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Abstract

A robust and sensitive reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the quantitative determination of Levosulpiride in human plasma. The study aimed to facilitate accurate and reliable bioanalytical evaluation of Levosulpiride for pharmacokinetic, bioavailability, and bioequivalence assessments. The chromatographic method was optimized using a Quality by Design (QbD) approach, with a mobile phase consisting of ammonium formatter buffer and methanol (28.19:71.81 v/v), and detection at 287 nm using a Waters Bridge C18 column. The method demonstrated a retention time of approximately 5.51 minutes and exhibited excellent linearity over the concentration range of 10–50 µg/mL. Bioanalytical validation was performed in accordance with ICH guidelines, confirming the method’s selectivity, specificity, accuracy, precision, robustness, and stability. The method showed high recovery rates (~99.94%) and low relative standard deviations, indicating its reproducibility and reliability. Stability studies, including freeze-thaw, short-term, and long-term storage conditions, confirmed the stability of Levosulpiride in plasma samples. This validated RP-HPLC method offers a simple, cost-effective, and sensitive analytical tool suitable for routine quality control, pharmacokinetic studies, and clinical monitoring of Levosulpiride, with significant improvements over previously reported methods in terms of efficiency and simplicity.

Keywords

Levosulpiride, RP-HPLC, bioanalytical method, method validation, human plasma, Quality by Design, pharmacokinetics, bioavailability, bioequivalence, stability studies, drug recovery, chromatographic optimization

Introduction

In the development of modern pharmaceuticals, accurate and reliable measurement of drug concentrations in biological matrices is essential for determining pharmacokinetic profiles, assessing bioavailability, and establishing bioequivalence. Bio analytical methods, particularly those utilizing high-performance liquid chromatography (HPLC), play a critical role in this domain by enabling the quantitative determination of drugs and their metabolites in biological fluids such as plasma, urine, and saliva.

Bioavailability refers to the rate and extent to which an active pharmaceutical ingredient becomes available at the site of action after administration, while bioequivalence signifies the absence of a significant difference in the bioavailability between two pharmaceutical products under similar conditions. Reliable assessment of these parameters requires precise analytical techniques and validated methodologies. The complexity of biological matrices such as blood plasma presents significant challenges in drug analysis. Hence, method development involves the careful selection of extraction procedures, chromatographic conditions, and detection settings. Commonly employed extraction techniques include protein precipitation, liquid-liquid extraction (LLE), and solid-phase extraction (SPE), each offering specific advantages and limitations depending on the physicochemical properties of the analyte and the matrix involved. Sample preparation is a crucial step that aims to isolate the drug from interfering substances while maintaining its stability and recoverability. The quantification of drug levels in plasma is often used as a surrogate marker for tissue concentrations, enabling the monitoring and optimization of therapeutic regimens. In this context, Levosulpiride—a substituted benzamide antipsychotic and prokinetic agent—requires robust analytical techniques to ensure precise measurement in plasma samples for clinical and pharmacokinetic studies. Bioanalytical method validation ensures that the method is suitable for its intended purpose, emphasizing parameters such as selectivity, accuracy, precision, sensitivity, and stability. Full, partial, and cross-validations are conducted depending on the study design and regulatory requirements. RP-HPLC (Reverse Phase High-Performance Liquid Chromatography) has become a preferred analytical tool due to its reproducibility, sensitivity, and suitability for complex matrices. The present study focuses on the development and validation of a robust and sensitive RP-HPLC method for the estimation of Levosulpiride in human plasma. The validated method is expected to be applicable for routine drug analysis in bioavailability and bioequivalence studies, ensuring compliance with regulatory standards and contributing to the safe and effective therapeutic use of Levosulpiride.

Drug Profile

Name

Levosulpiride

Description

Levosulpiride, sold under the brand name Neoprad is a substituted benzamide antipsychotic, reported to be a selective antagonist of dopamine D2 receptor activity on both central and peripheral levels. It is an atypical neuroleptic and a prokinetic agent. Levosulpiride is also claimed to have mood elevating properties.

