{"id":55302,"date":"2024-03-20T11:02:30","date_gmt":"2024-03-20T11:02:30","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=55302"},"modified":"2024-04-01T19:32:55","modified_gmt":"2024-04-01T19:32:55","slug":"method-development-and-validation-for-estimation-of-bedaquiline-and-in-tablet-dosage-form-by-hptlc-and-rp-hplc-study","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no1\/method-development-and-validation-for-estimation-of-bedaquiline-and-in-tablet-dosage-form-by-hptlc-and-rp-hplc-study\/","title":{"rendered":"Method Development and Validation for Estimation of Bedaquiline and in Tablet Dosage form by Hptlc and Rp-Hplc Study"},"content":{"rendered":"\n

Introduction<\/strong><\/p>\n\n\n\n

BDQ\nChemically known as (1R, 2S) 1(6Bromo-2methoxy-3quinolyl) 4-dimethylamino-2(1-\nnaphthyl) 1-phenylbutan-2-ol. The structure of BDQ is shown in fig1. A novel\nbactericidal anti-mycobacterial drug called bedaquiline. Multidrug-resistant\ntuberculosis is treated with this diarylquinoline anti-mycobacterial medication\nby inhibiting mycobacterial ATP synthase, which generates energy for\nMycobacterium tuberculosis; it inhibits the proton pump. Bedaquiline causes\nbacterial death. Its molecular formula is C32<\/sub>H31<\/sub>BrN2<\/sub>O2<\/sub>,\nand its molecular weight is 555.504 gm per mol1,2<\/sup>. A recent\nliterature review reported that LC-MS and MS had been employed to analyze\nBedaquiline and Rifabutin in human plasma3,4<\/sup>. A Liquid\nchromatography-Mass spectroscopy method has been reported to be useful in\ndetermining bedaquiline concentrations in human serum5-7<\/sup>. Bedaquiline\nwas enantio-separated by polar stationary phases made of polysaccharides8<\/sup>.\nA pharmacokinetic study on bedaquiline9,10 \n<\/sup>and the physical and chemical sustainability of bedaquiline in\nhydrolytic systems at various pH levels were evaluated by isolating eight\nvariation analog diastereomers by HPLC. They assessed the degradation kinetics\nof bedaquiline in the solution with UV spectrophotometric methods11,12<\/sup>.\nHPLC can simultaneously determine the levels of bedaquiline and endearing in\nplasma samples13<\/sup>. We have reported on the simultaneous\nenumeration and quantification by reversed Phase HPLC14<\/sup> of\nbedaquiline (TMC-207), moxifloxacin, and pyrazinamide formulation. As a result\nof the literature review, we concluded that it is essential to develop\nanalytical methods that are more sensitive, least expensive, take up less time,\nand use less drug quantity to estimate bedaquiline bulk and its tablet dosage\nform. As a result, the goal of this work is to provide a precise, accurate, and\nrepeatable method for calculating the concentration of bedaquiline utilizing\nHPTLC, HPLC, and UV spectrophotometric techniques. <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 1: Structure for Bedaquiline.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Experimental<\/strong><\/p>\n\n\n\n

Chemicals and reagents<\/strong><\/p>\n\n\n\n

Sirturo tablets labeled\nto contain Bedaquiline 100 mg. methanol (HPLC grade),\nglacial acetic acid, sodium acetate, ethanol, chloroform, and toluene were purchased\nfrom Loba chem (Mumbai,\nIndia). Throughout the project,\ncalibrated glassware was used. All the reagents used in this study were AR\nGrades.<\/p>\n\n\n\n

Instrumentation<\/strong><\/p>\n\n\n\n

In TLC, Silica gel 60\n(F254) is coated on an aluminum sheet, and in HPTLC, a Sample applicator\n(Linomat-V) with a 100L Hamilton syringe. The plates were developed in a 20 cm\nx 10 cm CAMAG twin trough tank that had been soaked with the extraction solvent\nfor 30 minutes prior to production. An RP-HPLC system with a serial dual\nplunger pump (micro-volume ten microliters on the primary side, five\nmicroliters on the secondary side) and a PDA detector with an injector were\ndeveloped. As computation, Shimadzu lab solution software was used. A novel UV-\nspectrophotometry using an Ultra Violet visible double beam spectrophotometer\nhas spectral widths of a nanometer and wavelengths accurate to 0.5 nanometer.\n10 mm matched quartz cells in pairs were used. The digital scale Shimadzu\nAUX220 is utilized for weighing purpose15<\/sup>.<\/p>\n\n\n\n

