Showing posts with label Introduction. Show all posts
Showing posts with label Introduction. Show all posts

The Introduction And Spread Of Japanese Beetles Biology Essay

Japanese Beetles (Popillia japonica) are a destructive pest of turf, landscape, agriculture, and ornamental plants. They were introduced into the United States sometime in the early 1900s on a shipment of iris bulbs. They were first identified in a nursery near Riverton, New Jersey in 1916 during a routine inspection. They are also found in their native Japan, China, Russia, Portugal, and Canada. They have become an important pest as both larvae and adults destroy roots and defoliate plants. The Japanese beetle easily established in the U.S. after its introduction. As with a lot of introduced pest they were lacking in natural enemies and found an abundant food source and favorable climate. Efforts have been made to control these invasive pests. In 1918, the USDA and New Jersey authorities attempted to eradicate the infestation. But after only two years the populations were so well established the efforts were deemed unproductive and were stopped. The best strategy for eradication is early detection, isolation and the total elimination of populations. The annual cost of control in the U.S. exceeds $460 Million (Shetlar 2008).

What are the implications of the introduction of Japanese beetles? And how can their spread across the United States be stopped in their tracks? This paper will be a thorough look into how Japanese beetles have spread across the U.S. and how to cope with this pesky pest.

Japanese beetles are polyphagous generalists and have a wide range of over 300 hosts. They are important pests of fruit, vegetables and field crops. Japanese beetles have one generation per year. A percent of the grubs may take two years to mature based on the soil conditions if they are cold and wet. The larvae are white colored grubs and are typically C Shaped around 1 inch in length. The third instar larvae overwinter in the soil below the frost line. The grubs develop in the soil feeding on the roots of plants. They can destroy lawns and turf grass. They are considered the most destructive pest of turf-grass in the United States. The adults are gregarious and collect together on a single plant. Single plants and trees may be completely defoliated. The adults are ½ inch in length and are metallic green with bronze-colored elytra. The males are typically smaller in size than the females. The adults skeletonize the leaves of their hosts by eating the material between the veins. The veins are often left untouched after feeding and the leaves will crumple and die. They feed on the upper and outermost parts of the plants and work their way down the plant (Cook and Gray 2010).

Japanese beetles are found throughout the Midwest and eastern United States and are spreading west ward. The furthest west they are expected to reach on their own is central Nebraska because of the unfavorable climatic conditions. There are areas in Western states that are suitable for their survival if they hitchhike to the area. The movement of these pests is a major concern to horticulture and farmers. They have been introduced into California three times and successfully eradicated. Material such as nursery commodities, sod, soil, compost and manure from infected states are now prohibited without a PPQ inspection. The USDA closely monitors the movement of cargo planes into the unaffected areas. The beetles are known to fly onto the cargo plane when the doors are open during times of adult beetle flight (Hamilton et al. 2007).

Airports are potential vectors for Japanese beetle infestation. A State Plant Health Director which is an APHIS employee arranges a monitoring survey to determine the risk status of an airport. Inspectors survey the airport to determine the potential risk. Monitoring surveys will determine the threat of entry into the aircraft and the population level in surrounding areas. The Indianapolis Airport is one of many airports where Japanese beetles pose a threat. Traps around the airport over a 4 year period showed that the relative distribution of adult beetles along the trap line were stable and at the same level in the same areas. Conclusions were drawn that the environment around the airport has considerable influence on the distribution of beetles and potential threat of a hijacker flying on board. The soil type, land use, location of host and nonhost plants, soil moisture all effected the movement of the Japanese beetles. A change in the environment such as land use and irrigation could affect the Japanese beetle distribution and be a viable practice to decrease the Japanese beetles’ opportunities to fly onto open airplanes (Hamilton et al. 2007). By manipulating the land around the airport the risk could be minimized.

In 1917 the former U.S. Bureau of Entomology established the Japanese Beetle Laboratory in 1917. They studied the biology of the pest in its new environment and worked on developing methods for control (Potter and Held 2002). They studied the pest-host interaction attempting to gain an understanding of this invasive pest. Entomologists realized this introduction had the potential of becoming a devastating pest that would lead to high economic costs.