Structure

 

 

 

Fig No.1.1 Structure of Levosulpiride

CAS NO

23672-07-3

Molecular Formula

C15H3N3O4S

Molecular Weight

341.43 g/mol

Mono isotopic Mass

340.92 g/mol

IUPAC Name

N-[[(2S)-(-)-1-Ethylpyrrolidin-2-yl]methyl]-2-methoxy-5-sulfamoylbenzamide

Category

Levosulpiride is an a antipsychotic used mainly in the treatment atypical neuroleptic and a prokinetic agent.

Colour

White Powder

pka

10.24 (strongest acidic)

8.39 (Strongest Basic)

Melting point

183-186 °C

Storage

Store at room temperature away from light and moisture.

Uses

Levosulpiride is used in the treatment of: Psychoses, particularly negative symptoms of schizophrenia, anxiety disorders, dysthymia,

Literature Survey:

Sr. No.

Reference

Type of Work Done

Conclusion

1

Nivedeetha Halekote Shivaraju, et.al.

Stability Indicating RP-HPLC Method For The Simultaneous Estimation Of Pantoprazole And Levosulpiride In Pharmaceutical Dosage Form

Mobile Phase: 10 mM ammonium acetate (pH 4.0 adjusted using acetic acid): Acetonitrile in the ratio of 20:80% v/v Column: C18 column Kromasil (250 x 4.6 mm, 5μ)m Flow rate: 1.0 ml/min. Detection Wavelength: 241 nm. Temp: 25° C

2

Raghu Khimani, et.al.

Development and Validation of HPLC Method for determination of Ilaprazole and Levosulpride

Mobile Phase: 10 mM Ammonium acetate in water, pH 5.0: Diluent in proportion of ratio 45:55%V/v Column: C18 (250 x 4.6 mm, 5 mcm) Flow rate: 1.0 ml/min.  Temp: 25° C

3

K.SENTHILNATHAN, et.al.

Method Development And Validation For The Simultaneous Estimation Of Esomeprazole And Levosulpiride By Using RPHPLC In Its Bulk And Pharmaceutical Dosage Form

Mobile Phase: Buffer and Acetonitrie in the ratio of 32;68 Column: ODS (150mm 4.6mm, 5μ) Flow rate: 1.0 ml/min. Detection Wavelength: 290 nm. Temp: 25° C

4

Daniel Gonzalez, et.al.

Simultaneous determination of Levosulpiride and sulfamethoxazole in dried plasma and urine spots

Levosulpiride-sulfamethoxazole (TMP-SMX) is an antimicrobial drug combination commonly prescribed in children and adults. The study objectives were to validate and apply an HPLC-MS/MS method to quantify TMP-SMX in dried plasma spots (DPS) and dried urine spots (DUS), and perform a comparability analysis with liquid matrices

MATERIAL AND METHOD

  1. Preparation of Phosphate Buffer Solution

6.8 gm of Potassium dihydrogen orthophosphate was dissolved in sufficient water (HPLC grade) with aid of sonicator. Then add triethylamine or orthophosphoric acid was used to adjust the pH to 5.

  1. Preparation of stock solutions

10 mg of Levosulpiride diluted with 10mL Methanol in volumetric flask to get concentration of 1000 µg/ml. From the resulting solution 0.1 ml was diluted to 10 ml with Methanol to obtain concentration of 10 µg/ml of Levosulpiride.

  1. Selection of detection wavelength

From the standard stock solution further dilutions were done using water and scanned over the range of 200-400 nm and the spectra were overlain. It was observed that drug showed considerable absorbance at 287 nm.

RESULT & DISCUSSION

According to the above prescribed, the RP-HPLC method was developed and validated as per ICH guideline with the QbD approach. It is first stated the application of Quality by design approach to Bioanalytical method development and validation. Optimized mobile phase by design expert software was used to estimate drug in human plasma. Details are as follows.

Optimized Chromatographic Conditions

Mobile phase: Ammonium format buffer: Methanol (28.19:71.81v/v), pH of buffer: 3, Analytical column: Cis column Waters XBridge (4.6x 250mm id. particle size 5µm), UV detection: 287 nm, Injection volume: 10 µL., Flow rate: 1.00 mL. min, Temperature: Ambient, Run time: 10 min.

