@article {2158, title = {Effect of Plant Stage and Solvent Extraction on Catechin Contents in Borrasus flabellifer L. Male Flower}, journal = {Pharmacognosy Journal}, volume = {15}, year = {2023}, month = {December 2023}, pages = {1036-1041}, type = {Original Article}, chapter = {1036}, abstract = {

Borrasus flabellifer L. is a plant in the Arecaceae (Palmae) family with numerous bioactivities such as diuretic promotion, antioxidant, and antimicrobial effects. In this study, we are interested in the phytochemical compounds present in B. flabellifer L. male flowers. We investigated the effects of plant stage and solvent used for extraction. Initially, both the young and mature male flowers of this plant were extracted using water or ethanol. Several standard compounds, including gallic acid, catechin, caffeic acid, and quercetin, were utilized to screen the phytochemicals in these plant flowers. After analyzing the samples using High-Performance Liquid Chromatography (HPLC) method, only catechin was detected in all extracts, while the others were not detected (did not match the peak spectrum pattern compared with standard). Using a detection wavelength and curve parameters with an R2 value of 0.9999, catechin detection via HPLC was performed at 280 nm within the range of 13.12 to 210.00 {\textmu}g/mL detection. Regarding the stage of the male flower, the young ones extracted with water provided the highest catechin content at 6.65 {\textmu}g/mg extract, while the others fell within the range of 4.02 to 4.44 {\textmu}g/mg extract. Therefore, it has been confirmed that catechin can be found in B. flabellifer L., and water should be used as the solvent for extraction for further applications of this plant.

}, keywords = {Borrasus flabellifer L., Catechin, HPLC, Male flower, Mature, Young}, doi = {10.5530/pj.2023.15.190}, author = {Chayut Fongsuk and Patnaree Wongmanit and Kanoktip Pansuksan} } @article {1616, title = {Comparison of Colchicine Content in Gloriosa superba L., Root after Pretreatment by Thai Traditional Medicine Methods}, journal = {Pharmacognosy Journal}, volume = {13}, year = {2021}, month = {July 2021}, pages = {942-948}, type = {Research Article}, chapter = {942}, abstract = {

Introduction: In Thai traditional medical practice, the roots of Gloriosa superba L. must be treated prior to use as Thai herbal remedies. Colchicine is a major active chemical constituent in the roots. It was reported that lethal doses of colchicine were 7-26 mg. There have been two pieces of evidence that indicated 5 different pretreatment methods of Gloriosa superba L., roots. Therefore, this research aimed to compare the amount of colchicine before and after pre-treated by 5 different methods and compare colchicine content after pretreatment between methods. Methods: Provide five different locations of fresh Gloriosa superba L., roots. Each sample divided into 6 groups. The first group would be treated as a control, the second to the sixth group was pre-treated by roasting, burning, boiling, stewing, and grilling which follow the methods from two pieces of evidence. Subsequently, quantitative analysis by High Performance Liquid Chromatography. Results: The amount of colchicine in roots before and after pre-treated by roasting, burning, boiling, stewing, and grilling methods were significantly reduced by 45.61{\textpm}10.61\% (p=0.000), 37.35{\textpm}14.15\% (p=0.000), 44.55{\textpm}8.60\% (p=0.000), 76.40{\textpm}10.70\% (p=0.000), and 30.35{\textpm}11.31\% (p=0.000), respectively. The stewing method was the greatest decrease amount of colchicine which significantly different from other methods. (p=0.000). Conclusion: All 5 methods were significantly reduced amounts of colchicine in the roots of Gloriosa superba L. They were effective methods to pre-treat the roots of Gloriosa superba L. prior to use as Thai herbal remedies.

}, keywords = {Colchicine, Gloriosa superba L., HPLC, Pretreatment methods, Thai traditional medicine}, doi = {10.5530/pj.2021.13.121}, author = {Sasithorn Tandhavadhana} } @article {1681, title = {Quantification of Andrographolide in Andrographis paniculata (Burm.f.) Nees, Myricetin in Syzygium cumini (L.) Skeels, and Brazilin in Caesalpinia sappan L. by HPLC Method}, journal = {Pharmacognosy Journal}, volume = {13}, year = {2021}, month = {November 2021}, pages = {1437-1444}, type = {Research Article}, chapter = {1437}, abstract = {

Introduction: Andrographolide, myricetin, and brazilin are bioactive compounds from Andrographis paniculata, Syzygium cumini, and Caesalpinia sappan plants that have potential as medicinal ingredients. Objectives: To determine the levels of andrographolide in A. paniculata herb extract (APE), myricetin in S. cumini leaf extract (SCE), and brazilin in C. sappan wood extract (CSE) as marker compounds for extract quality control using the HPLC method. Methods: The separation was carried out on a reverse-phase C18 column (150 x 4.6 mm; 5 μm). The isocratic was prepared from methanol - water (50:50 v/v); 0.1\% orthophosphoric acid - methanol (60:40 v/v); and 0,3\% acetic acid - acetonitrile (85.5: 14.5 v/v) as mobile phase with flow rate 1 mL/min for andrographolide, myricetin, and brazilin determination, respectively and detection using UV detector at a wavelength of 254 nm, 369 nm, and 280 nm, respectively. Results: The linear regression for andrographolide was y = 14113x + 5948.8 (r2= 0.9994); myricetin was y = 87766x {\textendash} 138895 (r2=0.9996); and brazilin was y = 18520x {\textendash} 42668 (r2=0.9992). The andrographolide content in APE was found to be 14.4686 \%. The myricetin content in SCE was found to be 0.3190 \%. The brazilin content in CSE was found to be 2.1280 \%. Conclusion: The described HPLC method was successfully used for the analysis of the APE, SCE, and CSE. This method can be used for the identification and quantification of andrographolide, myricetin, and brazilin in herbal raw materials or herbal products containing these three extracts.

