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Hplc Method For Analysis Of Piroxicam In Gel Biology

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Abstraction: A simple, precise method utilizing HPLC method was developed for the finding of Feldene in gel. A reversed stage HPLC system was used consisting of C18 column with the dimension size of 150mm ten 4.6mm. 55 volumes of methyl alcohol and 45 volumes of phosphate buffer ( 0.05M, pH 7 ) are used as the nomadic stage. The flow rate was 1ml/min and the wastewater was monitored at 254nm. The keeping clip was found to be about 6.0minutes. The stock solution of Feldene was prepared and the standard solutions runing from 5 to 20I?g/ml were prepared with phosphate buffer ?? methyl alcohol ( 60 ?? 40, v/v ) . This was injected into HPLC and the chromatograms were obtained. Test solution was prepared from the marketed merchandise Feldene gel and injected into HPLC. From the arrested development equation, which was obtained from the standard concentrations, the concentration of Feldene gel was determined. Validation methods are performed to show the truth of the method, preciseness of the method and the one-dimensionality.

This method has shown one-dimensionality and the correlativity coefficient was found to be 0.999, the truth of the method was good in the scope of 91 % – 99.03 % , and the preciseness which is expressed as % RSD was found to be 4.44 % , the method was non precise to reiterate. The proposed method was successfully applied to the quantitative finding of Feldene in topical dose regimen gel.

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Cardinal words: Piroxicam, HPLC, Feldene gel

Introduction:

Piroxicam [ 4-hydroxy-2-methyl-N- ( 2-pyridyl ) -H-1, 2-benzothiazine – 3 – carboxamide-1, 1-di-oxide ] is a non-steroidal anti-inflammatory and analgetic agent. It exhibits a decrepit acidic 4-hydroxy proton and a decrepit basic pyridyl nitrogen.Its molecular expression is C15H13N3O4S. Molecular weight was 331.35 [ 25 ] .

Figure 1 Structure of Piroxicam

hypertext transfer protocol: //www.chemspider.com/RecordView.aspx? rid=1bd5eb75-1933-4241-a3fd-81ee8b83a4bd

Piroxicam is a non-steroidal anti-inflammatory agent from the oxicam category [ 14 ] . NSAIDS are the drugs with analgetic and antipyretic effects which have in higher concentration anti inflammatory effects [ 26 ] . Piroxicam is used in the intervention Rheumatoid Arthritis, Osteoarthritis, ancylosing spondylitis and besides used in acute hurting in musculoskeletal upsets and in ague urarthritis [ 6 ] . Piroxicam Acts of the Apostless by cut downing the endocrines that cause redness and hurting in the organic structure. The anti-inflammatory consequence of Feldene may ensue from the reversible suppression of Cox, doing the peripheral suppression of prostaglandin synthesis. The prostaglandins are produced by Cox-1 enzyme, ensuing in the break of production of prostaglandins. It besides inhibits the migration of leucocytes into sites of redness and prevents the formation of thromboxane A2, aggregating agent, by the thrombocytes [ 14 ] . Piroxicam has a drawn-out continuance of action that is one dosage can supply hurting alleviation throughout the twenty-four hours [ 25 ] . Piroxicam is really somewhat soluble in H2O and somewhat soluble in intoxicant. The other names for Piroxicam are Candyl, Cycladol, Feldene, Novo-pirocam, and Piroflam. Piroxicam is a white crystalline pulverization [ 12 ] .

Piroxicam is good absorbed by unwritten path. When Feldene is administered throught the unwritten path, the drug plama concentrations peak within three to four hours. The drawn-out half life of 50hrs, has steady province plasma concentrations throughout the twenty-four hours on one dosage. With the nutrient there is a minor hold in the rate of soaking up but non the extent of soaking up after unwritten disposal. The coincident disposal of alkalizers with the Feldene has no consequence on plasma drug concentration when administered orally. The evident volume of distribution of Feldene is 0.14L/kg, 99 % of the plasma Feldene was bound to plasma proteins. Metamorphosis of the piroxicam drug occurs by the hydroxylation at the 5th place of the pyridyl side of the concatenation and a sequence of reactions occurs which involve hydrolysis of the amide linkage, decarboxylation, N- demethylation and the ring contraction. And the drug gets excreted in piss and the in the fecal matters. [ 25 ] .

