Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms. Ravi Kiran B*, Santosh L, kartick C, laasya P, Ravi L. Andhra University, Visakhapatnam –530045, Andhra Pradesh, India. *corresponding author:
[email protected]
Abstract A rapid, specific and accurate isocratic HPLC method was developed and validated for the assay of Quetiapine Fumarate in pharmaceutical dosage forms. The assay involved an isocratic –elution of Quetiapine Fumarate in Grace C18 column using mobile phase composition of buffer and acetonitrile in the ratio of 50:50 (v/v).The wavelength of detection is 252 nm .The proposed method can be used for routine quality control samples in industry in bulk and in finished dosage forms and for the stability studies as the method separates Quetiapine Fumarate from its degradation products. Drug Profile Quetiapine Fumarate [Figure-5.01] is recently approved third generation antidepressant Active pharmaceutical Ingredient very much different from first and second generation tri- and tetra-cyclic by its pharmaceutical activity. Its IUPAC name is [2-(Dimethylamino) -1-(4-methoxyphenyl) ethyl] cyclohexanol hydrochloride [15]. It is used as a medical agent; a viable solution for depression and other nervousness related disturbances. It was originally discovered in pharmaceutical corporation Wyeth® and commercially released in market in 1993; however it is now marketed under Pfizer®, another pharmaceutical corporation. In one of the statistics released in 2007, Quetiapine Fumarate is found to be one of the top ten prescribed anti-depressant pills in the United States of America, which is an indicative of its popularity.
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Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
OH N H
OMe
Figure-5.01 : Molecular Structure of Quetiapine Fumarate
5.1.2 Physical and Chemical Properties Quetiapine Fumarate is an Active pharmaceutical agent prescribed as hydrochloride salt of Venlafaxine; it is crystalline in nature, colored as white to offwhite powder. Its Molecular formula is C17H27NO2·HCl. Its melting point is 2170C. Ionization constant value is 9.4 and stable under normal room ambient conditions Due to its solubility in water and other good tableting properties it is formulated as orally administered pill [6-7]. 5.1.3 Chemistry and Pharmaceutical Activity On the basis of many clinical trials, it is found that Venlafaxine has no affinity for brain muscarinic. It is absorbed almost completely (92 % to be precise) after oral intake
and
gets disintegrated substantially in the liver [8]. This drug appears to
disown many side effects often related with other tricyclic anti-depressants. In humans, Venlafaxine (VEN) is metabolized N-desmethylvenlafaxine (NDV); O,Ndesmethyl venlafaxine (DDV) and O-desmethyl venlafaxine (ODV). Among all these metabolic sub-structures, ODV is chemically active at higher concentrations and increasingly contributes to the therapeutic effects of Quetiapine Fumarate since they are viable inhibitors of neuronal serotonin [9].
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Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
Anti-depressant action of Quetiapine Fumarate is considered to be related with its capacity to efficiently control action of neurotransmitters in the central nervous system (CNS) of our body [10]. It inhibits the reintake of serotonin, norepinephrine and dopamine [3-5]. 5.1.4 Review of Past Analytical Methods of Quetiapine Fumarate Mandrioli R, et al. [11] has reported a HPLC method for Quetiapine Fumarate and its major precursor ODV in biological fluids, utilizing a stationery-phase column (C8, 150 × 4.6 mm ID, 5 μm) A hyphenated and coupled HPLC method with electro spray ionization with mass spectrometry as detector (HPLC-ESI/MS) method has been reported by Liu W, et al. [12] for estimation of Quetiapine Fumarate and one of its major precursor ODV enantiomers in human plasma Ebenezer B, et al. [11] this research group has evaluated gastrointestinal stability of Quetiapine Fumarate in vitro simulated gastric (SGF) and intestinal (SIF) fluids utilising a stability-indicating HPLC method. Kirchherr H and Kuhn-velten WN [12] have discussed an innovative multitasking simultaneous determination of 48 depressing agents with a new approach of single sample and triple injection. Juan H, Zhiling Z and Huande L [15] have introduced a procedure for simultaneous measurement four popular anti-depressants including Quetiapine Fumarate. The compounds were charged in the electro spray ion source of the mass spectrometer and were detected in the new SIR mode.
