Phytochemical and anti-bacterial activity of epidermal glands extract of Christella parasitica (L.) H. Lev.

Objective:To study the morphology,biochemistry and bioactivity of the epidermal glands of the glandular morphotype of Christella parasitica(C.parasitica)(L.) H.Lev.Methods: Morphological studies on epidermal glands were earned out by using light microscope and scanning electron microscope.To prepare the extract,the shade-dried fronds of glandular morphotype were soaked in acetone.For antibacterial studies paper disc method was followed by using various pathogenic bacteria.Results:Detailed micromorphological,phytochemical and bioactivity studies on a medicinal fern C.parasitica(L.) H.Lev.showed its inlraspecific variation in antibacterial activity.The presence or absence of the epidermal glands was the key factor for antibacterial activity in the morphovariants of this species.The epidermal glands were orange-coloured,stalked and elongated ones of about 84.2μm×45μm,and distributed on the undersurface of cosla,coslules and veins in croziers,young and mature leaves.Frequency of glands varied from 15/cm on costa in mature leaves to 140/cm on costules in croziers.The acetone extract of the glands showed antibacterial activities and also toxic effect against mosquito larvae and tadpoles of frog.Preliminary phytochemical analysis and HPLC studies of the gland extract showed the presence of various kinds of terpenoids,alkaloids,tannins,saponins and flavonoids in it.Conclusions:The present study shows that epidermal glands of the glandular morphotype of C. parasitica(L.) H.Lev.have several bioactive compounds and such rare moiphovariant should be conserved in nature.The next step is to isolate the pure compounds and to screen the bioactivity of individual compounds of the epidermal glands.

Sensitive and rapid determination of amantadine without derivatization in human plasma by LC–MS/MS for a bioequivalence study

A highly sensitive, rapid and rugged liquid chromatography-tandem mass spectrometw （LC-ESI-MS/MS） method was developed for reliable estimation of amantadine （AMD）, an antiviral drug in human plasma. The analyte and internal standard （IS）, amantadine-d6 （AMD-d6）, were extracted from 200μL plasma by solid phase extraction on Phenomenex Strata-X-C 33 ~t cartridges. Chromatography was performed on Synergi^TM Hydro-RP C18 （150 mm×4.6 mm, 4 μm） analytical column using a mixture of acetonitrile and 10 mM ammonium formate, pH 3.0 （80：20, v/v） as the mobile phase. Detection and quantitation was done by multiple reaction monitoring in the positive ionization mode for AMD （m/z 152.1→ 135.1 ） and IS （m/z 158.0 → 141.1） on a triple quadrupole mass spectrometer. The assay was linear in the concentration range of 0.50-500 ng/mL with correlation coefficient （r^2） 〉 0.9969. The limit of detection of the method was 0.18 ng/mL The intra-batch and inter-batch precisions were 〈 5.42% and the accuracy varied from 98.47% to 105.72%. The extraction recovery of amantadine was precise and quantitative in the range of 97.89%-100.28%. IS-normalized matrix factors for amantadine varied from 0.981 to 1.012. The stability of AMD in whole blood and plasma was evaluated under different conditions. The developed method was successfully applied for a bioequivalence study with 100 mg of AMD in 32 healthy volunteers. The reproducibility of the assay was determined by reanalysis of 134 subject samples.

Chromatographic finger print analysis of steroids in Aerva lanata L by HPTLC technique

Objective:To determine the chemical profile and steroids composition of the medicinally important plant Aerva lanata(A.lanata) L.Methods:Preliminary phytochemicul screening was done by the method as Harborne described.HPTLC studies were canied out as Harborne and Wagner et al described.The Ethyl acetate-ethanol-water(8:2:1.2) was employed as mobile phase for glycosides.Results:The desired aim was achieved using Chloroform-acetone(8:2) as the mobile phase.The methanolic extract of stem,leaves,root,flower and seeds of A.lanata showed the presence of 30 different types of steroids with 30 different Rf values from 0.04 to 0.97. Maximum number(11) of steroids has been observed in leaves followed by root(10).Conclusions: HPTLC profile of steroids has been chosen here to reveal the diversity existing in A.lanata.Such finger printing is useful in differentiating the species from the adulterant and act as biochemical markers for this medicinally important plant in the pharma industry and plant systematic studies.