Commercially Piroxicam is available in the signifier of capsules, tablets, injectables, gel. Piroxicam is good absorbed by unwritten disposal but has been associated with a figure of side effects on tummy, sickness, indigestion, diarrhea, irregularity and some nephritic 1s. And besides it can do stomachic mucosal harm which may ensue in ulceration or shed blooding [ 1 ] . In order to cut down these inauspicious effects a topical preparation has been prepared [ 4 ] . It besides has the extra advantage of avoiding hepatic first-pass metamorphosis and supplying the controlled bringing of the drug for an drawn-out period [ 21 ] . Several analytical processs for the sensing and besides quantification of Feldene in pharmaceutical readyings and in biological fluids have been used [ 23 ] . These involve Quantitative finding of Feldene by thin bed chromatography-matrix-assisted optical maser desorption ( MALDI ) TOF mass spectroscopy [ 3 ] . UV spectrophotometric method UV sensing is most normally employed, as it is sufficiently sensitive to observe degrees associated with common unwritten dose regimens [ 7 ] , spectrofluorimetric method [ 5 ] , Chemiluminescence method [ 15 ] , Piroxicam has been quantified in capsules, suppositories, injection solutions, tablets, suspensions by capillary zone cataphoresis and micellar electro kinetic capillary chromatography [ 10 ] , A colorimetric finding of Feldene in capsules has been used [ 8 ] , Liquid chromatography has besides been reported [ 13 ] , HPLC with amperometric sensing of Feldene in human plasma and tissues was reported [ 2 ] , there are many methods for the finding of Feldene in plasma by HPLC by utilizing different nomadic stages [ 23 ] , And there is an HPLC method for the finding of Feldene in pharmaceutical merchandises like capsules, tablets, unctions, suppositories, Ophthalmic suspensions [ 9 ] . And there was besides an hplc method for the quantitative finding of Feldene in a new preparation piroxicam-I?-cyclodextrin [ 7 ] .

There are so many HPLC methods for quantitative finding of Feldene in all pharmaceutical readyings [ 23 ] , but at that place was a less literature about quantitative finding of Feldene in gel, there was a literature about stableness survey of Feldene gel by utilizing HPMC and ACUPEC HV-505 bases [ 21 ] , So a simple, rapid and sensitive method has to be developed for topical dose regimen ( gel ) . When Feldene gel is applied to the tegument where the redness is there it gets absorbed through the tegument into the underlying tissues [ 27 ] . A Feldene gel is used to alleviate hurting and redness in the applied country.

In general, high-performance liquid chromatography has been the most employed method to measureA piroxicam [ 1 ] . The HPLC has the major betterment over column chromatography sing the sensing methods which are used [ 19 ] . These methods are extremely automated and highly sensitive, It is used to analyse, place, sublimate & amp ; quantify the compounds [ 19 ] . There are two discrepancies in usage in HPLC depending on the comparative mutual opposition of the dissolver and the stationary stage, they are normal stage and the reversed stage HPLC. High public presentation liquid chromatography is fundamentally a extremely improved signifier of column chromatography. Alternatively of a dissolver being allowed to drip through a column under gravitation, it is forced through under high force per unit areas of up to 400 ambiances which makes it much faster [ 19 ] . The conventional diagram of an HPLC was given below, this shows how the HPLC plants.

Figure 2: The conventional diagram of HPLC was shown below:

HPLC requires a pump to travel the liquid nomadic stage through the system, a column to execute the separation, a sensor and informations aggregator to enter a chromatogram, and a fraction aggregator to roll up the end product. When sample is injected into watercourse of nomadic stage the gesture of analytes in the sample depends on the physical and chemical attractive forces with the stationary stage in the column and base on ballss through the sensor and this sends signals to the processing unit and besides to the show [ 19 ] .

The clip taken for a peculiar analyte to go through through the column to the sensor is known as itsA keeping clip. The sum of analyte can be related to the country under the extremum obtained [ 19 ] .