Preparation of stock\nsolutions<\/strong><\/p>\n\n\n\n

Standard Solution<\/strong><\/p>\n\n\n\n

One hundred milligrams of\nBDQ were accurately weighed and transferred into One hundred milligram\nvolumetric flasks. Methanol adds to\ndissolve. After ten minutes\nof sonicating, the volume of the solution was topped up with methanol to\nachieve 1000 mg per ml of BDQ concentration. <\/p>\n\n\n\n

Sample solution<\/strong><\/p>\n\n\n\n

Weighing 10 tablets accurately and (Sirturo100\nmg tablets), approximately the exact weight of the powdered tablet was identified.\nFor dissolving the tablet powder, methanol was added to a flask containing one\nhundred milligrams of BDQ. Following this, the solution was sonicated for ten minutes.\nMethanol was used to makeup and then filter paper (Whatman -41) was used for\nfiltering purposes. The finished solution contained 1000 \u03bcg per ml.<\/p>\n\n\n\n

HPTLC Method<\/strong><\/p>\n\n\n\n

1, 1.5, 2.0, 2.5,\nand 3.0 ml of the 1000 \u00b5g\/ml standard stock solution were pipetted into a 10 ml\nvolumetric flask. The final concentration was 100, 150, 200, 250, and 300\nmicrograms per ml. Solutions were spotted on a pre-coated aluminum Plate 60\nF254 using a CAMAG1000 \u00b5l sample applicator syringe. The plate for 20 mins placed\nin an oven at the temperature of 110o<\/sup>C before spotting the samples\nfor activation. There was a constant flow rate of 0.1L per second, with a space\nbetween two bands of 10mm, and a constant application rate of 0.1L per\nsecond. Previously, ethanol: chloroform: toluene (6:3.5:0.05 v\/v\/v) was\npre-saturated for 30 min in a twin trough chamber. To a distance of eight\ncm, it took 15 to 20 minutes to develop the plate. As a result of the\ndevelopment of the plate, it has been dried through passing air from the dryer.\nIn order to scan the spots at 10 mm per second, they were slit at 5 mm by 0.45\nmm as they developed at 240 nm16<\/sup>.<\/p>\n\n\n\n

HPLC Method<\/strong><\/p>\n\n\n\n

Chromatographic\nconditions <\/strong><\/p>\n\n\n\n

The stationary phase C18 column was used in this method, and Buffer (Sodium acetate at pH-4.5)\nsolvent phase. The samples flow at a speed of 1ml per min. Whattmann filter\npaper was used to filter the mobile phase. A detector (PDA) detects the\ncompounds at 240nm. Under vacuum, Acetonitrile was mixed, degassed, and passed to 0.45\nmicron pore to filter.<\/p>\n\n\n\n

Mobile Phase Preparation<\/strong><\/p>\n\n\n\n

About 700 ml of sodium acetate buffer (pH range 4.5) and\n300 ml of methanol in an ultrasonic water bath were used to mix and degas the\nmixture for 5min. A 0.45 pore filter was used to separate the filtrate, which\nwas transferred under vacuum into a volumetric flask containing 1 litre of\nfiltrate.<\/p>\n\n\n\n

Preparation of Calibration\nCurve solution<\/strong><\/p>\n\n\n\n

A 10 ml standard flask was filled with 0.5 ml of standard\nstock solution (1000 g\/ml). Methanol (100g per ml) was then added to the\nsolution, and from above said concentration (100 g per ml), 0.5, 1.0, 1.5, 2.0,\nand 2.5 ml pipetted out and transferred to the separate volumetric flask of\n10ml capacity. Final concentrations ranged from 5 to 25 g per ml.<\/p>\n\n\n\n

