Densitometer Research Articles

Application of the green analytical procedure index to the simultaneous analysis of co-formulated tinidazole and ciprofloxacin in pure form, tablet dosage form, and human plasma using an environmentally friendly micellar high-performance thin-layer chromatographic technology

In individuals with chronic refractory pouchitis and other gastrointestinal disorders, the combination of ciprofloxacin (CIP) and tinidazole (TIN) is beneficial and safe. As a result, a green micellar high-performance thin-layer chromatographic (HPTLC) approach for the immediate analysis of TIN and CIP in pure, spiked human plasma, and co-formulated tablet dosage form has been developed. It is rapid, extremely easy, sensitive, cost-effective, and environmentally friendly. The stationary phase was Merck aluminum HPTLC plates covered with silica gel 60 F254, while the mobile phase was acetone–ethanol–2% watery sodium dodecyl sulfate (3:4:2, V/V). For quantification of both medications, the densitometric scanner was set at 310 nm. For TIN and CIP, this chromatographic separation yielded symmetric, compact peaks with RF values of (0.22 ± 0.009) and (0.42 ± 0.007), respectively. At 310 nm, the separated spots were densitometrically scanned. For TIN and CIP, the detection thresholds were 6.7 ng/band and 25.03 ng/band, respectively. For TIN and CIP, the quantification limits were 20.3 ng/band and 75.25 ng/band, respectively. The approach was validated according to International Council for Harmonisation (ICH) principles and then used to determine the researched medicines in their various pharmaceutical dosage forms, and human plasma yielding an exceptional percent of recovery. In terms of precision and accuracy, the results were in great accordance with the published approach. This method is suitable for the sequential analysis of the two drugs in pure form, tablet dosage forms, and spiked human plasma due to its simplicity, speed, greenness, robustness, and low cost.

Simultaneous densitometric determination of β-sitosterol and lupeol through validated HPTLC method in different plant parts of Uraria picta (Jacq.) Desv. ex DC. – A dashmool species

Abstractβ-sitosterol (BS) and lupeol (LU) exhibit a number of biological activities and are the important bioactive marker compounds in pharmaceutical science. In the present study, a simple, precise, accurate and validated high performance thin layer chromatographic (HPTLC) method was developed for simultaneous quantification of these two compounds in leaves, stem and roots of Uraria picta, a critically endangered medicinal plant and one of the important constituents of ten plants ayurvedic formulation called Dashmoola. Standards and test samples were applied on TLC aluminum plate precoated with 0.2 mm layer of silica gel 60F254. The plate was run in a twin glass chamber comprising toluene: methanol: chloroform (8:1:1, v/v/v) as a mobile phase. The plates were immersed in anisaldehyde-sulfuric reagent and then heated at 105 °C for 5 min in CAMAG TLC plate heater for appearance of bands. Densitometric scanning was performed at λmax = 525 nm using tungsten light source in CAMAG TLC Scanner4 armed with WinCATS software. RF values of BS and LU standards and those of test samples were found to be 0.53 ± 0.01 and 0.63 ± 0.01 respectively. The method was further validated by following the International Conference of Harmonization (ICH) guidelines. For BS and LU, the linear regression data for the calibration plots revealed a satisfactory linear association with r2 = 0.995 and 0.998, respectively. Linear range for both BS and LU was 200–600 ng/band. Accuracy of the method was evaluated by performing recovery study at three different levels by standard addition methods with an average recovery of 99.86% and 101.07%. The results revealed that the leaf samples of U. picta contained highest concentration of BS (0.150 ± 0.02%) while its root samples confined the highest concentration of LU (0.149 ± 0.01%). The developed method can be applied for routine and quality control analysis in different herbal formulations containing U. picta species.

