1

NOVEL SEPARATION AND QUANTITATIVE DETERMINATION OF LEVOFLOXACIN,

PRULIFLOXACIN, GATIFLOXACIN, SPARFLOXACIN, MOXIFLOXAXIN AND BALOFLOXACIN

FLUOROQUINOLONE ANTIBACTERIALS IN PHARMACEUTICAL DOSAGE FORMS BY RP-HPLC

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05/02/2023 VIGNAN PHARMACY COLLEGE 2

Aim and Objectives

Aim: The core AIM of the present study is to develop a novel, rapid,

precise and accurate RP-HPLC method for simultaneous separation and quantification of six fluoroquinolones OF LEVOFLOXACIN (LEVO), PRULIFLOXACIN (PRFX), GATIFLOXACIN (GATI), SPARFLOXACIN

(SPAR), MOXIFLOXAXIN (MOXI) AND BALOFLOXACIN (BALO) for the the day to day analysis.

The author felt that a novel single method for separation and quantification of all the above said drugs on single chromatographic system without any minor changes in detection wavelength and mobile phase composition.

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To develop rapid, sensitive and economical analytical method based on HPLC for separation and

estimation of six fluoroquinolones pharmaceutical dosage forms.

To develop method with shorter run time and better sensitivity.

Reducing the solvent consumption to make it more eco-friendly.

Avoid the column damage by minimizing the buffer strength and pH of mobile phase

than reported methods.

To validate the method for different parameters like Accuracy, Precision, Linearity,

specificity, Robustness International Conference on Harmonization ICH Q2(R1)

guidelines..

To apply the developed RP-HPLC method in the analysis of pharmaceutical

formulations.

Objectives

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Table 1. Commercial brand names of LEVO, PRFX, GATI, SPAR, MOXI and BALO used for the present study.

Brand nameFormulation Labeled amount (mg) Manufacturer

GlevoTablets Levofloxacin -500 mg

Glenmark Pharmaceuticals Ltd.,

Mumbai, India.

Pruflox Tablets Prulifloxacin- 600 mg Cipla Ltd., Mumbai, India.

Segat Tablets Gatifloxacin- 400 mg Secure health care Inc. India.

SparcipTablets Sparfloxacin – 100 mg Cipla Ltd., Mumbai, India.

Moxicip Tablets Moxifloxacin-400 mg Intralabs, Bangalore, India.

Balox-100 Tablets Balofloxacin -100 mg Lupin Ltd., Mumbai, India.

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TITLE AUTHOR JOURNAL CONDITIONS RESULTS Reference

Simultaneous quantification of

linezolid, rifampicin, levofloxacin, and

moxifloxacin in human plasma using high-

performance liquid chromatography with UV

Baietto L et.al

Therapeutic Drug Monitoring: Vol. 31 No.1 pp.104-109.

The method is based on a

simple organic protein

precipitation that rapid

sample preparation and a direct

injection into the HPLC

Limit of quantification

0.234µg/mL,0.312µg/mL,

0.156µg/mL.,0.622µg/mL

LEVO,LZD,MOXI,RFP respectively.

1

Liquid chromatography method for the simultaneous

determination of levofloxacin, pazufloxacin, gatifloxacin,

moxifloxacin and trovafloxacin in human

plasma.

Joana sousa et.al

Journal of Chromatogra

phy B, Vol.930

pp.104-111.

Column:Lichro CART purspher Star C18 column (55mmX4mm,

3µm)Mobile phase: 0.1% aqueous

formic acid adjusted to pH 3.0

with triethyl amine, acetonitrile, methanol.

Flow rate: 1mL/min.

Detection wave length 260 nm.

Correlation Coefficient 0.9923

Range 0.005-5 µg/mL, 0.02-5µg/mL and 0.04-5µg/mL for

GAT,LEV,PAZ and MOX, TRO respectively

Precision: Did not exceed 7.32 %.

LOQ: 0.005 µg/mL2

Table 2. REVEW OF THE PAST WORK ON THE ANALYTICAL METHODS

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TITLE AUTHOR JOURNAL CONDITIONS

RESULTS Reference

Determination of the newer quinolones

levofloxacin and moxifloxacin in plasma

by high-performance liquid

chromatography with fluorescence detection

Schulte S.et.al

J. Chromatogr Sci.Vol.44 No.4 pp.205-208.

Moxi. Used as internal standard

Liquid –liquid extraction from human plasma.

Used fluorescence

detection

Levo:Linearity 0.1-15µg/mL.

