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Development and validation of an HPLC-UV method for accelerated stability study and pharmacokinetic analysis of venlafaxine

Abstract

A reverse phase high performance liquid chromatography method has been developed and validated for accelerated stability study and determination of pharmacokinetic parameters of venlafaxine HCl. The chromatographic separation was carried out using ODS analytical column (250 × 4.6 mm i.d., 5 µm particle size). The mobile phase included acetonitrile, methanol and potassium dihydrogen phosphate buffer (30:30:40; pH 6.1) at a flow rate 1.5 mL min−1. UV-Visible detector was used at wavelength of 227 nm to monitor elutions. Retention time observed was 2.745 min. The method was validated for linearity, accuracy, precision, sensitivity and robustness. Accelerated stability study of venlafaxine HCl capsules was carried out at 40 and 50 ºC under 75% RH level. Suggested method was successfully applied for the pharmacokinetic analysis of venlafaxine hydrochloride tablets. Each of ten albino rabbits (≈ 1.2 kg each) was orally administered with 5 mg dose of venlafaxine HCl. The method was proved to be linear (R2 >0.998), accurate (98.25-99.27%), sensitive (LOD: 35ngmL−1; LOQ: 105 ng mL−1) and robust (RSD<1%). The drug showed stability at accelerated conditions of temperature and humidity. The main pharmacokinetic parameters of tested products were as follows: tmax was 2.5h, Cmax was 56.5 µg mL−1, t1/2 was 8.2 h, AUC0-36 was 845.9 µg h mL−1. The developed method is suitable to apply for quality control analysis and pharmacokinetic studies.

Keywords:
Venlafaxine; HPLC; Stability; Pharmacokinetics; Chromatography


INTRODUCTION

Venlafaxine (IUPAC Name: 1-[2-Dimethylamino-1-(4-methoxy-phenyl)-ethyl]-cyclohexanol, (Molecular formula: C17H27NO2, Mw: 277 g mol−1) (Figure 1) is a third generation, structurally novel phenethyl bicyclic antidepressant (Younus et al., 2015Younus M, Arif MF, Richards MP, Kumar DB. Determination of venlafaxine and modafinil in individual tablet dosage forms using single RPHPLC method. Trop J Pharm Res. 2015;14: 239-45.). It was first introduced in 1993 by Wyeth. Venlafaxine is an important inhibitor of neuronal serotonin, norepinephrine reuptake and dopamine reuptake (Seshadri, Manohari, Kunchithapatham, 2013Seshadri VC, Manohari PJ, Kunchithapatham J. Formulation and characterization of mouth dissolving tablets containing benzodiazepine. Indo Am J Pharm Res. 2013;3(12):1457-74.). In the gastrointestinal tract, absorption rate is about 92% but as a result of metabolism, its bioavailability is just 12.6% (Sánchez-Mata et al., 2000Sánchez-Mata MC, Cámara-Hurtado M, Díez-Marqués C, Torija-Isasa ME. Comparison of high-performance liquid chromatography and spectrofluorimetry for vitamin C analysis of green beans (Phaseolus vulgaris L.). Eur Food Res Technol. 2000;210(3):220-25.). In liver, venlafaxine is rapidly metabolized by the action of CYP2D6 into an active metabolite ortho-desmethylvenlafaxine (Mw: 263 g). Venlafaxine is highly water soluble drug (500 mg mL−1) (Asafu-Adjaye et al., 2007Asafu-Adjaye EB, Faustino PJ, Tawakkul MA, Anderson LW, Lawrence XY, Kwon H, et al. Validation and application of a stability indicating HPLC method for the in vitro determination of gastric and intestinal stability of venlafaxine. J Pharm Biomed Anal. 2007;43(5):1854-59.).

FIGURE 1
Chemical structure of venlafaxine (1-[2-dimethylamino-1-(4-methoxy-phenyl)-ethyl]-cyclohexanol).