Physicochemical and Phytochemical Evaluation of Siddha formulation Saaranai chooranam

Background: Siddha system of medicine depends largely on herbal for the treatment of diseases which was practiced at village levels and now becomes an important medicine in worldwide, According to the Akasthiyar -2000-Part– III textbook, Saaranai Chooranam is a herbal drug that is indicated for Raththa kothippu (Systemic Hypertension) Saaranai is the Tamil name for Trianthema portulacastrum, which belongs to the family Aizoaceae. Aim & Objective: This study primarily aims to evaluate the Physicochemical and Phytochemical evaluation of Saaranai Chooranam, Preliminary Phytochemical analysis such as High Performed Thin Layered Chromatography and Powder Microscopy of Saaranai Chooranam. Methodology: The Physicochemical analysis of Saaranai Chooranam is carried out using standard procedures. Results: Preliminary Phytochemical screening found, the presence of phytochemicals such as Proteins, Terpenoids, Alkaloids, Carbohydrates, and Tannins. High-performance thin-layer chromatography fingerprinting revealed the presence of many phytochemicals with different Rf values and densitometric scans of the plates showed numerous bands and peaks. The Powder Microscopy reveals the presence ofTracheidal fibre, Vessel with bordered pits, Calcium oxalate crystal, Stone cell, Group of sclereids with brownish content, Paracytic stomata and surrounding subsidiary cells, Rosette Calcium oxalate crystal, Sclereid with narrow lumen, Thick walled cells, Tracheidal fibre and Trichome. Physicochemical analysis revealed the values of total ash (17.86%), acid-insoluble ash (1.34%), water-soluble ash (13.62), sulphated ash (25.72%), pH (6.05 in 4% alcohol solution), volatile oil (0.5%), foaming index (111.11%) and swelling index (4ml). Conclusion: This study is an effort to explore the different Physico and Phytochemical compounds of Saaranai Chooranam effective in the management of Raththa kothippu (Systemic hypertension).

Kinetic Study of Hydrolytic degradation of Rivaroxaban by HPTLC

The main objective of a research involved kinetic study of hydrolytic degradation of Rivaroxaban and validation of stability indicating HPTLC Method for estimation of rivaroxaban as per ICH guidelines.The chromatographic separation was carried on Merck TLC aluminium sheets of silica gel 60 F254 (10×10cm) with mobile phase of Acetonitrile: Ethyl acetate (3:7 v/v) with densitometric scanning at 249nm. Acceptable peak parameters were obtained at the retardation factor of about 0.75±0.02. The proposed method was validated according to ICH guidelines. The results were acceptable for linearity and range, accuracy, precision, robustness, detection limit and quantitation limit. During stress degradation study, it was observed that the rivaroxaban is more sensitive to hydrolytic conditions. Degradation product peak was well resolved when the sample exposed to alkali catalysed hydrolysis for 72 hrs. was spotted on TLC plate and studied by the developed method. Such a product peak was not observed at 48 hrs. exposure, though there was reduction in drug peak area. This prompted us to study degradation kinetics under alkaline hydrolytic conditions, and the results of the degradation kinetic study indicates that the reaction follows first order kinetics. Interestingly there was no product peak under acidic hydrolytic condition. Results of thermal, oxidative and photolytic degradation studies show reduction in drug peak area.The developed method describes simple, sensitive, and specific stability indicating HPTLC method for estimation of rivaroxaban. This optimised method can be considered as good alternative for routine analysis of the rivaroxaban stability.

Method development and validation for quantification of six bioactive compounds (andrographolide, columbin, piperine, gallic, paracoumaric and oleanolic acids) by HPTLC.

During the spread of pandemic diseases, immunity boosting herbal drugs are taken as a preventive medicine. Kapacurak Kuṭinīr Cūraṇam is a Siddha drug used for flu like viral infections, cold and fever. Developing an analytical method to estimate the content of active phytoconstituents in such antiviral immune boosting drug will be useful in the phyto pharmaceutical industry. A precise, reliable and sensitive ordinary phase high performance thin layer chromatography (HPTLC) method has been developed and validated for identification and simultaneous estimation of six bioactive components namely like andrographolide, columbin, gallic acid, ρ-coumaric acid, piperine and oleanolic acid from any Indian traditional medicine, medicinal plant, drugs and food materials etc. The separation was achieved on silica gel 60F254 TLC plates using toluene: ethyl acetate: formic acid (7:3:0.5, v/v) as mobile phase. The gallic acid, ρ-coumaric acid, piperine markers were estimated using the densitometric scanning in absorption mode at 254nm. The densitometric scanning was done after derivatization (vanillin-sulphuric acid reagent) at λ=520nm for andrographolide, columbin and oleanolic acid. The linear regression analysis data for the calibration plots showed a correlation coefficient in the concentration range 1-5μg per band for the bioactive markers with respect to area. The method was validated for accuracy, precision, limit of detection (LOD), and quantitation of limit (LOQ). Developed method was accurate, precise and fast to ensure the quality of Kapacurak Kuṭinīr Cūraṇam.