Moxi:0.2-7µg/mLLoq 0.05µg/mL and

0.2µg/mL for Levo and Moxi respectively.

3

Development and validation of a HPLC

method for simultaneous

quantitation of gatifloxacin,

sparfloxacin and moxifloxacin using

levofloxacin as internal standard in human

plasma

Srinivas N et.al

Biomed Chromatogr

Vol. 22 No. 11 pp.1288-

1295.

Phosphate buffer pH 2.5:Acetonitrile (80:20 v/v)Flow rate : 1mL/min.

Kromasil, C18 column

Run time (total) 18.0

min.

Linearity-10-100 µg/mL.

Correlation Coefficient 0.999

Limit of quantitation observed to be

10µg/mL. Runtime:GFC,SFC,MFC

(internal standard) 10.8,12.8.17.0.6.0

Respectively.

4

REVEW OF THE PAST WORK ON THE ANALYTICAL METHODS (continued)

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However, few RP-HPLC methods so far have been reported. But the methods available hitherto are poorly validated, uneconomical and consume longer runtimes.

Infact no method for the determination of all drugs in single chromatographic wavelength and with out changing the detection wavelength.

Keeping in view the complete evaluation of the above reported methods, the author developed a novel RP-HPLC method which is considered to be accurate, precise, rapid with shorter runtime as well as economical for the separation and estimation of the above said fluoroquinolones in tablet dosage forms.

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EXPERIMENTAL Materials and methods Table 3. Materials used in the present study

S.No. Materials Procured from

1. Levofloxacin, Gatifloxacin Aristo Pharmaceuticals Pvt.Ltd., Bombay.

2. Prulifloxacin, Balofloxacin Hetero Labs Ltd., Hyderabad

3. Sparfloxacin Anant Pharmaceuticals, Kamal, Haryana.4. Moxifloxacin Torrent Pharmaceuticals, Ahmedabad.5. Acetonitrile HPLC grade Thermo Fisher Scientific India Pvt. Ltd., Mumbai.

6. Water HPLC grade Merck Specialties Pvt. Ltd., Mumbai.7. Methanol HPLC grade Merck Specialties Pvt. Ltd., Mumbai.

8. Dipotassium hydrogen phosphate Thermo Fisher Scientific India Pvt. Ltd., Mumbai.

9. Potassium dihydrogen phosphate Glaxo Smith Kline Pharmaceuticals Ltd., Mumbai.

10. O-Phosphoric acid RFCL Ltd., New Delhi.

11. Triethylamine Merck Pharmaceuticals Private Limited, Mumbai.

12. Concentrated Hydro chloric acid Qualigens fine chemicals, Mumbai.

13. Sodium Hydroxide S.D Fine-Chem. Ltd., Mumbai.

All the chemicals and reagents used in the present study were of Anal R grade and solvents were of HPLC grade.

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Table 4. Instruments used in the present study

S.No. Instrument Name of the company and model

1. HPLC

Shimadzu LC-20AT Prominence Liquid Chromatograph with Shimadzu SPD-20A Prominence UV-Vis detector,Welchrom C18

Column (4.6 X 250 mm, 5μm), with Rheodyne manual loop injector (20 μL) and Spinchrom data acquisition software.

2. UV-Vis spectrophotometer

UV-Visible Spectrophotometer (Systronics model 2203). The UV-VIS spectrophotometer achieves a resolution of 1 nm with matched quartz cells of 1 cm path length.

3. Weighing balance Essae vibra AJ (0.001g), Essae-Teraoka Ltd.

4. pH meter Elico LI120 pH meter, Elico India Ltd.

5. Ultrasonicator Ultrasonic bath sonicator, PCI ltd., Mumbai.

6. Vacuum pump Single Stage Vacuum Pump.

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Preparation of reagents and standards Preparation of phosphate buffer pH 3.1o Phosphate buffer with 10 mM was prepared by dissolving 6.056 g KH2PO4 in 445

mL of HPLC grade water. To this said solution 55 mL of 0.1M H3PO4 was added to adjust the pH 3.1.

Preparation of mobile phaseo The above stated prepared phosphate buffer (pH 3.1) 500 ml (70%) and

acetonitrile 200 ml (30%) were mixed completely in the proportion of 70: 30 v/v. Preparation of standard stock solution

o 1 mg/mL of LEVO, PRFX, GATI, SPAR, MOXI and BALO were prepared. Further dilution was made based on the required concentrations.