UV \u2013 Visible Spectrophotometric Method<\/strong><\/p>\n\n\n\n

Preparation of Stock solution for calibration<\/strong><\/p>\n\n\n\n

In a standard flask of 250 ml, 5.0 ml of the stock\nsolution (1000 g per ml) was pipetted and transferred into the standard flask. The solution\nwas made up to volume using methanol (20 g per ml). Using a pipette, 4, 8, 12,\n16, and 20 ml solutions were pipetted from above the concentration (250g per\nml) into 10 ml standard flasks. The resulting concentrations were 8 to 40 g per\nml.<\/p>\n\n\n\n

Validation <\/strong><\/p>\n\n\n\n

The method validation was conducted per International Conference on Harmonization Guidelines15-16<\/sup>.<\/p>\n\n\n\n

Linearity and Range<\/strong><\/p>\n\n\n\n

     HPTLC calibration curve of\nBDQ was prepared using methanol solutions containing 100, 150, 200, 250, and\n300 mg per ml. To calculate the regression equations for the calibration curve,\npeak areas were plotted against BDQ concentrations. There were five\ndeterminations in each response. To calibrate BDQ with RP\u2013HPLC, various\nconcentration ranges, such as 5 to 25\u00b5g\/ml, has been made by diluting the stock\nsolution (standard). Under operating chromatographic conditions, an aliquot\n(10\u00b5l) of each solution was injected. UV-visible spectrophotometry was\nperformed in methanol with BDQ solutions containing 8, 16, 24, 32, and 40 mg\nper ml of BDQ and measured at 240 nm. By plotting absorbance versus\nconcentration, calibration curves are constructed.<\/p>\n\n\n\n

LOQ and LOD<\/strong><\/p>\n\n\n\n

We calculated the detection and quantification limit, by SD and calibration curve. ICH guidelines estimated LOD and LOQ. LOD=3.3\u00d7\u03c3\/S and LOQ=10\u00d7\u03c3\/S, where \u03c3 is the standard deviation ‘S’ is the slope.<\/p>\n\n\n\n

Content Estimation (Assay)<\/strong><\/p>\n\n\n\n

For content estimation, Sirturo100 mg tablets were used. The sample stock solutions were prepared for each HPTLC, HPLC, and UV method from the standard stock. Samples and standards were spotted for HPTLC. A chromatogram was recorded after injecting 20 liters of standard and sample solutions. Standard and sample absorbance were measured for the UV method. The purity percentage was calculated using peak area and absorbance.<\/p>\n\n\n\n

Accuracy                                                                             <\/strong><\/p>\n\n\n\n

BDQ recovery was calculated at three levels (50, 100, and 150%) to determine the method’s accuracy. We added known quantities of standard solutions of BDQ 100, 200, and 300g per ml to a pre-analyzed sample solution containing 15\u00b5g per ml BDQ before spotting 10\u00b5L of each solution on an activated silica plate. A known amount of standard BDQ 7.5, 15, 22.5\u00b5g per ml was mixed with pre-analyzed sample solution containing 15\u00b5g BDQ for performing reverse phase HPLC. To pre-analyze sample solutions containing 30\u00b5g per ml, known amounts of BDQ standard solution were added.<\/p>\n\n\n\n

Precision<\/strong><\/p>\n\n\n\n

A procedure’s use circumstances will determine how much\nintermediate precision is required. For the HPTLC method, 200\u00b5g per ml was used\nfor the triplicate determination. For the HPLC method, 15\u00b5g per ml\nconcentration solution was used. The peak area was measured after the injection\nof 20\u00b5l of solution. The UV method found 24 \u00b5g per ml concentration solution\nabsorbance at 240 nm.<\/p>\n\n\n\n

Ruggedness<\/strong><\/p>\n\n\n\n

In\nthis definition, reproducibility means that test results can be replicated no\nmatter where they are performed, what analyzers are used, what instruments are\nused, and how long it takes to run the experiment.<\/p>\n\n\n\n