If the extremum is smaller so the concentration will be smaller and frailty versa. If there are two different substances in the mixture ( X and Y ) so it is hard to detect the concentration from the country under the extremum, because one sample absorbs UV visible radiation at one moving ridge length and the other at different moving ridge length. Frequently a series of trials are performed on the analyte and a figure of test tallies may be processed in order to happen the HPLC method which gives the best extremums [ 19 ] .

In this paper an HPLC method has been used for the quantitative finding of Piroxicam in gel. In the old documents for the quantitative finding of Feldene in pharmaceutical readying different nomadic stages used like ethanoate buffer, methanol H2O etc [ 23 ] , the nomadic stage selected in this proposed method was methanol: Phosphate buffer ( 0.05M pH 7 ) 55:45 v/v, the wastewater was monitored ate 254nm. A nomadic stage has to be buffered to ionise the analyte, because when an analyte gets ionized it becomes less hydrophobic, hence keeping clip will acquire reduced [ 16 ] . And this method was performed in a strictly isocratic manner. Isocratic methods are used when a individual analyte has been widely considered and the compound is being run to corroborate its individuality [ 19 ] . There are two methods of HPLC normal and reversed stage [ 19 ] , the one which is used in this survey was reversed stage hplc. In this the dissolver used will be polar solvent illustration methyl alcohol and a non polar stationary stage is used like silicon oxide, to do it non polar long hydrocarbon ironss was attached to its surface with either 8 or 18 C atoms in them and the column size used in this will be same like normal stage, a typical column has an internal diameter of 4.6 millimeter ( and may be less than that ) , and a length of 150 to 250 millimeters [ 19 ] . The flow rate of nomadic stage was set to be at 1ml/min. And the injection volume was 20I?l.

Before this method, one method has been tried but this does non give good consequences. In the old method the keeping clip has been changed for inter twenty-four hours runs, it has been changed from 7 to 8 and so to 13. So little alterations has been done to the old method, the nomadic stage used was the same but the proportions used are different 40 volumes of methyl alcohol and 60 volumes of phosphate buffer 0.05M are used, in the present method methyl alcohol composing has been changed to diminish the keeping clip, and the moving ridge length used was 240nm, this has been changed to 254nm because in many diaries different wave lengths are used so this moving ridge length was used for a test tally, it gave good consequences so it has been used for farther surveies. The keeping clip was found to be around 6 proceedingss after the method has been changed and it was changeless for inter twenty-four hours runs. The proof of any new method has to be carried out to look into whether the proposed method is giving good consequences or non [ 20 ] . So proof has to be carried out for the hplc method for truth, preciseness, Limit of sensing, Limit of quantification, one-dimensionality, specificity, hardiness, one-dimensionality, scope [ 20 ] .

Aim: To develop a new method for the quantitative finding of Feldene in gel which have less literature.

MATERIALS AND METHOD:

Apparatus:

Analysis was performed by utilizing analytical balance, pH metre, the HPLC used is equipped with a series 200 binary pump with variable individual moving ridge length UV/VIS sensor set at 254nm, Column used is of SNiXF13, C18 column and the dimensions of the column is 150mm ten 4.6mm with a flow rate of 1ml/min ( Isocratic ) . The nomadic stage consists pH 7, 0.05M phosphate buffer: methyl alcohol ( 60:40 v/v ) , the injection volume was 20 I?l and the sample injection was manual.

Materials:

Piroxicam manufactured by sigma Aldrich, UK. Phosphate buffer ( pH 7.0, 0.05M ) this is prepared by fade outing 7.8g of Na dihydrogen phosphate ( analytical class, purchased from sigma Aldrich, UK ) in 1000ml of H2O and pH adjusted with 1.0 N NaoH ( analytical class purchased from sigma Aldrich, UK ) . Methanol HPLC class was manufactured by Fischer UK ltd. Company. Feldene gel was manufactured by Pfizer research lab and the per centum pureness of Feldene in gel was 0.5 % w/w in 60g of gel. Bi distilled H2O.