The most vital steps during drug development are pharmaceutical analysis and stability studies that are required to determine and assure the identity, potency and purity of drug and formulated form of drug (Chan et al., 2004Chan CC, Lee YC, Lam H, Zhang XM. Analytical method validation and instrument performance verification, John Wiley & Sons; 2004.). The common analytical techniques such as HPLC (Vu et al., 1997Vu RL, Helmeste D, Albers L, Reist C. Rapid determination of venlafaxine and O-desmethyl venlafaxine in human plasma by high-performance liquid chromatography with fluorimetric detection. J Chromatogr B Biomed Sci Appl. 1997;703(1-2):195-201.; Titier et al., 2003Titier K, Castaing N, Scotto-Gomez E, Pehourcq F, Moore N, Molimard M. High performance liquid chromatographic method with diode array detection for identification and quantification of the eight new antidepressants and five of their active metabolites in plasma after overdose. Ther Drug Monit. 2003;25(5):581-87.; Waschgler et al., 2004Waschgler R, Moll W, König P, Conca A. Quantification of venlafaxine and O-desmethyl venlafaxine in human serum using HPLC analysis. Int J Clin Pharmacol Ther. 2004;42(12):724-28.), LC-MS (Goeringer, McIntyre, Drummer, 2001Goeringer KE, McIntyre IM, Drummer OH. Postmortem tissue concentrations of venlafaxine. Forensic Sci Int. 2001;121(1-2):70-75.; Juan, Zhiling, Huande, 2005Juan H, Zhiling Z, Huande L. Simultaneous determination of fluoxetine, citalopram, paroxetine, venlafaxine in plasma by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-MS/ESI). J Chromatogr B. 2005;820(1):33-39.; Bhatt et al., 2005Bhatt J, Jangid A, Venkatesh G, Subbaiah G, Singh S. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for simultaneous determination of venlafaxine and its active metabolite O-desmethyl venlafaxine in human plasma. J Chromatogr B. 2005;829(1-2):75-81.) and CE (Rudaz et al., 2000Rudaz S, Stella C, Balant-Gorgia AE, Fanali S, Veuthey JL. Simultaneous stereoselective analysis of venlafaxine and O-desmethylvenlafaxine enantiomers in clinical samples by capillary electrophoresis using charged cyclodextrins. J Pharm Biomed Anal. 2000;23(1):107-15.), are utilized for pharmacokinetics and metabolism studies of venlafaxine. A number of HPLC methods (Makhija, Vavia, 2002Makhija SN, Vavia PR. Stability indicating LC method for the estimation of venlafaxine in pharmaceutical formulations. J Pharm Biomed Anal. 2002;28(6):1055-59.; Bernardi et al., 2009Bernardi LS, Oliveira PR, Murakami FS, Borgmann SH, Arend MZ,Cardoso SG. Development and validation of a stability-indicating LC method for the determination of venlafaxine in extended-release capsules and dissolution kinetic studies. J Chromatogr. Sci. 2009;47(9):770-76.; Kaur et al., 2010Kaur J, Srinivasan KK, Joseph A, Gupta A, Singh Y, Srinivas KS, et al. Development and validation of stability indicating method for the quantitative determination of venlafaxine hydrochloride in extended release formulation using high performance liquid chromatography. J Pharm Bioallied Sci. 2010;2(1):22-6.) have been reported in literature for pharmacokinetics and stability studies of venlafaxine (Liu et al., 2011Liu W, Dai YC, Deng N, Liu XR, Luo Y. Development and validation of a HPLC-MS/MS method for the determination of venlafaxine enantiomers and application to a pharmacokinetic study in healthy Chinese volunteers. Biomed Chrom. 2011;25(3):412-16.; Baldania et al., 2008Baldania SL, Bhatt KK, Mehta RS, Shah DA, Tejal RG. RPHPLC estimation of venlafaxine hydrochloride in tablet dosage forms. Indian J Pharm Sci. 2008;70(1):124-128.; Gursharanjit, Ghos, Dave, 2008Gursharanjit S, Ghosh B, Dave D. Screening of venlafaxine hydrochloride for delivery. AAPS Pharm SciTech. 2008;9(3):791-797.). Determination of pharmacokinetics parameters for native drug and formulated form of the drug is utmost important to get real information about the bioavailability of newly developed/formulated drug. Therefore, we are interested to develop and validate HPLC-UV method for evaluation of venlafaxine hydrochloride for accelerated stability and pharmacokinetic parameters.