Preparation of spiked plasma samples

To each Eppendrof tube add required amount of Levosulpiride. Add 100 µL of plasma to it. Then add 100 µL acetonitrile as a precipitating agent to it and allow vortex for 5 min. To get Proper mixture of above solution, centrifuge it for 15 min at 2000 rpm. Subject it under nitrogen gas for concentration and separate supernatant in Eppendrof tubes. 20 µL of the supernatant was the final resulting sample.

System Suitability Parameter

Following are the parameters when taken results of samples of spiked plasma.

Table 1.1: Peak Properties of Chromatogram of Levosulpiride Esylate (10u/ml)

Sr. No.

Parameters

Observation

1.

Retention time

5.513 min

2.

Peak area

84852

3.

Theoretical plates

19145

Figure 1.2: Chromatogram of Levosulpiride Esylate spiked in human plasma (10µg/ml)

Bioanalytical Validation

  1. Selectivity

For selectivity, the blank samples of the plasma were obtained from six different persons. Each blank sample was tested for interference in the Levosulpiride peak. The plasma and Levosulpiride peak were well resolved. It was found that the peak from blank plasma does not interfere with the peak of Levosulpiride. Hence developed method is selective and the peak obtained at 5.5min is only because of Levosulpiride Esylate.

Blank (Spiked Plasma)

Figure 1.3: A typical chromatogram of blank human plasma

Sample 1

Figure 1.4: Chromatogram of Levosulpiride Esylate spiked in human plasma (10µg/ml)

  1. Linearity

Linearity of the Bioanalytical method is the capability to obtain responses which are directly proportional to the concentration of drug molecule. Linearity of Levosulpiride Esylate was employed from 10µg/mL. to 50µg/mL. The concentration range was found to be linear and results of slope and correlation coefficient were found to be acceptable.

Table 1.2: Results of Linearity

Sr. No

Concentrati on (µg/mL)

Peak Area

Peak Properties

Retention Time

Asymmetric Factor

Theoretical Plates
  1.  

10

84352

5.514

1.151

19163
  1.  

20

161504

5.514

1.153

19147
  1.  

30

251456

5.514

1.157

19200
  1.  

40

344408

5.514

1.151

19108
  1.  

50

421260

5.514

1.155

19197
  1. Precision

Table 1.3: System Precision results for Levosulpiride by RP-HPLC

Sr. No

Concentration µg/ml

Intraday Precision Peak Area

Interday Precision Peak Area

Repeatability
  1.  

10

84627

84536

84124
  1.  

10

84122

85641

84279
  1.  

10

84794

85647

84877
  1.  

10

84261

85982

84934
  1.  

10

85552

85461

85655
  1.  

10

84764

85344

84165

Average

84686.67

85435.17

84672.33

Standard Deviation

460.05

447.91

546.00

RSD%

0.543

0.524

0.644
  1. Specificity

In a selectivity study, we have demonstrated that the peak of Levosulpiride Esylate is resolved efficiently in human plasma.amount of drug vered was found to be 99.94%.

Table 1.4: Recovery results of Levosulpiride by RP-HPLC

Sample

Label Claim (mg)

Amount of Drug Found (mg)

Recovery (%)

Retention Time (min)

Capsule

100

99.94

99.94

5.514
  1. Repeatability

Table 1.5: Repeatability results for Levosulpiride by RP-HPLC

Sr. no

Concentration ug/ml

Peak Area
  1.  

10

84124
  1.  

10

84279
  1.  

10

84877
  1.  

10

84934
  1.  

10

85655
  1.  

10

84165

Mean

84672.33

Std. Dev.

546.00

%RSD

0.644
  1. Robustness

Mobile Phase

Table 1.6: Results of Robustness (Mobile Phase) for Levosulpiride by RP-HPLC

Sr. No

Parameter

Response

Buffer: Methanol

(V/V)

Retention Time (min)

1

30:70

5.590

2

29: 71 (Optimized)

5.564

3

28:72

5.524

Relative Standard Deviation

0.488.