In 1998, the National Plant Board initiated the first version of the U.S. Domestic Japanese Beetle Harmonization Plan. This established a plan for the movement of commodities that host Japanese beetles such as nursery stock and other regulated commodities. This program has two objectives to establish a framework that encourages states to assess the Japanese beetle risk and infestations status. The second objective is to provide a more uniform adoption and implementation of pest risk improvement measure to reduce the risk of pest introduction to receiving states. Nursery stock will requires a phytosanitary certificate for moving to states that consider Japanese beetle a quarantine pest (National Plant Board 2011). Movement is restricted from quarantined states, all but Florida, Mississippi, and Louisiana in the east are restricted. Infestations west of the Mississippi River are eradicated before they become established, although partial infestation occurs in Arkansas, Iowa, Kansas, Minnesota, Missouri, and Oklahoma.

Areas that are at risk for introduction have implemented protocols to eradicate this pest if they are introduced. If the Japanese beetle becomes established in Oregon and generally disperses throughout the state, the economic impact to all crops, commodities, and other related businesses could be over $34 million (Hamilton et al. 2007). The absence of natural enemies has allowed the Jap beetles to establish and spread in areas with little predator control. The indigenous host plants that Japanese beetles use as foods source are highly susceptible due to the lack of coevolutionary history with the beetle leaving the plants defenseless.

Japanese beetles are classified as an A1 quarantine organism for EPPO and of quarantine significance for CPPC, JUNAC, and OIRSA. They are at low risk of establishing in the Mediterranean area, and the risk is also low in the UK, Ireland and continental Europe north of 53N, because of the naturally cool summers. The area of concern is between latitudes 43N and 53N east to longitude 30E. If they are introduced and established they could cause massive damage and have significant economic loss. The EPPO makes a suggestion that countries prohibit the importation of plants with roots from countries where P. Japonica is established. If material is imported it should be planted in inorganic growing medium or in treated medium and kept under conditions which prevent infestation (Potter and Held 2002). The removal of soil from the roots of nursery plants will remove the larvae, in some cases the soil should be treated. The Japanese beetle was established in Terceira Island where in 1970 it escaped from a U.S. air base (National Plant Board 2011).

Japanese beetles release an aggregation pheromone and often are found in mating clusters. Invasive pests can benefit native populations through the plant stress-mediated interactions. Grapes are among their preferred hosts. This is an interesting interaction between an invasive species benefiting a native species. The Japanese beetle facilitates the green June bug feeding on grapes by biting through the skin providing access to the GJB and by eliciting yeast-mediated fermentation volatiles that the GJB uses in host-finding and aggregation. The GJB are usually unable to break through the tough skin of grapes so they benefit from the Japanese beetle feeding. This allows the GJB to expand its range into the fruit-growing region. Green June bugs were able to feed on grape plants before but the Japanese beetle injury leads to an additive increase in feeding and aggregation by breaking apart the fruit and attracting the GJB to the plants. In this case Japanese beetles have elevated the economic impact of an indigenous species (Hammonds et al. 2009). If these beetles are introduced and established into California this could potentially threaten the grape industry.

Japanese beetles have favored hosts that they prefer to feed on but will feed successfully on a list of non-preferred hosts. The preferred hosts successfully attract the beetle to feed. Since they attract other beetles to food sources, killing beetles or hand picking them off plants will reduce the populations. Some commercial traps bated with pheromones will also attract them. Grubs have trouble surviving in dry soils, so irrigation should not be used if an infestation is suspected. Bacillus papillae, a bacterial milky spore disease can kill the buried grubs. The bacteria can be applied to the soil and the spores will remain in the soil to infect further inhabitants (Shetlar 2008).

The insect-plant system is a complex interaction that understanding will benefit management strategies. Japanese beetles are generalists that feed on over 300 species making them an important pest of horticultural crops. They are attracted to hosts by green leaf odors and color instead of specific host stimuli. These generalists avoid nonhost plants because they sense deterrents in these plants such as cucurbitacins, a bitter triterpenes characteristic of cucurbits (Potter and Held 2002).