}, keywords = {Andrographis paniculata, Caesalpinia sappan, HPLC, Marker compounds, Quality control, Syzygium cumini}, doi = {10.5530/pj.2021.13.182}, author = {Eem Masaenah and Berna Elya and Heri Setiawan and Zahra Fadhilah and Varda Arianti} } @article {1183, title = {Anti-Inflammatory Activity and Quantitative Analysis of Major Compounds of the Mixtures of Derris scandens (DZSS) Formula}, journal = {Pharmacognosy Journal}, volume = {12}, year = {2020}, month = {June 2020}, pages = {828-834}, type = {Research Article}, chapter = {828}, abstract = {

Background: The mixtures of Derris scandens (DZSS) formula is a Thai traditional medicine, which consists of 4 medicinal plants, including Derris scandens (Roxb.) Benth. (D) Zingiber cassumunar Roxb. (Z), Suregada multiflora Baill. (S) and Siphonodon celastrineus (S). The DZSS formula has been used in an oral dosage form for the treatment of muscle pain. However, the phytochemical profiles and the pharmacological activities of the combined DZSS formula have not been clearly investigated. Objective: This study aimed to investigate the phytochemical profiles and the anti-inflammatory activity of the DZSS formula. Materials and Methods: The ethanolic extracts of the formula (50\% and 95\% ethanolic extracts) were prepared by using soxhlet extraction and which were analyzed by using HPLC. The anti-inflammatory activity of the DZSS formula was tested for its inhibitory effect against nitric oxide (NO) production in Raw 264.7 cells macrophage.The cytotoxic effect of the formula was determined by using the MTT assay. Results: The 95\% ethanolic extract of the DZSS formula exhibited a pronounced anti-inflammatory activity with the IC50 of 40.08 {\textpm} 2.78 μg/mL. The 95\% ethanolic extract possessed a more potent anti-inflammatory activity than that of the 50\% ethanolic extract and with no cytotoxicity. HPLC analysis indicated that the 95\% ethanolic extract also had a higher yield of genistein and compound D, which are obtained the amount of 0.71 {\textpm} 0.00 and 18.89 {\textpm} 0.24 mg/g extract, respectively. Conclusion: Our data suggest that the 95\% ethanolic extract of the DZSS formula possessed a significant anti-inflammatory activity but which is still required to investigate about biological activity in vivo and clinical study.

}, keywords = {Anti-inflammatory activity, Compound D, Derris scandens, genistein, HPLC, Nitric Oxide (NO)}, doi = {10.5530/pj.2020.12.119}, author = {Orapan Ayameang and Ruchilak Rattarom and Catheleeya Mekjaruskul and Wanida Caichompoo} } @article {1234, title = {Bioactivity and Extraction Method with Ultrasonication of Nelumbo nucifera Linn. Anti Aging Drinks}, journal = {Pharmacognosy Journal}, volume = {12}, year = {2020}, month = {August 2020}, pages = {1097-1103}, type = {Research Article}, chapter = {1097}, abstract = {

Introduction: Nelumbo nucifera Linn. Flowers health drinks currently are one of the popular demand globally in Thailand and all over the world. There are plenty of substandard or over claimed of antioxidant content labeled at health drink bottles. The consumers do not obtain enough health benefits anti-oxidant contents which are destroyed by heat during their production process. Methods: This study aimed to evaluate the optimization of extraction including water, 40 \% and 50\% ethyl alcohol using maceration and ultra-sonication method to obtain the highest antioxidant activity compared with DPPH, FRAP and ABTS methods were investigated including total phenolic and flavonoid contents by HPLC in different period of time. The analysis of acute toxicity in white wistar rats by oral administrating of Nelumbo nucifera Linn. flowers extract was testing for after 24 h and 14 d. Results: It was found that Nelumbo nucifera Linn. flower extract drink consisted of high contents of gallic acid, catechin and rutin by HPLC method. Nelumbo nucifera Linn flowers Highest Total Phenolic compound in Nelumbo nucifera Linn. flower extracts with sonicating with 50\% ethanol was 0.954639 {\textpm} 0.109672. Highest Total flavonoids in Nelumbo nucifera Linn. flower extracts macerating in 50 \% ethanol at day 5th was 1.100275 {\textpm} 0.777271.and the sonication with 40\% ethanol was 0.394283+/- 0.51175. Conclusions: The result of acute toxicity analysis showed no toxicity. Nelumbo nucifera Linn. flower extract drinks which are safe as a health drink for consumers. The further market analysis with sensory test should be essential for further research.

}, keywords = {Acute toxicity, Flavonoids, Flower extract drink, HPLC, Nelumbo nucifera Linn., Phenolic content, Ultra sonication Extraction}, doi = {10.5530/pj.2020.12.155}, author = {Buavaroon Srichaikul} } @article {1076, title = {Qualitative and Quantitative Assay of Hydroxycinnamates of Prunus spinosa L.}, journal = {Pharmacognosy Journal}, volume = {12}, year = {2020}, month = {February 2020}, pages = {157-161}, type = {Research Article}, chapter = {157}, abstract = {

Background: Blackthorn (Prunus spinosa L.) is a plant commonly found in the Russian Federation on the roadsides, forest margins, and meadows. Despite lack of recognition by the official medicine, blackthorn fruits possess antioxidant properties and are used in homeopathic preparations. They may also demonstrate antibacterial and anticancer potential due to hydroxycinnamic acids. The aim of present study was to identify and assay hydroxycinnamates in fruits of P. spinosa cultivated in Moscow Region. Materials and Methods: Fresh and dried fruits of P. spinosa, gathered from plants cultivated in Moscow Region in the harvest maturity stage, were used in the study. Qualitative composition of hydroxycinnamic acids was assessed by high performance liquid chromatography, using reversed phase C18 column. Total hydroxycinnamic acids (THA) content was assessed spectrophotometrically. Results: Similar chromatographic profiles were obtained for both fresh and dried blackthorn fruits, the two most abundant compounds being epicatechin (2.91\%) and chicoric acid (2.90\%). Fruits gathered in Chekhovsky District had lower content of hydroxycinnamates (0.798 {\textpm} 0.89) than those coming from Klinsky District (0.886 {\textpm} 0.92). However, the THA content in dried fruits grown in both districts was found to be similar (0.540 {\textpm} 0.71 and 0.557 {\textpm} 0.74, respectively). Conclusions: It can be concluded that blackthorn fruits can be considered as a source of hydroxycinnamic acids, as both fresh and dried fruits contain at least eleven hydroxycinnamates. It was found that the dried fruits have similar content of hydroxycinnamic acids, independently of their origin. Future research should be aimed at drying method optimization.

