HPTLC Quantification Research Articles

Quality Control of Triphala Churna

Article Quality Control of Triphala Churna Arun Kumar Singh Parihar 1, Umakant Sahu 1, Kamleshwar Singh Karbhal 1, Shrikant Rambhau Inchulkar 1, Nagendra Singh Chauhan 1,*, and Mayank Krishna Kulshreshtha 2 1 Drugs Testing Laboratory AvamAnusandhan Kendra, Raipur, Chhattisgarh 492010, India 2 Government Ayurved College, Bilaspur, Chhattisgarh 495001, India * Correspondence: chauhan.nagendra@gmail.com; Tel. +919406558176 Received: 22 April 2024; Revised: 3 July 2024; Accepted: 15 July 2024; Published: 23 July 2024 Abstract: Objectives: Triphala Churna (TC) is a tridoshashamak Ayurvedic rasayana used for treating various diseases such as skin diseases, diabetes, and as a mild laxative. Methods: In this study, TC was prepared and quality standardized standardized using various parameters such as macroscopical evaluation, physical properties, physicochemical properties, and phytochemical analysis (HPTLC fingerprinting and quantification) methods to assess its quality and purity. Results: The qualitative and quantitative phytochemical assays via HPTLC revealed concentrations of Gallic acid as 3.62 ± 0.05%, 3.67 ± 0.04%, 1.52 ± 0.03%, and 2.96 ± 0.02% for Amlaki (Emblica officinalis), Haritiki (Terminalia chebula), Vibhitki (Terminalia bellerica), and TC, respectively. Conclusion: This study provides valuable insights into establishing robust quality control measures and developing reliable assays for both Triphala Churna (TC) and its individual components.

HPTLC quantification of quercetin from leaf extracts of C. occidentalis L. and its inhibitory activity against protease enzyme of P. falciparum

HPTLC quantification of quercetin from leaf extracts of C. occidentalis L. and its inhibitory activity against protease enzyme of P. falciparum

A validated HPTLC quantification of artemisinin from different extracts of Artemisia annua L. and its inhibitory activity against serine hydroxyl methyltransferase (SHMT)

The present study aimed to analyze the artemisinin content in various extracts of Artemisia annua L. using a validated HPTLC densitometry method and evaluate the inhibitory activity of artemisinin against serine hydroxyl methyltransferase (SHMT) of Plasmodium falciparum using in silico approach. High performance thin layer chromatography revealed that the petroleum ether extract of A. annua had the highest artemisinin content (0.71% ± 0.02 w/w), methanol extract (0.25% ± 0.02 w/w), chloroform extract (0.14% ± 0.01 w/w), and water extract (0.06% ± 0.01 w/w). Densitometry result indicated a well-defined and symmetrical peak in the petroleum ether extract, indicating excellent separation of high-purity artemisinin. In silico study of artemisinin with SHMT enzyme portrayed an excellent binding affinity (–9.1 kcal/mol) and convenient interactions compared with control. Moreover, both artemisinin and control compound showed good absorption, distribution, metabolism, excretion and toxicity properties. In conclusion, the study suggested that the petroleum ether extract of A. annua is the most suitable and efficient solvent for extracting high-quantity artemisinin. The findings of the molecular docking analysis further supported the potential inhibitory activity of artemisinin against the SHTM of P. falciparum. This research contributes to the understanding of artemisinin’s antimalarial properties and optimized method of its production efficiently.