Fast and sensitive LC–MS/MS method for the simultaneous determination of lisinopril and hydrochlorothiazide in human plasma

A sensitive and rapid liquid chromatography-tandem mass spectrometry(LC–MS/MS) method has been developed for the simultaneous determination of lisinopril(LIS) and hydrochlorothiazide(HCTZ) in human plasma using their labeled internal standards(ISs). Sample pre-treatment involved solid phase extraction on Waters Oasis HLB cartridges using 100 μL of plasma, followed by liquid chromatography on Hypersil Gold C_(18)(50 mm×3.0 mm, 5 μm) column. The analytes were eluted within 2.0 min using acetonitrile-5.0 m M ammonium formate, p H 4.5(85:15, v/v) as the mobile phase. The analytes and ISs were analyzed in the negative ionization mode and quantified using multiple reaction monitoring. The method showed excellent linearity over the concentration range of 0.50–250.0 ng/m L for both the analytes. The intra-batch and inter-batch precision(% CV) was ≤5.26% and their extraction recoveries were in the range of 96.6%–103.1%. Matrix effect evaluated in terms of IS-normalized matrix factors ranged from 0.97 to 1.03 for both the analytes. The validated method was successfully applied to determine the plasma concentration of the drugs using 10 mg lisinopril and 12.5 mg hydrochlorothiazide fixed dose formulation in 18 healthy Indian volunteers.

Preliminary phytochemical and antimicrobial studies on a spike-moss Selaginella inaequalifolia(hook.& grev.) Spring

Objective:To screen the anti-cancer spike-mosses for the presence of various bioactivities and to identify the important bioactive chemicals present in Selaginella inaequalifolia(S. inaequalifolia)(Hook.& Grev.) Spring.Methods:Preliminary phytochemical screening was done by following the method of Brindha et al.Antimicrobial study was carried out by disc diffusion method.Results:Results of preliminary phytochemical screening on five different extracts(petroleum ether,benzene,chloroform,ethanol and distilled water) of the spike-moss S.inaequalifolia show the presence steroids,triterpenes,phenolic group,tannin,sugars and catechin.Alkaloids,amino acids,anthraquinone and reducing sugar did not show any positive result.Among the five different extracts,ethanol and chloroform extracts show the presence of maximum number(4 each) of compounds.The results on antimicrobial studies show that all the three microbes[Staphylococcus aureus(S.aureus),Escherichia coli(E.coli) and Candida albicans (C.albicans)]tested are resistant to the ethanol extract and susceptible to petroleum ether extract.The petroleum ether extract shows maximum inhibition with 45 mm of inhibition zone in C.albicans.The inhibition zone in S.aureus and E.coli are 26 mm and 22 mm respectively. Conclusions:The present study shows S.inaequalifolia having potent antibacterial and anticandidal activities.

The effect of leaves extracts of Clitoria ternatea Linn against the fish pathogens

Objective:To investigate the antimicrobial activity of Clitoria ternatea(C.ternatea) against the fish pathogens viz.,Pseudomonas aeruginosa(P.aeruginosa),Escherichia coli(E.coli),Klebsiella pneumonia(K.pneumonia),Bacillus subtilis(B.subtilis),Aeromonas formican(A.formicans)s, Aeromonas hydrophila(A.hydrophila) and Streptococcus agalactiae(S.agalactiae )isolated from diseased Tilapia(Oreochromis niloticus).Methods:The extracts of C.ternatea was tested against P.aeruginosa,E.coli,K.pneumonia,B.subtilis,A.formicans,A.hydrophila and S.agalactiae by the agar well diffusion method.Results:Different extracts of C.ternatea showed inhibitory effects against P.aeruginosa,E.coli,K.pneumonia,B.subtilis,A.formicans,A.hydrophila and S. agalactiae.Ethyl acetate extracts of C.ternatea showed maximum of zone of inhibition against A. formicans(18 mm),A.hydrophilia(19 mm),B.subtilis(19 mm) and P.aeruginosa(21 mm) next to that ethanol extract of C.ternatea showed A.formicans(18 mm) and E.coli(14 mm) followed by Acetone extract showed maximum zone of inhibition S.agalactiae(19 mm) and K.pneumonia(17 mm).Conclusions:The antimicrobial activities of all the four plant extracts are comparable and their potential as alternative in the treatment of infectious by these microorganisms was present in the fish.Susceptibility testing is conducted on isolates using drugs selected on the basis of their importance to human medicine and use in fish production.