}, keywords = {Blackthorn, Herbal raw material, HPLC, Hydroxycinnamic acids, Prunus Spinosa L., Spectrophotometry}, doi = {10.5530/pj.2020.12.23}, author = {Sergey Kondrashev and Nadezhda Nesterova and Alexey Luzin and Vitaliy Kochanov and Anna Luzina and Alexey Matyushin} } @article {1186, title = {Simultaneous Quantification of Lupeol, Stigmasterol and β- Sitosterol in Extracts of Adhatoda vasica Nees Leaves and its Marketed Formulations by a Validated RP-HPLC Method}, journal = {Pharmacognosy Journal}, volume = {12}, year = {2020}, month = {June 2020}, pages = {850-856}, type = {Research Article}, chapter = {850}, abstract = {

Background: Adhatoda vasica Nees (Acanthaceae) is a well-known medicinal plant used for the treatment of respiratory disorders such as asthma and bronchitis. Objective: To develop a simple and precise RP-HPLC method for the simultaneous assessment of lupeol, stigmasterol and β-sitosterol of various extracts of Adhatoda vasica Nees. Materials and Methods: The compounds were separated on RP-Phenomenex C18 (250mm{\texttimes}4.6mm; 5μ) column with a mobile phase comprising of 0.1\%v/v formic acid in water and methanol (28:82\%v/v) splashed at a flow of 0.8mL/min with PDA detector at 208nm. Results: The retention time of lupeol, stigmasterol and β-sitosterol was found to be 16.89, 18.26 and 20.72 minutes respectively. The amount of lupeol was abundant in hexane extract (0.952\%w/w) and formulation III (23.72ng/g) whereas, stigmasterol (0.285\%w/w) and β-sitosterol (8.649\%w/w) was highly abundant in chloroform extract and formulation I stigmasterol (2.57ng/g) and β-sitosterol (0.98ng/g). The optimized method was validated for different parameters and all the validated constraints were within the limits as per ICH guidelines. The proposed method was linear over the concentration range of 12.5-200μg/mL with correlation coefficients greater than 0.997. The LOD and LOQ values of lupeol, stigmasterol and β-sitosterol were found to be 0.66, 5.64 and 12.8μg/mL and 2.01, 17.10 and 36.62μg/mL respectively. Conclusion: To conclude, the developed method for the simultaneous estimation of lupeol, stigmasterol and β-sitosterol was simple, precise, accurate and thus reliable for the quality control investigations of crude drugs and its herbal formulations.

}, keywords = {Adhatoda vasica Nees, HPLC, Lupeol, Simultaneous quantification, Stigmasterol, β-sitosterol}, doi = {10.5530/pj.2020.12.122}, author = {Nandhini S and Ilango K} } @article {829, title = {HPLC Quantification of Hydroxycinnamic and Organic Acids of Canadian Goldenrod (Solidago canadensis L.)}, journal = {Pharmacognosy Journal}, volume = {11}, year = {2019}, month = {February 2019}, pages = {400-404}, type = {Original Article}, chapter = {400}, abstract = {

Background: Canadian goldenrod (Solidago canadensis L.) is a medicinal plant widely used in traditional medicine across the world for several hundred years. According to literature data, S. canadensis contains various groups of biologically active substances, including tannins, flavonoids, etc. The aim of the study was to identify and quantify hydroxycinnamic and organic acids in aerial parts of Canadian goldenrod, as these groups of substances demonstrate a broad spectrum of therapeutic activities. Materials and Methods: Ethanolic extracts of S. canadensis, gathered in Central Russia, were analyzed using highperformance liquid chromatography (HPLC). Hydroxycinnamic acids (HCA) determination was carried out by HPLC method with UV detection at 330 nm using HCA reference standards (RS). Organic acids (OA) determination was performed in the same manner, utilizing UV detection at 210 nm and corresponding OA RS. Results: It was established that S. canadensis HCA composition is represented by cichoric, caffeic, chlorogenic, quinic and ferulic acids. The total HCA content in was 1.16 g {\textpm} 10.7 mg / 100 g. Main OA, found in S. canadensis, are ascorbic, citric, tartaric, succinic, gallic, malic, oxalic and fumaric acids, with the total OA content of 426.5 mg {\textpm} 6.4 mg / 100 g. Conclusion: The described HPLC method was successfully used for analysis of S. canadensis aerial parts ethanolic extracts. The method can be utilized for HCA and OA identification and quantification in both herbal raw material and herbal medicinal products containing Canadian goldenrod.

}, keywords = {Canadian goldenrod, HPLC, Hydroxycinnamic acids, Organic acids, Solidago canadensis}, doi = {10.5530/pj.2019.11.62}, author = {Fidan Suleymanova and Olga Nesterova and Alexey Matyushin} } @article {802, title = {Pharmacognostic Specifications, Quercetin and Quercitrin Quantification in Bauhinia malabarica Leaf}, journal = {Pharmacognosy Journal}, volume = {11}, year = {2019}, month = {January 2019}, pages = {155-160}, type = {Original Article}, chapter = {155}, abstract = {

Introduction: Bauhinia malabarica Roxb. is a tropical tree that found throughout Thailand. Leaves have a sour taste and have been used in Thai remedies for wound healing, diuretic, dysentery and emmenagogue. Objective: This study aimed to focus on pharmacognostic specification and quantitative analysis of quercetin and quercitrin in B. malabarica leaves. Methods: Various methods such as macroscopic and microscopic evaluations of B. malabarica leaf were studied along with physico-chemical parameters and quantitated quercetin and quercitrin using RP-HPLC. Results and Conclusion: Whole plant, structures of dried powder crude drug, cross section of midrib and leaf measurement were established. Paracytic stomata and multicellular trichome were found on lower epidermis. B. malabarica leaves from 15 sources throughout Thailand were examined the pharmacognostic specification according to WHO guideline. Physico-chemical parameters showed that loss on drying, total ash, acid insoluble ash and water content should not be more than 8.00, 7.08, 1.79 and 8.28 \% of dry weight while ethanol and water soluble extractive values should not be less than 13.78 and 16.47 \% of dry weight respectively. Quercetin and quercitrin were the markers for quantitative analysis using RP-HPLC with diode array detector in B. malabarica ethanolic extract. Quercetin and quercitrin contents were found to be 0.18 g and 0.38 g in 100 g of dried crude drug. Method validation was determined according to ICH guideline. All results were in the acceptable range and could be used for identity, safety, efficacy and quality of B. malabarica leaves in Thailand.