Although, corn and soybeans plants are not a preferred host of the Japanese beetle, they can do considerable damage defoliating soybeans and feeding on silks of corn plants which can reduce the pollination and kernel set. Both the grubs and adults can cause injury in corn. The grubs are able to feed on the root hairs of the plant which interferes with the plant taking up water and nutrients. If fields are heavily infested they can experience reduced plant stands. The adults beetle feed on the silks and this interferes with pollination. In soybean fields, they can also cause damage in both the larvae and adult stage. The grubs also feed on the root hairs of the soybean plants, this usually and economic important issue. The adults can cause extensive damage to the soybean plants, if this happens during the reproductive stages during the plant life a loss in yield will occur. They will feed on the flowers and defoliate the leaves. Management can be difficult. Adults are very mobile and can quickly move from fields. Fields should be scouted to determine the percent defoliation and to correctly identify the species. If the standard economic thresholds are met for corn or soybeans the fields should be sprayed with insecticide (Cook and Gray 2010).

The larvae have limited mobility so the oviposition site is very important for survival of the offspring. They have a strong preference to oviposit in grass the chemical cues stimulate females to lay eggs in the soil (Szendrei and Issacs 2005). Japanese beetles have an interesting mating behavior. The egg load and body size may influence their mating frequency. They have a promiscuous mating system in which both males and females mate repeatedly during their lives during this type the females moves to the soil to lay her eggs. The males are attracted to the virgin females by sex pheromones released (Tigreros and Switzer 2010). They seek out non-virgin females on food plants where they tend to aggregate. There is a last male advantage for fertilization in sperm competition. The males will mount unpaired females or try to remove the male from the paired female. Males may stay on the female for a couple hours “guarding” the female. Males will choose larger females over smaller females and guard these longer. This could be because larger females have more eggs and may oviposit more frequently (Switzer et al. 2008).

Management systems are multifaceted and can be complicated when trying to treat both the grub and adult stage. They both damage plants through different modes of feeding. The adults are mobile and move from place to place while the grubs are restricted to the area where the female laid her egg. They need to be laid by a suitable host for feeding if they are to survive.

Cultural controls can help decrease the larval populations in grasses. Avoiding irrigation during peak beetle flights this can make it difficult for the beetles to oviposit. One of the interesting discoveries was concerning geraniums. Beetles that feed on petals of geraniums roll over on their backs and remain paralyzed for 24 hours. They will typically recover but under field conditions are often preyed on by predators. Some type of substance in these plants paralyzes the beetle. The plants do not deter feeding and could potentially be used as a control agent for Japanese beetles if the volatile could be identified (Flores 2010).

The Japanese beetle problem should be treated using an IPM strategy to fight the infestation. A couple of management tips to follow are to avoid irrigation in areas where grubs are feeding and to avoid planting plants that are preferred hosts of Japanese beetles. Spot treat areas with insecticide if infestation is heavy, treat lawn with pesticides when the soil is wet for better absorption. Being aware of the life cycle of this pest and ways to manage them can help decrease expanding populations. Infestations are usually a threat in areas where there is a history of feeding, so scout and be aware of the potential threat.

Since the accidental introduction of the Japanese beetle into the United States they have slowly spread throughout the country threatening indigenous plants and ecosystems. The Japanese beetles are here to stay. They are voracious generalists that aggregate and can quickly defoliate plants and trees. Their grubs can destroy turf grass leaving irritating bald areas across your lawn. Comprehensive strategies restricting the movement of these mobile pests are in place and should be stringently followed. Stopping the spread of these beetles is very important for areas in the West such as Oregon and California where preferred host plants would be ravaged by their feeding. The combination of their mobility and reproductive capabilities makes Japanese beetles an invasive pest that will need to be closely monitored and quarantined in the present and future.

Cook, K.A. and M.E. Gray. 2010. Japanese Beetle Popillia japonica Newman. University of Illinois Extension. http://ipm.illinois.edu/fieldcrops/insects/japanese_beetles/

Flores, Alfred. 2010. Geraniums could help control devastating Japanese beetles. U.S. Department of Agriculture.

Hamilton, R.M., R.E. Foster, T.J. Gibbs, C.S. Sadof, J.D. Holland, and B.A. Engel. 2007. Distribution and Dynamics of Japanese Beetles along the Indianapolis Airport Perimeter and the Influence of Land Use on Trap Catch. Environment Entomology 36 (2): 287-296.

Hammons, D.L., S.K. Kurtural, M.C. Newman, & D.A. Potter. 2009. Invasive Japanese beetles facilitate aggregation and injury by a native scarab pest of ripening fruit. PNAS, 106 (10): 3686-3691.