Preparation of sample solution o Twenty tablets of LEVO, PRFX, GATI, SPAR, MOXI and BALO were separately

transferred into a mortar and ground to a fine powder. From this 1 mg/mL LEVO, PRFX, GATI, SPAR, MOXI and BALO were prepared in different calibration flasks. Further dilution was made based on the required concentration of each drug.

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Results and Discussion Method Development

o Selection of a common detection wavelength• In the present study drug solutions of 10 mcg/mL of LEVO, PRFX, GATI, SPAR, MOXI AND

BALO were prepared and scanned over a range of 200-400 nm. UV overlain spectra of these drugs showed that they absorbed appreciably at 293 nm, so that it was selected as the detection wave length for further study.

The optimum wavelength for detection was 293 nm at which much better detector responses for six drugs were obtained.

293nm

Figure 1. UV overlain spectra of the six fluoroquinolones compounds

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Method optimization Numerous trials were performed to obtain the better separations. From all trials, eventually better reproducibility of the results and good resolution good peak shape, short

runtime, minimal peak tailing were well identified, when 10 mM phosphate buffer (pH 3.1) : Acetonitrile 70:30, v/v was used and found best satisfactory results.

Parameter Optimized Chromatographic conditions

Instrument Shimadzu LC-20AT Prominence liquid chromatograph

Column Welchrom C18 column (4.6X250mm,5µm)

Detector Shimadzu SPD-20A prominence UV-VIS detector

Mobile phase 10mM phosphate buffer (pH 3.1) : Acetonitrile 70:30, v/v

Flow rate 1mL/min.

wave length UV at 293 nm.

Run time 10 minutes

Temperature Ambient temperature (250C)

Injection volume 20 µL

Method

LEVO PRFX GATI SPAR MOXI BALORetention time

(minutes) 3.613 4.230 4.707 5.497 5.880 6.253

Th.Pl (Efficiency) 12,261 12,554 13,155 14,761 14,912 15,916

Resolution - 4.508 2.866 4.604 2.056 2.017

Tailing factor 1.106 1.067 1.040 1.073 1.030 1.086

Table 5. Optimized chromatographic conditions

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Figure 2. Typical chromatogram showing the separation of LEVO, PRFX, GATI, SPAR, MOXI and BALO in synthetic mixture.

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Method validation

The developed analytical method was validated in pursuance of the guide lines of International Conference on Harmonization.

SPECIFICITY– The purpose of this study is to determine the effect of excipients and other additives that are

usually present in the formulations. The test results obtained were compared with the results of standard drug.

Name of the solution

Method LEVO PRFX GATI SPAR MOXI BALO

Mobile phase No peaks No peaks No peaks No peaks No peaks No peaks

Placebo No peaks No peaks No peaks No peaks No peaks No peaks

Separate injections

of individual standard solutions

Peak for LEVO at 3.613 minutes

Peak for PRFX at 4.230 minutes

Peak for GATI at 4.707 minutes

Peak for SPAR at 5.497 minutes

Peak for MOXI at 5.880

minutes

Peak for BALO at 6.253

minutes

Table 6. Results of specificity

Result: The present study shows that the ingredients are not interfering with the developed method.

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Linearity The purpose of this study is to verify that the detector response is directly proportional to the

concentration (amount) of analyte in the sample. Procedure: Standard drug solutions of LEVO, PRFX, GATI, SPAR, MOXI AND BALO were

prepared to get the final concentration of 2-10 µg/mL. The solutions were injected in to the HPLC system separately and the response of peak areas were measured at 293 nm.

calibration curve is constructed by plotting concentration on X-axis and the peak areas on y-axis.

The least square analysis method was adopted for achieving slope, intercept and correlation coefficient, regression data values.

MethodLEVO PRFX GATI SPAR MOXI BALO

S. No.

Conc.

μg/ML

Peak area, mV.s.

Conc.

μg/ML

Peak area, mV.s.

Conc.

μg/ML

Peak area, mV.s.

Conc.

μg/ML

Peak area, mV.s.

Conc.

μg/ML

Peak area, mV.s.

Conc.

μg/ML

Peak area, mV.s.