135
Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
Waschgler R, et al. [16] has described an HPLC method based on reverse phase technique for the estimation of Quetiapine Fumarate and its main active precursor ODV in human biological fluids. Its mode of detection is fluorescence at 227/300 nm (excitation/emission) without any interferences in serum. Frahnert C, et al. [18] this research group introduce reverse-phase technique based UV-HPLC method in human serum. This method is applicable for drug monitoring of about 30 anti-depressants and atypical antipsychotics to effectively analyze in biological fluids. Liu W, et al. [19] research group has introduced a hyphenated and coupled HPLC method with electro spray ionization with mass spectrometry as detector (HPLC-ESI/MS) method for simultaneous determination of Quetiapine Fumarate and three of its precursors ODV, NDV and DDV in human biological fluids. The procedure discussed is sensible and can be used as a pharma-kinetic model in clinical studies. Bhatt J, et al. [17] have reported a sensible hyphenated LC technique coupled with mass spectrometry as detector (LC-MS-MS) and applied for dual estimation of Quetiapine Fumarate and ODV in human biological fluids. Patel BN, et al. [21] have reported
a sensible hyphenated LC technique
coupled with mass spectrometry as detector (LC-MS-MS) for complete estimation of Quetiapine Fumarate and its active metastructure, ODV in human plasma. The method was successfully applied to establish bioequivalence of Venlafaxine in healthy Indian male member’s clinical settings.
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Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
Vu RL, et al. [22] has devised a rapid and sensitive HPLC technique for simultaneous measurement Quetiapine Fumarate (VEN) and its metastructure ODV in biological fluids. This method is reliable and is applied for regular clinical monitoring of the drug and its precursor. Matoga M, et al. [23] has reported a rapid specific and accurate RP-HPLC method for the complete determination of Quetiapine Fumarate and ODV in biological fluids. This method is fast with run tine less than 10 minutes. Numerous procedures have been discussed for the complete estimation of Quetiapine Fumarate in API, pharmaceutical and clinical analytes. The discussed procedures involve HPLC [9-16], HPLC-MS [17-19] and florometric detection [2024].
5.1.5 List of Important Brand Names of Quetiapine Fumarate Formulations The conditional or conventional release circulates the distribution of the pill into the human intestinal duct over a extended time than rapid release Venlafaxine formulations. This result in minimum peak plasma concentration, clinical studies have proved that the conditional release formulations has a lower incidents of patients effecting from nausea and other side effects. Quetiapine Fumarate is utilized essentially for the medication of stagnation, anguishes, social solitude, distract thinking and other related syndroms. Quetiapine Fumarate is also widely available in
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Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
Tablet and Capsule dosage form in Indian market as well. Various formulations available in market and there dosage forms are summarized as follows [Table-5.01]. Table-5.01 : List of Important Brand Names of Quetiapine Fumarate available in Indian Market Brand name
Formulation/Combination
Dosage strength
Dosage form
Name of the manufacturer
Dalium™
Quetiapine Fumarate
37.5/75 mg
Tablet/Kit
Piramal® Health care
Envelaf™
Quetiapine Fumarate
75/150 mg
Tablet/Kit
Pentacare® (Alkem)
Sentosa
Quetiapine Fumarate
75 mg
Tablet
Sarabhai Piramal® healthcare Ltd
Ven-OD™
Quetiapine Fumarate
75/150 mg
Tablet
Zydus®neurosciences
VenaxinSR™
Quetiapine Fumarate
25/37.5/ 75 mg
Tablet
Psycho® remedies
™
VenizXR™
Quetiapine Fumarate
75/150 mg
Capsule
Sun® pharmaceutical Ltd
VenlorXR™
Quetiapine Fumarate
37.5/75/150 mg
Capsule
Cipla®
5.2 Aim, Scope and Salient Features of Present Work can be Summarized as follows To introduce a superior stability-suitable HPLC method for assay of Quetiapine Fumarate using external standard method.