Standard readying:

Piroxicam ( 50mg ) was weighed out accurately into a 50ml volumetric flask, dissolved in nomadic stage ( 0.05M phosphate buffer ?? methyl alcohol, 60??40, v/v ) , Standard solutions runing from 5 to 20 I?g/ml were prepared, diluted to 10ml with nomadic stage ( phosphate buffer?? methyl alcohol, 60??40, v/v ) .These standard solutions were injected into HPLC which are used to build standardization graph. This standardization graph was obtained as optical density peak country versus concentration. A twosome of tallies are performed on the standard solutions to acquire the better extremums and the norm of both peak countries are taken for the standardization graph, standardization graph was constructed by taking peak country on X-axis and the concentration on Y-axis.

Sample solution:

An accurately weighed 500mg of Feldene gel was transferred to a 50.0 milliliter volumetric flask, dissolved with methyl alcohol, volume was so adjusted to the grade with methyl alcohol. The solution was sonicated in an supersonic sonicator for 20 min, from this 4ml of solution was transferred to 10ml volumetric flask, diluted to 10 milliliter with nomadic stage ( phosphate buffer: methyl alcohol, 60:40 ( v/v ) ) prior to analysis. And for ciphering truth different concentrations were taken, after the analysis of 4ml sample 5ml,6ml were taken from the sample solution and diluted to 10ml with the nomadic stage 60:40 v/v,

Frequently a series of trials are performed on the analyte and a figure of test tallies should be carried out in order to acquire best extremums [ 19 ] , the mean of the both peak countries were taken for truth and preciseness. By utilizing standardization curve, quantitative finding of Feldene in gel was determined.

Validation process:

Validation is required for any new method to guarantee that the proposed method is capable of giving good dependable and consistent consequences when performed by different operators in the same research lab and with the same equipment. A proof of the assay consisting of one-dimensionality, truth, preciseness of the method was performed for this proposed method with the same equipment in the same research lab [ 20 ] .

Preciseness:

The preciseness of the method was studied by the analysis of the sample for the multiple tallies. The preciseness was expressed as the % RSD. The preciseness value should non be more than 2 % [ 14 ] . If this method exceeds that so the method was non precise to reiterate once more or to follow in the regular research lab analysis.

% RSD= criterion divergence x 100

Mean

Accuracy:

Accuracy for this method can be performed by using the method in triplicate samples. The sample with different concentrations were taken and injected into the HPLC and the chromatograms were recorded from which the extremum country can be noted down and the concentration can be calculated by subjecting the peak country value in the topographic point Y in the arrested development equation obtained by the standardization graph of the standard solutions runing from 5I?g/ml to 25I?g/ml. The obtained value is considered as the sum found value and it is compared with the true value to cipher the per centum recovery. Therefore it can be defined as the measuring of grade of intimacy of measurings of the measure of its true value. This was expressed as the % recovery [ 11 ] .

% Recovery = sum found x 100

True value

One-dimensionality and scope:

The one-dimensionality is the method ‘s ability to acquire the consequences which are relative to the concentration of the analyte within a given scope [ 20 ] . Linearity is determined by ciphering arrested development line utilizing peak country versus analyte concentration [ 14 ] . The one-dimensionality of the responses of the sensor can be established by the graph constructed as peak versus concentration of the standard solutions and besides by finding the correlativity coefficient [ 20 ] . A series of standard solutions of Feldene runing from 5I?g/ml, 10I?g/ml, 15I?g/ml, 20I?g/ml, and 25I?g/ml were injected into HPLC and the peak countries were noted. These standard solutions are injected into HPLC for a twosome of times to acquire the best extremums and the norm of the peak countries were considered to build the standardization graph to happen out the one-dimensionality and besides scope of the responses.

RESULTS AND DISCUSSION:

Several methods are reported for the quantitative finding of Feldene in pharmaceutical readyings [ 9 ] , but at that place was a less literature about quantitative finding of Feldene in gel so, this has been chosen, two methods have been tried for the finding of Feldene, one method has shown good consequences so it has been considered for farther surveies. Piroxicam standard solutions concentration runing from 5I?g/ml – 25I?g/ml and the sample were injected into HPLC system, the Feldene extremum in the sample was identified by comparing with the piroxicam standard solutions and besides identified by the keeping clip which was found to be around 6.0 proceedingss. The appraisal of Feldene was carried out by HPLC utilizing nomadic stage holding a composing of 55 volumes of methyl alcohol and 45 volumes of 0.05M phosphate buffer ( 55:45v/v ) . The pH of the phosphate buffer was 7. The column used was c18 ( 150mm x 4mm dimension ) . Flow rate of nomadic stage was 1ml/min ; the wastewater was monitored at 254nm.