EXPERIMENTAL

Chemicals

Methanol (RCl Labscan), acetonitrile (Sigma-Aldrich) and acetic acid (Riedel-de Haën) were of analytical grade and no further purification was required.

HPLC Analysis

HPLC analyses were carried out using Agilent 1200 series instrument (Agilent, Germany) equipped with a degasser (G1322A), diode array detector VL (G 1315B), quaternary pump (G 1311A), column compartment (G 1316A) and an auto sampler (G 1329A). Separation and quantification, of both Standard (1.0 mg mL−1) and sample (0.75 mg mL−1) solutions of venlafaxine HCl in mobile phase, were performed on a Shim-pack ODS (250 × 4.6 mm i.d., 5 µm particle size) analytical column maintained at 35ºC.

Most efficient separation conditions were achieved by employing different compositions of mobile phase during HPLC method development. A promising separation of drug was obtained by using a mobile phase mixture composing of CH3CN, CH3OH and KH2PO4 buffer in ratio of 30:30:40 with pH 6.1 (Table I).

TABLE I
Different compositions of mobile phase at pH 6.1

Method validation

The validation of an analytical method authenticates the parameters of the method for application (Parejiya et al., 2014Parejiya PB, Movaliya VR, Barot BS, Modi D, Shelat PK, Shukla A. Quantitative determination of milnacipran HCl in rabbit plasma by HPLC and its application to pharmacokinetics study. J Liq Chromatogr Relat Technol. 2014;37(1):99-11.). The method used for the determination of venlafaxine HCl was validated for precision, accuracy, linearity, sensitivity and robustness. The developed method was validated according to the International Conference on Harmonization guidelines (ICH, 2005ICH Validation of analytical procedures: text and methodology. In International Conference on Harmonization Q2 (R1). Geneva; 2005.) and USP requirements (ICH, 1996ICH HTG . International Conference on Harmonisation; Guideline on Validation of Analytical Procedures: Definitions and Terminology Availability. Part VIII. Federal Register. FDA. USA, 1996;60:11260.)

Accuracy and precision

Accuracy of analytical method is the proximity of the test results achieved by the method to the actual value. The precision is the extent of concurrence among the results of individual tests if the method is repeatedly applied to various samplings of standards (Chhalotiya, Patel, Bhatt, 2010Chhalotiya UK, Patel HB, Bhatt KK. Development and validation of an ultra-performance liquid chromatography method for venlafaxine hydrochloride in bulk and capsule dosage form. Indian J Pharm Sci. 2010;72(6):814-18.). The accuracy and precision were calculated by using quality control samples which were prepared in three concentrations (50, 100, 200 µg mL−1) representing the whole range of standard curve (low, middle and high quality) of control sample. The response of each standard solution was measured in triplicate and the recovery percentage was calculated. The optimal chromatographic conditions were used to analyze the sample for ten times and it was followed calculation of mean.

