Physicochemical and marker based optimization of fermentation process of Drakshasava, an Ayurvedic polyherbal formulation

Drakshasava a classical herbal formulation containing Vitis vinifera L. (grapes) as a chief ingredient. It is one of the best asava preparation, referred in Ayurvedic Formulary of India (AFI). The present study explores the changes brought by fermentation during the processing of Drakshasava with respect to physicochemical parameters using correlation marker based approach. The grapes were extracted to form decoction and further fermented with Woodfordia flowers along with prakshepadravyas (additional drugs in small quantity) and sugar source, as per AFI, part II. The fermented broth was analyzed daily for 14 days for various physicochemical parameters. Further marker based profiling was done by High Performance Thin Layer Chromatography (HPTLC) with respect to caffeic acid and kaempferol. The fermentation took 3 days lag period for generation of quantifiable alcohol (0.5%) on 4th day to 16.7% on 14th day. The admissible limit of alcohol for Drakshasava as per API (5-10%) has been reached on 9th day itself.The pH, specific gravity, solid content has been decreased from day 0 (5.12; 1.32; 28.25) to 14th day (4.35; 1.15; 22.81) respectively whereas phenolic content has been upgraded remarkably (0.040 to 0.165 g/100 mL) during the study. Kaempferol quantity was found to be decreased (0.186% to 0.0810%) after fermentation whereas caffeic acid quantity has been significantly increased in post fermented sample with respect to decoction (0.0168% to 0.0233%). The change brought in quantity of the phytochemicals, indicates the metabolic conversion during fermentation due to glycosylation or deglycosylation. The research limits the fermentation time for preparation of Drakshasava from 21 days to 8-9 days, saving time and money. The new optimized method adheres to the Ayurvedic principles and opens the scope for better quality industrial research and large scale production

Physicochemical and marker based optimization of fermentation process of Drakshasava, an Ayurvedic polyherbal formulation

120-127Drakshasava a classical herbal formulation containing Vitis vinifera L. (grapes) as a chief ingredient. It is one of the best asava preparation, referred in Ayurvedic Formulary of India (AFI). The present study explores the changes brought by fermentation during the processing of Drakshasava with respect to physicochemical parameters using correlation marker based approach. The grapes were extracted to form decoction and further fermented with Woodfordia flowers along with prakshepadravyas (additional drugs in small quantity) and sugar source, as per AFI, part II. The fermented broth was analyzed daily for 14 days for various physicochemical parameters. Further marker based profiling was done by High Performance Thin Layer Chromatography (HPTLC) with respect to caffeic acid and kaempferol. The fermentation took 3 days lag period for generation of quantifiable alcohol (0.5%) on 4th day to 16.7% on 14th day. The admissible limit of alcohol for Drakshasava as per API (5-10%) has been reached on 9th day itself.The pH, specific gravity, solid content has been decreased from day 0 (5.12; 1.32; 28.25) to 14th day (4.35; 1.15; 22.81) respectively whereas phenolic content has been upgraded remarkably (0.040 to 0.165 g/100 mL) during the study. Kaempferol quantity was found to be decreased (0.186% to 0.0810%) after fermentation whereas caffeic acid quantity has been significantly increased in post fermented sample with respect to decoction (0.0168% to 0.0233%). The change brought in quantity of the phytochemicals, indicates the metabolic conversion during fermentation due to glycosylation or deglycosylation. The research limits the fermentation time for preparation of Drakshasava from 21 days to 8-9 days, saving time and money. The new optimized method adheres to the Ayurvedic principles and opens the scope for better quality industrial research and large scale production

DENSITOMETRIC VALIDATION OF LAPACHOL IN TECOMELLA UNDULATA SEEM BARK BY HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY

Objective: To develop a validated high-performance thin layer chromatography (HPTLC) method for the quantitative estimation of Lapachol in a Soxhlet extracted bark of Tecomella undulata seem (T. undulata). Methods: The bark of T. undulata was extracted with chloroform by Soxhlet apparatus. The separation was achieved on a silica-gel 60 F254 HPTLC plate using toluene-ethyl acetate–glacial acetic acid (8.5:1.5:0.02 v/v/v) as a mobile phase. Densitometric analysis of Lapachol was carried out in absorbance mode at 254 nm.Results: The proposed method was accurate for the separation and resolution between peaks of the standard and Lapachol (5.04µg) with Rf value 0.77. Calibration curves were found to be linear over the concentration range (10-130µg) for Lapachol and correlation coefficient over (R2= 0.9973), indicating an excellent correlation between peak areas and concentrations of the marker compound. The experimentally derived LOD and LOQ for Lapachol were determined to be 0.028 µg and 0.086 µg respectively and the developed HPTLC-UV method showed lower %RSD and SEM, value indicating the method to be precise, accurate and robust.Conclusion: The study concludes that HPTLC-UV validation method can be very efï¬cient and promising technique for the identiï¬cation and quantitative analysis of Lapachol from T. undulata bark. The statistical analysis of data indicates that the developed method is reproducible and specific

Optimization of fermentation variables for Ayurvedic formulation, Drakshasava by Response Surface Methodology and its marker based validation studies

The concept of fermentation has been part of various Ayurvedic formulations since ages. Drakshasava is one of the self fermented ayurvedic medicine containing Vitis vinifera L. as major ingredient. However, lack of modern technology and long processing time of these classical formulations may interfere with growing needs of industries. To cope up with the current health care standards, optimization of existing Ayurvedic formulations with multi-marker approach is required. A statistical model has been developed to optimise the fermentation process of Drakshasava (without prashpeka dravya). The selected fermentation process parameters i.e., pH, incubation temperature and fermentation time were optimized by BBD of Response Surface Methodology. The combination of parameters was selected to achieve a required alcohol percentage. The method was validated by HPTLC with respect to non established biomarker piperine, a bioavailability enhancer required to prevent early metabolism of resveratrol (therapeutic active constituent of Drakshasava). The quadratic model was found significant with F and P value, 2.20 and 2.302, respectively. The optimum conditions obtained in the batch fermentation process were incubation temperature (30.59°C); pH (4.84) and fermentation period of 9 days with predicted concentration of 9.82% alcohol. The method was validated in accordance to ICH guidelines and found statistically reproducible and selective for the quantitative estimation of piperine in Drakshasava. The in-house fermented samples were found to have significant increase in piperine concentration (2.42 μg/mL), compared to decoction (0.11 μg/mL). The optimized method reduced the manufacturing time without interefering the Pharmacopoeial limits. It leads to standardization, better quality and consistency of Drakshasava

Formulation and Evaluation of Guggul Lipid Nanovesicles for Transdermal Delivery of Aceclofenac

Context. Most new drugs have low water solubility and liposome is an important formulation to administer such drugs; however, it is quite unstable and has negligible systemic absorption. Objective. Aceclofenac nanovesicles were made using guggul lipid for formulating stable transdermal formulation. Materials and Methods. Guggul lipid was formulated into vesicles along with cholesterol and dicetyl phosphate using film hydration method. The formulations were analyzed for physicochemical properties and stability. Then its skin permeation and anti-inflammatory activity were determined. Results. Both categories of vesicles (PC and GL) showed optimum physicochemical properties; however, accelerated stability study showed considerable differences. GL-1 was appreciably stable for over 6 months at 4°C. Corresponding gels (PCG-1 and GLG-1) showed Cmax values at 4.98 and 7.32 μg/mL along with the Tmax values at 4 and 8 hours, respectively. GLG-1 inhibited edema production by 90.81% in 6 hours. Discussion. PC liposomes are unstable at higher temperature and upon longer storage. The formulation with higher lipid content (GL-1) showed good drug retention after 24 hours and appreciable stability both at higher temperature and for longer duration. Guggul lipid being a planar molecule might be stacked in vesicle wall with cholesterol. Conclusion. The composition of the nanovesicle played an important role in stability and drug permeation. Guggul lipid is suitable for producing stable vesicles

ISOLATION AND ANTIDIABETIC ACTIVITY OF NEW LANOSTENOIDS FROM THE LEAVES OF PSIDIUM GUAJAVA L.