}, keywords = {Bauhinia malabarica Roxb, HPLC, Leaf measurement, Pharmacognostic specification, Quercetin, Quercitrin}, doi = {10.5530/pj.2019.1.26}, author = {Paphitchaya Thetsana and Chayanon Chaowuttikul and Chanida Palanuvej and Nijsiri Ruangrungsi} } @article {826, title = {Pharmacognostical, Physicochemical Standardization and In vitro Antioxidant Activity of Punica granatum Linn fruit}, journal = {Pharmacognosy Journal}, volume = {11}, year = {2019}, month = {February 2019}, pages = {272-277}, type = {Original Article}, chapter = {272}, abstract = {

Introduction: Punica granatum Linn. fruit (Family: Punicacea), known as Pomegranate is ethno-medicinally prescribed in various part of world for treatment of different diseases it is used as antioxidant, hepatoprotective, anticancer and antiparasitic agent. Method: The present study was thus undertaken to find out the necessary pharmacognostical standards for evaluating the fruit of P. granatum. Different assessment such as macroscopical characters, microscopical studies, physicochemical evaluations (loss on drying, moisture content by Karl Fischer titration, ash values, extractive values) and TLC/HPTLC finger print profiling were performed and the relevant quantitative and qualitative parameters were reported. Invitro antioxidant activity is also performed by HPLC-DPPH method. Results: Fruit of P. granatum are Reddish brown in color, Globular and Oval, smooth, 5.0 o 12.0 cm in diameter. Powdered fruit confirmed the presence of Stone cell, Endospermic cell, Group of stone cells, Nonlignified fiber, Starch grain and Lignified fibers and vessels. TLC of the extracts was also carried out in the current study. Physicochemical standards quantified include loss on drying (36.62 {\textpm} 4.17 \%), moisture content (32.15 {\textpm} 3.64 \%) total ash (8.58\% {\textpm} 1.06 \%), water soluble ash (7.15 {\textpm} 0.97 \%), acid insoluble ash (0.45 {\textpm} 0.03 \%). Safety profile of plant part was recognized by quantify microbial limit test, pesticide residue and heavy metals (Cd, As, Hg and Pb) evaluation. Here is no visible microbial growths were seen in sample. Pesticide residue and heavy metals were observed to be present within the acceptable limits. Conclusion: Scientific investigations do not yet exist to identify the exact plant part and to determine its quality and purity. These studies provided referential information for accurate identification and standardization of this herbal material. These analyses will also be useful to distinguish P. granatum from the closely associated to other species of Punica.

}, keywords = {DPPH, HPLC, Pharmacognostical, Punica granatum, Quality control}, doi = {10.5530/pj.2019.11.42}, author = {Mohd Amir and Niyaz Ahmad and Md Sarfaroz and Wasim Ahmad and Sayeed Ahmad and Mohd Mujeeb} } @article {848, title = {Reduction of Colchicine Content from Radix Gloriosae Superbae Preparata}, journal = {Pharmacognosy Journal}, volume = {11}, year = {2019}, month = {February 2019}, pages = {310-314}, type = {Original Article}, chapter = {310}, abstract = {

Introduction: Gloriosae Superbae Radix is a crude drug employed in Thai herbal remedies for several ailments. Colchicine is known as an active constituent in the roots. It was reported that 7-11 mg. of colchicine may causes lethal effect in human. In Thai traditional medical practice, the roots must be treated prior use to prepare Thai herbal preparations. However, pre-treated method has not yet been well established in any literature. This study aimed to document the pre-treated method and to quantitatively compare the amount of colchicine both before and after pre-treated. Methods: Firstly, interviewing with Thai traditional medicine experts and document the pre-treated methods of Gloriosae Superbae Radix to conclude the 2 commonly used methods. Two pre-treated methods (roasting and burning) were chosen for further study. Colchicine in Gloriosae Superbae Radix from 8 sources were analyzed before and after pre-treat by High Performance Liquid Chromatography (HPLC). Results: After pre-treat by roasting and burning, amount of colchicine in root were significantly decreased by 40.61{\textpm}9.55\% (p=0.000) and 26.79{\textpm}10.89\% (p=0.001), respectively. Comparison of colchicine contents of samples after roasting and burning, the amount of colchicine decreased non statistically insignificantly (p=0.110). Conclusion: Pre-treats of Gloriosae Superbae Radix by roasting and burning had significantly reduce colchicine content. Both methods have been proven to be the effective ways in preparing certain potentially toxic crude drugs before using in compounding into Thai herbal remedies.

}, keywords = {Colchicine, Gloriosae Superbae Radix, HPLC, Pre-treat method, Thai herbal remedy}, doi = {10.5530/pj.2019.11.46}, author = {Sasithorn Tandhavadhana and Chayan Picheansoonthon} } @article {951, title = {Validation of High-Performance Liquid Chromatography for Determination of Bromelain in Pineapple (Ananas comosus (L) Merr) Water}, journal = {Pharmacognosy Journal}, volume = {11}, year = {2019}, month = {September 2019}, pages = {901-906}, type = {Original Article}, chapter = {901}, abstract = {

Objective: The aim of the present study was to validate HPLC method for analysis bromelain levels in pineapple water and application that method to determine the storage time of pineapple water. Methods: The reversed phase of HPLC method was tested and optimized before it is validated. The composition and the flow rate of the mobile phase is the optimized parameter. The analytical parameters validated were detection limits, linearity, accuracy and precision. Pineapple water was stored for 8 h at 10{\textordmasculine}C and bromelain was determined using the validated HPLC method. Result: The optimum mobile phase composition was methanolwater (70:30) with a flow rate of 1 ml/min. The detector concentration-response was linear with coefficient of determination of 0.993. The accuracy of HPLC method at a recovery of 1 and 2\% bromelain was 106.37 {\textpm} 1.94\% and 98.12 {\textpm} 1.29\% (n = 5), respectively. The precision, expressed as the coefficients of variation (CV), at 1 and 2\% bromelain were 1.83 and 1.32\% (n = 5), respectively. Bromelain level at zero time was 81.53\%. After storage for 8 h at 10{\textordmasculine}C, bromelain levels in pineapple juice appeared to decrease not statistically significant (p \> 0.05), with to mean value of 78.46 {\textpm} 2.88\%. Conclusion: The HPLC method developed was valid to analyze accurately concentrations of bromelain in pineapple water and it can be used to study the shelf life of pineapple water based on bromelain content. The bromelain content in pineapple juice was not statistically significantly different after 8 h storage at 10{\textdegree}C.