HPTLC Method Development and quantification of marker compound Gallic acid and Piperine in Ayurvedic Polyherbal formulations: Avipattikar Churna

Ayurvedic medications are becoming more and more well-liked and accepted globally because they are inexpensive and have no negative side effects. There is a chance that original medications will be adulterated with substances that chemically or physically mimic raw pharmaceuticals because of the increasing demand for herbal raw materials for the production of various classical as well as phytoformulation. Avipattikar Churna is an excellent Ayurvedic formulation for treating health issues resulting from an imbalance of Pitta dosha, such as acidity, heartburn, and indigestion. These issues might arise from bad eating habits, a sedentary lifestyle, or a lack of physical activity. Avipattikar Churna is also beneficial for ailments affecting the digestive and excretory systems' ability to operate normally. It counteracts the gastrointestinal tract's acid secretion and encourages the synthesis of digestive enzymes, which facilitate food absorption. Acid dyspepsia, often known as indigestion, is a condition marked by burning and sour vomit, nausea, heartburn (retrosternal burning), and burning in the throat. Avipattikar Churna is beneficial in treating these symptoms. The standardisation process, which can be carried out using a variety of methods and advanced techniques, conforms the identity, quality, and purity of herbal pharmaceuticals. The HPTLC method is superior to other methods since it may be used with herbal medications and is inexpensive, simple to use, and repeatable. For the purpose of evaluating the quality of Avipattikar Churna and identifying any changes made to the drug's composition, an HPTLC method has been devised. The existence and quantity of the marker compound in the sample are confirmed by the overlap of all relevant spectra with the marker. A gramme of Avipattikar Churna (AVC) methanolic extract contained 2.51 mg of piperine and 2.70 mg of gallic acid, the marker compounds.

HPTLC Quantification of α-Glucosidase Inhibitor Mangiferin in Hydro-alcoholic extract of Salacia Species and Antidiabetic Poly-herbal Formulation

Salacia oblonga Wall. and Salacia reticulata Wight. (Family Celastraceae) are commonly known as Saptarangi and used in Ayurvedic medicine as potent antidiabetic agent. It is a woody climber, mainly habitat in Sri Lanka and Southern India. A large number of chemical constituents such as salacinol, neosalacinol, kotalanol, neokotalanol, and mangiferin were isolated from stem and root of saptarangi which shows various therapeutics activities. Chemical constituent presents in both species of Salacia are working as α-glucosidase inhibitor for diabetes management. Among them mangiferin is a plant natural polyphenol of C-glycosylxanthone structure and found in many plant species. Many chromatography techniques like LC-MS, HPLC are available to quantify mangiferin but no proper data available for simple, rapid, precise, economic HPTLC method. So, HPTLC method was developed with mobile phase ethyl acetate: formic acid: water (4:0.5:0.5 v/v/v) which confirmed the presence of mangiferin at 0.38±0.01 Rf value observed under 254nm. Mangiferin was present in 1.02% in Salacia oblonga root extract, 0.94% in Salacia reticulata stem extract and 0.42% in polyherbal formulation. The validated HPTLC method will be useful in standardization of different Ayurvedic formulations using mangiferin as a marker.

HPTLC method development and quantification of marker compound eugenol in ayurvedic polyherbal formulations: Avipattikar churna

The popularity and acceptance of ayurveda drugs are increasing day by day worldwide due to their low cost and no side effects. Due to high demand of herbal raw materials for the manufacturing of different classical as well as phytoformulation there is scope for adulteration of original drugs with chemically or physically resemble raw drugs. Standardization process conforms the identity, quality and purity of herbal drugs, which can be done by different procedures and by using sophisticated techniques. HPTLC method has advantages over other techniques due to its suitability for herbal drugs, low cost, reproducibility and it is easy to operate as well. An HPTLC method has been developed for the quality testing of Avipattikar churna (AVC) and to discriminate any kind of alteration in drugs composition.

HPTLC quantification, assessment of antioxidant potential and in vivo hypoglycemic activity of Scorzonera latifolia (Fisch. & C.A. Mey.) DC. and its major compounds