Application of an LC–MS/MS method for the analysis of amlodipine,valsartan and hydrochlorothiazide in polypill for a bioequivalence study

A sensitive and selective method has been proposed for the simultaneous determination of amlodipine(AML),valsartan(VAL) and hydrochlorothiazide(HCTZ) in human plasma by liquid chromatography–tandem mass spectrometry(LC–MS/MS). The analytes and their deuterated analogs were quantitatively extracted from100 μL human plasma by solid phase extraction on Oasis HLB cartridges. The chromatographic separation of the analytes was achieved on a Chromolith RP18 e(100 mm × 4.6 mm) analytical column within 2.5 min. The resolution factor between AML and VAL, AML and HCTZ, and VAL and HCTZ was 2.9, 1.5 and 1.4, respectively,under isocratic conditions. The method was validated over a dynamic concentration range of 0.02–20.0 ng/m L for AML, 5.00–10,000 ng/m L for VAL and 0.20–200 ng/m L for HCTZ. Ion-suppression/enhancement effects were investigated by post-column infusion technique. The mean IS-normalized matrix factors for AML, VAL and HCTZ were 0.992, 0.994 and 0.998, respectively. The intra-batch and inter-batch precision(% CV) across quality control levels was ≤ 5.56% and the recovery was in the range of 93.4%–99.6% for all the analytes. The method was successfully applied to a bioequivalence study of 5 mg AML + 160 mg VAL + 12.5 mg HCTZ tablet formulation(test and reference) in 18 healthy Indian males under fasting. The mean log-transformed ratios of C max, AUC0–120 h and AUC0-inf and their 90% CIs were within 90.2%–102.1%. The assay reproducibility was demonstrated by reanalysis of 90 incurred samples.

Simultaneous quantification of prodrug oseltamivir and its metabolite oseltamivir carboxylate in human plasma by LC-MS/MS to support a bioequivalence study

A simple, precise and rapid liquid chromatography-tandem mass spectrometry （LC-MS/MS） method has been developed and validated for the simultaneous determination of oseltamivir and oseltamivir carboxylate, a neuraminidase inhibitor, using their deuterated analogs as internal standards （ISs）. The method involved solid phase extraction of the analytes and ISs from 200 μL human plasma with no reconstitution and drying steps. The chromatographic separation was achieved on a Symmetry C18 （100 mm × 4.6 mm, 5 μm） column using 10 mM ammonium formate and acetonitrile （30：70, v/v） as the mobile phase in a run time of 2.0 min. Quantitation of analytes and ISs were done by multiple reaction monitoring on a triple quadrupole mass spectrometer in the positive ionization mode. The linearity of the method was established in the concentration range of 0.5-200 ng/mL and 2.0-800 ng/mL for oseltamivir and oseltamivir carboxylate respectively. The mean extraction recovery for oseltamivir （94.4%） and oseltamivir carboxylate （92.7%） from spiked plasma samples was consistent and reproducible. The application of this method was demonstratedby a bioequivalence study in 42 healthy Indian subjects with 75 mg oseltamivir phosphate capsules. The assay reproducibility was established by reanalysis of 151 incurred subject samples.

Prediction of influence of process parameters on tensile strength of AA6061/TiC aluminum matrix composites produced using stir casting

Stir casting was used to produce AA6061/15%TiC （mass fraction） aluminum matrix composites （AMCs）. An empirical relationship was developed to predict the effect of stir casting parameters on the ultimate tensile strength （UTS） of AA6061/TiC AMCs. A central composite rotatable design consisting of four factors and five levels was used to minimize the number of experiments, i.e., castings. The factors considered were stirring speed, stirring time, blade angle and casting temperature. The effect of those factors on the UTS of AA6061/TiC AMCs was derived using the developed empirical relationship and elucidated using microstructural characterization. Each factor significantly influenced the UTS. The variation in the UTS was attributed to porosity content, cluster formation, segregation of TiC particles at the grain boundaries and homogenous distribution in the aluminum matrix.