Krischik, Vera and Maser, Doree. 2010. Japanese Beetle Management in Minnesota. University of Minnesota Extension. http://www.extension.umn.edu/distribution/horticulture/dg7664.html

National Plant Board. 2011. U.S. Domestic Japanese beetle Harmonization Plan. http://nationalplantboard.org/docs/jbcolumn.pdf

O’Neill, B.F. A.R. Zangerl, E.H. DeLucia, and M.R. Berenbaum. 2008. Longevity and Fecundity of Japanese Beetle (Popillia japonica) on Foliage Grown under Elevated Carbon Dioxide. Environmental Entomology 37(2): 601-607.

Popillia japonica. Data Sheets on Quarantine Pests. 2010. Prepared by CABI and EPPO for the EU.

Potter DA, Held DW. 2002. Biology and management of Japanese. Annual Review of

Entomology 47:175-205.

Shetlar, David. 2008. Japanese Beetle Fact Sheet. The Ohio State University Extension. http://ohioline.osu.edu/hyg-fact/2000/2504.html

Switzer, P.V., Enstrom, P.C. and C.A. Schoenick. 2009. Behavioral Explanations Underlying the Lack of Trap Effectiveness for Small-Scale management of Japanese Beetles (Coleoptera: Scarabaeidae). J.of Ec. Entom. 102(3): 934-940.

Switzer, P.V., Enstrom, P.C. and C.A. Schoenick. 2008. Environmental Conditions Affect Sperm Competition Risk in Japanese Beetles (Coleoptera: Scarabaeidae). Ann. Entomol. Soc. 101(6): 1154-1161.

Szendrei, Zsofia & Rufus Isaacs. 2005. Do plants cues influence the oviposition behavior of Japanese beetles? The Netherlands Entomological Society 117: 165-174.

Tigreros, N. & Switzer, P.V. 2008. Effects of food deprivation, body size, and egg load on the mating behavior of female Japanese beetles. Ethology Ecology & Evolution 20: 89-99.

Tigreros, Natasha, R. Jadhav, K.A. Kowles, B.P. Nathan and P.V. Switzer. 2010. Physiological Status of male and Female Popillia japonica (Coleoptera: Scarabaeidae) Affects mating and Grouping Behavior. Environmental Entomology 39 (3): 892-897.

USDA. Department of Agriculture. 2010. Japanese Beetle Program Manual for Airports.



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Tile And Introduction Concept Of Physiological Properties Biology Essay

Biology » Tile And Introduction Concept Of Physiological Properties Biology Essay

Dissociation constant “pka” It shows the charge on the molecules. Partition coefficient shows the substance which is well partitioned in liquid and water both the parameters are required for understanding the behaviour of the drug, many substances would varied from its physical and chemical properties. So it’s important to determine ionic form of a present molecule is necessary.

Partition coefficient and PKa both in combination can be help full to determine distrubutation of drug compound in a system. Absorption execration and penetration. Affects both the parameters.

Dissociation constant

Ionisation of both acid and base

Pka is not dependent on the concentration for acid

Pka = -log 10 (ka) where ka = (H+) (A-) / (HA)

Concentration ionisation constant, measured by spectroscopy. It is temperature dependent constant

Log p and partition coefficient

It can be shown by

Partition coefficient P = (organic) / (aqueous)

Log P = log 10 (partition coefficient)

Where log D = distributation coefficient

{{54 Earll, Marks. 1999-2006 ;}}

There may be different choose of partition solvent

Octanol the most commonly used solvent

Through which partition or distrubutation ratio can be determined. It’s also determines the distrubutation or partition law. which shows inconsiderateness to concentration and temperature .Concentration studies shows that distribution ratio was clearly concentration dependent in case of” benzoic acid” between water and benzene

The octal water coefficient studies show the resemblance to the body system .so it is mostly used solvent for determination of partition coefficient. Its shows different quantitative structural activity relationship. It shows correlation of biological activity and chemical structure .its lead to high activity, application, implementation in the theory,

It can be shown by BR = a+Bb+cC+......

Where BR is Biological response, B, C are the molecular properties and log Kow, shows large fraction of QSARS relation BR = a+b log Kow

Biological response interferes with the large numbers of reaction to be undertaken in concerned with interaction of xenobiotic activity (in vivo), and in sense of chemical interaction in biological reaction (in vitro). Concepts of pka and partition coefficient studies is relatively important in drug discovery or formulation in pharmaceuticals .it is also showed in vivo chemical studies as Bioaccumulation and relates with the tissue partition coefficient.

