Compatibility Studies of Quercetin with Pharmaceutical Excipients Used in the Development of Novel Formulation

 

Rajendra Jangde* and Deependra Singh

University Institute of Pharmacy, Pt. Ravishankar Shukla University, Raipur-492010 C.G. India

 

ABSTRACT:

The aim of study was for the development of any novel formulation, assessment of drug-excipients compatibility using different techniques such as thermal and isothermal stress testing were used to assess the compatibility of drug with excipients., represents an important phase in preformulation stage. Differential scanning calorimetry (DSC) and Fourier transformation infrared spectroscopy (FTIR) and X-ray diffraction study (XRD) were the common methods for the study of compatibility. The potential physical and chemical interactions between the drug and excipients can affect the chemical nature, stability, bioavailability of drugs and subsequently, affects their therapeutic efficacy and safety. . In the present study drug excipients compatibility study of quercetin was conducted with different controlled release excipients. The drug and excipients mixtures were stored at 50°C for 4 weeks. The samples were then characterized using DSC, FTIR and UV spectrophotometric methods. The results show that quercetin was compatible with the all the excipients used in the study. The excipients used in the present study were definitely incorporated in the extended release quercetin formulation. On the basis of the results obtained from DSC, FTIR and XRD studies, all the excipients used were found to be compatible with the quercetin and can be used for the development of novel formulation.

 

 

INTRODUCTION:

Assessment of drug-excipients compatibility is very significant to identify product’s stability as well as its reproducibility with confirmed therapeutic efficiency [1]. Drug – excipients compatibility studies lays a foundation in careful selection of most appropriate excipients and helps in designing a chemically stable and effective dosage form [2, 3]. Incompatibility between drug and excipients can affect the chemical nature, stability and bioavailability of drug and subsequently affects their therapeutic efficacy and safety [4, 5]. Despite the importance of the issue, there is no universal accepted protocol for drug-excipients incompatibility testing [6,7]. Quercetin, a flavonol occurring in fruit and vegetables is a food component with proven beneficial impact on health [8]. It is one of the most potent antioxidants among polyphenols [9]. Quercetin has also been demonstrated to display the antiviral, antibacterial, anticarcinogenic and anti-inflammatory effects [10,11].

 

In recent years, nanostructured materials such as nanoparticles, liposomes etc., have been considered as potential carriers for hydrophobic drug delivery that may resolution the abovementioned complications [12]. Novel carrier has developed a significant part of research in the field of drug delivery because they have the ability to distribute a extensive assortment of drugs to variable parts of the body for sustained periods of time. Novel carrier systems are actively investigated as drug carriers to diminish drug toxicity and degradation, so as to deliver therapeutic agents to several sites of action and to promote a suitable, selective and specific targeted therapy [13].

 

Differential Scanning Calorimetry is widely used thermal technique in drug excipients compatibility assessment. Differential scanning calorimetry (DSC) technique involves the application of a heating or cooling signal to a sample and a reference. When the substance undergoes a thermal event, the difference in heat flow to sample and reference is monitored against time and temperature. As a result, energy associated with various thermal events (e g melting, glass transition temperature and crystallization etc.) Therefore, the use of analytical technique such as Fourier Transform Infrared (FTIR) Spectroscopy is another approach used incompatibility tests based on the hypothesis that same functional group change during drug-excipients interaction [14].The purpose of the present investigation was to evaluate the compatibility of quercetin with various pharmaceutical excipients to be used in the nanoparticle formulations utilizing the different analytical techniques such as differential scanning calorimetry, Fourier Transform Infrared Spectroscopy and X-ray diffraction study.

 

EXPERIMENTAL:

Quercetin, Phosphatidylcholine and cholesterol were purchased from Hi-media chemicals (Mumbai, India). HPLC grade solvents were purchased from Merck (Mumbai, India). All other materials and solvents used were of analytical grade.

 

Sample Preparation:

The quercetin, phospholipid and cholesterol mixtures prepared at 1:1:1 ratio. The quercetin, phospholipid and cholesterol were individually weighed in a 10 mL glass vial and mixed on a vortex mixer for 2 min. In each of the vials,10% of the water was added and the drug-excipient blend was further mixed. Each vial was sealed Teflon-lined screw cap and stored at 50°C for 4 weeks. These samples were periodically examined for any change of unusual color change.

 

Characterization by UV spectrophotometer:

The samples after 4 weeks were withdrawn from storage and analyzed by UV spectrophotometric. The drug content was determined at initial and stored samples in triplicate. An accurately weighed amount of the quercetin, phospholipid and cholesterol mixture were taken and suitably dissolved under sonication in pH 7.4 phosphate buffer and filtered through 0.45 (Millipore) filters. The sample was analyzed after making appropriate dilutions using UV spectrophotometer (Systronic 2202 double beam UV spectrophotometer) at 256 nm against blank.

 

Fourier transforms infrared radiation measurement (FT-IR):

FT-IR spectra’s were recorded on a FTIR spectroscopy using the instrument Shimadzu FT-IR 8400S in the frequency range of 400-4000 cm-1 with the resolution of 4 cm-1 using potassium bromide discs method . The drug and each selected excipients (1:1 w/w) were stored at 40 ± 2^oC and 75 ± 5 % RH for 1 month. Individual samples as well as the mixture of drug and excipients were ground, mixed thoroughly with potassium bromide for 3- 5mins in a mortar and compressed into disc by applying a pressure of 5 tons for 5 minute in hydraulic press. The concentration of sample in potassium bromide should be in the range of 0.2% to 1%. The pellets were placed in light path and spectrum was obtained and reviewed for evidence of any interactions.