}, keywords = {Bromelain, HPLC, Pineapple water, Storage}, doi = {10.5530/pj.2019.11.144}, author = {Novi Yantih and Alfadella Methananda and Yahdiana Harahap and Wahono Sumaryono and Lestari Rahayu} } @article {616, title = {Acute and Sub-Acute Antihypertensive Effects of Syzygium polyanthum Leaf Extracts with Determination of Gallic Acid using HPLC Analysis}, journal = {Pharmacognosy Journal}, volume = {10}, year = {2018}, month = {June 2018}, pages = {663-671}, type = {Original Article}, chapter = {663}, abstract = {

Context: Aqueous decoction of Syzygium polyanthum (ADSP) leaf is one of Malay traditionalremedial- preparations for hypertension. Intravenous ADSP reduced blood pressure of anesthetized Wistar-Kyoto (WKY) and Spontaneously-Hypertensive rats (SHR); however, acute and sub-acute effects of oral ADSP on conscious rats and its bioactive compound(s) are not comprehensively studied. This study aims to examine the acute and sub-acute effects of ADSP in comparison to methanol extract (MESP) on systolic blood pressure (SBP) of conscious WKY and SHR rats, as well as to identify their major phenolic-compound using highperformance- liquid-chromatography (HPLC). Methodology: For acute study, SHR and WKY rats were gavaged with single-dose of ADSP or MESP (2.00, 2.50 or 3.00 g/kg), vehicles, or losartan (0.01 g/kg). SBP was measured after 1, 3, 5, 6 and 24 hr-post-administration. For subacute experiment, SHR rats were gavaged once-daily with ADSP or MESP (2.50 g/kg/day), vehicles, or losartan (0.01 g/kg/day) for 3-week and their SBP was weekly-measured. Phenolic compounds were screened using ferric-chloride test, then gallic acid was determined using HPLC. Results: Acute administration of ADSP (2.50 to 3.00 g/kg) and MESP (2.00 to 3.00 g/ kg) significantly reduced SBP of SHR, but not of WKY rats. Repeated-daily-dose administration of MESP (2.5 g/kg/day) significantly reduced SBP of SHR after 2-week and sustained until 3-week, while ADSP (2.5 g/kg/day) significantly reduced SBP only after 3-week. Gallic acid was the major phenolic-compound in ADSP (157.09 \± 4.18 ppm) and MESP (134.04 \± 10.30 ppm). Conclusion: Oral ADSP and MESP that contain gallic acid possess acute and sub-acute antihypertensive effects in SHR, but MESP\’s effect was more prominent than ADSP

}, keywords = {Antihypertensive, Gallic acid, HPLC, Hypertension, Syzygium polyanthum}, doi = {10.5530/pj.2018.4.109}, url = {http://fulltxt.org/article/647}, author = {Azlini Ismail and Nurul Syahida Ramli and Mahaneem Mohamed and Wan Amir Nizam Wan Ahmad} } @article {577, title = {Detection and Quantification of Major Phytochemical Markers for Standardization of Talinum Portulacifolium, Gomphrena Serrata, Alternanthera Sessilis and Euphorbia Heterophylla by HPLC}, journal = {Pharmacognosy Journal}, volume = {10}, year = {2018}, month = {March 2018}, pages = {439-446}, type = {Original Article}, chapter = {439}, abstract = {

Background: High-performance liquid chromatography is one of the major analytical techniques used in the quality control of phytochemicals. Objective: This research article presents the development of HPLC method to detect and quantify the major marker components, kaempferol, and quercetin from four plant species. Materials and Methods: HPLC method was developed for the qualitative and quantitative analysis of plant extracts by using orthophosphoric acid and methanol (95:5) at 370 nm for kaempferol, methanol and orthophosphoric acid (60:40) at 262nm for quercetin. Results: Kaempferol was detected from the hydro alcoholic extracts of Talinum portulacifolium leaves (RT 13.720, concentration 1.08 mg/ml) and flowers of Gomphrena serrata (RT 13.758, concentration 2.13mg/ml). Kaempferol was reported for the first time from Gomphrena serrata. Quercetin was separated and identified from the hydro alcoholic extracts Alternanthera sessilis stems (RT 6.503, concentration 0.01mg/ml). The hydroalcoholic extract of Euphorbia heterophylla stems (RT 6.588, concentration 0.01mg/ml) was also evaluated for the presence of quercetin. Conclusion: The method developed is very useful tool for qualifying and quantifying the plant specimens as well as their extracts.

}, keywords = {Extracts., HPLC, Kaempferol, Marker, Plant specimens, Quality control, Quercetin}, doi = {10.5530/pj.2018.3.72}, url = {http://fulltxt.org/article/505}, author = {Mamillapalli Vani and Shaik Abdul Rahaman and Avula Prameela Rani} } @article {719, title = {Phytochemical Screening and Antimicrobial Studies in Leaf Extracts of Indigofera aspalathoides (Vahl.)}, journal = {Pharmacognosy Journal}, volume = {10}, year = {2018}, month = {August 2018}, pages = {1208-1215}, type = {Original Article}, chapter = {1208}, abstract = {

Introduction: In traditional Indian herbal medicine, the plant Indigofera aspalathoides (Vahl.) has been used to treat various human ailments. Methods and Results: Various phytochemical compounds (Rf value) such as Tannin (1.14), Flavonoid (1.14), Saponin (0.87), Sterol (0.88) and Phenol (0.86) were found in the leaf extract. The antimicrobial effects of the phytoconstituents were examined for three bacterial and fungal species. The highest anti-bacterial and antifungal activities were found in flavonoid compound, where the maximum zone of inhibition was recorded in Staphylococcus aureus 18mm compared to positive control chloramphenicol (24mm). 13mm observed in A. flavus and A. ochraceous where positive control streptomycin exhibits 18mm. GC-MS analysis revealed the presence of three major compounds in 8\α (2H)-Phenanthrenol, 7-ethenyldodecahydro-1,1,4\α, 7-tetramethyl-, acetate, [4\αs(4\α\α, 4b\α, 7\α, 8\α\α, 10\α\α)], Benzoic acid, 4-methyl- , 2-oxo-2- phenylethyl ester and Cyclohexanol, 5-methyl-2-(1-methylethyl)-, [1S-(1\α,2\α,5\α)] in the crude extracts. The preparative HPLC analysis proved the presence of single peak from the crude flavonoid compounds, where the absorption maximum was between 207-290nm exhibited by the UV spectrum analysis. FTIR spectrum confirmed the presence of amide group, phenol group, carboxylic acid, alkynyl, alkene and aromatic ring. NMR studies proved the presence of phenyl group, methyl group and H2C-CH group and their molecular weight recorded as 353 through the mass spectrum analysis. Conclusion: The identified compound considered as the vital compound to design the \“green antimicrobial drugs\”.