Abstract 14621: The Endothelial Lipase Loss of Function Mutation R450G Causes Hyperalphalipoproteinemia That Does Not Protect Against ASCVD

Background: High-density lipoprotein cholesterol (HDL-C) levels and cardiovascular risk are inversely related in epidemiological studies. However, some rare monogenetic mutations that raise HDL do not appear to confer atheroprotection. Methods: The LIPG gene, encoding Endothelial Lipase (EL), was sequenced in 337 patients with hyperalphalipoproteinemia (HDL-C>80mg/dL) for whom a causative variant in other genes affecting plasma HDL levels ( CETP, APOA1, ABCA1, LCAT, LPL, GALNT2 ) had been excluded. HDL particle size distribution was determined using a Lipoprint HDL subtraction test and further assessed by non-denaturing gradient gel electrophoresis followed by densitometric scanning. EL variants were cloned by site directed mutagenesis, expressed in HEK293 cells and compared to wild-type EL. Their expression in cell extracts and secretion in culture medium were measured by western-blot and ELISA. EL phospholipase enzymatic activity was assessed by measuring the production of non-esterified free fatty acids (FFA) using native and reconstituted HDL (rHDL) as substrates. Results: We identified the novel EL R450G missense mutation in three individuals (2 heterozygotes and one homozygote). Their HDL particles were larger than those of normolipemic controls (66±14% vs. 36±6% of large HDL1 particles; P=0.02). Six week treatment with Probucol 1g/day reduced the amount of HDL1 in those patients. Despite elevated circulating HDL levels, these patients presented with atherosclerotic cardiovascular disease. In vitro, the R450G variant was expressed and secreted normally from cells but displayed markedly reduced enzymatic activity compared with wild-type EL (Vmax = 6±4, vs. 32±5 nmol/Hr and Km = 0.01±0.07 vs. 0.30±0.15 mM, respectively; P<0.01, all). Conclusion: EL-R450G is a loss-of-function variant associated with hyperalphalipoproteinemia. Carriers of this mutation do not appear to be protected against atherosclerosis.

Determination of the effect of iatrogenic blood contamination on lactate dehydrogenase and creatine kinase activity in canine cerebrospinal fluid.

Lactate dehydrogenase (LDH) and creatine kinase (CK) have differential tissue activity and isoenzyme profiles. LDH and CK exist as 5 and 3 isoenzymes, respectively, in both serum and cerebrospinal fluid (CSF). Studies have demonstrated that measuring LDH, CK, and their isoenzymes in CSF has diagnostic and prognostic values for dogs and people with neurologic disorders. Iatrogenic blood contamination can distort the results of CSF analysis. The purpose of this study was to determine allowable thresholds of blood contamination (RBC/μL) for accurate measurement of LDH, CK, and their isoenzymes in canine CSF. Venous blood and CSF were collected from healthy dogs. Total LDH and CK activity were measured spectrophotometrically. Isoenzyme profiles were determined using gel electrophoresis and densitometric scanning. All samples were analyzed within 6hours of collection. A nonlinear mixed effects regression model was used to estimate the allowable thresholds of blood contamination for accurate measurement of LDH, CK, and their isoenzymes in canine CSF. The threshold of iatrogenic blood contamination for total LDH and total CK in healthy dogs are 6696 RBC/μL (95% CI 3879-11 187) and 5961 RBC/μL (95% CI 2939-12 085), respectively. LDH-1 is the most sensitive isoenzyme to iatrogenic blood contamination, while LDH-4 is the least sensitive. These results are important for the interpretation of LDH, CK, and their isoenzymes in canine CSF. Additionally, our methodology is translatable for determining thresholds of acceptable iatrogenic blood contamination in CSF for other diagnostic and prognostic biomarkers of neurologic disease.