{{55 Sangster. 1997 ;}}

Physiochemical properties serve to be the important factor in the drug penetration, solubility, permeability, ionisation, bioavailability, and also adsorption surface.

{{57 Pallasch, Thomas, J. January 22 1988 ;}}

Pka is an essential factor in determining solubility and lipophilicity of drug molecule for better solubility, drug need to be converted into suitable salt form. Traditional method for calculation of pka was potentiometric titration. It is available commercially, due to its high throughput (up to 30 compounds / day) with mini sample requirement method used was Yasuda Shedlovsky Method.

Spectral gradient analysis is recently used technique. UV absorption spectrum is used . Along the GLPKa Sirius Instrument.

Measurement of physiochemical properties can be done along with the properties

Solubility is an important factor in drug absorption. low drug solubility which is always associated with plasma binding, slow tissue distribution, drug-drug interaction .so overtaking factors to determine solubility of a particular drug .Methods such as Potentiometery Titration and Nephetrometric Assay can be implemented but this methods having high throughput, its solubility range is limited .so solubility in buffer solution is measured by LC/MS based solubility determination protocol

Partition coefficient and distrubitation coefficient are the two factors between two immussiable solvent phase (octonal and water) are mostly used to determine lipophilicity of organic compounds.

In a development of drug log D at PH 7.4 is widely used which shows an indication of lipophility of drug molecules at PH of blood plasma. It is necessary to determine the PH due to changing environment of biological system such as gastro instential tracks for proper permeability. Lipophility is one of the factors which are related to ADME properties. So drugs with low or high lipophilicity, shows drug with low lipophility have improper absorption through passive transport and with high lipophlic molecule may get fascinated inside the membrane. Other complication are also related to high lipophility is important. Indirect methods such as REVERSE PHASE HPLC have been proven to be high throughput method. It is an indirect method for lipophilicity determination but traditional methods such as conventional SHAKE AND FLASK method is still used to its simplicity and cost efficiency.

{{56 Yan, Zhengyin.Caldwell, Gary, W. 2004 ;}}

Fluconazole used in the form of semisolid formulation to be applied tropically. It is used as an antifungal agent. To determine partition coefficient is necessary for studies. Distribution of drugs, partition gives information on surface tension, hydrolysis, solubility, dissociation of substance along with structural formula. Partition coefficient of solute between octanol and water is widely used to predict drug pharmacokinetic properties ,techniques involved as traditional shake flask method, potentiometeric titration , liquid chromatography , counter current distribution ,

Drug shows toxic effect and also reduced systemic absorption due to the drug contacted with adsorption force which may be physical or chemical applied to them. Adsorption is important phenomenon in terms of interaction of drug and excipients. The is effected by many different factors such as ph, time of surface area, nature of adsorbent used. “Langmuir and freundlich” adsorption isotherm equation describes adsorption theoretically.

Fluconazole is a bis –triazole antifungal agents, it highly reactive against Cryptococcus neoforman and Candida species in vitro because of their higher infection of Candida species except C albuian they are highly susceptible to oral formulation so needs an alternative antifungal therapy. So partition coefficient of fluconazole is studied in different organic and phosphate buffer to identify the technique which is suitable for tropical use. physiochemical properties such as adsorption , partition coefficient are important to study for development of dosage form , so that behaviour of drug can be understood through which suitability and compatibility of drug can be predicated.{{59 Hajare,Ashok,A.Mali,Mahesh,N.Sarvagod,Sushil.Kurane,Sachin.Patwardhan,Shweta.and.Dange,Arun,S. April-June 2009;}}

Studies show that bioavailability of fluconzole is more concerned with IV and oral administration.

Pharmacokinetics of fluconazole

Forms of administration

1. Administration through intravenous injection by rapid bolus or infusion.

2. Oral administration through tablet, capsules or solution /suspension

Studies proven on voluntaries shows

Tissue penetration of fluconazole

Penetration in the tissue and fluid is important, fluconazole having low lipophility due to low affinity towards plasma proteins. It readily penetrates into body tissue by the means of passive diffusion. Long plasma life gives better distribution phase, which is necessary for fluconazole, available at the site of infection during implementing dosage. It has also properties to penetrate through CSF by the oral means.