}, keywords = {Antimicrobial activities, GC-MS, HPLC, Indigofera aspalathoides, MS, NMR}, doi = {10.5530/pj.2018.6.207}, author = {Tamilarasan Tamil Kumar and Seeni Mohamed Salique and Mohamed Hussain Muhammad Ilyas and Nooruddin Thajuddin and Annamalai Panneerselvam and Mohamed Khan Syed Ali Padusha and Hussain Syed Jahangir} } @article {693, title = {Quantitative and Optimization of Phenolic Acid Extracted from Pomegranate by High Performance Liquid Chromatography (HPLC)}, journal = {Pharmacognosy Journal}, volume = {10}, year = {2018}, month = {August 2018}, pages = {969-972}, type = {Original Article}, chapter = {969}, abstract = {

Objective: Pomegranate is scientifically known as Punica granatum L. which is a nutrient dense fruit rich in phytochemical compounds. Phenolic content is the main compound attribute for the most of the functional properties in pomegranate. The aim of this study is to quantify and optimize the composition of phenolic acids extracted from pomegranate extract by using High-performance Liquid Chromatography (HPLC). Method: The pomegranate extracted with three different methods by using two different solvents which is 50\% ethanol and water. The methods were blended (aril+ seed), Soaking (aril+ seed) and soaking + squeezed manually. HPLC-PDA was used as equipment to quantify and optimize the phenolic acids extracted from pomegranate. Result: Validation method of HPLC was analysed according to the percentage of recovery, LOD, LOQ and coefficient correlation. Result showed that GA was detected in all sample from different method of extraction applied while CA compound not detected in any of extraction method applied. FA compound was only detected in blended method by 50\% ethanol and water as solvent while EA compound was detected only in water extraction of all three methods applied. Conclusion: As a conclusion, according to the standard calibration data curve showed that this method proved to detect and quantify the targeted compounds. By comparing the data obtained from this study, it showed that water blended extract method is significantly higher content of targeted compound except for the CA compound. To the best of our knowledge, this sample can be a valuable source of antioxidant for better used in health benefits.

}, keywords = {Caffeic acid, Ellagic acid, Ferulic acid, Gallic acid, HPLC, Pomegranate extract}, doi = {10.5530/pj.2018.5.164}, author = {Jumli Mimie Noratiqah and Ridzwan Norhaslinda and Baig Atif Amin and Rohin Mohd Adzim Khalili} } @article {609, title = {Renin and Angiotensin Converting Enzyme Inhibition of Standardized Bioactive Fractions of Hyphaene thebaica L. Mart Growing in Egypt}, journal = {Pharmacognosy Journal}, volume = {10}, year = {2018}, month = {June 2018}, pages = {622-627}, type = {Original Article}, chapter = {622}, abstract = {

Introduction: Hyphaene thebaica L. fruit is known in Egypt for its antihypertensive activity. However a standardized herbal extract/fraction was never prepared. Methods: A biologically guided fractionation was carried out in-vitro for the 50\% and 70\% ethanol extracts of Hyphaene thebaica L. fruit using the angiotensin converting enzyme (ACE) inhibition and renin inhibition assays. A validated reversed phase HPLC method was developed for the standardization of the active fractions. Results: The ethyl acetate fraction of the 70\% extract contained higher percentages of the three bioactive markers chlorogenic acid, quercetin and apigenin (1.940 \± 0.140\%, 2.994 \± 0.349\% and 0.612 \± 0.0354\%, respectively) relative to the ethyl acetate fraction of the 50\% extract (1.384 \± 0.157\%, 0.342 \± 0.0834\% and 0.070 \± 0.00225\%, respectively). The butanol fraction of the 70\% extract was found to possess the highest antihypertensive activity (93.69 \± 5.695 \% renin inhibition activity at 0.5 mg/mL and IC50 of 0.001436 +0.00044 mg/mL for ACE inhibition activity). A standard calibration curve for the three compounds was established at a concentration range of 0.1-50 \μg/Ml, they showed good linearity with a correlation coefficient (R2) of (1.00, 1.00 and 0.999; respectively). A high degree of precision (relative standard deviation values \<5\%) was achieved. The limits of detection for the three compounds were 0.428, 0.368 and 0.849; respectively, while the limits of quantitation were 1.29, 1.11 and 2.57, respectively. Conclusions: Current results showed that the butanol fraction of the 70\% extract revealed the highest antihypertensive activity through ACE and renin inhibition mechanisms. In addition, recorded observations concerning linearity of the used bioactive markers offer a support for the possible utility of the tested fractions as potent standardized antihypertensive drugs.

}, keywords = {Antihypertensive, Doum, HPLC, Phenolics, standardization, Validation}, doi = {10.5530/pj.2018.4.102}, url = {http://fulltxt.org/article/640}, author = {Noha A Khalil and Amira Abdel Motaal and Khaled Meselhy and Soad M Abdel khalek} } @article {248, title = {Assessment of Biological Activity and Comparison of UPLC and RP-HPLC Chromatographic Profiles of Clausena excavata Burm.F.}, journal = {Pharmacognosy Journal}, volume = {9}, year = {2017}, month = {February 2017}, pages = {185-191}, type = {Research Article}, chapter = {185}, abstract = {

Clausena excavata Burm. F. (Rutaceae) has long been associated with medicinal benefits in folk medicine, particularly in the treatment of cancer and its related disorders in the eastern region of Thailand. In the the present study Ultra Pressure Liquid Chromatography (UPLC) analysis of methanolic leaf extracts of Clausena excavata were carried out using short (5 cm \× 2.1 mm I.D.) columns, containing 2 m particles at very high pressures (up to 16000 psi) to obtain high efficiency separations in very short run times. The results indicate that UPLC provided better resolution compared to routine RP-HPLC. The analysis time was drastically reduced (4 min.) which is proved to be a faster and more economical analytical tool than RP-HPLC. In addition to that, the present study also evaluates anti-fungal activity against dermatophytes Trichophyton rubrum and Trichophyton mentographytes by using an agar incorporation technique. The anti-fungal activity may be due to the presence of Angelecin and gallic acid in the plant extract.