Wavelength

Table 1.7: Results of Robustness (Wavelength) for Levosulpiride by RP-HPLC

Sr. No

Parameter

Response

Detection Wavelength

(nm)

Peak Area

1

391

84016

2

392 (Optimized)

84694

3

393

84219

Relative Standard Deviation

0.33369

pH of mobile phase

Table 1.8: Results of Robustness (pH) for Levosulpiride by RP-HPLC

Sr. No

Parameter

Response

pH of Buffer

(mmol/L)

Peak Area

1

2.8

85219

2

3 (Optimized)

85698

3

3.2

85348

Relative Standard Deviation

0.236
  1. Recovery

Table 1.9: Recovery results of Levosulpiride by RP-HPLC

Sr. No

Amount of Sample (µg/ml)

Amount of Drug Added (µg/ml)

Amount of Drug Recovered (µg/ml)

Recovery %

1

10

8

7.99

99.87

2

10

10

9.98

99.80

3

10

12

12.01

100.08
  1. Stability of Levosulpiride in Human Plasma

The objective of this study was to determine the stability of Levosulpiride in human plasma. We interpreted the storage conditions by studying freeze and thaw, short term, long term and stock solution stability.

  1. Freeze and Thaw Stability

The stability of Levosulpiride was determined in spiked plasma by three freeze-thaw cycles at - 20 deg * C = 1 deg * C After comparing the Stability samples with freshly prepared samples and concluded that the drug remains stable in the freeze/thaw cycles.

Table 1.10: Results of Freeze and Thaw Stability

Replicate No.

Actual Concentratio

LQC (8 µg/ml)

HQC (24 µg/ml)

Comparison Sample

Stability Sample

Comparison Sample

Stability Sample

1

67526

68952

203105

210132

2

68497

67963

206586

206988

3

67349

67198

201489

205266

Mean

67790

68037

203726

207462

SD

504.65

718.01

2126.76

2014.61

% RSD

0.744

1.055

1.043

0.9710
  1. Short Term Stability

The spiked plasma samples were estimated for 8 hours stored at room temperature. Comparing stability samples against the freshly prepared spiked quality control samples assessed stability..

Table 1.11: Results of Short-Term Stability

Replicate No.

Actual Concentratio

LQC (2 µg/ml)

HQC (60 µg/ml)

Comparison Sample

Stability Sample

Comparison Sample

Stability Sample

1

16433

16425

503947

500385

2

16513

16654

507976

517196

3

16351

16942

500159

501729

Mean

16432.33

16673.67

504027.3

506436.7

SD

66.13

211.52

3191.78

7627.7

% RSD

0.402

1.268

0.633

1.506
  1. Long Term Stability

A sample of spiked plasma was determined for 7 days stored at room temperature, comparing them against the freshly weighed stock solution assessed for stability.

Table 1.12: Results of Long-Term Stability

Replicate No.

Actual Concentratio

LQC (8 µg/ml)

HQC (24 µg/ml)

Comparison Sample

Stability Sample

Comparison Sample

Stability Sample

1

68462

67394

201894

214665

2

68116

69421

208962

217642

3

69923

67287

202119

214874

Mean

68833.6

68034

204325

215727

SD

783

981.7

3280

1356

% RSD

1.137

1.442

1.605

0.628

CONCLUSION

Our current experiment illustrates the development and validation of a simple, rapid, and very sensitive RP-HPLC method and Bioanalytical method developed for the determination of Levosulpiride in pure form, dosage forms, and in spiked human plasma. This developed experiment overcomes the drawbacks that have been found in the other reported method where no need to use the isocratic method, more retention time, and complex extraction for this simple method. Also, this method is money-saving as it needs less expensive instrumentations, solvents, and reagents. The high accuracy, precision, and sensitivity make this simple method be a reliable and reproducible method to be applied in quality control, Quality assurance as well as pharmacokinetic analysis of Levosulpiride Esylate

ACKNOWLEDGEMENT

I would like to express my sincere gratitude to my guide and mentor for their invaluable guidance, encouragement, and support throughout the course of this research work. I am thankful to the Principal and management of our esteemed institution for providing the necessary facilities to carry out this study. I also extend my heartfelt thanks to the faculty members of the Department of Pharmaceutical Chemistry for their technical assistance and constant inspiration. Lastly, I am grateful to my family and friends for their unwavering support and motivation throughout this project.