Scorzonera L. (Asteraceae) species have long been the topic of many phytochemical, analytical, and biological studies since various species of the Scorzonera genus have been widely used as food and medicinal purposes. Apart from its traditional use to relieve pain, promote wound healing, treat helminth infections or women infertility, Scorzonera latifolia (Fisch. & Mey.) DC is also used for its antidiabetic activity. We aimed to investigate antidiabetic activity of the aerial parts of the title plant and its major secondary metabolites (hyperoside, isoquercitrin, 7- O -methylisoorientin, isoorientin, swertisin, chlorogenic acid, 4,5- O -dicaffeoylquinic acid and hydrangenol-8- O - β -glucoside) isolated from aerial parts in alloxan-induced diabetic mice. Blood glucose levels were measured four times: before the treatment, after 1 st , 2 nd , and 4 th hours of sample treatments (100 mg/kg i.p.). S. latifolia extract displayed notable decline after 4 hours of administration. Among the metabolites; swertisin, 7- O -methyl-isoorientin, and hydrangenol-8- O - β -glucoside were associated with significant reduction on blood glucose level of alloxan-induced diabetic mice. Due to the strong relationship between oxidative stress and diabetes, antioxidant activity of S. latifolia was additionally tested. Furthermore, 4,5- O -dicaffeoylquinic acid, chlorogenic acid, hyperoside, and swertisin contents as major components of the extract were quantified by HPTLC-densitometry, as their biological effects can be attributed to their phenolic contents.

HPTLC Quantification and Estimation of Rutin and Kaempferol using aerial parts of Ixora javanica (blume) dc. and Ixora barbata roxb. ex. sm.

Plants extracts continue to be natural sources of various drugs which play an important role in combating several diseases and infections. Flavonoids are an important class of secondary metabolites with immense potential to be used as drugs, for treating several diseases. With an aim to discover more natural sources of flavonoids to be used as drugs, leaf and flower extracts of two species of Ixora were analyzed. Current study deals with phytochemical analysis and quantification of 2 flavonoids, Rutin and Kaempferol, present in Ixora javanica and Ixora barbata using HPTLC method. The present study involves detection, quantification and comparison of both the above flavonoids present in their leaf and flower extracts. The solvents used for extraction were also compared to suggest the best solvent to be used to obtain maximum amount of flavonoids. Leaf and flower extracts of both plants were made using petroleum ether, 85% methanol and 90% ethanol solvents. HPTLC analysis indicated the presence of Rutin and Kaempferol in the extracts. The highest amount of Rutin was recorded in 85% methanolic extract of flowers of Ixora javanica (1298.6µg/ml). This was followed by 85% methanolic extract of leaf of Ixora barbata (1183.5µg/ml). The lowest amount of Rutin was observed in 90% ethanolic extract of leaf of Ixora barbata (10.9µg/ml). The highest amount of Kaempferol was observed in 90% ethanolic extract of leaf of Ixora javanica (527.8µg/ml). The lowest amount of Kaempferol was observed in 90% ethanolic extract of leaf of Ixora barbata (58.2µg/ml).

Development and Evaluation of a Polyherbal Tablet for Polycystic Ovarian Syndrome (PCOS)

The main objective of the present study was to develop and evaluate polyherbal tablet for the treatment of Polycystic Ovarian Syndrome (PCOS). For the formulation of tablet, the herbs were selected based on the concepts of pathophysiology and Ayurvedic perspective of the disease. The extracts of <em>T. foenum graceum, C. longa, B. aristata, S. asoka, B. variegata</em> and Guggul purified in cow’s urine was used for the preparation of polyherbal tablet. The physicochemical, phytochemical evaluation and quantification of markers by HPTLC of crude drugs and extracts were performed. The guggul was used as binder in different proportion for the preparation of tablet. Pre-formulation study of the powder blend and post compression evaluation of tablet was done by various parameters like weight variation, friability, hardness, thickness, diameter, disintegration time, in-vitro dissolution and accelerated stability study. The physicochemical evaluations of crude drugs yielded results which were in accordance with the monograph’s standard values. The phytochemical analysis of the extracts revealed the presence of variety of phytoconstituents. From the results of HPTLC quantification the amount of curcumin (7.14%) in <em>C. longa</em>, diosgenin (40.6%) in <em>T. foenum graceum ,T. foenum graceum</em>, berberine hydrochloride (4.83%) in <em>B. aristata</em>, catechin (3.48%) in S. asoka, lupeol (0.11%) in <em>B. variegata</em> and Z-guggulsterone (0.140%) and E-guggulsterone (0.146%) in purified guggul was found. The micrometrics of the powder blend of all the formulations showed good flow properties. Formulation F3 showed better results compared to other formulations in post compression evaluation. Accelerated stability study showed that the formulation F3 was stable during the course of study.