DEVELOPMENT AND VALIDATION OF STABILITY INDICATING HPTLC METHOD FOR DETERMINATION OF IGURATIMOD IN BULK AND PHARMACEUTICAL DOSAGE FORM

Objective: The objective of the work was to develop and validate stability indicating HPTLC method for the estimation of Iguratimod. Methods: The method employed HPTLC aluminium pre-covered silica gel 60 GF254 plates (10 cm × 10 cm with 250 μm layer thickness) as stationary phase while the solvent system was n-Hexane: Ethyl Acetate (5:5 v/v) with densitometric scanning at 256 nm. Sample was applied as a band of 8 mm width using Camag 100 μl sample syringe (Hamilton, Switzerland) using a linomat 5 applicator (Camag, Switzerland). Migration distance was 80 mm. Further the sample was subjected for stress conditions under acid and base hydrolysis, oxidation, thermal, neutral and photolytic conditions. Method validation done according to ICH Q2 (R1) guidelines. Results: Retention factor (Rf) of the drug was 0.41±0.02. The linearity of the method was found to be within the concentration range of 200-1200 ng/band with R2= 0.983. Limit of detection and limit of quantification were found to be 34.69 and 105.12 ng/band respectively. The % mean recovery was found to be 100.38±0.83. Stress results showed that there is degradation in acid and base conditions but two degradant peaks were observed only under alkaline stress condition Conclusion: The developed method found to be accurate, simple and precise. Method is successfully employed for quantification of the drug under various stress conditions.

Phytochemical analysis of Clerodendron infortunatum Linn. leaves in various seasons by different solvent and extraction techniques

Clerodendron infortunatum Linn is widely used for the treatment of malaria and vermifuge, scorpion and snake bites, tumors, inflammation, bronchitis and in wound healing. The present study was carried out to estimate the variation in the amounts of Gallic acid, Rutin and Quercetin in Clerodendron infortunatum Linn.leaves extracts collected in different seasons using high-performance thin layer chromatography (HPTLC).In this study, leaves of C. infortunatum were subjected to methanol extractions using Soxhlet method and cold maceration. The leaves were collected in different seasons in the months of December (Sample A -Shishir rutu-Winter season) and July (Sample B -Varsha Rutu -Rainy season). The methanolic extracts were subjected to HPTLC study & using CAmag HPTLC system, employing a mixture of toluene:ethyl acetate:formic acid:methanol(3:6:1.6:0.4 ) as the mobile phase with densitometric scanning at 254nm. Quantity of Quercetin is found in both samples, extracted by Soxhlet method. Its quantity is higher in the winter season. Quantity of Gallic acidis found in both samples. Its quantity is the same in Sample B by Soxhlet extraction method & maceration method respectively. Quantity of Rutin is found in both the samples extracted by Soxhlet & maceration methods. It is higher in the sample-A. The present HPTLC method is used for the detection and quantization of Gallic acid, Quercetin and Rutin of C. infortunatum leaves. It needs to study the season wise and region wise variation of phytochemicals in useful parts of all other plants by different extraction methods.

Simultaneous Estimation of Stigmasterol and Lupeol in Adenanthera pavonina using HPTLC Method

Adenanthera pavonina , a well-known medicinal plant has been used traditionally to treat diarrhea and inflammation. Its medicinal properties result from its active phyto-constituents, including non-protein amino acids, sterols, dulcitol, and fixed oil. Stigmasterol (STIGMA) and lupeol (LPL) are significant phytosterols in the plant’s bark as they possess anti-inflammatory activities that may be used in multiple therapies. To design an accurate HPTLC method for the simultaneous estimation of lupeol and stigmasterol from the bark of Adenanthera pavonina. The STIGMA and LPL were isolated using silica gel (60–120 #) column chromatography and estimated using HPTLC. The TLC aluminium plates were pre-coated with silica gel 60F254; the mobile phase comprised toluene and methanol in the ratio of 92:8v/v, and the compounds were detected using the vanillin-phosphoric acid reagent by densitometric scanning at 550 nm. The R f value for STIGMA and LPL was 0.430.02and0.550.02 with linearity in the range of 100– 500 ng/spot and 400-2000 ng/spot, respectively. The mean % recovery was in the range of 93.8–97.2 % for STIGMA and 94.9 –95.6 % for LPL with LOD and LOQ values of 11.74 and 35.58 ng/spot for STIGMA and 90.44 and 274.06 ng/spot for LPL, respectively. Our HPTLC method was accurate and precise for simultaneous estimation of STIGMA and LPL from the bark of Adenanthera pavonina, thereby making it effective in quality control investigations of the herbal formulations. Keywords: Adenantherapavonina; HPTLC; Stigmasterol; Lupeol

Simultaneous Analysis and Efficient Separation of Anabolic Androgenic Steroids in Dietary Supplement by a Validated HPTLC Method.

