gel 60 F254 plates using chloroform- tolune-methanol-glacial acetic acid (7: 1.5: 1.5: 0.5 by volume) as a developing system and UV detection at 243nm. In addition, visible- spectrophotometric method was developed for determination of amlodipine besilate in presence of azilsartan medoxomil through formation of yellowish orange colored product after reaction of amlodipine besilate with anisaldehde in acid medium with λmax at 443 nm.
 Results: UHPLC method was linear over the concentration ranges of 2-20 μg/ mL and 4-40 μg/ mL while HPTLC method was linear over the concentration ranges of 0.2 -4.0 μg/ spot and 0.5-8.0 μg/ spot for amlodipine besilate and azilsartan medoxomil, respectively. The visible spectrophotometric method was found to be valid over the concentration range of 10–80 μg/mL for amlodipine besilate.
 Conclusion: The proposed three techniques are rapid, accurate and precise, thus can be effectively applied for the routine estimation of both drugs in bulk and in their combined formulations.

Validated Stability Indicating HPTLC, UHPLC and UV-Spectrophotometric Techniques for the Determination of Bepotastine Besilate in Presence of Its Oxidative Degradate

The study is aimed at developing methods which have a complete validation as stipulated in the ICH guidelines and to be applied for the determination of Bepotastine besilate (BB) in pure form and in pharmaceutical formulations in the presence of its oxidative degradation product. High performance thin layer chromatography (HPTLC), Ultra high performance liquid chromatography (UHPLC) and different spectrophotometric methods (first derivative, first derivative of ratio spectra and ratio difference are developed for simultaneous determination of bepotastine besilate in laboratory-prepared mixtures of bepotastine besilate with its oxidative degradate and in pharmaceutical formulations were used in the study design. Firstly, HPTLC was performed and separation occurred on silica gel 60 F254 plates, with butanol: ammonia (8:2, v/v) as a developing system. UHPLC in which separation occurred on a Kinetex C 18 column using methanol- 0.1% O-phosphoric acid - acetonitrile (70:20:10, by volume) as mobile phase, followed. And lastly was UV/Vis spectrophotometry which included first derivative determination of the drug at 252.6 nm, first derivative of ratio of peak amplitudes at 233.4, 250 and 275.6 nm and the ratio difference with the amplitude difference between (240 nm and 260 nm). Result showed that HPTLC method was applicable over the concentration range of 0.5-5 μg / band, while UHPLC method was linear over the concentration 2- 12 μg/mL and spectrophotometric methods were linear over the concentration range 20-120 μg/mL for bepotastine besilate. The proposed three techniques are quite accurate and precise. They can be used for routine analysis of bepotastine besilate in pharmaceutical formulation and stability indicating methods.

Pharmacognostic standardization and chromatographic fingerprint analysis on triterpenoidal constituents of the medicinally important plant Artocarpus heterophyllus by high-performance thin layer chromatography technique

Background: Artocarpus heterophyllus commonly known as Kathal in Hindi and Jackfruit in English has a wide horizon of medicinal possessions. The plant is found in India and in its tropical regions. Objective: Although the plant and its extracts are renowned for its ethnic medicinal values diversely in India, yet organized data somewhere lack in reverse pharmacognostical approach of this plant that shows that plant have not been completely explored for its therapeutic potency. Materials and Methods: In the present study, the folklore potential of this plant has been explored by generating down its pharmacognostical standards along with measurement of its active therapeutic constituent ursolic acid and lupeol via. High-performance thin layer chromatography (HPTLC), evidence from organized data search says that ursolic acid and lupeol is ubiquitous to A. heterophyllus. The plant was also subjected to spectroscopic-based estimation of tannins (gallic acid and tannic acid) and flavonoids (quercetin and rutin). Antimicrobial testing was also performed. Results: Microscopic features revealed the presence of anomocytic type of stomata, collateral open type vascular bundle in which fascicular cambium is present, calcium oxalate crystal and covering trichomes were key features in leaves. Methanolic extract of leaves of the plant was subjected to HPTLC. HPTLC studies revealed that both ursolic acid and lupeol are present in appreciable amount. Plant showed good antibacterial activity which may be due to the high amount of tannins as the tannins has the ability to disintegrate the bacterial cell wall. Conclusion: The data generated could be significantly used as a reference for the authentication and quality control of A. heterophyllus.