}, keywords = {Angelecin, Clausena, HPLC, Rutaceae, UPLC}, doi = {10.5530/pj.2017.2.30}, url = {http://phcogj.com/fulltext/297}, author = {Guntupalli Chakravarthi and Alavala Rajasekhar Reddy and Maddi Ramaiah} } @article {222, title = {Exploring a novel edible mushroom Ramaria subalpina: Chemical characterization and Antioxidant activity}, journal = {Pharmacognosy Journal}, volume = {9}, year = {2017}, month = {December 2016}, pages = {30-34}, type = {Original Article}, chapter = {30}, abstract = {

Background: During macrofungal inventorisation from an unexplored subalpine forest of Sikkim Himalayas, a new edible mushroom, Ramaria subalpina was found and documented in consultation with the forest dwellers. Objective: The aim of the present study was to understand the antioxidative potentiality and bioactive constituents of a novel taxon, Ramaria subalpina, that is highly prized by the ethnic people of subalpine Sikkim Himalayas, India, for its flavor, texture and gastronomic delicacy since time immemorial. Methods: Chemical composition and antioxidant properties of methanolic extract of dried basidiocarps were assessed using HPLC and several in vitro assay systems. Results: Methanolic extract had phenolics in highest amount, among which pyrrogallol was identified. It also showed potent antioxidant activity. Conclusion: The present study suggests that Ramaria subalpina has strong medicinal prospects. This novel mushroom can safely be added to the world\’s edible mushrooms list.

}, keywords = {antioxidant activity, DPPH, Ferrous ion chelating, HPLC, India.}, doi = {10.5530/pj.2017.1.6}, author = {Krishnendu Acharya and Kanad Das and Soumitra Paloi and Arun Kumar Dutta and Manoj Emanuel Hembrom and Somanjana Khatua and Arvind Parihar} } @article {355, title = {Pharmacognostic Studies and HPLC Analysis of Roots of Helicteres isora (L.)}, journal = {Pharmacognosy Journal,}, volume = {9}, year = {2017}, month = {May 2017}, pages = {523-527}, type = {Original Article}, chapter = {523}, abstract = {

Background: The juice of roots of Helicteres isora Linn. has been widely used as an antidiabetic in traditional medicine. Objective: The present study deals with pharmacognostical studies and determination of oleanolic acid from the roots of H. isora by new HPLC method. Materials and methods: Detailed study of morphological, microscopical characteristics, physicochemical parameters and phytochemical screening of roots were carried out. The sapogenins were isolated from the roots of H. isora. RP-HPLC method was developed and validated for estimation of oleanolic acid from the sapogenins of roots of H. isora. Results: Detailed quality control parameters of roots of H. isora were reported. Total content of oleanolic acid was 0.075\%w/w from roots of H. isora determined by HPLC. Conclusion: The present study is useful for accurate identification and authentication of roots of H. isora. The HPLC method for determination of oleanolic acid from the roots of H. isora is efficient, precise, reliable and sensitive and can be adopted for routine analysis.

}, keywords = {Helicteres isora, HPLC, Oleanolic acid, Sapogenins}, doi = {10.5530/pj.2017.4.84}, url = {/files/PJ-9-4/10.5530pj.2017.4.84}, author = {Pinal A. Harde and Mamta B. Shah} } @article {347, title = {Simultaneous-HPLC Quantification of Phenolic Acids in Traditionally used Ayurvedic Herb Diplocyclos palmatus (L.) Jeffry}, journal = {Pharmacognosy Journal}, volume = {9}, year = {2017}, month = {May 2017}, pages = {483-487}, chapter = {483}, abstract = {

Introduction:The present study deals with the simultaneous HPLC-quantification of phenolic acid(s) in the aerial parts of Diplocyclos palmatus (Cucurbitaceae) and evaluation of their bioactivity potential through in vitro antioxidant assay\’s. Method: The HPLC elution was done using C18 column using gradient (binary phases) solvent system at a flow rate of 0.6 ml/min. Total phenolic and, flavonoid contents were determined and the antioxidant potential was estimated by four assay\’s viz. DPPH radical scavenging assay, ferric reducing power assay, total antioxidant capacity and 2-deoxy ribose assay. Results: The species is rich in three phenolic acids, among which gallic acid (1708 ug/g) is in maximum concentration followed by caeffic acid (437 ug/g) and protocateuchic acid (337.7 ug/g). Total phenolic content was higher (10.5 mg/g) than flavonoid content (3.78 mg/g) and TAC was found at 0.137 mg/g ASE (ascorbic acid equivalent). IC50 of D. palmatus extract for scavenging of hydroxyl radical by 2-deoxy ribose and DPPH was at concentration of 125.61 \± 0.834 (\μg/ml) and 353.71 \± 0.663 (\μg/ml) respectively. In vitro antidiabetiv potential, via inhibition of alpha amylase enzyme through starch iodine and 3,5- DNS assay reveals the IC50 of extract at 146.31 \± 0.415 ug/ml and 286.23 \± 0.671 ug/ ml respectively. Conclusion:\ The species (aerial part) was rich in phenolic acid with potential bioactivity, identified leads will be useful\ in further chemical characterization and pharmacological validation.

}, keywords = {Anti diabetic, Anti oxidant, Diplocyclos palmatus, HPLC, Phenolic acid}, doi = {10.5530/pj.2017.4.78}, url = {/files/PJ-9-4/10.5530pj.2017.4.78}, author = {Ankita Misra and Pushpendra Kumar Shukla and Bhanu Kumar and Abhishek Niranjan and AKS Rawat and Sharad Srivastava} } @article {108, title = {Estrogenic Potential of Flemingia vestita Benth Tubers in Ovariectomized Rat Model}, journal = {Pharmacognosy Journal}, volume = {8}, year = {2016}, month = {09/2015}, pages = {44-49}, type = {Original Article}, chapter = {44}, abstract = {