1. 0 0 0 0 0 0 0 0 0 0 0 0

2. 2 112.736 2 40.124 2 243.422 2 105.342 2 90.102 2 126.302

3. 4 225.134 4 81.234 4 467.872 4 208.569 4 180.864 4 245.953

4. 6 329.242 6 122.324 6 707.7 6 299.802 6 270.56 6 364.604

5. 8 442.668 8 161.537 8 929.68 8 402.045 8 363.98 8 487.255

6. 10 556.325 10 199.213 10 1169.0 10 499.25 10 452.436 10 609.906

Table 7. Results relating to Linearity data

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Parameter LEVO PRFX GATI SPAR MOXI BALO

Detection wavelength(λmax) UV at 293 nm UV at 293 nm UV at 293 nmUV at 293

nm

UV at 293 nm UV at 293 nm

Linearity range (µg/mL) 2-10 µg/mL 2-10 µg/mL 2-10 µg/mL 2-10 µg/mL 2-10 µg/mL 2-10 µg/mL

Regression equation (Y =aX + b) Y = 55.365X+0.860

Y = 0.02X+0.639

Y= 8.363X+0.578

Y= 43.07Xy = 45.336x -

0.3556Y =

60.729X+2.024

Slope(a) 55.365 20.02 58.363 43.07 45.336 60.729

Intercept(b) 0.8607 0.6391 0.5785 0 -0.3556 2.0243

Standard error of slope (Sa) 0.32184 0.1537 0.24642 0.7290 0.0220 0.2464

Standard error of intercept (Sb) 1.9488 0.9310 1.49213 2.4277 0.00798 1.4922

Standard error of estimation (Se) 2.6927 1.2864 2.06167 1.5001 0.02322 2.0618

Regression coefficient (R2) 0.9999 0.9998 0.9999 0.9999 0.9999 0.9999

% Relative standard deviation* 0.9804 1.1022 0.5239 0.104 1.050 1.0957

Percentage range of errors

0.05 significance level

0.01 significance level

1.02904

1.6138

1.15695

1.8144

0.54992

0.86242

0.12232

0.19183

0.36373

0.57042

1.150090

1.803643

Table 25. Regression analysis data relating to LEVO, PRFX, GATI, SPAR, MOXI and BALO

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Figure 3. Standard chromatogram relating to LEVO (10µg/mL)

Figure 4. Standard chromatogram relating to PRFX (10 µg/mL)

Figure 5. Standard chromatogram relating to GATI (10 µg/mL)

3.613 min

4.230 min

4.707 min

CHROMATOGRAMS

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Figure 6. Standard chromatogram relating to SPAR (10 µg/mL)

Figure 7. Standard chromatogram relating to MOXI (10 µg/mL)

Figure 8. Standard chromatogram relating to BALO (10 µg/mL)

5.497 min

5.880 min

6.253 min

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0 2 4 6 8 10 120

100

200

300

400

500

600

f(x) = 55.3647 x + 0.860666666666702R² = 0.999864848056459

Series1Linear (Series1)

Concentration, µg/mL.

peak

are

a, m

V.s

Figure 9. Linearity plot pertaining to LEVO

0 2 4 6 8 10 120

50

100

150

200

250

f(x) = 20.0199142857143 x + 0.639095238095237R² = 0.999764121630718

Series1Linear (Series1)

Concentration, µg/mL.

peak

are

a, m

V.s

Figure 10. Linearity plot pertaining to PRFX

0 2 4 6 8 10 120

100

200

300

400

500

600

700

f(x) = 58.3628714285714 x + 0.578476190476181R² = 0.999928699028504

Series1Linear (Series1)

Concentration, µg/mL.

peak

are

a, m

V.s

Figure 11. Linearity plot pertaining to GATI

0 2 4 6 8 10 120

50100150200250300350400450500

f(x) = 43.0716181818182 xR² = 0.999977905679732

Series1Linear (Series1)

Concentration of standard Sparfloxacin(µg/mL)

peak

are

a of

Spa

rflo

xaci

n

Figure 12. Linearity plot pertaining to SPAR

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Figure 13. Linearity plot pertaining to MOXI

0 2 4 6 8 10 120

100

200

300

400

500

600

700

f(x) = 60.7291428571429 x + 2.02428571428572R² = 0.999934133925654

Series1Linear (Series1)

Concentration, µg/mL.

