New spectrophotometric methods for the determination of Loratadine
Mukthinuthalapati Mathrusri Annapurna*, Pappala Heshitha
GITAM School of Pharmacy, GITAM (Deemed to be) University, Visakhapatnam, India-530045.
*Corresponding Author E-mail: mmukthin@gitam.edu
ABSTRACT:
Loratadine is a second-generation antihistamine that is used to treat allergic rhinitis and hives. New spectrophotometric techniques have been proposed for the estimation of Loratadine in pharmaceutical dosage forms. A double-beam SHIMADZU Model UV-1800UV-VIS spectrophotometer was used for the present study. Loratadine has shown linearity over the concentration range 0.5-50 µg/mL in borate buffer (pH 9.0), phosphate buffer (pH 5.8), phosphate buffer (pH 7.8), and distilled water, and the methods were validated as per ICH guidelines. The proposed methods are simple, and economical and can be successfully applied for the assay of Loratadine in pharmaceutical dosage forms.
KEYWORDS: Loratadine, Spectrophotometric, Borate buffer, Phosphate buffer, First derivative spectroscopy, Validation.
INTRODUCTION:
Loratadine has a role as H1- receptor antagonist, and cholinergic antagonist. Loratadine is a piperidine drug derivative that is related to azatadine which has long-acting and non-sedating effects. It is chemically Ethyl 4-(8-chloro-5,6-dihydro-11H-benzo [5,6] cyclohepta [1,2-b] pyridin-11-ylidene)-1-piperidine carboxylate with molecular formula C22H23ClN2O2 and a molecular weight 382.88 g/mol. Loratadine blocks the H1-histamine receptor and prevents the symptoms that are caused by histamine activity on capillaries, bronchial smooth muscle, and gastrointestinal smooth muscle. Loratadine was quantitatively determined by different authors using spectrophotometry2-5and in the present study the authors have proposed new spectrophotometric techniques for the assay of Loratadine in pharmaceutical dosage forms and validated as per ICH guidelines.
Figure 1: Chemical structure of Loratadine
MATERIALS AND METHODS:
Shimadzu Model No. UV-1800 double beam UV-VIS spectrophotometer with quartz cells was used for the entire study, and all the solutions were scanned 200-400 nm. Buffer solutions such as borate buffer (pH 9.0), and phosphate buffer (pH 5.8, 7.8) were prepared as per IP 2022. Loratadine stock solution was prepared by dissolving 25mg of Loratadine in 25mL volumetric flask with methanol (1000µg/mL), working standard solutions were prepared in methanol (100µg/mL) and further dilutions were prepared by diluting the working standard solutions with respective buffers as per the proposed methods requirement. Loratadine is available as tablets with the brand names Lorinol Loratadine Tablets (Micro Labs Limited, Label claim: 10 mg), Welllora tablets (Wellona pharma, Label claim: 10 mg), Clardin tablets (Morepen Laboratories, Label claim: 10 mg), Loratec tablets (John Lee pharmaceuticals, Label claim: 10 mg), respectively in India.
Method validation:
Zero order spectroscopy (Do):
A series of Loratadine solutions 0.5 - 50 µg/mL were prepared from working standard solution on dilution with borate buffer (pH 9.0) (Method I), phosphate buffer (pH 5.8) (Method II), phosphate buffer (pH 7.8) (Method III) and distilled water (Method IV) and scanned (200-400 nm) against their reagent blanks. The zero-order spectrum so obtained has shown maximum absorbance (λmax) at 247 nm for Methods I, II, III, and IV. The absorbance of all the solutions was noted at λmax, and a calibration curve was drawn by taking the concentration on the X-axis and the corresponding absorbance on the Y-axis for all six methods respectively. Precision studies were performed by calculating the percentage relative standard deviation of independent assays of 3 determinations of the test concentration, whereas accuracy studies were carried out by the standard addition method.
First order spectroscopy (D1):
The individual zero-order absorption spectra of Loratadine so obtained were converted into their first-order derivative spectra with the help of inbuilt software of the instrument for all four methods. The resultant derivative spectrum has shown minima and maxima in all four methods and therefore the amplitude was selected for the construction of calibration curves for Methods I, II, III, and IV.