Results<\/strong><\/p>\n\n\n\n

Precision, accuracy, and reproducibility were optimized for HPTLC, HPLC, and UV methods. Toluene: Acetonitrile, methanol: water, and methanol: chloroform ratios were used for HPTLC. Observe spreading and tailing spots. To overcome this problem, we used ethanol-chloroform-toluene (6:3.5:0.5). There was a sharp, symmetrical peak with Rf 0.63 (Fig 2). Different trials for HPLC were performed using different ratios of mobile phases (Water: Acetonitrile (55: 45) & Methanol: H2<\/sub>O 50: 50), Sodium acetate buffer (3.0): methanol 70%:30%), and it has flowed in range of 0.8, 1.0, 1.2ml per min. There was no tailing of peak and theoretical plate numbers within the limit (above 2000). In the end, sodium acetate buffer: & methanol (70:30%) was used. Chromatograms showed sharp peaks, and system suitability parameters were within limits. Fig 3 shows the optimized chromatogram. The solvent for the UV method was selected from the solubility study. A concentration solution of 10g per ml was prepared and scanned between 200-400nm. Fig 4 shows the UV spectrum at 240nm with maximum absorbance.<\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 2: Optimized Chromatogram by HPTLC method<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 3: Optimized Chromatogram by HPLC method.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 4: UV Spectrum of Bedaquiline<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Linearity<\/strong><\/p>\n\n\n\n

The\nrange of analytes in a sample depends on the concentrations of analytes at\ntheir highest and lowest concentrations. In the case of HPTLC, a linear\ncorrelation was observed between peak areas and concentrations in the range (of\n100 to 300g\/ml) when the HPTLC method was used. The correlation coefficient\ncame out to be r2<\/sup>=0.9992. A linear concentration value for the\nHPLC method was 0.9994 \u00b5g per ml, with a range from 5-25 \u00b5g per ml, and the r2<\/sup>\nvalue was 0.9994 \u00b5g per ml. There was linearity in concentrations between\n8 and 40 micrograms\/ml of UV solution for the method, and a correlation\ncoefficient value of 0.9997 was identified. Figures 5-7 show the calibration\ncurves for HPTLC, HPLC, and UV methods. Table 1 shows linearity reports.    <\/p>\n\n\n\n\n\n
\"\"<\/td>\n\n

Figure 5: Calibration Curve for Bedaquiline by HPTLC method.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 6: Calibration Curve for Bedaquiline by HPLC method.<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n\n
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Figure 7: calibration curve for Bedaquiline by UV method<\/strong><\/p>\n

<\/p>\n

Click here to view Figure<\/a><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Table 1: Reports for Linearity study<\/strong>.<\/p>\n\n\n\n\n\n\n
\n

Parameters<\/strong><\/p>\n<\/td>\n

\n

HPTLC<\/strong><\/p>\n<\/td>\n

\n

HPLC<\/strong><\/p>\n<\/td>\n

\n

UV<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Range( \u00b5g\/ml)<\/p>\n

y = mx + c<\/p>\n

r2<\/p>\n

Slope (m) Intercept (c) LOD (\u00b5g\/ml)<\/p>\n

   LOQ(\u00b5g\/ml)<\/p>\n<\/td>\n

\n

150-300\u00b5g\/ml<\/p>\n

y = 16.773x + 60.979 0.9992<\/p>\n

16.773<\/p>\n

60.979<\/p>\n

0.9081<\/p>\n

0.1640<\/p>\n<\/td>\n

\n

5-25 \u00b5g\/ml<\/p>\n

y = 20387x + 5289.3 0.9994<\/p>\n

20387<\/p>\n

5289.3<\/p>\n

0.0070<\/p>\n

0.0212<\/p>\n<\/td>\n

\n

8-40 \u00b5g\/ml<\/p>\n

y = 0.014x + 0.0055 0.9997<\/p>\n

0.014<\/p>\n

0.0055<\/p>\n

0.1791<\/p>\n

0.5428<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

LOD and LOD <\/strong><\/p>\n\n\n\n

It was determined that the LOD and LOQ were 0.9081 \u00b5g per ml, 0.1640\u00b5g per ml, 0.0070\u00b5g per ml, 0.0212\u00b5g per ml and 0.1791\u00b5g per ml, 0.542\u00b5g per ml for HPTLC, HPLC and UV methods respectively. The report is shown in table 1.<\/p>\n\n\n\n

Content Estimation <\/strong><\/p>\n\n\n\n

It was determined that BDQ is pure in tablet formulation by an assay (content estimation). An estimate of BDQ concentration from a calibration curve was selected and quantified. The tablet formulation Sirturo (100 mg tablets) was used for identification, and the purity of analytes present in the formulation was analyzed from 100.37 – 100.78% for the proposed methods. The % RSD values were 0.2048, 0.3678, and 1.4235 for the proposed methods. Table 2 shows the result.<\/p>\n\n\n\n