A Study on HPTLC Quantification for Quality Control of Myricetin a Nutraceutical from Different Plant Parts of Myrica esculenta Linn

The present study was performed to report the quantification of myricetin present in different extracts (i.e. ethanol, methanol, petroleum ether, chloroform and aqueous extract) of root, leaf, bark, stem and seed of Myrica esculenta using TLC densitometric method. Densitometric scanning was performed at 366 nm for the determination and quantitation of standard compounds. The compact spots were achieved at Rf 0.67 corresponding to myricetin. The content of myricetin present in different solvent extracts of M. esculenta was 0.2159 ± 0.002% w/w, 0.3139 ± 0.003% w/w, 0.0184 ± 0.004% w/w, 0.0139 ± 0.003% w/w and 0.2120 ± 0.005% w/w in ethanolic, methanolic, petroleum ether, chloroform and aqueous extract of M. esculenta stem, stem, bark, leaf and leaf plant parts, respectively. The highest myricetin content was found in the order of methanolic stem extract>ethanolic stem extract>aqueous leaf extract>petroleum ether bark extract>chloroform leaf extract of M. esculenta. The developed HPTLC method was a precise, simple, convenient and accurate for screening of large number of active compounds present in various solvent extracts. Thus, the study concluded that the developed HPTLC method for quantification of myricetin can be used for a routine quality check and analysis of any formulation and composition containing M. esculenta.

RP-HPLC and HPTLC Identification and Quantification of Flavonoids in the Leaves Extract of Bauhinia acuminata Linn. (Caesalpiniaceae)

Aims: The present investigation is aimed at the identification and quantification of flavonoids in the methanol extract of leaves of Bauhinia acuminata Linn. (Caesalpiniaceae) by a validated HPTLC method and confirmation of the same by RP-HPLC.
 Methodology: CAMAG HPTLC system with VisionCATS software and HPLC Agilent Infinity 1260 were employed for the study. Pre-coated silica gel 60 F254 plates were used as stationary phase and Toluene : Ethyl acetate : Formic acid (5:4:0.2, v/v/v) was used as mobile phase in HPTLC while in HPLC Thermo hypersil BDS C18 column and Methanol and 0.4% Phosphoric acid (65 : 35) were used as stationary and mobile phase respectively.
 Results: Among different standards used in HPTLC, only quercetin was found to be present and further quantitatively analyzed. The method shows good correlation coefficient (R2 ≥ 0.99) and linearity in the range of 0.5 to 5.0 µg/band. The concentration of quercetin in the test extract was found to be 0.514 µg/mg of the extract. The LOD and LOQ values were 0.84 µg/band and 2.59 µg/band respectively. The RP-HPLC study also represented good % RSD of Retention time (0.025) and Area (0.09) which signifies the high precision and repeatability. The assay result (4.99 %) also indicated good presence of the quercetin in the extract.
 Conclusion: The methods were rapid, cost effective and may be employed for quality-control and standardization of quercetin in different plant extracts.

Application of approach for development of hptlc identification and quantification methods for determination of phenolic compounds and terpenoids of several Thymus L. species