Linearity

The linearity of analytical method shows the direct proportionality of obtained test results to the concentration of analyte in sample within the given range (Chhalotiya, Patel, Bhatt, 2010Chhalotiya UK, Patel HB, Bhatt KK. Development and validation of an ultra-performance liquid chromatography method for venlafaxine hydrochloride in bulk and capsule dosage form. Indian J Pharm Sci. 2010;72(6):814-18.). The linearity was evaluated by linear regression analysis. Quantitative analytical results are greatly affected by the quality of the calibration curve (Parejiya et al., 2014Parejiya PB, Movaliya VR, Barot BS, Modi D, Shelat PK, Shukla A. Quantitative determination of milnacipran HCl in rabbit plasma by HPLC and its application to pharmacokinetics study. J Liq Chromatogr Relat Technol. 2014;37(1):99-11.). The calibration curve was obtained with seven concentrations of reference standard venlafaxine HCl solution (10, 50, 100, 200, 300, 400 and 500 µg mL−1) of the target analyte concentrations for the chromatographic method. 20 µL of every concentration was injected in the chromatographic column through auto-sampler and allowed to separate.

Sensitivity

For HPLC method, the limit of detection (LOD) and limit of quantification (LOQ) were calculated based on the standard deviation of the response and the slope of calibration curve. Seven concentration levels except the blank were used to generate the calibration curve of venlafaxine HCl in the mobile phase. This curve was analyzed statistically to calculate the concentration of analyte in sample.

Robustness

For the HPLC method, the robustness was determined by the analyses of the samples under a variety of conditions making small changes in the parameters like column temperature, buffer pH and mobile phase composition.

Accelerated stability study

A novel and validated HPLC-UV method was applied to carry out accelerated stability studies of venlafaxine HCl capsules packed in gelatin shells placed in environment chamber (AC520, Perani, Italy). The study was carried out at 40 ºC and 50 ºC with 75% relative humidity during six months at various time intervals to determine any physical changes as well as their drug content. F-test was used to compare stability of two data replicates sets and calculate their respective precisions.

Pharmacokinetic study

In vivo trial was conducted to determine the rate of absorption, distribution, metabolism and elimination of venlafaxine from the body. Ten male white albino rabbits (≈ 1.2 kg each) were fasted overnight but allowed free access to water prior to the experiment.

All animals testing and experimental procedures were carried out under the guidelines mentioned in Good Laboratory Practice (GLP) regulations as described by United States Food and Drug Administration (USFDA) and Organization for Economic Co-operation and Development (OECD) Test Guidelines 425 (Up and Down Procedure) (Chen et al., 2006Chen Z, Meng H, Xing G, Chen C, Zhao Y, Jia G, et al. Acute toxicological effects of copper nanoparticles in vivo. Toxicol Lett. 2006;163(2):109-120.). Before starting experiment, approval was taken from ethical committee of University of Sargodha, Sargodha, Pakistan (Ref. No. 20A28IEC UOS). The jugular vein of each rabbit was cannulated the day before drug administration. Venlafaxine HCl (5 mg) was orally administered to the rabbits. After administration, blood samples (3 mL) collected from the jugular vein cannula at 0.5, 1, 1.5, 2, 2.5, 4, 6, 8, 10, 12, 24 and 36 h were kept in heparin containing tubes. The plasma samples were separated by centrifugation (3000 rpm, 5 min) using a refrigerated table top centrifuge and kept frozen at 0 ºC until analysis.

The developed HPLC-UV method was used to analyze the plasma concentration of venlafaxine HCl. HPLC-UV analysis outcomes were plotted as mean plasma drug concentration vs. time. The pharmacokinetic parameters including tmax, Cmax, and AUC were calculated by non-compartmental analysis by Kinetica 5.0® computer software method.