1. To study forced degradation behavior of Quetiapine Fumarate under various stress conditions such as acidic, basic, oxidative, photolytic and thermal. 2. To satisfactorily separate and resolve all major degradents if formed from major peak. 3. To develop assay method which is free from interference from excepients. 138
Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
4. To develop method which is less tedious and easy to perform and can easily be adapted in quality control laboratories for routine analysis. 5. To perform complete validation of developed method and assess its system suitability parameters. Literature survey revealed that most of the HPLC methods used hyphenated techniques with detectors such as mass spectrometry, flourimetry, Electro spray mass spectrometric techniques all these methods have high sensitivity, but most of them highly expensive and are not easily available in small and medium sized quality control laboratories. There is urgent scope to evolve a totally reliable and affordable HPLC procedure for the regular quality control applications of drug in its API and Branded formulations. 5.3 Experimental 5.3.1 Chemicals and Reagents Quetiapine Fumarate (99.89 %) purity is used in these experiments. Acetonitrile, methanol and tetrahydrofuran (grade of HPLC) was procured from Qualigens®. Milli-Q® water was procured from Ranbaxy® fine chemicals limited (RFCL). Chemicals such as potassium dihydrogen orthophosphate, disodium hydrogen orthophosphate, ammonium acetate, sodium acetate, ammonium dihydrogen orthophosphate, ortho phosphoric acid, acetic acid used belong to grade of HPLC. 5.3.2 Instrumentation The HPLC system consisted of a Shimadzu® Class VP Binary pump LC10Atvp, SIL-10Dvp auto sampler, CTO-10Avp column temperature oven, PDA-UV Detector. All the components of the system are controlled using SCL-10Avp System controller. Data acquisitions were done using LC-solution software. The other
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Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
important
instruments
used
which
includes
Shimadzu®
microbalance
(ModelNo:CPA225D) and ultrasonicator is make of Systronics® (Model No 289-A) and Eppendorf™ micro-pipette of capacity range of 100 to 1000 µL) and Hamilton® syringe of 10 µL is employed for sample injection in these experiments pH meter of Elico® make, water bath(Life-scan®) for maintaining constant temperature. UVVisible spectrophotometer and lab-oven of make Lab-India®. Graphite mortar for grinding was also used. 5.4 Method Development and Optimization For starting method development, a comprehensive study for optimization of chromatographic conditions was taken up. This is done by taking one chromatographic parameter at a time. 5.4.1 Choice of Mobile Phase The pH of the buffer moves the analyte equilibrium to either neutral or ionic form. It is analytically better to select pH for buffer; so that analytes ionization constant value is at the minimum two units away, so each analyte be in a predominantly in single form (>90%) which could be either neutral or ionic during the chromatographic run. This ideally decreases the impact of secondary ionization equilibrium on retention of analyte, which otherwise has significant effect. Ionic and neutral forms of any sample analyte have substantial dissimilarities in their apparent polarity and hence moves through the column with variable velocities. The development of HPLC methods close to pKa value is not recommended because the analyte will co-exist in both neutral and ionic forms in ratio (50:50) which in turn leads to secondary equilibrium; these methods are not rugged and cannot be easily applicable to different laboratories, since any small 140
Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
variation in the chromatographic conditions in mobile phase pH, leads to major differences in the retention and other parameters of analyte. Every analyte has its specific pKa value, thus pH of the buffer selected should be at the minimum two units away, such that 99 % of the analyte will be either in neutral or ionized form. The mobile phase components consists of (65:35, v/v) of acetonitrile and sodium acetate with 0.1 % tri-ethylamine. After finalizing buffer, we proceed to mobile phase selections. For selection of mobile phase from buffer, solvents like acetonitrile (ACN), methanol (MeOH), tetrahydrofuran (THF) used, elution strength in reverse phase chromatography depends on both organic solvents percentage and variety (e.g., ACN, MeOH, THF). The essential aim of process development is to obtain suitable and equilbrated run time for all the samples in isocratic conditions, preferably 2 < k > 10, where k is capacity factor. Acetonitrile is selectively chosen as organic solvent for the mobile phase. ACN – buffer can be used at low wavelengths of detection. ACN – buffer has lower viscosity resulting in lower column pressure and higher plate numbers. Besides acetonitrile; methanol and THF were also implemented, but they were less preferred, because they have higher UV absorbance and slower column equilibration. Tri-ethyl amine is popular mobile phase additive applied with buffer reduce peak tailing, it acts as a tailing-suppressing reagent by reducing the association activity of free-silanol groups, thus reducing the peak tailing. The HPLC procedure was standardized with an idea to innovate a stabilitysuitable assay method. Numerous permutations and combinations, at pH values from 3.0-11.0 were tested. Mobile phase components consist of (65:35, v/v) of acetonitrile
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Development and Validation of a Reversed-phase HPLC Method for the Determination of Quetiapine fumarate in Pharmaceutical Dosage Forms.
and sodium acetate with 0.1 % of tri-ethyl amine operated on isocratic mode was finally selected. Diluent is prepared in the same way as mobiles phase which consist of (50:50, v/v) methanol and 0.1 % ortho phosphoric acid. 5.4.2 Choice of Column During method development process various columns such as such as C8, Ccyano, and C-phenyl were utilized. In reverse phase chromatography; compounds usually follow the following trend of cyano