Assay of Loratadine Pharmaceutical dosage form:
10mg of Loratadine tablet (equivalent to 10 mg of Loratadine) was pipetted into a 10 mL volumetric flask containing methanol and sonicated. The dilution of concentration 10µg/mL was prepared in borate buffer (pH 9.0) (Method I), phosphate buffer (pH 5.8) (Method II), phosphate buffer (pH 7.8) (Method III), and distilled water (Method IV) from the stock solution and the assay was carried out and the percentage recovery was calculated.
RESULTS AND DISCUSSION:
Two different techniques – Zero-order (D0) and First-order derivative spectroscopy (D1) have been developed for the determination of Loratadine in tablets in four different reagents such as borate buffer (pH 9.0) phosphate buffer (pH 5.8, 7.8), and distilled water.
Zero order spectroscopy (D0):
Loratadine has shown absorption maxima at 247 nm for Methods I, II, III, and IV. It obeys Beer-Lambert’s law (Table 1) over the concentration range of 0.5-50 in borate buffer (pH 9.0), phosphate buffer (pH 5.8, 7.8), and distilled water. The linear regression equations were found to be y=0.0424x-0.0067 (0.9999), y=0.0426-0.0042 (0.9999), y=0.0439x-0.0208 (0.9994), y=0.0403x-0.0127 (0.9997) for borate buffer (pH 4.0), phosphate buffer (pH 5.8, 7.8) and distilled water respectively. The absorption spectrum of Loratadine (Do) is given in Figure 2.
Table 1: Linearity of Loratadine (Zero order spectroscopy)
|
Conc. (µg/ml) |
Absorbance |
|||
|
Methods |
I |
II |
III |
IV |
|
0.5 |
0.0176 |
0.0162 |
0.0073 |
0.0175 |
|
1 |
0.0434 |
0.0406 |
0.0356 |
0.0296 |
|
2 |
0.0795 |
0.0881 |
0.0636 |
0.0647 |
|
5 |
0.1972 |
0.2101 |
0.1905 |
0.1795 |
|
10 |
0.4092 |
0.4123 |
0.4106 |
0.3616 |
|
20 |
0.8311 |
0.8365 |
0.8189 |
0.8093 |
|
40 |
- |
- |
- |
1.6001 |
|
50 |
2.1171 |
2.1297 |
2.1902 |
2.0014 |
*Mean of three replicates
|
|
|
|
Method I (Borate buffer pH 9.0) |
Method II (Phosphate buffer pH 5.8) |
|
|
|
|
Method III (Phosphate buffer pH 7.8) |
Method IV (Distilled water) |
Figure 2: Absorption spectra of Loratadine (10 µg/mL) (D0)
|
|
|
|
Method I (Borate buffer pH 9.0) |
Method II (Phosphate buffer pH 5.8) |
|
|
|
|
Method III (Phosphate buffer pH 7.8) |
Method IV (Distilled water) |
Figure 3: Calibration Curve of Loratadine (D0)
First-order derivative spectroscopy (D1)
The overlay first-order derivative spectra of Loratadine in borate buffer (pH 9.0) (Method I), phosphate buffer (pH 5.8) (Method II), phosphate buffer (pH 7.8) (Method III), and distilled water (Method IV) shown in Figure 3.
The derivative spectrum has shown both maxima and minima and therefore the amplitude was taken against concentration and calibration curves were drawn. Loratadine obeys Beer-Lambert’s law (Table 2) over the concentration range of 10-50 µg/mL for borate buffer (pH 9.0), phosphate buffer (pH 5.8, 7.8), and distilled water respectively and the linear regression equations that are found to be y= 0.002x-0.0015, y= 0.0016x+0.0044, y=0.002x-0.0037, y=0.0016x-0.0133 respectively.
