HPTLC Determination of Swertiamarin and Amarogentin in Swertia Species from the Western Himalayas

JPC – Journal of Planar Chromatography – Modern TLC - A high-performance thin-layer chromatographic (HPTLC) method has been established for simple and rapid quantification of two...     

Simultaneous densitometric determination of artemisinin, artemisinic acid and arteannuin-B in Artemisia annua using reversed-phase thin layer chromatography

A rapid and simple RP-TLC method for simultaneous quantification of pharmacologically important sesquiterpene artemisinin (AM) together with its precursors arteannuin-B (AB) and artemisinic acid (AA) in the inflorescence part of Artemisia annua plant has been developed. The RP-TLC of sesquiterpenes was performed on RP-18 F254 S thin-layer chromatographic plates by developing in mobile phase, containing 0.2% TFA in water/ACN (35:65, v/v). The densitometric determination of AM, AB and AA was carried out after derivatization with anisaldehyde reagent at 426 nm in absorption-reflectance mode.     

Identification and quantification of anthocyanins,flavonoids, and phenolic acids in flowers of Rhododendron arboreum and evaluation of their antioxidant potential

The current study aimed to investigate the anthocyanins, non-anthocyanins (flavonoids and phenolic acids), and free radicals scavenging potential in the flowers of Rhododendron arboreum using ultra high performance liquid chromatography with ion mobility quadrupole time of flight tandem mass spectrometry. A total of 25 constituents including nine anthocyanins, six phenolic acids, and ten flavonoids were identified in the flower extract. The major anthocyanins identified were cyanidin-3-O-β-galactoside ( 1 ), cyanidin-3-O-α-arabinoside ( 4 ), and cyanidin-3-O-rhamnoside ( 8 ), while quercetin glycosides were the main identified flavonoids in R. arboreum flowers. Additionally, ultra high performance liquid chromatography methods were developed and validated for the quantification of nine compounds (anthocyanins, flavonoid glycosides, and phenolic acids); five of them were quantified using internal standards. The extracts were analyzed for total phenolics (123.6 mg GAE/g), anthocyanin content (1.76% w/w), and evaluated for antioxidant properties against 2,2-diphenyl-1-picrylhydrazyl radical (IC₅₀: 102.06 and 96.92 μg/mL) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical cation (112.25 and 45.59 μM TE/g) assays. The profiling of R. arboreum for anthocyanins is reported for the first time. The findings suggest that the flowers are a promising source of bioactive constituents and could be used as functional food, antioxidants, and nutraceuticals.     

Steroidal alkaloids from Holarrhena antidysenterica (L.) WALL

Chemical investigations on the stem bark of Holarrhena antidysenterica resulted in the isolation of a new steroidal alkaloid designated as holadysenterine (1), together with three known steroidal alkaloids, conessine (2), isoconessimine (3) and kurchessine (4). Their structures were elucidated on the basis of 1D- and 2D-NMR techniques and high-resolution mass spectrometry.     

Stability-Indicating LC–PDA Method for Determination of Picrosides in Hepatoprotective Indian Herbal Preparations of Picrorhiza kurroa

A liquid chromatography method was developed and validated for the determination of picroside-I and picroside-II in herbal preparations containing Picrorhiza kurroa as one of the ingradients. Resolution of picrosides was achieved on a reversed phase (C-18) endcapped bidentate column by using a mobile phase of acetonitrile: water (25:75 v/v). The detection of picrosides was carried out at 270 nm. The method was validated for precision, accuracy and robustness according to the International Conference on Harmonization (ICH) guidelines and is applicable for the quality control of preparations containing P. kurroa. Analysis of samples in forced degradation proved it to be applicable for stability evaluation. The linear regression analysis data showed good linear relationship (r ² = 0.9999 ± 0.0010 for picroside-I and 0.9997 ± 0.0012 for picroside-II) in the concentration range of 0.4–4.0 μg. The limit of detection and quantification for picroside-I and picroside-II were recorded to be 28.1 and 73.1 ng and 85.2 and 221.5 ng, respectively. Satisfactory recovery results were observed from the herbal preparations (97.5–100.5%). Intra- and inter- day precision of the method was acceptable, with relative standard deviation (%RSD) values in the range of 0.04–1.16% and 0.03–0.27%, respectively.     