REFERENCES

  1. Surve, S.H.R.E.E.N.LD.H.I., Patwari, A.R.P.IT., Patel, J.LT.E.N., Rathod, I.S.H.W.A.R.S.L.N.H. and Chhabria, M.A.H.E.S.H., 2013. HPTLC and HPLC method development and validation for simultaneous estimation of rabeprazole sodium and levosulpiride in bulk and its pharmaceutical dosage form. Int J Pharm Pharm Sci, 5(3), pp.65-9,
  2. JHANWAR, B., SINGH, B., SAINI, G. and VERMA, S., 2017. Method development and validation for simultaneous estimation of levosulpiride and rabeprazole sodium: A new analytical Q-absorbance ratio approach. Asian Journal of Pharmaceutical and Clinical Research, 10, pp. 16-22.
  3. Agarwal, N.A.N.D.A.K.I.S.H.O.R.E. and Jagdigsh, B., 2012. Development and validation of stability indicating RP-HPLC method for simultaneous estimati…[23:12, 08/07/2025] Bhushan Tekwade: 7. Karunakaran, K., Navaneethan, G. and Elango, K.P., 2012. Development and validation of a stability-indicating RP-HPLC method for simultaneous determination of paracetamol, tramadol HCl and domperidone in a combined dosage form. Tropical Journal of Pharmaceutical Research, 11(1), pp.99-106.
  4. Pereira, F.J., Rodríguez-Cordero, A., López, R., Robles, L.C. and Aller, A.J.. 2021. Development and validation of an RP-HPLC-PDA method for determination of paracetamol, caffeine and tramadol hydrochloride in pharmaceutical formulations. Pharmaceuticals, 14(5), p.466.
  5. Brahmankar, D. (1995) 'Biopharmaceutics and Pharmacokinetics-A Treatise', 2nd edition. Vallabh prakashan, Delhi, India..
  6. Beckett, A., Stenlake, J. (1997) 'Practical Pharmaceutical Chemistry, 4th editi… [23:13, 08/07/2025] Bhushan Tekwade: 15. European Medicines Agency (EMEA). Guideline on bioanalytical method validation. (2011) 'Published by Committee for Medicinal Products for Human Use (CHMP), London.
  7. Gaikwad, H., Borde, S., Suralkar, M., Choudhari, V. and Kuchekar, B., 2014. Development and validation of RP-HPLC method for simultaneous determination of tramadol hydrochloride, paracetamol and dicyclomine hydrochloride by using design of experiment software (DOE). Int J Pharm Sci, 4(6), pp.792-801.
  8. Guidance for industry bio analytical method validation, (2001), US FDA, May, 1-20.
  9. Sinha, M., 2014. RP-HPLC METHOD DEVELOPMENT AND VALIDATION OF TREMADOL HYDROCHLORIDE IN BULK FORM BY ION-PAIR LIQUID CHROMATOGRAPHY. Journal of Drug Delivery and Therapeutics, 4(1), pp.63-65.
  10. Indian Pharmacopoeia, 2007. The Indian Pharmacopoeia commission Gaziabad: Govt. of India ministry of health and family welfare, India.
  11. Kamble, R.M., Singh, S.G. and Singh, S., 2011. Validated RP-HPLC method for simultaneous estimation of paracetamol and tramadol hydrochloride in a commercial tablet. Journal of Pharmacy Research, 4(11), pp.4038-4040.
  12. Jain, D., Kachave, R.N. and Bhadane, R.N., 2010. Simultaneous estimation of tramadol hydrochloride, paracetamol and domperidone by RP-HPLC in tablet formulation. Journal of liquid chromatography & related technologies, 33(6), pp.786-792.
  13. Modi, N., Dresser, M., Wang, B., Gupta, S. (2006) 'Dapoxetine pharmacokinetics and tolerability in hypertensive men', European Urology Supplements, Vol.5 no 2, pp. 173176.
  14. Musteata, F., Pawliszyn, J. (2007) 'Bioanalytical applications of solid-phase micro extraction', Trends in Analytical Chemistry, Vol. 26, pp. 36-45.
  15. Natraj, K., K. Reddy, D. Kumar, Kumar, S. (2011) 'Simple validated UV-Spectrophotometric method for estimation of dapoxetine, Journal of Chinese Pharmaceutical Sciences, Vol.4, no 3, pp. 105-107.
  16. Raynord P. (1994) 'Liquid Chromatography for the Analyst', 4th edition, Marcel Dekker Inc, New York.
  17. Rohith, T., Ananda, S. (2012) 'A validated chiral liquid chromatographic method for the enantiometric separation of dapoxetine hydrochloride', International journal of Advanced Research in Pharmaceutical and Bio Sciences, Vol. 2, ?? 3, pp. 311-319.
  18. Shah, V. (2007) The history of bioanalytical method validation and regulation: Evolution a guidance document on bioanalytical methods validation', The American Association of Pharmaceutical Scientist, Vol. 9, pp. E43-E47.
  19. Sethi, P. D. (2001) 'High performance liquid chromatography: Qualitative analysis of pharmaceutical formulation', 1" edition CBS publication and distributors, New Delhi.
  20. Shargel, L., Pongs, W., Andrew, B. (2005) 'Applied Biopharmaceutics and Pharmacokinetics', 5th edition, Mc Graw-Hill, New York.
  21. Snyder, L. R., Kirkland, J. J., Dolan, J. W. (2012) 'Introduction to Modern Liquid Chromatography, 3rd edition, John Wiley, New Jersey.