Peak

are

a, m

V.s

Figure 14. Linearity plot pertaining to BALO

0 2 4 6 8 10 120

50100150200250300350400450500

f(x) = 45.335857142857 x − 0.355619047618575R² = 0.999969165357194

Series1Linear (Series1)

concentration, µg/mL.

peak

are

a, m

V.s

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Drug ParameterRecovery level*

80% 100% 120%

LEVO (M3)Mean % Recovery ± SD* 100.10 ± 0.3649 100.05 ± 0.1604 100.85 ± 0.1536

%RSD 0.225

PRFX (M3)Mean % Recovery ± SD* 100.154 0.2459 99.0973±0.7108 100.569 0.2968

%RSD 0.419

GATI (M3)Mean % Recovery ± SD* 100.046± .2430 100.02 ±0.1633 99.791±0.0637

%RSD 1.156

SPAR (M3)Mean % Recovery ± SD* 99.702±0.461 100.456±0.402 100.232±0.423

%RSD 0.290

MOXI (M3)Mean % Recovery ± SD* 100.153±0.2380 100.456±0.1612 99.6308±0.2089

%RSD 0.202

BALO (M3)Mean % Recovery ± SD* 100.216±0.2020 100.083±0.1100 100.839±0.2594

%RSD 0.190

Table 8. Results relating to Accuracy (recovery) method

Regarding the accuracy of the proposed method by adding the known amount of pure standard drugs to the pre-analyzed samples at three levels 80%, 100% and 120% .

Accuracy

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Precision To check the reproducibility of the method. Intra-day precision (repeatability) was studied by repeating the assay 6 times in the same day. Inter-day precision (Intermediate precision) was studied by repeating the assay on 3

consecutive days triplicate on each day and percentage RSD(Coefficient of variation (CV) were calculated.

The % RSD for all six drugs were showed less than 2% which clearly indicates that the present method is said to be highly precise.

Precision Study

Method LEVO PRFX GATI SPAR MOXI BALO

% RSD % RSD % RSD % RSD % RSD % RSD

Intra-Day 0.3380 0.3554 0.1294 0.1145 0.3465 0.1964

Inter-Day 0.5087 0.4298 0.1347 0.1721 0.5033 0.3296

Table 9. Results relating to intraday and inter-day precision

•The % RSD for the assay 6 drugs for six replicate samples were< 2.0 %.•The individual assay results for all six drugs were found between 95.0% to 105.0%

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LOD and LOQ Limit of Detection is the lowest concentration in a sample that can be detected, but not

necessarily quantified under the stated experimental conditions.

The limit of quantitation is the lowest concentration of analyte in a sample that can be determined with acceptable precision and accuracy.

LOD and LOQ were calculated using following formula

LOD= 3.3(SD)/S and LOQ= 10 (SD)/S,

where SD=standard deviation of response S= slope of the calibration curve.

PARAMETERMethod

LEVO PRFX GATI SPAR MOXI BALO

LOD µg/mL 0.116 0.152 0.084 0.186 0.0018 0.112

LOQ µg/mL 0.348 0.460 0.255 0.558 0.0055 0.339

Table 9. Results relating to LOD and LOQ

Robustness To decide the robustness of the proposed method the experimental conditions such as flow

rate (± 0.1 ml/min), detection wavelength (±5 nm) and mobile phase composition (±2%) were deliberately altered to known their effect on the peak area, peak tailing as well as number of theoretical plates.

Parameter Flow rate (±2 mL/min) Detection wave length(±5 nm) Mobile phase compostion (± 5%)

Used 0.8 mL/min 1mL/min(optimized) 1.2 mL/min 288 nm 293 nm

(optimized) 298 nm 65:35,v/v 70:30,v/v(optimized) 75:25,v/v

Retentiontime (min)LEVO 3.848 3.613 3.325 3.613 3.613 3.613 3.596 3.613 3.712PRFX 4.390 4.320 4.290 4.321 4.320 4.320 4.180 4.290 4.390GATI 4.769 4.707 4.695 4.706 4.707 4.707 4.603 4.707 4.807SPAR 5.499 5.497 5.427 5.496 5.497 5.497 5.477 5.497 5.523MOXI 6.024 5.880 5.438 5.881 5.880 5.878 5.720 5.880 5.970BALO 6.855 6.253 6.239 6.253 6.253 6.253 6.200 6.253 6.335

Plate count$

LEVO 12,562 12,261 12,054 12,242 12,261 12,274 12,106 12,261 12,282PRFX 12,598 12,554 12,454 12,555 12,554 12,559 12,588 12,554 12,564GATI 13,168 13,155 13,135 13,153 13,155 13,155 13,165 13,155 13,195SPAR 14,898 14,761 14,661 14,761 14761 14,762 14,757 14,761 14,797MOXI 15,064 14,912 14,726 14,892 14,912 14,928 14,838 14,912 14,786BALO 15,994 15,916 15,816 15,923 15,916 15,918 15,929 15,916 15,987