Reversed phase-HPLC for rapid determination of polyphenols in flowers of rose species

A rapid, simple, sensitive, robust, and improved HPLC method was developed and validated for determination of 10 polyphenols, namely gallic acid, catechin, epicatechin, rutin, m-coumaric acid, quercitrin, myricetin, quercetin, apigenin, and kaempferol in fresh flowers of Rosa bourboniana and R. brunonii and in both fresh flowers and marc (left after industrial distillation of rose oil) of R. damascena. Six polyphenols, gallic acid, rutin, quercitrin, myricetin, quercetin, and kaempferol, were detected and quantified in all extracts. The chromatographic separation of 10 polyphenols was achieved in less than 16 min by RP-HPLC (Phenomenex, Luna C18 (2) column, 5 microm, 250 mm x 4.6 mm) using linear gradient elution of water and acetonitrile (0.02% trifluroacetic acid) with a flow rate of 1 mL/min at lambda 280 nm. Standard calibration curves were linear in the range of 0.39-500 microg/mL. Good results were achieved with respect to repeatability (RSD <3%) and recovery (98.6-100.8%). The method was validated for linearity, accuracy, repeatability, LOD, and LOQ.     

Online antioxidant activity and ultra-performance LC-electrospray ionisation-quadrupole time-of-fight mass spectrometry for chemical fingerprinting of Indian polyherbal formulations

A HPLC–DAD–DPPH method was developed for evaluating the 1, 1-diphenyl-2-picryl hydrazyl free radical scavenging activity of ethylacetate extracts of different polyherbal formulations (draksarista, draksava, lohasava and arvindasava) by using RP-18e column. The ethylacetate extract from polyherbal, ‘draksarista’ exhibited maximum free radical scavenging activity (99.9 ± 0.38%) followed by draksava (99.8 ± 0.34%), lohasava (98.5 ± 0.30%) and arvindasava (42.3 ± 0.34%) at 100 μg mL^? 1. Simultaneously, ultra-performance liquid chromatography coupled with electrospray ionisation-quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) was used to study chemical composition of the ethylacetate extracts of formulations. The characteristic electrospray mass ionisation reveals the dominance of polyphenols and their glycosides in the four polyherbal formulations.     

Simultaneous determination of sugars and picrosides in Picrorhiza species using ultrasonic extraction and high-performance liquid chromatography with evaporative light scattering detection

Sugars play a critical role in regulating overall cellular metabolism in high altitude growing plants. These plants are shown to have high levels of sugars to enhance their tolerance to abiotic stresses such as drought and freezing temperature. In the present study, a simple, sensitive, selective and reliable HPLC method based on ultrasonic extraction and evaporative light scattering detection (ELSD) has been developed and validated for the simultaneous determination of important sugars (xylose, xylitol, mannitol, glucose and sucrose) and picrosides (picroside-I and picroside-II) in two species Picrorhiza kurroa and P. scrophulariiflora. The analysis was carried out on a Zorbax amino column (250 mm x 4.6 mm i.d., 5 microm) with isocratic elution of acetonitrile:water (78:22, v/v). The method was validated for accuracy, precision, limit of detection and quantification according to International Conference on Harmonization (ICH) guidelines. The drift tube temperature of the ELSD system was set to 81 degrees C and nitrogen flow rate was 2.0 standard liter per minute (SLM). The regression equation revealed a good linear relationship (r(2)=0.9997+/-0.0012) within test ranges. The limit of detection and quantification for seven analytes in ELSD were less than 0.98 and 2.95 microg, respectively. The method showed good reproducibility for the quantification of seven analytes in Picrorhiza species with intra- and inter-day variation of less than 2.0%.     

Structural characterization,photoluminescence, computational studies and bioassay of newly synthesized N-(3-oxo-3-morpholino-1-phenyl-propyl) benzo sulfonamide with multifunctional application

Research on Chemical Intermediates - N-(3-oxo-3-morpholino-1-phenyl-propyl) benzo sulfonamide (BMS) (C19H22N2O4S) has been synthesized and characterized by FT-IR, 1H and 13C NMR, and ESI-mass...     

A rapid RP-HPTLC densitometry method for simultaneous determination of major flavonoids in important medicinal plants

A simple, sensitive, selective, precise, and robust high-performance TLC (HPTLC) method was developed and validated for determination of flavonoids in herbal extracts Bauhinia variegata, Bacopa monnieri, Centella asiatica, Ginkgo biloba, Lonicera japonica, Rosa bourboniana, Rosa brunonii, and Rosa damascena. The HPTLC of flavonoids was performed on RP-18 F(254) TLC plates with dual run, water (5% formic acid)/methanol (70:30) and water (5% formic acid)/methanol (50:50) as mobile phases. Densitometric determination of flavonoids was performed at lambda = 280 nm in reflectance/absorbance mode. The linear regression analysis data for the calibration plots showed a good linear relationship with r(2 )= 0.998 +/- 0.0003 in the concentration range of 150-800 ng/spot for apigenin and rutin and 200-1000 ng/spot for quercetin, luteolin, and quercitrin with respect to peak area. The average recovery for apigenin, quercetin, rutin, luteolin, and quercitrin was 97-99.8% indicating the excellent reproducibility. Statistical analysis of the data showed that the method is reproducible and selective for determination of flavonoids.