Table 1: Optimised Chromatographic Conditionss

Parameter

OPTIMIZED CONDITION

Chromatogram

HPLC with 200 series binary pump with variable individual wavelength US/VIS sensor

Column

150mmx 4.6mm

Mobile stage

Methanol and 0.05M phosphate buffer 55:45v/v

Detection

254nm

Injection Volume

20I?l

The Chromatograms obtained for the standard solutions and sample solutions when injected into HPLC were shown below.

Figure3: Chromatogram demoing the extremum of standard solution holding the concentration of 5I?g/ml.

Figure 4: Chromatogram demoing extremum of the trial solution.

The standard solutions of piroxicam concentrations runing from 5I?g/ml to 25I?g/ml were injected into HPLC a twosome of times in order to find the better extremum and the norm of the extremums were taken to build the standardization graph.

Table 2 shows the mean peak countries of the standard solutions.

Concentration ( mcg/ml )

Average of peak country

5

2603863.86

10

4495002.98

15

5853247.93

20

8137025.215

25

10188667.99

The 4ml volume taken from the trial solution has been analyzed for 8 times, 5ml trial solution for 3 times and the 6ml trial solution for 3 times and the norm of these peak countries were taken to find the concentration of Feldene in Feldene gel.

Table 3 shows the peak countries of the trial solution.

Volume of sample taken

Average of 8 peak countries

4ml

785101.73

Table 4 shows the peak countries of the trial solution.

Volume of sample taken

Average of 3 peak countries

5ml

936166.91

6ml

1166749.405

The Validation of the hplc method has been carried out. To describe the preciseness of the method the sample has to be run for 10 times in the HPLC. The selected method shown good one-dimensionality and truth but the method was non precise because the method exceeded the normal bounds for the preciseness, it has shown the % RSD value of 4.44, the normal value is 2 % . For truth different concentrations of sample solutions were taken and these are injected into hplc for triplicate and the norm of these values are considered to cipher the truth, the truth was good with the value of 91.9 % – 99.03 % recovery of the drug.

One-dimensionality: Under the optimal conditions described above, the one-dimensionality of the Feldene in the standard solutions holding concentrations 5I?g/ml to 25I?g/ml were injected into hplc for twosome of times, the mean values of the peak countries were taken to plot the graph. Graph was plotted by taking peak country on y-axis and the concentration on x- axis. This is shown in figure 5.There is a one-dimensionality between 5I?g/ml, 10I?g/ml, 20I?g/ml, 25I?g/ml, the correlativity coefficient ( r2 ) value was found to be 0.993 and the arrested development equation was y = 37623x + 61207 where Y is the peak country of piroxicam trial solution and ten is concentration of Feldene in the trial solution. There might be some job with the injections with the 15I?g/ml concentration of standard solution. The value of 15I?g/ml has been somewhat varied from the one-dimensionality so the point has been left out to acquire the one-dimensionality. If that point has been left out the correlativity coefficient value was found to be 0.999, and the arrested development equation was y= 37623x+71625, the values after the point has been left out and before that has been shown in the tabular array 5, the graph with the point left out was shown figure 6.

Table 5 shows the arrested development equation and the correlativity coefficients values.

Parameters

Actual values

The values after the point which has been left out

Concentration ( mcg/ml )

5I?g/ml – 25I?g/ml

5,10,20,25I?g/ml

Regression equation

Y = 37623x + 61207.

y= 37623x+71625

Correlation coefficient

0.993

0.999

Figure 5: The graph with the concentrations runing from 5I?g/ml to 25I?g/ml

Figure 6: The graph with left out concentration of standard solution ( 15I?g/ml )

Statistical analysis: To prove the one-dimensionality of the standard concentrations a statistical simple additive arrested development trial has been performed on the standard concentrations. The end product was attached in the appendix.

Coefficientsa

Model

Unstandardized Coefficients

Standardized Coefficients

T

Sig.