Determination of lercanidipine in human plasma by an improved UPLC–MS/MS method for a bioequivalence study

An improved and reliable ultra-performance liquid chromatography/tandem mass spectrometry （UPLC- MS/MS） method has been developed and validated for the determination of lercanidipine in human plasma. Plasma samples with lercanidipine-d3 as an internal standard （IS） were prepared by solid phase extraction on Phenomenex Strata-X cartridges using 100 μL of human plasma. Chromatographic analysis was performed on UPLC BEH C18 （50 mm × 2.1 mm, 1.7 μm） column under isocratic conditions. Linear calibration curves were obtained over a wide dynamic concentration range of 0.010-20.0 ng/mL. Matrix effect was assessed by post-column infusion, post-extraction spiking and standard-line slope methods. The mean extraction recovery was 〉 94% for the analyte and IS. Inter-batch and intra-hatch precision （% CV） across five quality controls was 〈 5.8%. Bioequivalence study was performed with 36 healthy sub- jects after oral administration of 10 mg of lercanidipine and the assay reproducibility was evaluated by reanalysis of 133 incurred samples.

Determination of cilostazol and its active metabolite 3,4-dehydro cilostazol from small plasma volume by UPLC-MS/MS

A simple,rapid and sensitive ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） method has been developed for the simultaneous determination of cilostazol and its pharmacologically active metabolite 3,4-dehydro cilostazol in human plasma using deuterated analogs as internal standards（ISs）.Plasma samples were prepared using solid phase extraction and chromatographic separation was performed on UPLC BEH C18（50 mm × 2.1 mm.1.7 μm） column.The method was established over a concentration range of 0.5-1000 ng/mL for cilostazol and 0.5-500 ng/mL for 3.4-dehydro cilostazol.Intra- and inter-batch precision（%CV） and accuracy for the analytes were found within 0.93-1.88 and 98.8-101.7% for cilostazol and 0.91-2.79 and 98.0-102.7% for the metabolite respectively.The assay recovery was within 95-97% for both the analytes and internal standards.The method was successfully applied to support a bioequivalence study of 100 mg cilostazol in30 healthy subjects.

SPE-UPLC-MS/MS assay for determination of letrozole in human plasma and its application to bioequivalence study in healthy postmenopausal Indian women

A rapid and sensitive ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） method is described for determination of letrozole in human plasma.Following solid phase extraction（SPE） of letrozole and letrozole-d4 on Orochem DVB-LP cartridges,chromatography was performed on Acquity UPLC BEH C_（18）（50 mm × 2.1 mm.1.7 μm） column using methanol-0.1%formic acid in water（85：15,v/v） as the mobile phase.Detection was carried out on a triple quadrupole mass spectrometer with an electrospray source,operated under positive ionization mode.Quantitation of letrozole and letrozole-d4 was done using multiple reaction monitoring（MRM） following the transitions at m/z286.2→217.0 and m/z 290.2→221.0,respectively.The calibration plots were linear through the concentration range of 0.10-100 ng/mL（r^2 ≥ 0.9990） using 100 μL human plasma.The extraction recovery of letrozole ranged from 94.3%to 96.2%and the intra-batch and inter-batch precision was ≤ 5.2%.The method was successfully applied to a bioequivalence study of letrozole after oral administration of2.5 mg tablet formulation to 16 healthy postmenopausal Indian women.The assay reproducibility was also established through incurred sample reanalysis（ISR） of 74 subject samples.

Liquid chromatography-tandem mass spectrometry method for simultaneous determination of albendazole and albendazole sulfoxide in human plasma for bioequivalence studies

An improved high performance liquid chromatography-tandem mass spectrometry（LC-MS/MS） method has been developed for sensitive and rapid determination of albendazole（ABZ） and its active metabolite,albendazole sulfoxide（ABZSO）,in the positive ionization mode.The method utilized solid phase extraction（SPE） for sample preparation of the analytes and their deuterated internal standards（ISs） from100 μL human plasma.The chromatography was carried out on Hypurity C_（18） column using acetonitrile-2.0 mM ammonium acetate,pH 5.0（80：20,v/v） as the mobile phase.The assay exhibited a linear response over the concentration range of 0.200-50.0 ng/mL for ABZ and 3.00-600 ng/mL for ABZSO.The recoveries of the analytes and ISs ranged from 86.03%-89.66%and 89.85%-98.94%,respectively.Matrix effect,expressed as IS-normalized matrix factors,ranged from 0.985 to 1.042 for the both analytes.The method was successfully applied for two separate studies in healthy subjects using single dose of 400 mg conventional tablets and 400 mg chewable ABZ tablets,respectively.