RESULTS AND DISCUSSION

Optimization of solvent system

The mobile phase A (Table I) and standard solution of venlafaxine HCl were used to develop and validate an assay method for accelerated stability study of venlafaxine HCl. The pH and composition of mobile phase was varied to optimize the system. Figure 2 shows a standard chromatogram obtained from the analysis of a reference standard for venlafaxine HCl. The typical retention time (2.7 min with ḱ value 2.1) of venlafaxine HCl standard solution in mobile phase indicates that there is no drug-excipients interaction in the formulation (Kaur et al., 2010Kaur J, Srinivasan KK, Joseph A, Gupta A, Singh Y, Srinivas KS, et al. Development and validation of stability indicating method for the quantitative determination of venlafaxine hydrochloride in extended release formulation using high performance liquid chromatography. J Pharm Bioallied Sci. 2010;2(1):22-6.), showing the efficiency of our validated method. The venlafaxine HCl peak was confirmed by spiking with the standard. The column is considered very efficient if the number of theoretical plates is more than 1500 (Sánchez-Mata et al., 2000Sánchez-Mata MC, Cámara-Hurtado M, Díez-Marqués C, Torija-Isasa ME. Comparison of high-performance liquid chromatography and spectrofluorimetry for vitamin C analysis of green beans (Phaseolus vulgaris L.). Eur Food Res Technol. 2000;210(3):220-25.) that was found to be 1849.6 in our study. The tailing factor was found as 0.68, which is considered acceptable for chromatographic separations. The consistency of analytical results has great significance as it is required for proper analysis of pharmacokinetics results.

FIGURE 2
Standard Chromatogram of venlafaxine HCl.

Method validation

Linearity

The method was significantly linear for the drug in the concentration range of 10-500 µg mL-1 (Figure 3). The plots represented the highest linearity (R2 = 0.998) revealing the good in vitro drug release of optimized formulation. The linearity was further confirmed by residual analysis, as a residual plot is a graph that shows the residuals on the vertical axis and the independent variable on the horizontal axis. If the points in a residual plot are randomly dispersed around the horizontal axis, a linear regression model is appropriate for the data; otherwise, a non-linear model is more appropriate. As shown in the Figure 4, the random pattern indicates that a linear model provides a decent fit to the data. Table II shows the calibration curve data. The correlation coefficient for venlafaxine HCl obtained in this phase was 0.998 (Table IV).

FIGURE 3
Linearity Curve for venlafaxine HCl.

FIGURE 4
Residual analysis of calibration curve.

TABLE II
Linearity data for venlafaxine HCl
TABLE III
Intra-day and inter-day precision data for quantification of venlafaxine HCl
TABLE IV
System suitability parameters for venlafaxine HCl

Precision and accuracy

The validity of the planned method was assessed by recovery studies of standard addition method. Excellent recoveries (98.25-99.27%) were achieved for venlafaxine HCl indicating the method to be accurate. Percentage RSD was used to determine precision for intra-day and inter-day precision as shown in the Table III. The repeatability of newly developed method is proved by very low value of % RSD (Table V) confirming the method to be sufficiently precise under the same operating conditions.

TABLE V
Robustness of HPLC-UV method

Sensitivity

The sensitivity of developed method for quality control analysis of any drug is proved by LOD and LOQ values. For venlafaxine HCl, LOD and LOC were found to be 35 and105 ngmL-1, respectively (Table IV) that were higher than reported in literature (Chhalotiya, Patel, Bhatt, 2010Chhalotiya UK, Patel HB, Bhatt KK. Development and validation of an ultra-performance liquid chromatography method for venlafaxine hydrochloride in bulk and capsule dosage form. Indian J Pharm Sci. 2010;72(6):814-18.).

Robustness

In all the deliberately varied chromatographic conditions, the chromatogram for system suitability solution showed satisfactory resolution. RSD<1% with no significant changes in chromatographic parameters proves the robustness of newly developed method (Table V).

Accelerated stability study of venlafaxine HCl capsules

Figure 5 clearly shows the result of accelerated stability study of venlafaxine HCl capsules at 40 ºC and 50 ºC with 75% RH, at various time intervals. The concentration of venlafaxine HCl (the active ingredient) decreased slowly with R2 = 0.9774 and R2 = 0.9762 at 40 and 50 ºC, respectively. The F-test results indicated that there is no significant difference in stability of venlafaxine HCl capsules at 40 and 50 ºC with 75% RH. Stability study of the optimized formulation proved reliability of the drug tablets formulated by newly developed and validated HPLC-UV method, even after exposing to stress conditions of temperature and humidity.