Bacillus

Std. Mistake

Beta

1

( Constant )

712650.863

94363.372

7.552

.017

concentration

376232.610

5565.248

1.000

67.604

.000

a. Dependent Variable: country

In this the ‘t ‘ value was found to be 67.604 and the P value was found to be 0.000 which provinces that there is a additive relationship between peak country and the concentration that is if there is a little alteration in the concentration there will be alteration in the peak country as the concentration increases peak country additions.

Model Summaryb

Model

Roentgen

R Square

Adjusted R Square

Std. Mistake of the Estimate

1

1.000a

1.000

.999

87994.298467

a. Forecasters: ( Constant ) , concentration

B. Dependent Variable: country

The correlativity coefficient value was 1.000 that is 100 % fluctuation in the peak country was explained by the arrested development theoretical account.

Analysis of variance

Model

Sum of Squares

df

Mean Square

F

Sig.

1

Arrested development

3.539E13

1

3.539E13

4570.291

.000a

Residual

1.549E10

2

7.743E9

Entire

3.540E13

3

a. Forecasters: ( Constant ) , concentration

B. Dependent Variable: country

In this the F value was found to be 4570.291 and the value of P value was found to be 0.000 which is less than 0.001 this provinces that peak country linearly depends on the concentration that is as the concentration increases the peak country will increase.

Preciseness: The method preciseness is evaluated by inter and intraday repeatability [ 20 ] . In this method intraday repeatability was performed. For the intraday repeatability the sample with same concentration has to analyze for many times ( 10 times ) in the same twenty-four hours, in this method the sample has been analysed for 8 times. The preciseness is expressed as % RSD and the % RSD for the samples was 4.44 % , it exceeds the normal bound of the preciseness that is 2 % this indicates that the method may non be precise to reiterate once more. The preciseness was satisfactory. Necessitate to be performed once more to look into whether the method was non satisfactory or if there are any jobs with the injections made earlier.

Accuracy: The truth of this method was checked at three volumes incorporating three different concentrations. The truth is expressed as % recovery, and the % recovery is the scope of 91.5- 99.03 % . The proof of this method shows that the truth is within the bounds, which shows that this method is capable of demoing good truth.

Table 6: Percent Recovery of the trial solution.

Volume of sample taken

True value

Sum found

% recovery

4ml

0.5 %

0.474 %

94.8 %

5ml

0.5 %

0.459 %

91.9 %

6ml

0.5 %

0.495 %

99.03 %

The proof methods tested in this method were accuracy, preciseness and the one-dimensionality. Accuracy and one-dimensionality were good but the preciseness was satisfactory. May be that was non performed right or may be due to the machine duplicability jobs. If any farther survey has to be carried out, the method for the preciseness has to be performed once more because as this method gives good consequences for truth and one-dimensionality, the preciseness has to be checked once more. In one-dimensionality, 15I?g/ml concentration of standard solution has to be analysed once more because it was somewhat varied from the standardization curve. This may due to some jobs in the injections or may be due to the drosss in the volumetric flask of that concentration or may be in the pipette tip.

There are so many methods reported for quantitative finding of Feldene in pharmaceutical readyings [ 9,23 ] , but there was less literature about the finding of Feldene in topical dose signifiers, there was a literature about hplc method to find the measure of Feldene in unctions but there is no literature for gel. Hence Gel has been used for the quantitative finding of Feldene. Commonly HPLC method was used to find the measure of Feldene in the pharmaceutical readyings, hence HPLC was selected as the method to analyze the measure of Feldene in the topical dose signifier gel. So the method has been tried for the quantitative finding of the marketed merchandise Feldene gel. Different nomadic stages are used to quantify the Feldene in pharmaceutical dose signifiers, the largely used nomadic stage was methanol and the phosphate buffer, in this the nomadic stage has to be buffered to cut down the keeping clip that is when an analyte gets ionised the analyte becomes less hydrophobic, if the analyte is less hydrophobic the keeping clip will be decreased. This method was performed in isocratic manner because when a individual analyte has to be analysed so this manner was used. There the parametric quantities used were given in the below tabular array.