ASSESSMENT OF CHROMATOGRAPHIC SEPARATIONS FOR ANTIHYPERTENSIVE AND ANTIDIABETIC DRUGS

Context: The focus of this review is to compile the different chromatographic methods that were reported earlier for the analysis of different antihypertensive and antidiabetic drugs. Objective: The magnitude of chemical entities investigated and entering into the medicinal field for various health-related ailments is escalating year after year. The drugs are either innovative entities or fractional structural variation of the preexisting chemical molecule. These drugs may exhibit unexpected toxicities after Phase IV of clinical trials, resulting in their withdrawal from the market. Under these circumstances, analytical measures for these drugs may not be accessible in the pharmacopeias. The main aim of this work is to compile the different analytical techniques for the quantification of various antihypertensive drugs and antidiabetic drugs. Methods: The present work is to thoroughly study the literature for the application of different analytical techniques such as high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectroscopy/tandem mass spectroscopy (LC-MS/MS) for the quantification of antihypertensive drugs and anti-diabetic drugs. Results: The present study attempts to collate various analytical techniques that were developed and validated for the estimation of few important antidiabetic and antihypertensive drugs either in pure, individually or combined with other pharmaceutical dosage form by HPLC, LC-MS/MS, and high-performance thin-layer chromatography techniques. Conclusion: Different chromatographic methods are considered to be rapid tools for qualitative and quantitative analysis of newer chemical entities in pharmaceuticals. The amount of these newer chemical entities which are reaching the pharmaceutical market is increasing day by day nevertheless there exists a lag in establishing the standard protocols for the identification, impurity profiling, related substance and assay method. Hence, the present review compiles the different analytical methods that were reported in the literature and thus helps the researchers and chemists to make use of the analytical techniques for the quantification and validation of various antidiabetic and antihypertensive drugs.

Separation and identification of polar polyphenols in oily formulation using high-performance thin-layer chromatography and mass spectroscopy techniques

In cosmetics, there is some interest to solubilized polar antioxidant molecules in oil. This is a matrix that can indeed protect these fragile molecules against oxidation and that makes possible the polar antioxidants transfer through the epidermis in order to be delivered within the dermis. Varying the oil formulation allows the achievement of a high content of antioxidants in a natural vegetable oil using an oleo-eco extraction process. However, it was necessary to characterize this content using quantitative and efficient methods such as High-Performance Thin-Layer Chromatography technique. The aim of this work was to develop a method of antioxidant’s characterization with this technique without a preliminary re-extraction of the antioxidants.

Characterization of the Phenolic Compound, Gallic Acid from Sansevieria roxburghiana Schult and Schult. f. Rhizomes and Antioxidant and Cytotoxic Activities Evaluation.

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 mg/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.SUMMARY Sansevieria roxburghiana has been selected for in vitro antioxidant and cytotoxicity screeningEthyl acetate fraction of methanol extract of S. roxburghiana exhibited effective antioxidant and antiproliferative activitiesThe activity of ethyl acetate fraction may be due to the presence of phenolic compound which is identified by high-performance liquid chromatography and high-performance thin layer chromatography techniques. Abbreviations used: %: Percent, ºC: Celsius, mg: Microgram, ml-Microlitre, ANOVA: Analysis of variance, DMSO: Dimethyl sulfoxide, g: Grams, IC50: Concentration yielding 50% inhibition, Kg: Kilogram, mg: Milligram, min: Minutes, ml: Milliliter, HPLC: High-performance liquid chromatography, HPTLC: High-performance thin layer chromatography, DPPH: 1,1-diphenyl-2-picrylhydrazyl, ABTS: 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, GAE: Gallic acid equivalents, SRME: Methanol extract of S. roxburghiana, ROS: Reactive oxygen species, SRPE: Petroleum ether fraction of S. roxburghiana, SREA: Ethyl acetate fraction of S. roxburghiana, SRAQ: Aqueous fraction of S. roxburghiana, DMEM: Dulbecco's Minimum Essential Medium, FBS: Fetal bovine serum, OD: Optical density, TPC: Total phenolic content, SRBU: Butanol fraction of S. roxburghiana.