The percentage RSD in accuracy (Table 3) and precision studies (Table 4-5) for all the methods were found to be less than 2 indicating that the methods are precise and accurate. The assay was and the percentage recovery was calculated (Table 6). The optical characteristics of the method are shown in Table 7.
|
|
|
|
Method I (Borate buffer pH 9.0) |
Method II (Phosphate buffer pH 5.8) |
|
|
|
|
Method III (Phosphate buffer pH 7.8) |
Method IV (Distilled water) |
Figure 4: Overlay of first-order derivative absorption spectra of Loratadine (D1)
|
|
|
|
Method I (Borate buffer pH 9.0) |
Method II (Phosphate buffer pH 5.8) |
|
|
|
|
Method III (Phosphate buffer pH 7.8) |
Method IV (Distilled water) |
Figure 5: Calibration Curve of Loratadine (D1)
Table 2: Linearity of Loratadine (First derivative spectroscopy) (Max: Maxima; Min: Minima)
|
Conc. (µg/mL) |
Method I |
Method II |
|||||
|
Max (239.18 nm) |
Min (262nm) |
Amplitude |
Max (238.64 nm) |
Min (260 nm) |
Amplitude |
||
|
10 |
0.0051 |
0.0138 |
0.0189 |
0.0062 |
0.0045 |
0.0105 |
|
|
20 |
0.0096 |
0.0281 |
0.0377 |
0.0127 |
0.0136 |
0.0263 |
|
|
30 |
0.0187 |
0.0404 |
0.0591 |
0.0228 |
0.0193 |
0.0421 |
|
|
40 |
0.0264 |
0.0524 |
0.0788 |
0.0324 |
0.0268 |
0.0592 |
|
|
50 |
0.0348 |
0.0636 |
0.0984 |
0.0429 |
0.0307 |
0.0736 |
|
|
Conc. (µg/mL) |
Method III |
Method IV |
|||||
|
Max (240 nm) |
Min (280 nm) |
Amplitude |
Max (238.67 nm) |
Min (256.33 nm) |
Amplitude |
||
|
10 |
0.0067 |
0.0085 |
0.0152 |
0.0027 |
0.0071 |
0.0098 |
|
|
20 |
0.0102 |
0.0249 |
0.0351 |
0.0109 |
0.0137 |
0.0246 |
|
|
30 |
0.0248 |
0.0316 |
0.0564 |
0.0172 |
0.0211 |
0.0383 |
|
|
40 |
0.0388 |
0.0377 |
0.0765 |
0.0252 |
0.0276 |
0.0528 |
|
|
50 |
0.0429 |
0.0544 |
0.0973 |
0.0356 |
0.0318 |
0.0674 |
|
Table 3: Accuracy of Loratadine Pharmaceutical dosage form
|
Zero order spectroscopy |
||||||
|
Spiked Conc. (µg/mL) |
Formu lation (µg/mL) |
Total Conc. (µg/mL) |
*Conc. obtained (μg/mL) [% Recovery] (RSD) |
|||
|
I |
II |
III |
IV |
|||
|
0.5 (50%) |
10 |
15 |
14.73 [98.25] (0.06) |
14.83 [98.87] (0.06) |
14.94 [99.65] (0.04) |
14.97 [99.83] (0.02) |
|
1 (100%) |
10 |
20 |
19.99 [99.97] (0.03) |
19.85 [99.28] (0.04) |
19.70 [98.50] (0.03) |
19.95 [99.75] (0.03) |
|
1.5 (150%) |
10 |
25 |
24.54 [98.18] (0.02) |
24.56 [98.24] (0.03) |
24.75 [99.01] (0.03) |
24.88 [99.54] (0.02) |
|
First Derivative spectroscopy |
||||||
|
Spiked Conc. (µg/mL) |
Formu lation (µg/mL) |
Total Conc. (µg/mL) |
Conc. obtained (μg/mL) [% Recovery] (RSD) |
|||
|
I |
II |
III |
IV |
|||
|
0.5 (50%) |
10 |
15 |
14.45 [96.33] (0.53) |
15.56 [103.75] (0.60) |
14.95 [99.66] (0.33) |
14.88 [99.23] (0.50) |
|
1(100%) |
10 |
20 |
19.65 [98.25] (0.42) |
19.81 [99.06] (0.63) |
19.55 [97.75] (0.39) |