FIGURE 5
Stability curve showing the concentration of active ingredient in venlafaxine HCl capsules vs number of days at 40 ºC & 75 % RH(■) and 50 ºC & 75 % RH (♦).

Application of developed method (Pharmacokinetic study)

The precision, sensitivity and reproducibility of HPLC-UV method were good for pharmacokinetics studies. The pharmacokinetic parameters were evaluated by single oral dose of 5mg venlafaxine hydrochloride capsule (dose as per body weight) to rabbits. The mean plasma concentration profile in oral single dose study is shown in Figure 6. Table VI presents the pharmacokinetic parameters. The Cmax and AUC0-36 values are 56.5 and 845.9 µg h mL-1, respectively for venlafaxine tablet. These values corresponded to good bioavailability. The results indicate that the newly developed HPLC-UV method can also be used for the bioavailability study of venlafaxine HCl.

FIGURE 6
Mean plasma concentration-time curve after a single oral dose of 5 mg venlafaxine HCl.

TABLE VI
Non compartmental pharmacokinetic analysis after a single oral dose (5 mg) administration

CONCLUSIONS

Newly developed and validated HPLC-UV method was successfully applied for accelerated stability and pharmacokinetic study of venlafaxine HCl. The method was proved to be efficient, robust, time conserving and offers good accuracy and precision when it is used to monitor full pharmacokinetic profile of venlafaxine HCl in rabbits. The main advantage of the method is that it uses small processing volume used for analysis and can therefore be employed for routine quality control analysis.