 

Differential Scanning Calorimetry (DSC):

Differential Scanning Calorimetry (Perkin Elmer Jade, California, USA, department of pharmaceutical sciences dibrugarh university, Assam, India) was used for thermal analysis of drug and mixture of drug and excipients in a 1:1w/w ratio. Individual samples (drug and excipients) as well as physical mixture of drug and excipients were weighed to about 5mg in DSC aluminum  pan. The sample pan was crimped for effective heat conduction and scanned in the temperature range of 50-300°C. Heating rate of 20°C min-1 was used and the thermogram obtained was reviewed for evidence of any interactions.

 

X-Ray Diffractometry (XRD):

The solid state of the drugs was investigated by X-ray powder difractometry with Bragg- Brentano geometry at a wavelength of 1.5406. Powder X-ray difractograms were recorded in a diffraction angle (2θ) range of 20-400 using a step size of 0.030 under an exposure time of 8s.

 

RESULTS AND DISCUSSION:

Drug content estimation by UV spectroscopy:

The drug and excipients mixture was physically observed at different intervals. No Characteristic color change was observed. The assay of the drug excipients mixtures were found good. Assay value of 99.12 to 100.1 was observed at initial. Good correlation was observed with the samples of drug excipients mixtures stored at 50°C for 4 weeks. This clearly indicates the stable nature of the quercetin with excipients (Table 1).

 

Table1: It shows drug content of quercetin after storage at 50 °C for 4 weeks.

Time	Quercetin	Quercetin-phospholipid	Quercetin -cholesterol
Initial 500C for 4 weeks	98.45 99.52	97.46 99.01	96.45 98.78

 

Fourier transforms infrared radiation measurement (FTIR):

The use of FTIR technique allows, pointing out the implication of the different functional groups of guest and host molecules by analyzing the significant changes in the shape and position of the absorbance bands. Data obtained from FT-IR spectrophotometric study clearly indicates insignificant changes in spectra obtained from physical mixture of quercetin and excipients. Spectra obtained from pure quercetin were found 3411 cm^-1 and 1663.1 cm^-1 for O-H str. and C=O respectively. (Figure 1) Spectrograph of both the quercetin and excipients physical mixture shows 3140 cm^-1, 3220.24 cm^-1 and 3310.71cm^-1 for O-H str. group. Spectrography of C=O str. shows peak at 1650.45 cm^-1, 1750.26 cm^-1 for quercetin, phospholipid and cholesterol respectively. Which indicates no interference between them (Table 2). There was no major changes in peaks of ketone (C=O) and Hydroxyl (O-H) in reference to the observed value of quercetin.

(A)	(B)
Fig. 1:  FTIR spectrum of (A) Pure quercetin and (B) Mixture of quercetin with cholesterol

 

 

Table 2: It shows FTIR spectroscopy data of quercetin and phospholipid

S. No	F.G	Standard (cm-1)	Quercetin (cm-1)	Physical mixture (cm-1)
1	O-H	3100-3400	3411	3140
2	C=O	1650-1700	1663.1	1650.45
3	C-OH	1350-1400	1383.1	1370
4	C-H	680-850	677	750

 

Differential scanning calorimetry (DSC):

The DSC thermogram corresponding to quercetin, phospholipid and physical mixture. Quercetin exhibits a characteristic endothermic peak at 260.44°C corresponding to its melting temperature. A broad endothermic band from 60-120°C was observed for the amorphous phospholipid, which was related to loss of water molecules i.e., dehydration process. The DSC thermogram for the solid complex of quercetin, phospholipid and cholesterol showed (Figure 2) an endothermic peak at 263.77°C associated with the formation of complex in the solid state. As such there is no interaction between quercetin, phospholipid and cholesterol.

 

Fig. 2: DSC thermogram of Pure quercetin, Cholesterol and Physical mixture of quercetin and Cholesterol

 

X-Ray Diffractometry (XRD):

X-ray diffraction study (XRD) was performed to analyze crystalline or amorphous nature of the quercetin. XRD studies of quercetin, cholesterol, physical mixture of excipients the crystalline peak for quercetin was clearly evident whereas in deformed peak for quercetin was observed (Figure 3). This change in diffraction pattern supported the conversion of crystalline drug to amorphous form.

 

 

Fig.3: XRD  spectrum of Pure quercetin, Cholesterol and Mixture of quercetin with cholesterol

 

 

CONCLUSIONS:

The compatibility of quercetin with excipients was studied by different analytical techniques like Differential Scanning Calorimetry, Fourier Transform Infrared Spectroscopy and X-ray diffraction study. However, techniques such as FTIR and X-ray diffraction techniques after storage of the mixture of quercetin and individual excipients under stressed conditions should be taken in conjunction with DSC results to reach any definite conclusion. In the present study, results of DSC along with FTIR and XRD (for IST) were successfully employed to assess the compatibility of quercetin with the excipients. No concrete evidence of interaction was observed between quercetin and the excipients like phospholipid, cholesterol. No characteristic color change was observed during the storage at 50 °C for 4 weeks. The UV analysis results of this mixture evident the chemical stability of quercetin as the assay was within the acceptable range. Hence, this data’s demonstrates the potentiality of the excipients for the successful development of a novel formulation.

 

ACKNOWLEDGEMENT:

The author, Dr. Deependra Singh is thankful to UGC-MRP41-748-2012, UGC-RA-70371/2012, DST-FIST and SAP. The authors are thankful to Director, University Institute of Pharmacy, Pt. Ravi Shankar Shukla University for providing facilities and guidance.

 

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Received on 12.05.2014                Modified on 07.07.2014

Accepted on 12.08.2014                © RJPT All right reserved