Reference

  1. Surve, S.H.R.E.E.N.LD.H.I., Patwari, A.R.P.IT., Patel, J.LT.E.N., Rathod, I.S.H.W.A.R.S.L.N.H. and Chhabria, M.A.H.E.S.H., 2013. HPTLC and HPLC method development and validation for simultaneous estimation of rabeprazole sodium and levosulpiride in bulk and its pharmaceutical dosage form. Int J Pharm Pharm Sci, 5(3), pp.65-9,
  2. JHANWAR, B., SINGH, B., SAINI, G. and VERMA, S., 2017. Method development and validation for simultaneous estimation of levosulpiride and rabeprazole sodium: A new analytical Q-absorbance ratio approach. Asian Journal of Pharmaceutical and Clinical Research, 10, pp. 16-22.
  3. Agarwal, N.A.N.D.A.K.I.S.H.O.R.E. and Jagdigsh, B., 2012. Development and validation of stability indicating RP-HPLC method for simultaneous estimati…[23:12, 08/07/2025] Bhushan Tekwade: 7. Karunakaran, K., Navaneethan, G. and Elango, K.P., 2012. Development and validation of a stability-indicating RP-HPLC method for simultaneous determination of paracetamol, tramadol HCl and domperidone in a combined dosage form. Tropical Journal of Pharmaceutical Research, 11(1), pp.99-106.
  4. Pereira, F.J., Rodríguez-Cordero, A., López, R., Robles, L.C. and Aller, A.J.. 2021. Development and validation of an RP-HPLC-PDA method for determination of paracetamol, caffeine and tramadol hydrochloride in pharmaceutical formulations. Pharmaceuticals, 14(5), p.466.
  5. Brahmankar, D. (1995) 'Biopharmaceutics and Pharmacokinetics-A Treatise', 2nd edition. Vallabh prakashan, Delhi, India..
  6. Beckett, A., Stenlake, J. (1997) 'Practical Pharmaceutical Chemistry, 4th editi… [23:13, 08/07/2025] Bhushan Tekwade: 15. European Medicines Agency (EMEA). Guideline on bioanalytical method validation. (2011) 'Published by Committee for Medicinal Products for Human Use (CHMP), London.
  7. Gaikwad, H., Borde, S., Suralkar, M., Choudhari, V. and Kuchekar, B., 2014. Development and validation of RP-HPLC method for simultaneous determination of tramadol hydrochloride, paracetamol and dicyclomine hydrochloride by using design of experiment software (DOE). Int J Pharm Sci, 4(6), pp.792-801.
  8. Guidance for industry bio analytical method validation, (2001), US FDA, May, 1-20.
  9. Sinha, M., 2014. RP-HPLC METHOD DEVELOPMENT AND VALIDATION OF TREMADOL HYDROCHLORIDE IN BULK FORM BY ION-PAIR LIQUID CHROMATOGRAPHY. Journal of Drug Delivery and Therapeutics, 4(1), pp.63-65.
  10. Indian Pharmacopoeia, 2007. The Indian Pharmacopoeia commission Gaziabad: Govt. of India ministry of health and family welfare, India.
  11. Kamble, R.M., Singh, S.G. and Singh, S., 2011. Validated RP-HPLC method for simultaneous estimation of paracetamol and tramadol hydrochloride in a commercial tablet. Journal of Pharmacy Research, 4(11), pp.4038-4040.