REFERENCES

  • Asafu-Adjaye EB, Faustino PJ, Tawakkul MA, Anderson LW, Lawrence XY, Kwon H, et al. Validation and application of a stability indicating HPLC method for the in vitro determination of gastric and intestinal stability of venlafaxine. J Pharm Biomed Anal. 2007;43(5):1854-59.
  • Baldania SL, Bhatt KK, Mehta RS, Shah DA, Tejal RG. RPHPLC estimation of venlafaxine hydrochloride in tablet dosage forms. Indian J Pharm Sci. 2008;70(1):124-128.
  • Bernardi LS, Oliveira PR, Murakami FS, Borgmann SH, Arend MZ,Cardoso SG. Development and validation of a stability-indicating LC method for the determination of venlafaxine in extended-release capsules and dissolution kinetic studies. J Chromatogr. Sci. 2009;47(9):770-76.
  • Bhatt J, Jangid A, Venkatesh G, Subbaiah G, Singh S. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for simultaneous determination of venlafaxine and its active metabolite O-desmethyl venlafaxine in human plasma. J Chromatogr B. 2005;829(1-2):75-81.
  • Chan CC, Lee YC, Lam H, Zhang XM. Analytical method validation and instrument performance verification, John Wiley & Sons; 2004.
  • Chen Z, Meng H, Xing G, Chen C, Zhao Y, Jia G, et al. Acute toxicological effects of copper nanoparticles in vivo. Toxicol Lett. 2006;163(2):109-120.
  • Chhalotiya UK, Patel HB, Bhatt KK. Development and validation of an ultra-performance liquid chromatography method for venlafaxine hydrochloride in bulk and capsule dosage form. Indian J Pharm Sci. 2010;72(6):814-18.
  • Goeringer KE, McIntyre IM, Drummer OH. Postmortem tissue concentrations of venlafaxine. Forensic Sci Int. 2001;121(1-2):70-75.
  • Gursharanjit S, Ghosh B, Dave D. Screening of venlafaxine hydrochloride for delivery. AAPS Pharm SciTech. 2008;9(3):791-797.
  • ICH HTG . International Conference on Harmonisation; Guideline on Validation of Analytical Procedures: Definitions and Terminology Availability. Part VIII. Federal Register. FDA. USA, 1996;60:11260.
  • ICH Validation of analytical procedures: text and methodology. In International Conference on Harmonization Q2 (R1). Geneva; 2005.
  • Juan H, Zhiling Z, Huande L. Simultaneous determination of fluoxetine, citalopram, paroxetine, venlafaxine in plasma by high performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-MS/ESI). J Chromatogr B. 2005;820(1):33-39.
  • Kaur J, Srinivasan KK, Joseph A, Gupta A, Singh Y, Srinivas KS, et al. Development and validation of stability indicating method for the quantitative determination of venlafaxine hydrochloride in extended release formulation using high performance liquid chromatography. J Pharm Bioallied Sci. 2010;2(1):22-6.
  • Liu W, Dai YC, Deng N, Liu XR, Luo Y. Development and validation of a HPLC-MS/MS method for the determination of venlafaxine enantiomers and application to a pharmacokinetic study in healthy Chinese volunteers. Biomed Chrom. 2011;25(3):412-16.
  • Makhija SN, Vavia PR. Stability indicating LC method for the estimation of venlafaxine in pharmaceutical formulations. J Pharm Biomed Anal. 2002;28(6):1055-59.
  • Parejiya PB, Movaliya VR, Barot BS, Modi D, Shelat PK, Shukla A. Quantitative determination of milnacipran HCl in rabbit plasma by HPLC and its application to pharmacokinetics study. J Liq Chromatogr Relat Technol. 2014;37(1):99-11.
  • Rudaz S, Stella C, Balant-Gorgia AE, Fanali S, Veuthey JL. Simultaneous stereoselective analysis of venlafaxine and O-desmethylvenlafaxine enantiomers in clinical samples by capillary electrophoresis using charged cyclodextrins. J Pharm Biomed Anal. 2000;23(1):107-15.
  • Sánchez-Mata MC, Cámara-Hurtado M, Díez-Marqués C, Torija-Isasa ME. Comparison of high-performance liquid chromatography and spectrofluorimetry for vitamin C analysis of green beans (Phaseolus vulgaris L.). Eur Food Res Technol. 2000;210(3):220-25.
  • Seshadri VC, Manohari PJ, Kunchithapatham J. Formulation and characterization of mouth dissolving tablets containing benzodiazepine. Indo Am J Pharm Res. 2013;3(12):1457-74.
  • Titier K, Castaing N, Scotto-Gomez E, Pehourcq F, Moore N, Molimard M. High performance liquid chromatographic method with diode array detection for identification and quantification of the eight new antidepressants and five of their active metabolites in plasma after overdose. Ther Drug Monit. 2003;25(5):581-87.
  • Vu RL, Helmeste D, Albers L, Reist C. Rapid determination of venlafaxine and O-desmethyl venlafaxine in human plasma by high-performance liquid chromatography with fluorimetric detection. J Chromatogr B Biomed Sci Appl. 1997;703(1-2):195-201.
  • Waschgler R, Moll W, König P, Conca A. Quantification of venlafaxine and O-desmethyl venlafaxine in human serum using HPLC analysis. Int J Clin Pharmacol Ther. 2004;42(12):724-28.
  • Younus M, Arif MF, Richards MP, Kumar DB. Determination of venlafaxine and modafinil in individual tablet dosage forms using single RPHPLC method. Trop J Pharm Res. 2015;14: 239-45.

Publication Dates

  • Publication in this collection
    16 Mar 2020
  • Date of issue
    2020

History

  • Received
    09 Nov 2017
  • Accepted
    28 June 2018
Universidade de São Paulo, Faculdade de Ciências Farmacêuticas Av. Prof. Lineu Prestes, n. 580, 05508-000 S. Paulo/SP Brasil, Tel.: (55 11) 3091-3824 - São Paulo - SP - Brazil
E-mail: bjps@usp.br