Objective: This study investigates the potential estrogenic activity of the ethanolic extract of Flemingia vestita Benth tubers using ovariectomized rat model. Materials and Methods: The ethanolic extract of F. vestita tubers has been standardized using validated HPLC method in terms of its genistein content (8.43 \± 0.05 mg/g of extract). Three to four week old young albino Wistar female rats were ovariectomized and treated for 14 days post ovariectomy with the standardized ethanolic extract at three different dose levels (100, 250, 500 mg/kg body weight) with a positive control of Estradiol valerate (1 mg/kg/day). The parameters evaluated were uterine weight, uterine glycogen, G6PDH, LDH, 17\β-estradiol, progesterone, total cholesterol, triglycerides, HDL and histo architecture of uterus. Results: Treatment with the ethanolic extract of F. vestita tubers showed dose dependent increase in uterine weight, glycogen levels, G6PDH levels, estrogen and progesterone levels when compared with the ovariectomized control. Amongst three dose levels, high dose of plant extract showed significant increase in the uterine weight (p \< 0.001), uterine glycogen content (p \< 0.001), 17-\β estradiol and progesterone levels (p \< 0.001), G6PDH and LDH levels (p \< 0.001) as well as significant decrease in HDL and triglycerides levels (p \< 0.001) compared to ovariectomized control. Histopathological evaluation of uteri sections revealed that the high dose of the plant show increase in the endometrial response as indicated by proliferation of endometrial glands and luminal epithelium of the ovariectomized rats. Conclusion: Thus, these data suggests that ethanolic extract (500 mg/kg body weight) of F. vestita tubers may exhibit good estrogenic activity in ovariectomized rat model.

Key words: Estrogenic activity, Flemingia vestita, genistein, HPLC, ovariectomized rats.

}, keywords = {Estrogenic activity, Flemingia vestita, genistein, HPLC, ovariectomized rats.}, doi = {10.5530/pj.2016.1.10}, author = {Sunita Shailajan and Suman Kumaria and Suhas Pednekar and Sasikumar Menon and Hiranjit Choudhury and Archana Matani} } @article {216, title = {Revelation of β-sitosterol from Benincasa hispida Seeds, Carissa congesta Roots and Polyalthia longifolia Leaves by High Performance Liquid Chromatography}, journal = {Pharmacognosy Journal}, volume = {8}, year = {2016}, month = {September 2016}, pages = {610-613}, type = {Original Article}, chapter = {610}, abstract = {

Background: Fruit juice of Benincasa hispida (BH) is regarded as Vrindamadhava that is recommended for internal use in snake\’s bites. The fruits have been reported to contain good amount of proteins, enzymes, Vitamin B1 and C1 flavonoid C-glycoside, terpenes, phenolic acids and free sugars such as glucose, rhamnose, mannitol, uronic acid, astilbin, catechin, naringenin, pectic polysaccharides and even some trace metals. Polyalthia longifolia (PL) has been well known for its ayurvedic preparations such as Sitaphala kula, Kaphapitashama, Anulomak and Krimighna. PL leaves has been reported to contain phytoconstituents like polylongine, Aporphine-Noxide alkaloids, allo-aromadendrene, caryophyllene oxide, \β-caryophyllene, \β-selinene, \α-humulene, \α-pinene and camphene. Carissa congesta (CC), known as Bengal currant, have yielded 2-acetyl phenol, carissone, carindone and Des-n-methylnoracronycine. \β-sitosterol, an important phytoconstituent recognized from all these selected plants has been well known for its ethnopharmacological importance. In this new study, the research team members has focused on determining the percentage of the \β-sitosterol present in the BH seeds, CC roots and PL leaves by subjecting the extract to High Performance Liquid Chromatography (HPLC). Materials and Methods: BH, CC and PL plants were shade-dried and extracted by suitable extraction methods. In HPLC, peaks obtained in the extracts were compared with the standard by matching their retention time. Results: The amounts of \β-sitosterol present in the BH seeds, CC roots and PL leaves extracts obtained at 254 nm were found to be 36.00, 7.46 and 3.21\% w/w respectively. Conclusion: Thus, BH, PL and CC extracts were said to contain \β-sitosterol as a key constituent.

}, keywords = {Benincasa hispida, Carissa congesta, HPLC, Polyalthia longifolia, β-sitosterol.}, doi = {10.5530/pj.2016.6.15}, author = {Gaurav Mahesh Doshi and Pratip Kashinath Chaskar and Hemant Devidas Une} } @article {87, title = {Quantitative Determination and Variation Tendencies of Flavonoids in Five Selaginella Plant Drugs}, journal = {Pharmacognosy Journal}, volume = {7}, year = {2015}, month = {Nov-Dec 2015}, pages = {378-382}, type = {Original Article}, chapter = {378}, keywords = {Amentoflavone content, HPLC, Selaginella, Total flavonoids content, UV-Vis spectrophotometry, Variation tendencies}, doi = {10.5530/pj.2015.6.11}, author = {Weifang Long and Qi Ding and Yujie Chen and Jiqing Hu and Luyang Li and Fei Zhang and Dingrong Wan} } @article {1488, title = {Chromatographic studies on Benincasa hispida (thunb.) Cogn. Seed extract scrutinized by HPLC and HPTLC}, journal = {Pharmacognosy journal}, volume = {6}, year = {2014}, month = {8th April 2014}, pages = {42-48}, type = {Original Article}, abstract = {

Background: Benincasa hispida (Thunb.) Cogn. is an extensive climbing annual herb in an agricultural country like India. Lupeol, a constituent of this species, has been reported to possess good amount of pharmacological potential. Objective: In the current studies, the research team focused on determining the percentage of the lupeol present in the extract of Benincasa hispida seeds by chromatographic techniques. Materials and Methods: Shade-dried seeds of Benincasa hispida were subjected to soxhlet extraction followed by scrutinization of the lupeol contents by HPTLC and HPLC methods after carrying out preliminary phytochemical screening for the constituents present in the extract. Results: The extraction yield was found to be 1.2\% (w/w). Phytochemical screening of the extract revealed the presence of carbohydrates, glycosides, alkaloids, fixed oils and fats, tannins phenolic compounds, steroids and flavonoids. The amount of lupeol present in the seeds extract was found to be 0.47\% w/w (HPTLC)and 6.85\% w/v (HPLC) by HPTLC quantification and HPLC analysis respectively. Research studies showed a peak which coincided with the peak of standard lupeol signifying the presence of lupeol in the extract. Conclusion: The extract contains significant amount of lupeol.

Key words:Benincasa hispida, Soxhlet extraction, HPLC, HPTLC, lupeol.

}, keywords = {Benincasa hispida, HPLC, HPTLC, Lupeol, Soxhlet Extraction}, author = {Hemant D. Une, and Gaurav M. Doshi} }