  12. Jain, D., Kachave, R.N. and Bhadane, R.N., 2010. Simultaneous estimation of tramadol hydrochloride, paracetamol and domperidone by RP-HPLC in tablet formulation. Journal of liquid chromatography & related technologies, 33(6), pp.786-792.
  13. Modi, N., Dresser, M., Wang, B., Gupta, S. (2006) 'Dapoxetine pharmacokinetics and tolerability in hypertensive men', European Urology Supplements, Vol.5 no 2, pp. 173176.
  14. Musteata, F., Pawliszyn, J. (2007) 'Bioanalytical applications of solid-phase micro extraction', Trends in Analytical Chemistry, Vol. 26, pp. 36-45.
  15. Natraj, K., K. Reddy, D. Kumar, Kumar, S. (2011) 'Simple validated UV-Spectrophotometric method for estimation of dapoxetine, Journal of Chinese Pharmaceutical Sciences, Vol.4, no 3, pp. 105-107.
  16. Raynord P. (1994) 'Liquid Chromatography for the Analyst', 4th edition, Marcel Dekker Inc, New York.
  17. Rohith, T., Ananda, S. (2012) 'A validated chiral liquid chromatographic method for the enantiometric separation of dapoxetine hydrochloride', International journal of Advanced Research in Pharmaceutical and Bio Sciences, Vol. 2, ?? 3, pp. 311-319.
  18. Shah, V. (2007) The history of bioanalytical method validation and regulation: Evolution a guidance document on bioanalytical methods validation', The American Association of Pharmaceutical Scientist, Vol. 9, pp. E43-E47.
  19. Sethi, P. D. (2001) 'High performance liquid chromatography: Qualitative analysis of pharmaceutical formulation', 1" edition CBS publication and distributors, New Delhi.
  20. Shargel, L., Pongs, W., Andrew, B. (2005) 'Applied Biopharmaceutics and Pharmacokinetics', 5th edition, Mc Graw-Hill, New York.
  21. Snyder, L. R., Kirkland, J. J., Dolan, J. W. (2012) 'Introduction to Modern Liquid Chromatography, 3rd edition, John Wiley, New Jersey.

Photo
Asha Chopde
Corresponding author

Madhav University Pindwara Sirohi Rajasthan 307026

Photo
Babu Anmulwad
Co-author

Madhav University Pindwara Sirohi Rajasthan 307026

Photo
Amrut Bhandari
Co-author

Loknete Shri Dadapatil Pharate college of pharmacy Mandavgan Pharata Tal Shirur Dist Pune

Photo
Vidya Barhate
Co-author

Loknete Shri Dadapatil Pharate college of pharmacy Mandavgan Pharata Tal Shirur Dist Pune

Photo
Chavan Vishal
Co-author

Loknete Shri Dadapatil Pharate college of pharmacy Mandavgan Pharata Tal Shirur Dist Pune

Asha Chopde*, Babu Anmulwad, Amrut Bhandari, Vidya Barhate, Chavan Vishal, Bio-analytical (RP-HPLC) Method Development and Validation for Levosulpiride from Human Plasma, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 7, 1886-1896. https://doi.org/10.5281/zenodo.15878983

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