Using sports supplements is common among athletes. The presence of anabolic steroids in sports supplements as a hormonal contaminant can increase production efficiency. Since anabolic steroids cause health problems and result in positive doping tests in athletes, it is important to investigate their presence in the supplement preparations consumed by athletes. This paper aims to simultaneously determine ten anabolic steroids by high-performance thin-layer chromatography (HPTLC) method in sports supplements. Chromatographic analysis was conducted on glass silica gel 60F254 plates. The extracts loaded on silica gel plates are subjected to programed multiple development (PMD) to separate anabolic androgenic steroids (AASs). Densitometric scanning is carried out at the wavelength of 245 and 366nm. The method was validated according to the ICH guidelines. Spots at retardation factor (Rf) 0.72 (elution system 1), 0.4 (elution system 1), 0.29 (elution system 2), 0.25 (elution system 2), 0.1 (elution system 1), 0.65 (elution system 2), 0.59 (elution system 1), 0.44 (elution system 1), 0.8 (elution system 3), and 0.82 (elution system 3) values were recognized as 19-nor androstenedione, 19-nortestosterone, methyl testosterone, clostebol, stanozolol, trenbolone enanthate, oxymetholone, oxandrolone, testosterone enanthate, and nandrolone decanoate, respectively. The linear ranges were 25 - 250 μg/mL for oxymetholone, 7 - 50 μg/mL for 19-nor androstenedione, 19-nortestosterone, and oxandrolone, and 3 - 20 μg/mL for methyl testosterone, clostebol, stanozolol, trenbolone enanthate, testosterone enanthate, and nandrolone decanoate. The developed method is validated by acceptable precision (CV < 20%) and good accuracy (94% < R < 114%). The value of limit of detection (LOD) for all derivatives was in the range of 0.02 - 0.16 μg/spot (20-160 μg/g of supplement), while limit of quantitation (LOQ) was found to be in the range of 0.06 - 0.5 μg/spot (60 - 500 μg/g of supplement). Fifty sports supplement samples as real sample were collected and analyzed. None of the samples screened positive using the HPTLC method. In the present study, the fast, cheap, and simple HPTLC method could be used for the multi-residue analysis of ten anabolic androgenic steroids in sports supplements.

Urolithiasis: HPTLC Method for Quantitative Detection of Rutin and Quercetin in an Herbal Plant

Anti-inflammatory, Hypoglycemic, Hepatoprotective, Antihyperlipidemic, Anti-Ulcerative, Cardioprotective Stimulant, Sedative, Hypnotic, Anticonvulsant Activity, Memory Retention, Stroke Preventive Activity, Antimicrobial Activity, Antimycobacterial, Antiviral, Larvicidal, Antiparasitic, Chemopreventative, Chemo-modulatory, Anticancer, Cytoprotective activities were found in a variety of herbal plants, including Ocimum basilicum. The purpose of this study was to design and develop a new HPTLC method that was accurate, precise and cost-effective for simultaneous measurement of rutin and quercetin in a hydroalcoholic extract of Ocimum basilicum seeds. The mobile phase was Toluene: Ethyl Acetate: Methanol: formic acid (6:4:3:1, v/v/v/v/v) and densitometric scanning was performed at 254 nm. Merck TLC aluminium sheets of silica gel 60 F254, (10 x 10 cm) with a thickness of 250 mm was used as stationary phase. At a wavelength of 254 nm, rutin and quercetin were detected. The constituents were resolved satisfactorily, with Rf values of 0.25 ± 2.01 for rutin and 0.80 ± 0.64 for quercetin, respectively. Linearity (300-1300 ng/spot for rutin and quercetin) was used to test the method’s accuracy and reproducibility. For both analytes, intra- and inter-day precision, as evaluated by coefficient of variation, was less than 3%. For rutin, the detection and quantification limits were 46.52 and 140.96 ng/spot, respectively, and for quercetin, they were 81.79 and 247.84 ng/spot. The proposed approach was found to be precise, accurate, repeatable, and specific, and it could be used to measure quercetin and rutin in samples at the same time.

