Identification, Validation and Standardization of Bioactive Molecules Using UPLC/MS-QToF, UHPLC and HPTLC in Divya-Denguenil-Vati: A Penta-Herbal Formulation for Dengue Fever.

For the last fifty years, Dengue has been one of the most common mosquito-borne arboviral infections which has spread over the tropical and subtropical world. Divya-Denguenil-Vati (DNV) has been formulated by blending five specific herbs for effective resolution of Dengue fever. In the present study, we aimed to identify, develop, validate, and standardize methods for Divya-Denguenil-Vati (DNV), on UHPLC and HPTLC analytical platforms, with rapid, sensitive, accurate and rugged attributes. At first, 97 phyto-constituents were identified by UPLC/MS-QToF in Divya-Denguenil-Vati. UHPLC method was then developed and validated for simultaneous determination of gallic acid, 5-HMF, protocatechuic acid, magnoflorine, methyl gallate, berberine, rutin, ellagic acid, ß-ecdysone and rosmarinic acid in DNV. Four selected markers, gallic acid, rosmarinic acid, magnoflorine and rutin were further developed and validated on HPTLC. Analytical processes were validated as per ICH Q2 (R1) guidelines; and were found linear (r 2 > 0.99), sensitive, precise (%RSD < 5%), and accurate, as indicated by high recovery values (88-105%). The limit of detection and quantification were also established for these phyto-metabolites, with their respective RSDs within 5% limits. Finally, these validated methods were employed to test twenty six different commercial batches of DNV. The quality, reproducibility and consistency of DNV have been well established using these developed and reliable analytical tools. These analytical strategies successfully set a path forward for robust quality evaluation and standardization of Divya-Denguenil-Vati, and other related herbal formulations. Supplementary Information: The online version contains supplementary material available at 10.1007/s10337-022-04183-7.

Standardization and validation of phytometabolites by UHPLC and high-performance thin layer chromatography for rapid quality assessment of ancient ayurvedic medicine, Mahayograj Guggul.

Mahayograj Guggul is an ancient ayurvedic medicine, prescribed for various joint disorders like arthritis, gout, and rheumatism. The present research was envisaged to develop a simple, sensitive, and comprehensive analytical method for standardization of Mahayograj Guggul. The analysis was conducted for gallic acid, protocatechuic acid, vanillic acid, cinnamic acid, piperine, guggulsterone-E, and guggulsterone-Z by high-performance thin-layer chromatography, and additionally ferulic acid, ellagic acid, and picroside I by ultra high-performance liquid chromatography. These developed methods were validated as per international guidelines, and were found linear (r2  > 0.99), sensitive, precise (relative standard deviation < 5%), and accurate with recovery values (85-105%). The limit of detection and quantification were in the range of 0.11-23.6 and 0.33-71.51 µg/g. Gas chromatography tandem mass spectrometry was used to develop Mahayograj Guggul fingerprinting profile and to identify mid-polar or nonpolar compounds. Proximate analysis was used to ascertain the functional groups present in Mahayograj Guggul. Ultra high-performance liquid chromatography and gas chromatography tandem mass spectrometry were further employed to authenticate quality reproducibility in the active ingredients of Mahayograj Guggul in six commercial batches. Taken together, these analytical methods provide a scientific basis and reference for quality control evaluation of Mahayograj Guggul and similar traditional broad-spectrum formulations.

Development, validation, and uncertainty measurement of multi-residue analysis of organochlorine and organophosphorus pesticides using pressurized liquid extraction and dispersive-SPE techniques.

Simple and efficient multi-residue analytical methods were developed and validated for the determination of 13 organochlorine and 17 organophosphorous pesticides from soil, spinach and eggplant. Techniques namely accelerated solvent extraction and dispersive SPE were used for sample preparations. The recovery studies were carried out by spiking the samples at three concentration levels (1 limit of quantification (LOQ), 5 LOQ, and 10 LOQ). The methods were subjected to a thorough validation procedure. The mean recovery for soil, spinach and eggplant were in the range of 70-120% with median CV (%) below 10%. The total uncertainty was evaluated taking four main independent sources viz., weighing, purity of the standard, GC calibration curve and repeatability under consideration. The expanded uncertainty was well below 10% for most of the pesticides and the rest fell in the range of 10-20%.