The aim of this work was to improve the approaches for standardization of Thymus L. species by development of HPTLC identification of phenolic compounds (flavonoids and phenylpropanoids) and terpenoids (essential oils and triterpenoids) and quantitative determination of rosmarinic acid and flavonoids for six Thymus L. species of Ukraine flora, as well as determination of chromatographic profiles of Thymus L. species’ extracts obtained using different extraction methods.
 Materials and methods. Objects were: samples of dry herb of Th.serpyllum, Th.vulgaris, Th.Marschallianus, Th.Pallasianus, Th.calcareus, and Th.moldavicus of Ukraine origin collected in 2019-2021 years or acquired commercially on Ukraine market. Instruments: CAMAG HPTLC Herbal System, visionCats 2.5. The analytical grade reagents were used. Reference substances were purchased from Extrasynthese, Sigma Aldrich. Chromatography was performed on HPTLC plates Si 60 F254, Merck according to the developed methods.
 Results. A new approach for quality control of different Thymus L. species of Ukraine flora included the development of HPTLC methods for identification of main groups of bioactive substances of these species, such as flavonoids and phenylpropanoids, essential oils, triterpenoids; development of quantification method of rosmarinic acid and assay of total flavonoids, expressed as luteolin-7-O-glucoside. The characteristic HPTLC fingerprints of six Thymus L. species in three mobile phases of different polarities that cover a wide range of bioactive substances were established. The content of rosmarinic acid in different Thymus L. species samples was in the range of 0,11-0,72 %: Th.moldavicus – 0,11 %; Th.Marschallianus – 0,19-0,27 %; Th.serpyllum – 0,38 %; Th.vulgaris – 0,51 %; Th.calcareus – 0,56 %; T.Pallasianus – 0,72 %. The total flavonoids content, expressed as luteolin-7-O-glucoside, was in the range of 0,8-2,72 %: Th.moldavicus – 0,8 %; Th.serpyllum – 0,87 %; Th.vulgaris – 1,06 %; Th.Pallasianus – 1,28 %; Th.Marschallianus – 1,89 %; Th.calcareus – 2,72 %.
 Conclusions. The proposed scientific approach for quality evaluation of Thymus L. species using HPTLC allows to determine comprehensive information of chemical composition and content of active substances of multiple samples in parallel, in a cost and time-efficient manner

Colchicine (a high-priced alkaloid) accumulation and HPTLC quantification in different stages of in vitro developed tuber of Gloriosa superba L.

BackgroundGloriosa superba L. belongs to Colchicaceae, which is an important medicinal plant containing high-priced alkaloid colchicines and other potent phytochemicals. Due to its extensive importance at the industrial level, this plant is overexploited. Moreover, indiscriminate harvesting for raw material leads to a decline in the population of this plant in the natural environment. Thus, the present study deals with the optimization of colchicine accumulation from the different intervals of in vitro and in vivo tubers of Gloriosa.ResultTo obtain in vitro tuberization, shoot tip explants were inoculated on Murashige and Skoog medium prepared with 3 mg/l BA and 0.5 mg/l Kn + 1 mg/l 2, 4-D followed by 2 mg/l BA + 0.2 mg/l NAA. In the high-performance thin-layer chromatography study (HPTLC), the linearity range of colchicine was set at a concentration range of 100–1000 ng/spot with a regression value (r) of 0.99. Its Rf value (0.25) was recorded at 254 nm. The colchicine amount in the in vivo tuber was 7.75 ± 0.25% dry weight, while the nearby amount of 7.7 ± 0.40% dry weight of colchicine was produced from 2 weeks in vitro old tuber. This value was followed by the 4th-week old tuber with 6.35 ± 0.17% dry weight and then a gradual decrease in its accumulation.ConclusionThe significant results for the accumulation of colchicine at different stages were observed. Hence, this strategy of colchicine production creates a new possibility for improved production of colchicine under in vitro conditions which will be helpful to various pharmaceutical industries without damaging the plants from the natural environment.Graphical abstract

Validation of Wet and Dry Drug Collection Principle in the Preparation of Vasa Ghrita Through Quantitative Estimation of Vasicine