Correlation to studied, fluconazole posses good pharmacologic properties due to rapid absorption due to water solubility, high oral bioavailability, and volume of distributation is high due to the relatively high plasma concentration along with high oral bioavailability in feeding and fasting state. In gastroinstenial disease it can be also admistred due to proper absorption, which shows extreme distbutation of fluconazole in body .Due to its low affinity for plasma proteins. {{58 Brammer, K.W.Farrow, P.Rand.Faulkner, J.K. March-April 1990 ;}}

Buffer Solutions

Method: Part one

First prepare four different solutions A, B, C and D in following concentration

Solution A, 30ml of sodium acetate (CH3COONA), with 0.1m was added with 30ml of acetic acid (CH3COOH) with 0.1m.

Solution B was also prepared by adding 20ml of sodium acetate (CH3COONA) with 50ml of acetic acid (CH3COOH) with 0.1m of both.

Solution C, prepared by adding 10ml of sodium acetate (CH3COONA) with 50ml of 0.1m acetic acid (CH3COOH) .

Lastly Solution D was prepared with 50ml of sodium acetate (CH3COONA) with 100ml of 0.1m, CH3COOH.

Next step was to measure the ph by PH meter of solution A, B and C along with its temperature, values are noted.

Part second:

It involves the titration of 50ml of solution D with HCL along the addition of 2ml of HCL each time and measurement of PH to be obtained until the total volume up to 90ml. Reading of PH was obtained by titertaion of solution D with 0.1m HCL.

TABLE

52ml

4.28

54ml

4.17

. . 56ml

4.08

58 ml

3.98

60 ml

3.83

62 ml

3.65

64 ml

3.35

66 ml

2.78

68 ml

2.33

70 ml

2.17

72 ml

1.93

74 ml

1.84

76 ml

1.78

78 ml

1.72

80 ml

1.65

82 ml

1.63

84 ml

1.65

86 ml

1.59

88 ml

1.55

90 ml

1.54

Part third

It involves the titration of 50ml of solution D with 0.1m of NAOH. Adding 2ml of 0.1m NAOH each time along with notification of PH value until the total 90ml volume, 20 different reading where obtained for Alkali.

TABLE

52ml

4.61

54 ml

4.45

56 ml

4.56

58 ml

4.62

60 ml

4.67

62 ml

4.75

64 ml

4.82

66 ml

4.84

68 ml

5.06

70 ml

5.01

72 ml

5.11

74 ml

5.16

76 ml

5.26

78 ml

5.39

80 ml

5.72

82 ml

5.78

84 ml

6.22

86 ml

9.88

88 ml

11.14

90 ml

11.47

Note: each time tip of PH meter is to be cleaned with distilled water in order not to vary the readings.

Readings of both acid and alkali were obtained to plot a graph and calculate the value of pKA and Ka along with the buffer capacity of both. Experiment was performed in the group of three.

Method

To determine partition coefficient of fluconazole .Two different solution where prepared, solution A and Solution B

Part first

Preparation of solution A

Phosphate buffer of PH 7.4, in which 0.1w/v fluconazole was dissolved for preparing stocking solution A.

Solution A was used to prepare different range of calibration of each 1ml containing 5,10,15,20,25,30,35,and 40 ug/ml of fluconazole in phosphate buffer of PH 7.4, seven readings where obtained each of standard lammad max 260nm, with the help of phosphate buffer PH7.4 , it was used as a blank reading to calibrate the absorbance for each time .Reading noted are shown as follows.

Concentration of fluconazole Absorbance of lammad max 260nm

5 0.007

10 0.021

15 0.06

20 0.043

25 0.075

30 0.083

35 0.103

40 0.092

Part second

Preparation solution B was prepared with 0.01% w/v of fluconazole dissolved in 1-octanol. Second part involves the preparation of partitioning solution in four different funnels. First separating funnel should be checked for the leakage, to fit the tap at its bottom, so that after loading the solution it should not be leaked. Four different samples were prepared as follows

First solution involves 5ml solution B with 20ml of 1- octane and 25ml of phosphate buffer.

Second solution, 10ml solution B with 15ml of 1- Octanol and 25ml of phosphate buffer.

Third solution , 15ml of solution B with 10ml of 1- octanol and 25ml of phosphate buffer.

Fourth solution, 20ml of solution B with 5ml of 1-octanol and 25ml of phosphate buffer.