Studies on isozymic variation among the South Indian species of Sphaerostephanos

Objective:To explore the identity and phylogenetic relationships among the three medicinally important species of Sphaerostephanos from South India using isozymic profile.Methods:The young fronds were homogenized with 3.5 mL of ice-cold homogenizing buffer in a pre-chilled pestle and mortar.The supernatant was subjected to electrophoresis as described by Anbalagan poly acrylamide gel electrophoresis.Staining solutions for isoperoxidase was prepared as per Smila method for the detection of isoenzymes.Results:A total of six different bands in five different positions with different molecular weight/Rf values and four active zones have been observed in the isoperoxidase enzyme system of Sphaerostephanos.Only one band with MW/Rf 0.399 is common to two different species i.e.Sphaerostephanos arbuscula(S.arbuscula) and Sphaerostephanos unitus(S.unitus).Among the remaining four bands,two bands(R_f.0.23,0.47)are present in Sphaerostephanos subtruncatus(S.subtruncatus) and one distinct band has been observed individually in S.arbuscula(R_f.0.507) and S.unitus(R_f.0.56).Conclusions:The present preliminary molecular study through isozymic analysis shows the identity of all the three species and the present results confirm distinctness of these three species based on macromicromorphology,phytochemistry and cytology.

Influence of Metallic Substitutions on the Optical and Mechanical Properties of NLO Benzoyl Glycine Crystals

Benzoyl glycine （BG） is a promising organic nonlinear optical （NLO） crystal, whose second harmonic generation （SHG） efficiency is much higher than that of KDP （potassium dihydrogen phosphate）. Single crystals of pure, Cu2＋ and Cd2＋ doped BG were grown by slow evaporation technique. Optically transparent and defect free single crystals of size up to 10 mm×15 mm×10 mm were harvested in a period of 40-60 days. The growth conditions of pure and doped crystals of BG were optimized and the grown crystals were confirmed by single crystal XRD （X-ray diffraction）. The grown crystals were characterized by FTIR （Fourier transform infrared spectroscopy）, optical absorption and microhardness studies. The microhardness studies confirm that BG has a moderate VHN （Vickers hardness number） value in comparison to the.other organic NLO crystals. The efficiency of frequency doubling was measured for the using Nd：YAG laser and the results were discussed.

Highly sensitive LC–MS/MS method to estimate doxepin and its metabolite nordoxepin in human plasma for a bioequivalence study

A selective, sensitive and rugged liquid chromatography–tandem mass spectrometry(LC–MS/MS) assay has been developed for the simultaneous determination of doxepin(Dox) and its pharmacologically active metabolite, nordoxepin(NDox) in human plasma. The analytes and their internal standards(IS)were extracted from 500 m L of human plasma by liquid-liquid extraction using methyl tert-butyl ether.Chromatographic separation was achieved on Hypurity C8 column(100 mm ? 4.6 mm, 5 mm) using a mixture of acetonitrile-methanol(95:5, v/v) and 2.0 mM ammonium formate in 93:7(v/v) ratio. Detection was accomplished by tandem mass spectrometry in the positive ionization and multiple reaction monitoring acquisition mode. The protonated precursor to product ion transitions studied for Dox, NDox,and their corresponding ISs, propranolol and desipramine, were m/z 280.1-107.0, 266.0-107.0,260.1-116.1 and 267.1-72.1, respectively. A linear dynamic range of 15.0–3900 pg/mL for Dox and 5.00–1300 pg/mL for NDox was established with mean correlation coefficient(r2) of 0.9991 and 0.9993, respectively. The extraction recovery ranged from 86.6%–90.4% and 88.0%–99.1% for Dox and NDox, respectively. The intra-batch and inter-batch precision(% CV) across quality control levels was r 8.3% for both the analytes. Stability evaluated under different storage conditions showed no evidence of degradation and the % change in stability samples compared to nominal concentration ranged from 4.7% to12.3%. The method was successfully applied to a bioequivalence study of 6 mg doxepin hydrochloride orally disintegrating tablet in 41 healthy Indian subjects under fasting and fed conditions.