Table 7: Optimised conditions for chromatograph

Parameters

Optimised conditions

Mobile stage

Methanol: 0.05M pH 7 phosphate buffer ( 40:60 v/v )

Column size

250mm x4.6 millimeter dimension

Detection

240nm

Flow rate

0.8ml/min

With the usage of these parametric quantities the consequences were non good ; the keeping clip was changed for twenty-four hours to twenty-four hours, it was changed from 7minutes to 9minutes and so to 13minutes and the chromatograms was besides non good. And there was no one-dimensionality in the standard concentrations. When the standardization graph has been plotted there is no one-dimensionality a v- molded standardization curve was obtained. This method was repeated for so many times but the consequences were same. So the method has been changed, the method changed was mentioned in the tabular array below.

Table 8: Optimised chromatographic conditions:

Parameters

Optimised conditions

Mobile stage

Methanol: 0.05M pH7 Phosphate buffer 55:45 v/v

Column size

150mm ten 4.6mm dimension

Detection

254nm

Flow rate

1ml/min

After the alterations in the method has been done, there are alterations in the keeping clip, it was found to be around 6.0 proceedingss, if the methyl alcohol proportions increases the keeping clip will be decreased so it the proportion has been increased and the consequences found were good that is the keeping clip has been changed from 13 proceedingss to 6 proceedingss and this was changeless which was non changeless earlier. And the chromatograms obtained were good. When the standardization graph was plotted the graph was additive with the consequences obtained from the changed method. With the changed method the consequences were good and the proof methods were performed on the method to cognize its truth and preciseness, the consequences for preciseness was satisfactory and the truth was good and the one-dimensionality was good.

In the antecedently reported methods quantitative finding of Feldene in a new preparation Feldene -I?- cyclodextrin the method was accurate and precise. And the several other methods are reported many other pharmaceutical preparations they were accurate and precise, the method proposed here was accurate but the preciseness it has to be repeated once more, as there was no clip to reiterate the method it was non done if any hereafter work has to be carried out on this method, so foremost the preciseness has to be performed for inter and intraday. This proposed method was the new method for the finding of Feldene of Feldene in gel. If this method was repeated once more for preciseness, and if the preciseness was good after the repeat of the method so it said to be a development of new method with good truth and preciseness for the quantitative finding of the Feldene in the pharmaceutical readying gel.

Decision:

High public presentation liquid chromatography is one of the most sophisticated tools of analysis at present. In this the appraisal of Feldene is done by reversed stage HPLC. In this method the nomadic stage used was 45 volumes of 0.05M pH7 phosphate buffer and 55 volumes of methyl alcohol. The sensing is carried out utilizing UV/VIS sensor at 254nm. The keeping clip for Feldene was found to be around 6.0 proceedingss. Linearity scope for Feldene was found to be 5I?g/ml, 10I?g/ml, 20I?g/ml, and 25I?g/ml. The quantitative appraisal of Feldene in Feldene gel was carried out by Reversed stage HPLC. A statistical trial, simple additive arrested development theoretical account, was performed on the standard concentrations runing from 5I?g/ml to 25I?g/ml. The proof of the assay consisting of one-dimensionality, truth and preciseness were carried out. The values of Relative criterion divergence value was found to be 4.4 % which was more than 2 % indicating that the preciseness was satisfactory for this method. The per centum recovery was found to be 91.5 % to 99.03 % for Feldene in the topical dose signifier ( Feldene gel ) .

The consequences obtained on the proof process for the preciseness does non run into the demands. The truth and the one-dimensionality were good. It says that the method was accurate, and the one-dimensionality was good. The method was found to hold satisfactory application to utilize in the everyday research lab analysis with a high grade of truth. A farther work has to be carried out to reiterate the preciseness once more.

Further Work: The proof method preciseness has to be carried out once more, to look into whether the proposed method was non satisfactory or there are any jobs during the injections of the trial solution to the HPLC. If preciseness was satisfactory after the repeat so the developed method for the quantitative finding of Feldene in the topical dose regimen, Feldene gel ( 0.5 % w/w of Feldene which was present in 60g of gel ) can be used in the everyday research lab analysis with a high

Cite this Hplc Method For Analysis Of Piroxicam In Gel Biology

Hplc Method For Analysis Of Piroxicam In Gel Biology. (2016, Dec 09). Retrieved from https://graduateway.com/hplc-method-for-analysis-of-piroxicam-in-gel-biology-essay/

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