Four solutions are needed to be shaken gently as soon as they are prepared. Shaking for about 30-60min is needed. After shaking check weather two clear layers are appeared in the funnel. Time taken was about 55min of shaking for getting the two different layers. After getting the layers separate attach funnel to the holder stand. Amongst the two different layers, lower layer was water and upper layer was octanol. This layer where separated in two different 50ml tubes as octanol and water. Total four pairs, each of octanol and water. Last part was to recoding the absorbance on UV visible spectrophotometer of lammad 260nm of water and octanol phase. This can be shown as follows

Reading

Organic Buffer

0.041 0.172

0.096 0.150

0.106 0.044

0.133 0.166

Lab Practical Results:

Ph of three solutions where noted as follows

Solution A ph 4.82.

Solution B ph 4.41.

Solution C ph 4.13.

Calculate pka=ph+log [base/acid] pka = -log ka , so Ka = antilog pka

Concentration of base = volume /total volume* molarity.

Solution A

Pka = 4.82

Ka = 1.5*105

Solution B

Pka = 4.01

Ka =1.0*105

Solution C

Pka = 3.42

Ka =2.6* 105

Plot graphs of pH against volume of 0.1M HCl and pH against volume of 0.1M NaOH on the same sheet of graph paper and calculate the buffer capacity with respect to both acid and alkali.

Graph

C:\Users\hp\Desktop\pras.png

Buffer capacity:

Buffer capacity of acid = 15.05ml

Buffer capacity of base =27.02ml

The concentration of fluconazole in the water and octanol phase, Cwat, Corg, respectively, for each composition of the partitioned mixture from the standard curve.

Graph

C:\Users\hp\Desktop\prashant cc.png

Calibration curve, slope is obtained as follows

Slope of the line is 0.0027

Now concentration of fluconazole in organic and buffer

For the solution 1

Concentration of fluconazole in buffer = 15.18

Concentration of fluconazole in octanolol =63.70

For the solution 2

Concentration of fluconazole in buffer = 35.55

Concentration of fluconazole in octanolol =55.55

For the solution 3

Concentration of fluconazole in buffer = 39.25

Concentration of fluconazole in octanolol =16.29

For the solution4

Concentration of fluconazole in buffer = 49.25

Concentration of fluconazole in octanolol =61.48

Calculate the partition coefficient Kow of fluconazole at each of the partitioned mixture,

Kow = Corg/Cwat

Kow =0.172/0.041 = 4.19

Kow =0.150/0.096 =1.56

Kow =0.044/0.106 =0.41

Kow =0.166/0.133 =1.24

Express the average value of the partition coefficient Kow from four experiments, and compare it with values from literatures.

Average value 1.85

, standered range shows the lipophilicity value in the range between -0.89 to 2.21.so average value falls in the range of standard value.

{{60 Alimuddin,Muhammad.Grant,Daniel,Bulloch,Daryl.Lee,Noelle.Peacock,Martin.and.Dahl,Russell. May 6 , 2008;}}

Calculate the distance from the active ser oxygen to the C- terminal carbonyl carbon of the peptide?

Ans: Distance between His 57 and Ser195: 8.30 A

Asp 102 and His 57 :6.47A

C:\Users\hp\Desktop\prashant 1.png

Calculate the distance from nitrogen to the peptide carbonyl oxyg5en, what do these imply?

Ans: 2.82A, shows hydrophobic properties

Peptide specification

C:\Users\hp\Desktop\manaoj 4.png

Identify the location of loops and amino acids type. Compare these to the equivalent residue in trypsin .pdb?

Ans: Amino acids: PRO, SER, GLY, ALA, VAL.

Location: AA- Amino Acid

A A ATOM X, Z, Y ATOM CORDINATE ATOM B FACTOR

PRO 225 C 26.376 9.625 45.862 9.51

SER 221 O 23.446 4.945 48.704 9.61

SER 214 CA 28.692 7.173 36.496 8.10

GLY 216 CA 28.971 5.629 43.557 12.57

ALA 185 CA 22.264 10.587 49.513 7.42

VAL 188 CA 17.455 6.480 46.705 7.05

C:\Users\hp\Desktop\prashant05.png

Calculated the surface in the context of residue 16-245.from chymotrypsin?

Ans : The surface context residue from 16 to 245 from chymotrypsin can be drawin as follows

Area : 8021

Volume:28018

C:\Users\hp\Desktop\prashant mol.png

(Due to some errors of software it shows two images)



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