A HPTLC method for the quantitative determination of Piperine and Capsaicin in Rasam, A South Indian spice soup

Rasam is a popular spice soup of South Indian traditional food. Rasam is prepared traditionally by incorporating spcies such as turmeric, coriander, black pepper, garlic, chili pepper, curry leaves, cumin, mustard, and asafoetida in tamarind juice. All ingredients used in the preparation of rasam are known for various medicinal uses which, makes rasam a traditional food with a lot of functional ingredients. There are reports to suggests that a regular chronic consumption of traditional foods with functional ingredients may prevent numerous diseases. There was no scientific literature available on the phytochemical composition of rasam. We, herein aim to estimate the quantity of piperine and capsaicin present in the standardized rasam by HPTLC method. Rasam was prepared as per reported method and the sample solution was prepared by extracting rasam with diethyl ether. The mobile phase used was toluene: ethyl acetate (7:3, v/v). Densitometric scanning was performed in absorption-reflection mode at at 527 nm. Linear range was 1 to 5 μg for both piperine and capsaicin. The amount of piperine and capsaicin from the standardized rasam was found to be 0.234 and 0.335 % w/v respectively. This TLC procedure may be used effectively for identity, quality evaluation as well as quantitative determination for piperine and capsaicin in rasam.

Standardization of Dooshivishahari Agada through HPTLC

Ayurveda medicines are a time-tested and valuable resource for healing, even today, globally. Hence the quality control standards for the Ayurvedic formulations is the need of time because of commercialization. Despite the availability of modern equipment and techniques, not all Ayurvedic formulations have been standardized as per modern day protocol. Dooshivishari agada (DVA), a herbo-mineral preparation; mentioned in the text of Asthangha hrudaya in the context of treatment of poisoning, specifically for dooshivisha (cumulative poison). High-performance thin-layer chromatography (HPTLC) is a validated analytical tool for qualitative and quantitative analysis of chromatographic information. In this present study, an effort has been made to standardize DVA through HPTLC. HPTLC photo documentation of DVA showed 8, 11, and 10 spots under short UV, long UV, and under white light after derivatization respectively. Densitometric scan at 254 nm revealed 3 high peaks corresponding to 3 different compounds in the ethanol extract, compounds, at 366 nm and 620 nm there were three high peaks. These Physico-chemical constants, TLC photo documentation, the unique Rf values, and densitogram obtained at different wavelengths can be used as a fingerprint to identify DVA.

Marker Based Standardization of Herbal Sunscreen Formulation by Using Validated High-Performance Thin Layer Chromatography Method

Plant based products are worldwide used and trusted for the care and cure of the health. Herbal sunscreen formulations are gaining popularity for their effectiveness and are devoid of side effects. The present investigation is an effort made to develop herbal sunscreen cream containing methanolic extracts of different plants and its standardization for the presence of bioactive compounds. The leaves of Cymbopogon citratus (Stapf), fruit peel of Punica granatum (Linn), flowers of Butea monosperma (Lam.) and leaves of Neolamarckia cadamba (Roxb.) were selected for the preparation of sunscreen cream. The standardization of herbal formulations is very important to determine its quality based on the concentration of their active phytoconstituents. The herbal cream F-5 (2 % w/w) imparted its sun-protective and antioxidant property. It shows free radicals scavenging activity due the presence of flavonoids and phenolic compounds. High performance thin layer chromatography method is used to determine the quality and quantity of the sun protective phytoconstituents present in the product. The method was validated according to ICH guidelines for the estimation butrin (BT), isobutrin (IBT), quercetin (QC), apigenin (API), chlorogenic acid (CA) and gallic acid (GA) using the optimized solvent systems. The estimation of bioactive markers was carried out on silica gel precoated thin layer chromatography plates with 60F254 as the stationary phase and Camag TC scanner III for densitometric scanning. The average Rf values for the markers were found to be 0.46 for BT, 0.57 for IBT, 0.50 for QC, 0.57 for API, 0.66 for CA and 0.42 for GA. The developed HPTLC method was linear with correlation coefficient 0.999 for BT, 0.998 for IBT, QC, API and 0.9966 for CA and 0.9989 for GA. Limit of detection (LOD) and limit of quantification (LOQ) were recorded. The developed analytical method for quantitative determination of phytoconstituents was found efficient, simple, accurate, and validated.