Deterioration of Early Holocene coral reef due to sea level rise along west coast of India:Benthic foraminiferal testimony

A total of 103 surface sediment samples collected from the water depth range of 15-3300 m along Vijaydurg-Karwar stretch of central west coast of India were analyzed for foraminiferal content. Relict benthic foraminiferal assemblage was noted within 50--135 m water depth. The relict benthic foraminiferal assemblage that includes Amphistegina, Operculina and Alveolinella in sediment samples within the water depth of 85-- 135 m indicates presence of coral reef at this depth during Early Holocene. The presence of barnacle fouling on Relict foraminifera at 60--90 m confirms the paleo-shoreline. The shallow depth zone is characterized by presence of agglutinated relict foraminifera. The agglutinated forms indicate freshwater influx, which eventually increased the sea level and subsequently deteriorated the paleo-coral reef.

An improved LC–MS/MS method for the quantification of alverine and para hydroxy alverine in human plasma for a bioequivalence study

A highly sensitive and selective high performance liquid chromatography–tandem mass spectrometry method was developed and validated for the quantification of alverine（ALV） and its active metabolite, para hydroxy alverine（PHA）, in human plasma. For sample preparation, solid phase extraction of analytes was performed on Phenomenex Strata-X cartridges using alverine-d5 as the internal standard. The analytes were separated on Symmetry Shield RP18（150 mm×3.9 mm, 5 μm） column with a mobile phase consisting of acetonitrile and10 mM ammonium formate（65：35, v/v）. Detection and quantitation was done by electrospray ionization mass spectrometry in the positive mode using multiple reaction monitoring. The assay method was fully validated over the concentration range of 15.0–15,000 pg/mL for ALV and 30.0–15,000 pg/mL for PHA. The intra-day and inter-day accuracy and precision（% CV） ranged from 94.00% to 96.00% and 0.48% to 4.15% for both the analytes. The mean recovery obtained for ALV and PHA was 80.59% and 81.26%, respectively. Matrix effect,expressed as IS-normalized matrix factor ranged from 0.982 to 1.009 for both the analytes. The application of the method was demonstrated for the specific analysis of ALV and PHA for a bioequivalence study in 52 healthy subjects using 120 mg ALV capsules. The assay reproducibility was also verified by reanalysis of 175 incurred subject samples.

A review:Advances in microbial remediation of trichloroethylene (TCE)

Research works in the recent past have revealed three major biodegradation processes leading to the degradation of trichloroethylene. Reductive dechlorination is an anaerobic process in which chlorinated ethenes are used as electron acceptors. On the other hand, cometabolism requires oxygen for enzymatic degradation of chlorinated ethenes, which however yields no benefit for the bacteria involved. The third process is direct oxidation under aerobic conditions whereby chlorinated ethenes are directly used as electron donors by microorganisms. This review presented the current research trend in understanding biodegradation mechanisms with regard to their field applications. All the techniques used are evaluated, with the focus being on various factors that influence the process and the outcome.

Antibacterial activity of leaves and inter-nodal callus extracts of Mentha arvensis L

Objective:To determine the anti-bacterial efficacy of chloroform,ethanol,ethyl acetate and water extracts of inter-nodal and leaves derived calli extracts from Mentha arvensis(M.arvensis) against Salmonella typhi(S.typhi),Streptococcus pyogenes(S.pyogenes),Proteus vulgaris(P. vulgaris) and Bacillus subtilis(B.subtilis).Methods:The inter-nodal and leaves segments of M.arvensis were cut into 0.5-0.7 cm in length and cultured on Murashige and Skoog solid medium supplemented with 3%sucrose,gelled with 0.7%agar and different concentration of 2,4-Dichlorophenoxyacetie acid(2,4-D) either alone or in combinations.The preliminary phytochemical screening was performed by Brindha et al method.Antibacterial efficacy was performed by disc diffusion method and incubated for 24 h at 37℃.Results:Maximum percentage of callus formation(inter-nodal segments 84.3±0.78;leaves segments 93.8±1.27) was obtained on Murashige and Skoog’s basal medium supplemented with 3%sucrose and 1.5 mg/L of 2,4-D.The ethanol extracts of leaves derived calli showed the maximum bio-efficacy than other solvents.The leaves and stem derived calli extracts on Proteus sp.showed that the plants can be used in the treatment of urinary tract infection associated with Proteus sp.Through the bacterial efficacy studies,it is confirmed that the in vitro raised calli tissue was more effective compared to in vivo tissue.Conclusions:The bio-efficacy study confirmed that the calli mediated tissues showed the maximum zone of inhibition.The present study paved a protocol to establish high potential cell lines by in vitro culture.