Principle of wet and dry drug collection is applied for collection of drugs while preparing any formulations i.e. wet state of drug is taken in double to its dry drug quantity but in case of exception, drug should be taken in wet state and also not taking in double to its prescribed quantity. Here, Vasa is mentioned as an exceptional drug for wet and dry drug collection principle. Hence, Vasa Ghrita were selected as a dosage form to validate this principle. Aim of present study was to validate the wet and dry drug collection principle in the preparation of Vasa Ghrita through quantitative estimation of Vasicine. Three samples of Vasa Ghrita were prepared as per the reference of Charaka Samhita from wet and dry state with equal quantity of Vasa and wet state and double quantity of Vasa. On comparing the data of three samples highest yield (94.16%) was found in Vasa Ghrita prepared from wet and equal quantity. The percentage of Vasicine was more in dry Vasa Panchanga (0.1914%) than wet state (0.0999%) in HPTLC quantification. Equal quantity and wet state of Vasa Ghrita showed more percentage of Vasicine (0.0032%) than dry state and equal quantity (0.0028%) while the percentage of Vasicine in wet state double quantity was in nearer range (0.0035%) to wet state and equal quantity of Vasa Ghrita. So, it can be concluded that Vasa should be used in wet state without taking in double quantity in Vasa Ghrita.

Comparative study of antioxidant and anticancer activities and HPTLC quantification of rutin in white radish (Raphanus sativusL.) leaves and root extracts grown in Saudi Arabia

AbstractThe nutrient contents ofRaphanus sativusL. (white radish) leaves (RSLs) and roots are known to have promising vital effects. We comparatively investigated the leaves and roots ofR. sativusgrown in Saudi Arabia to estimate the total phenol and flavonoid contents using the standard colorimetric methods. The antioxidant activity of RSLs andR. sativusroots (RSRs) were measured by 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) assays. Both extracts’ cytotoxic activity was assessed by MTT assay against several human cancer cell lines (A549, HepG2, MDA-MB-231, and MCF-7). High-performance thin-layer chromatography was used to identify and quantify the biomarker compound rutin in both extracts. The total phenolic content in RSLs (125.3 mg of gallic acid equivalent [GAE]/g) was higher than that in roots (95.8 of GAE/g), whereas the flavonoid content (44.5 mg of quercetin equivalent [QE]/g) was almost double compared in RSR extract (24.4 of QE/g). Similarly, RSLs exhibited a higher antioxidative activity than RSRs in both DPPH (IC50216.8 vs 359.7 µg/mL) and ABTS (IC50326.7 vs 549 µg/mL) models. RSLs also demonstrated the highest antiproliferative efficiency against all cell lines, with IC50values of 217–453 µg/mL. The reversed-phase-high-performance thin-layer chromatography results showed the presence of rutin (5.2 µg/mg) only in RSLs. Our study indicates RSLs as a promising source of bioactive compounds compared with roots.

Isolation, identification, and HPTLC quantification of dehydrodeoxycholic acid from Persian Gulf sponges

This study aims to investigate the chemical constituents of sponges Dysidea avara (D. avara) and Axinella sinoxea (A. sinoxea), grown up in the Persian Gulf, as well as dehydrodeoxycholic acid (DHCA) content in methanolic extracts of the selected sponges. The chromatography-mass spectrometry (GC–MS) fingerprint of bioactive compounds from methanolic extracts of the selected marine sponge samples was investigated. Based on molecular docking results, among chemical compounds found in marine sponges, DHCA has anti-inflammatory and antipsoriatic properties. They also indicated that DHCA generated stable complexes with 1w81, 3bqm, and 3k8o receptors (psoriasis-related targets) with a binding energy (BE) of −9.26, −10.62, and −7.59 kcal mol−1, respectively. DHCA is isolated from the methanolic extracts of marine sponge samples on chromatographic plates was quantified after derivatization with anisaldehyde reagent by the validated HPTLC method. In-situ HPTLC-DPPH was also calculated to evaluate the free radical-scavenging activity (FRSA) of DHCA. In-silico ADME (Absorption, Distribution, Metabolism, Excretion) predictions revealed that the compound had minimum toxicity and acceptable human intestinal absorption (HIA), as well as low skin permeability. These can potentially be employed as lead compounds to develop a novel antipsoriatic drug.