20.35 [101.7] (0.91) |
|
1.5 (150%) |
10 |
25 |
24.45 [97.80] (0.31) |
12.43 [99.45] (0.74) |
25.45 [101.8] (0.29) |
25.35 [101.4] (0.58) |
*Mean of three replicates
Table 4: Precision study of Loratadine Pharmaceutical dosage form (Zero order spectroscopy)
|
Intraday precision |
||||
|
Conc. (µg/mL) |
Absorbance |
|||
|
I |
II |
III |
IV |
|
|
10 |
0.4092 |
0.4123 |
0.4106 |
0.3616 |
|
10 |
0.4095 |
0.4128 |
0.4104 |
0.3612 |
|
10 |
0.4091 |
0.4121 |
0.4101 |
0.3617 |
|
10 |
0.4096 |
0.4126 |
0.4105 |
0.3615 |
|
10 |
0.4090 |
0.4122 |
0.4102 |
0.3618 |
|
10 |
0.4093 |
0.4125 |
0.4109 |
0.3611 |
|
*Mean ± S.D (% RSD) |
0.4092± 0.0002 (0.05) |
0.4124 ± 0.0002 (0.03) |
0.4140 ± 0.0002(0.69) |
0.3614 ± 0.0002(0.07) |
|
Inter-day precision |
||||
|
Conc. (µg/mL) |
Absorbance |
|
||
|
I |
II |
III |
IV |
|
|
10 |
0.4092 |
0.4123 |
0.4106 |
0.3616 |
|
10 |
0.4098 |
0.4127 |
0.4102 |
0.3612 |
|
10 |
0.4095 |
0.4125 |
0.4109 |
0.3619 |
|
Mean ± S.D (% RSD) |
0.4095 ± 0.0003 (0.07) |
0.4125 ± 0.0002 (0.04) |
0.4105 ± 0.0003 (0.08) |
0.3615 ± 0.0003 (0.09) |
*Mean of three replicates
Table 5: Precision study of Loratadine Pharmaceutical dosage form (First derivative spectroscopy)
|
Intraday precision |
||||
|
Conc. (µg/mL) |
Absorbance |
|||
|
I |
II |
III |
IV |
|
|
10 |
0.0189 |
0.0105 |
0.0152 |
0.0098 |
|
10 |
0.0187 |
0.0107 |
0.0154 |
0.0097 |
|
10 |
0.0185 |
0.0104 |
0.0156 |
0.0099 |
|
10 |
0.0188 |
0.0101 |
0.0154 |
0.0091 |
|
10 |
0.0184 |
0.0103 |
0.0125 |
0.0093 |
|
10 |
0.0186 |
0.0106 |
0.0151 |
0.0094 |
|
*Mean ± S.D (% RSD) |
0.0186 ± 0.0001 (1.00) |
0.0204 ± 0.0001 (1.15) |
0.0153 ± 0.0001 (1.19) |
0.0095 ± 0.0001 (1.04) |
|
Inter-day precision |
|
|||
|
Conc. (µg/mL) |
Absorbance |
|
||
|
I |
II |
III |
IV |
|
|
10 |
0.0189 |
0.0105 |
0.0152 |
0.0098 |
|
10 |
0.0184 |
0.0101 |
0.0154 |
0.0097 |
|
10 |
0.0188 |
0.0104 |
0.0156 |
0.0099 |
|
Mean ± S.D (% RSD) |
0.0187 ± 0.0002 (1.41) |
0.0103 ± 0.0002 (1.94) |
0.0154 ± 0.0002 (1.29) |
0.0098 ± 0.0001 (1.02) |
*Mean of three replicates
Table 6: Assay of Loratadine Pharmaceutical dosage form
|
Brand |
Zero order spectroscopy |
|||||||
|
Method I Borate buffer (pH 9.0) |
Method II Phosphate buffer (pH 5.8) |
Method III Phosphate buffer (pH 7.8) |
Method IV Distilled water |
|||||
|
*Observed amount (µg) |
% Recovery |
*Observed amount (µg) |
% Recovery |
*Observed Amount (µg) |
% Recovery |
*Observed Amount (µg) |
% Recovery |
|
|
I |
9.955 |
99.55 |
10.17 |
101.7 |
10.27 |
102.7 |
10.11 |
101.1 |
|
II |
9.824 |
98.24 |
10.22 |
102.2 |
10.15 |
101.5 |
10.14 |
101.4 |
|
|
First derivative spectroscopy |
|||||||
|
Method I Borate buffer (pH 9.0) |
Method II Phosphate buffer (pH 5.8) |
Method III Phosphate buffer (pH 7.8) |