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&gt;ethanolic stem extract&gt;aqueous leaf extract&gt;petroleum ether bark extract&gt;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.

Qualitative Detection of Triterpenes and Quantification of Betulinic Acid from Hexane Extract of Simarouba glauca Leaves by Gas Chromatography-Mass Spectrometry and High-Performance Thin Layer Chromatography

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.

Экспресс-методы селективного и неселективного определения индольных алкалоидов

In this work, we have considered the methodological features of the use of non-selective colorimetric determination of total content of indole alkaloids (with van Urk reagent) and qualitative-quantitative selective determination of the alkaloid composition by thin-layer chromatography (TLC) with densitometric scanning of chromatograms for related to cultivation a live collection of parasitic strains of Claviceps purpurea. The total content of ergoalkaloids, as well as the content of the target ergoalkaloid, was determined in 4 strains: for VKM F-3602 D, producent of ergocryptine, total content of alkaloids was 0,42±0,02 g/100 g with the content of α-ergocriptine from 71 to 79 %, for VKM F-2641 D ergotamine producent total content of alkaloids was 0,56±0,03 g/100 g with ergotamine content from 43 to 67 %, for VKM F-3410 D ergocristine producent total content of alkaloids was 0,42±0,02 g/100 g with an ergocristine content of 62 to 89 %, for A- 144 ergocristine producent, the amount of alkaloids was 0,42±0,04 g/100 g with an ergocristine content of 62 to 75 %. The solvent system methylene chloride: dioxane: ethanol: ammonia in the ratio 36:3:1:0.3 was used for separate of ergoalkaloids.

Assessment of the frequency of hormonal disorders of the gonads and bone mineral density among women treated for anorexia nervosa in adolescence.

There are very few studies evaluating the activity of gonads in women treated for anorexia nervosa (AN) in adolescence, and reports on the bone mineral density in such patients are divergent. Objective Assessment of the incidence of gonadal hormonal function disorders and reduction of bone mineral density in women treated for AN in adolescence. Out of 97 women who had participated in a survey study, 56 reported personally to the clinic. Their somatic condition, body weight, and BMI were evaluated, as well as levels of oestradiol, LH, FSH in blood serum were determined and DEXA scans were performed. The average period of hospitalisation until the time of the study was 7.08 ±4.47 years. BMI was as follows: 20.01 ±3.6 kg/m2. 25% women were still sick, 75% were considered cured. In 17.9% of the study subjects hypogonadotropic hy-pogonadism was diagnosed. Abnormal results of the densitometric scan were confirmed in 85.7% of the study subjects. In the group of women with normal body weight only 19% had normal levels of bone mineral density. 1. In almost 18% of women treated for AN in adolescence, disorders of the hormonal function of gonads persist, despite normal body weight in 50% of them. 2. Bone mineral density is reduced in most women treated for AN, which indicates the need to monitor the condition of the bones, an early introduction of prevention of osteopenia and osteoporosis since the onset of AN in order to prevent bone fractures in adulthood.

Standardization of Kirāmpu kutinīr Decoction and Extract Granules using Pharmacognostic, Physicochemical and HPTLC Studies

Introduction: Siddha being one of the ancient systems of medicine, has been counted on by most of the population during the outbreak of pandemics in recent times. The decoctions like nilavempu kutinir and kapacura kutinir have been authorized for prescription for conditions like dengue, Covid-19 etc. Kirampu kutinir (KK) is one such decoctions which has been found efficacious in Covid patients with respiratory tract infections. The current study is aimed at standardizing KK curnam and two of its dosage forms like Kutinir and extract granule employing pharmacognostic and chemical analyses. Methods: Kirampu kutintr (KK) and Kirampu kutinir extract granules (KEG) were analysed for pharmacopoeial standards following standard procedures. Results: The macroscopy and powder microscopy correlated with standards and matched with their characters. Both HPTLC photo documentation and densitometric scan revealed that the KEG comprises of the said ingredients. It also divulges that KEG and KK are of same bands and same pattern. Conclusion: The reported results will be supportive for standardization and future studies of KK.