Estimation of Withaferin-A by HPLC and standardization of the Ashwagandhadi lehyam formulation

Standardization is an important measurement for ensuring the quality control of herbal drugs. It has become essential to develop reliable, specific and sensitive quality control methods. Ashwagandhadi lehyam is an important Ayurvedic formulation containing Withania somnifera L., as one of the prime ingredient of formulation. The present study was undertaken to develop standardization parameters for Ashwagandhadi lehyam. Evaluation of various standardization parameters like organoleptic characters, Physico-chemical evaluation, HPTLC finger print profiling along with dominant bioactive markers and estimation of bioactive markers Withaferin-A by HPLC. The Rf value of Withaferin-A 0.35 and Withanolide-A 0.45 is in formulation and reference standards were found comparable under UV light at 254 nm and 540 nm. The HPLC chromatogram of Ashwagandhadi lehyam and standard Withaferin-A showed at Rt of 5.015 and 5.050 min. The percentage of Withaferin-A was 0.092% present in Ashwagandhadi lehyam formulation. Bioactive markers are characteristic to the ingredients or botanicals to identify the presence of ingredients in formulation easily. The presence of bioactive markers is possible and its verification through the HPTLC fingerprint profile and quantification of biomarker by HPLC are the best ways to identify evaluate the quality of the finished formulation in the course of development of a standardization protocol for quality control of Ayurvedic formulation.

HPTLC quantification of diosgenin in Dioscorea deltoidea: Evaluation of extraction efficacy, organ selection, drying method and seasonal variation

Dioscorea deltoidea (Dioscoreaceae) is an endangered species having tubers that possess anextensive range of therapeutic and economic value due to presence of bioactive compound diosgenin. Several industries have various necessities for compounds of specific classes and therefore, choosy extraction strategies for segregation of these classes ought to be produced.The current study aimed to screen potent part, identify best drying process, efficient extraction methods with solvent selection and seasonal variations on diosgenin content in D. deltoidea. The tuber contained the highest diosgenin content (1.15% DW) compared to leaf and stem. The highest yield of diosgenin was obtained using microwave-assisted extraction (MAE) with 50% ethanol (0.99% DW) and minimum incold maceration with water (0.46% DW). Shade drying was found to be the best drying method as compared to others in terms of diosgenin yield (1.43% DW) whereas sun drying produced low yield (1.12% DW). Estimation of yield and enriched diosgenin (1.19% DW) specified that winter (December) is the perfect time to harvest D. deltoidea.

Somatic embryogenesis in Leptadenia reticulata (Retz.) Wight and Arn along with assessment of shoot and callus cultures for HPTLC fingerprint and quantification of p-coumaric acid

Somatic embryogenesis in Leptadenia reticulata (Retz.) Wight and Arn along with assessment of shoot and callus cultures for HPTLC fingerprint and quantification of p-coumaric acid

<p><strong>Amelioration of cisplatin induced nephrotoxicity by <em>Phyla nodiflora</em> (L.) Greene</strong></p>

Phyla nodiflora (Verbenaceae), commonly called frog fruit and locally, jal bhuti , is the traditional folk medicine accepted in the Indian Medicine as well as the Traditional system of Chinese Medicine (TCM) for various treatments viz., urinary disorder, lithiasis, knee joint pain, diuresis and swelling. In the present study, we tried to standardise crude Phyla nodiflora ethanol extract (PNE) using HPTLC and also evaluated its protection against cisplatin induced nephrotoxicity in rats. HPTLC quantification of rutin and fingerprinting profile was performed. Serum kidney toxicity markers, renal tissue antioxidant and pro-inflammatory cytokine levels were assessed followed by DNA fragmentation assay and histopathological examination of renal tissue. Rutin concentration in ethanol extract was found to be 5.35% w/w. In HPTLC fingerprinting 12 peaks with R f ranges from 0.08 to 0.86 were confirmed. Serum biochemical parameters, renal tissue antioxidants and pro-inflammatory cytokines levels were found to be restored. The results demonstrated the nephroprotective activity of P. nodiflora ethanol extract (400 mg/kg, p.o.) as evidenced by protection of kidneys from cisplatin induced DNA fragmentation and damage tissue architecture.