Peak asymmetry#

LEVO 1.126 1.106 1.097 1.107 1.106 1.106 1.026 1.016 1.015PRFX 1.069 1.067 1.068 1.067 1.067 1.067 1.066 1.067 1.069GATI 1.052 1.042 1.048 1.043 1.042 1.042 1.142 1.042 1..69SPAR 1.077 1.073 1.086 1.073 1.073 1.074 1.187 1.073 1.172MOXI 1.134 1.030 1.021 1.038 1.030 1.036 1.168 1.030 1.184BALO 1.090 1.063 1.068 1.063 1.063 1.063 1.065 1.063 1.063

Table 10. Robustness data of LEVO, PRFX, GATI, SPAR, MOXI and BALO

Conclusion: The method remains unaffected due to deliberate changes to the analytical methodindicating that the method is Robust. 24

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Assay The developed method was finally successfully applied for quantification LEVO,

PRFX, GATI, SPAR, MOXI and BALO marketed formulation.

Formulation Labeled Amount Amount Found % Assay ± SD*LEVO tablets 500 mg 496.58 99.317±0.990PRFX tablets 600 mg 599.40 99.9±0.04GATI tablets 400 mg 399.60 99.9±0.02SPAR tablets 100 mg 99.45 99.45±0.010MOXI tablets 400 mg 399.77 99.495±0.056

BALO tablets 100 mg 99.67 99.68±0.09

Table 11. Results relating to assay for method M3

Result: Satisfactory results were obtained . The mean assay values for LEVO, PRFX, GATI, SPAR, MOXI and BALO were in good agreement with the label claim.

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Conclusion

The author first reported RP- HPLC method for LEVO, PRFX, GATI, SPAR, MOXI

and BALO fluoroquinolones can be estimated on single chromatographic system

without minor modifications in the detection wavelength.

The mobile phase composition used in this analysis is similar for the mentioned six

drugs.

The method was linear in the concentration range of 2-10 µg/mL.

Statistical analysis lucidly showed that this method had good sensitivity,

reproducibility, very fast, precise, accurate, highly efficient and suitable than the

existing methods so far utilized.

Sample preparation procedure is easy and inexpensive.

Therefore this method is aptly feasible for regular analysis of six fluoroquinolones

individually in quality control laboratories.

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1. Baietto L. Baietto L. D Avolio A. De Rosa FG. Garazzino S. Patanella S. Siccardi M. Sciandra M and Di Perri G. (2009) ‘Simultaneous quantification of linezolid, rifampicin, levofloxacin, and moxifloxacin in human plasma using high-performance liquid chromatography with UV’. Ther Drug Monit. Vol. 31 No.1 pp.104-109.

2. Joana Sousa. Gilberto Alves. Goncalo Campos. Ana Fortuna and Amilcar Falcao. (2013) ‘First liquid chromatography method for the simultaneous determination of levofloxacin, pazufloxacin, gatifloxacin, moxifloxacin and trovafloxacin in human plasma’. Journal of Chromatography B, Vol.930 pp.104-111.

3. Schulte S. Ackermann T. Bertram N. Sauerbruch T and Paar WD. (2006) ‘Determination of the newer quinolones levofloxacin and moxifloxacin in plasma by high-performance liquid chromatography with fluorescence detection’ J Chromatogr Sci.Vol.44 No.4 pp.205-208.

4. Srinivas N. Narasu L. Shankar BP. Mullangi R. (2008) ‘Development and validation of a HPLC method for simultaneous quantitation of gatifloxacin, sparfloxacin and moxifloxacin using levofloxacin as internal standard in human plasma’. Biomed Chromatogr Vol. 22 No. 11 pp.1288-1295.

References

05/02/2023 VIGNAN PHARMACY COLLEGE 28

8. Chaple D.R.and Sambhare A.G. (2010) ‘A validated stability indicating HPLC method for Prulifloxacin’ IJPT Vol. 1 No.2 pp.137-148.

9. Deepak Pokharkar. Varsha Jadhav. Sachin Gholve and Vilasrao Kadam. (2010) ‘Development and validation of spectrophotometric method for estimation ofPrulifloxacin in tablet dosage form’ International Journal of PharmTech Research Vol. 2 No.1 pp.960-963.

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ACKNOWLEDGEMENTS

• I oblige to pay my deep gratitude for the patience hearing of my presentation delivered by me with reference to power point.

• I thank one and all • Glenmark Pharmaceuticals Ltd., Mumbai, India. Hetero

laboratories Ltd., Hyderabad for providing standard samples.

• Management of Vignan pharmacy college.• Organizing committee.

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