Method IV Distilled water |
|||||
|
*Observed amount (µg) |
% Recovery |
*Observed amount (µg) |
% Recovery |
*Observed Amount (µg) |
% Recovery |
*Observed Amount (µg) |
% Recovery |
|
|
I |
10.15 |
101.5 |
9.937 |
99.37 |
9.950 |
99.50 |
9.928 |
99.28 |
|
II |
10.27 |
102.7 |
9.812 |
98.12 |
9.937 |
99.37 |
9.854 |
98.54 |
*Mean of three replicates
Table 7: Optical characteristics of Loratadine – Zero order spectroscopy
|
Parameters |
Methods |
||||
|
I |
II |
III |
IV |
||
|
Linearity range (µg/mL) |
0.5 – 50 |
0.5 – 50 |
0.5 – 50 |
0.5 – 50 |
|
|
λmax (nm) |
247 |
247 |
247 |
247 |
|
|
Molar extinction coefficient (litre/mole/cm-1) |
1.6234×104 |
1.4010×104 |
1.4437×104 |
1.3253×104 |
|
|
Sandell’s sensitivity (µg/cm2/0.001 absorbance unit) |
0.1816 |
1.2206 |
0.4965 |
3.3995 |
|
|
Slope |
0.0424 |
0.0426 |
0.0439 |
0.0403 |
|
|
Intercept |
0.0067 |
0.0042 |
0.0208 |
0.0127 |
|
|
Correlation coefficient |
0.9999 |
0.9999 |
0.9994 |
0.9997 |
|
|
Precision (%RSD) |
Intraday |
0.06 |
0.06 |
0.06 |
0.07 |
|
Interday |
0.009-0.073 |
0.007-0.048 |
0.006-0.085 |
0.007-0.018 |
|
|
Accuracy (% RSD) |
0.02-0.06 |
0.03-0.06 |
0.03-0.34 |
0.02-0.03 |
|
|
Assay (%) |
Brand I |
99.55 |
101.7 |
102.7 |
101.1 |
|
Brand II |
98.24 |
102.2 |
101.5 |
101.4 |
|
CONCLUSION:
All the spectrophotometric techniques were validated and found to be very simple, accurate, precise, and economical. These methods can be conveniently used for the routine analysis of Loratadine in Pharmaceutical formulations.
REFERENCES:
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2. Noor Jahan et al., Spectrophotometric determination of Loratadine in bulk and pharmaceutical dosage form. International Journal of Pharma Sciences and Research. 2018; 9(5): 65-73.
3. Georgeta Pavalachea et al., Determination of Loratadine in pharmaceuticals by a spectrophotometric method. Ovidius University Annals of Chemistry. 2015; 26(1): 27-31.
4. Prathap B et al., Analytical method development and validation for the estimation of Loratadine using UV spectrophotometer. World Journal of Pharmaceutical Research. 2023; 12(6): 701-714.
5. Heshitha P and Mathrusri Annapurna M. A review on analytical techniques for the assay of Loratadine. Acta Scientific Pharmaceutical Sciences. 2023; 7(12): 59-62.
6. International Conference on Harmonization, Q2A: Text on Validation of Analytical Procedures. 2005.
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Received on 10.07.2024 Revised on 22.09.2024 Accepted on 08.11.2024 Published on 24.12.2024 Available online from December 27, 2024 Research J. Pharmacy and Technology. 2024;17(12):5984-5990. DOI: 10.52711/0974-360X.2024.00908 © RJPT All right reserved
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