Objective: Diabetes mellitus is a chronic metabolic disease which affects our body's ability to use the energy found in food. Our study was planned to isolate chemical constituents from the leaves of Psidium guajava L. (Myrtaceae), to characterize their structures and to investigate their antidiabetic activity.Methods: The air-dried leaf powder was exhaustively extracted with methanol in a Soxhlet apparatus. The concentrated leaf extact was adsorbed on silica gel (60-120 mesh) for the preparation of a slurry. The dried slurry was chromatographed over silica gel column packed in petroleum ether. The column was eluted with petroleum ether, chloroform and methanol, successively, in order of increasing polarity to isolate the compounds. These natural constituents were tested for the antidiabetic activity in STZ-induced diabetic models.Results: Six new lanosterol-type triterpenoids characterized as lanost-7-en-3β-ol-26-oic acid (2), lanost-7-en-3β, 12β-diol-26-oic acid (3), lanost-7-en-3β, 12β, 29-triol-26-oic acid (4), lanost-cis-1,7,23-trien-3β, 12β, 18, 22α-tetraol-26-oic acid (5), lanosteryl-3β-O-D-xylopyranosyl-2′-p-benzaldehyde (7) and lanost-7-en-3β-ol-26-oic acid-3β-D-glucopyranoside (8) along with the known compounds arachidic acid (1) and β-sitosterol xylopyranoside (6) were isolated from the leaves. The compounds 2, 3, 4 and 8 exhibited significant antidiabetic activity against streptozotocin-induced diabetic rats.Conclusion: The leaves of P. guajava possessed antidiabetic lanostene-type triterpenoids

Enhancement of mangiferin by fungal endophytes isolated from Salacia chinensis L. and Salacia oblonga Wall.

Microbes residing in the internal tissues of a plant are called endophytes, and are known for producing phytochemicals such as taxol, podophyllotoxin, azadirachtin and vinca alkaloids. In this study, out of five isolates from Salacia species, two fungi Penicillium capsulatum and Aspergillus fumigatus have been evaluated and confirmed by polymerase chain reaction (PCR) amplification for their endophytic action to produce mangiferin. Mangiferin has been reported to possess protective properties, including antioxidant, antidiabetic and immunomodulatory. It has been reported that the content of mangiferin is 7-9% in Mangifera indica, and is also present in other plants like Swertia chirata, Salacia chinensis, and Hypericum aucheri. Therefore, an attempt was made to explore the biotechnological approach and regulation studies to increase the production of mangiferin in S. chinensis and S. oblonga. Endophytes were isolated, screened, and analyzed, to evaluate the mangiferin in fungal extracts in comparison with crude plant extracts. An HPLC analysis was used to determine the mangiferin content present in the fungal extract of S. chinensis stem (74.74 g/mL), followed by fungi extracts of S. oblonga root (33.75 g/mL) and S. chinensis root (30.50 g/mL), compared with the plant extracts. These results were confirmed by FTIR analyses.

Optimization of fermentation variables for Ayurvedic formulation, Drakshasava by Response Surface Methodology and its marker based validation studies

744-753The concept of fermentation has been part of various Ayurvedic formulations since ages. Drakshasava is one of the self fermented Ayurvedic medicine containing Vitis vinifera L. as major ingredient. However, lack of modern technology and long processing time of these classical formulations may interfere with growing needs of industries. To cope up with the current health care standards, optimization of existing Ayurvedic formulations with multi-marker approach is required. A statistical model has been developed to optimise the fermentation process of Drakshasava (without prakshepa dravya). The selected fermentation process parameters i.e., pH, incubation temperature and fermentation time were optimized by BBD of Response Surface Methodology. The combination of parameters was selected to achieve a required alcohol percentage. The method was validated by HPTLC with respect to non established biomarker piperine, a bioavailability enhancer required to prevent early metabolism of resveratrol (therapeutic active constituent of Drakshasava). The quadratic model was found significant with F and P value, 2.20 and 2.302, respectively. The optimum conditions obtained in the batch fermentation process were incubation temperature (30.59°C); pH (4.84) and fermentation period of 9 days with predicted concentration of 9.82% alcohol. The method was validated in accordance to ICH guidelines and found statistically reproducible and selective for the quantitative estimation of piperine in Drakshasava. The in-house fermented samples were found to have significant increase in piperine concentration (2.42 μg/mL), compared to decoction (0.11 μg/mL). The optimized method reduced the manufacturing time without interefering the Pharmacopoeial limits. It leads to standardization, better quality and consistency of Drakshasava

Biotransformation of lignan glycoside to its aglycone by Woodfordia fruticosa flowers: quantification of compounds using a validated HPTLC method

Context: Saraca asoca Linn. (Caesalpiniaceae) is an important traditional remedy for gynaecological disorders and it contains lyoniside, an aryl tetralin lignan glycoside. The aglycone of lyoniside, lyoniresinol possesses structural similarity to enterolignan precursors which are established phytoestrogens. Objectives: This work illustrates biotransformation of lyoniside to lyoniresinol using Woodfordia fruticosa Kurz. (Lythraceae) flowers and simultaneous quantification of lyoniside and lyoniresinol using a validated HPTLC method. Materials and methods: The aqueous extract prepared from S. asoca bark was fermented using W. fruticosa flowers. The substrate and fermented product both were simultaneously analyzed using solvent system:toluene:ethyl acetate:formic acid (4:3:0.4) at 254 nm. The method was validated for specificity, accuracy, precision, linearity, sensitivity and robustness as per ICH guidelines. Results: The substrate showed the presence of lyoniside, however, it decreased as the fermentation proceeded. On 3rd day, lyoniresinol starts appearing in the medium. In 8 days duration most of the lyoniside converted to lyoniresinol. The developed method was specific for lyoniside and lyoniresinol. Lyoniside and lyoniresinol showed linearity in the range of 250–3000 and 500–2500 ng. The method was accurate as resulted in 99.84% and 99.83% recovery, respectively, for lyoniside and lyoniresinol. Conclusion: Aryl tetralin lignan glycoside, lyoniside was successfully transformed into lyoniresinol using W. fruticosa flowers and their contents were simultaneously analyzed using developed validated HPTLC method

Comparative powder microscopy on the barks of four Ficus species

The genus Ficus constitutes one of the largest genera of medicinal plants containing more than 800 species occurring in tropical and subtropical areas. The more famous species are Ficus bengalensis (Bargad), Ficus religiosa (Pipal), Ficus racemosa (Goolar) and Ficus lacor (Pakar). These plants are reported to possess antidiabetic, antidiarrhoeal, antiepilepsy, anti-inflammatory properties and are used in many Ayurvedic and traditional formulations. The barks of these species are usually interchanged or adulterated with other species of ficus due to limited knowledge of identification and differentiation. The slide preparation method was optimized to visualize common as well as distinguishing characters of the bark powder. The photomicrographs were taken with Motic microscope moticam 3.0 MP, AE 2000. The variation in the bark powder lies in size and shape of the stone cells and sclereids, their occurrence, type of wall and lumen. Most of the herbal drugs in the industry are supplied in powdered form. So, there are more chances of adulteration as it is very easy to spoil a drug in the powdered state. Industry emphasizes for powder microscopy as it is effortless. Hence, a detailed powder microscopy evaluation was carried out with an aim to establish diagnostic features to differentiate between these four bark powders