Table 2: Content Estimation of Bedaquiline in marketed formulation by proposed methods.<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Methods<\/strong><\/p>\n<\/td>\n

\n

Standard peak area<\/strong><\/p>\n<\/td>\n

\n

Sample peak area<\/strong><\/p>\n<\/td>\n

\n

Mean (%)<\/strong><\/p>\n<\/td>\n

\n

SD<\/strong><\/p>\n<\/td>\n

\n

(%) RSD<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

HPTLC<\/strong><\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n<\/td>\n\n<\/td>\n\n<\/td>\n<\/tr>\n
\n

1<\/p>\n<\/td>\n

\n

3447.8<\/p>\n<\/td>\n

\n

3453.3<\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n<\/td>\n<\/tr>\n
\n

2<\/p>\n<\/td>\n

\n

3475.3<\/p>\n<\/td>\n

\n

3481.4<\/p>\n<\/td>\n

\n

100.37<\/p>\n<\/td>\n

\n

0.2056<\/p>\n<\/td>\n

\n

0.2048<\/p>\n<\/td>\n<\/tr>\n

\n

3<\/p>\n<\/td>\n

\n

3463.2<\/p>\n<\/td>\n

\n

3470.3<\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n<\/td>\n<\/tr>\n
\n

Average<\/p>\n<\/td>\n

\n

3462.1<\/p>\n<\/td>\n

\n

3475.3<\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n<\/td>\n<\/tr>\n
\n

HPLC<\/strong><\/p>\n<\/td>\n

\n

314092<\/p>\n<\/td>\n

\n

315223<\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n<\/td>\n<\/tr>\n
\n

1<\/p>\n<\/td>\n

\n

315324<\/p>\n<\/td>\n

\n

318401<\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n<\/td>\n<\/tr>\n
\n

2<\/p>\n<\/td>\n

\n

314962<\/p>\n<\/td>\n

\n

318230<\/p>\n<\/td>\n

\n

100.78<\/p>\n<\/td>\n

\n

0.3707<\/p>\n<\/td>\n

\n

0.3678<\/p>\n<\/td>\n<\/tr>\n

\n

3<\/p>\n

Average<\/p>\n<\/td>\n

\n

314792.6<\/p>\n<\/td>\n

\n

317284.6<\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n

\n<\/p><\/td>\n<\/tr>\n

 <\/td>\n\n

Absorbance of <\/strong>standard
<\/strong>solution<\/strong><\/p>\n<\/td>\n

\n

Absorbance of <\/strong>sample solution<\/strong><\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n<\/td>\n<\/tr>\n
\n

UV<\/strong><\/p>\n

1<\/p>\n

2<\/p>\n

3<\/p>\n

Average<\/p>\n<\/td>\n

\n

0.346<\/p>\n

0.353<\/p>\n

0.340<\/p>\n

0.346<\/p>\n<\/td>\n

\n

0.342<\/p>\n

0.357<\/p>\n

0.345<\/p>\n

0.348<\/p>\n<\/td>\n

\n<\/td>\n\n<\/td>\n\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n

Precision<\/strong><\/p>\n\n\n\n

A study of intermediate precision confirmed the method’s precision. The analyses of standards were carried out. The % of RSD of the intermediate precision analysis of Bedaquiline was found to be 0.7657, 0.0871, and 1.585 % for HPTLC, RP-HPLC, and UV, respectively. The reports are shown in table -3<\/p>\n\n\n\n

Table 3: Data for Intermediate Precision by proposed methods<\/strong><\/p>\n\n\n\n\n\n\n
\n

S.No<\/strong><\/p>\n<\/td>\n

\n

HPTLC<\/strong><\/p>\n<\/td>\n

\n

HPLC<\/strong><\/p>\n<\/td>\n

\n

UV<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

1<\/p>\n

2<\/p>\n

3<\/p>\n

4<\/p>\n

5<\/p>\n

6<\/p>\n

Avg SD<\/p>\n

%RSD<\/p>\n<\/td>\n

\n

3447.8<\/p>\n

3463.6<\/p>\n

3476.5<\/p>\n

3404.7<\/p>\n

3423.8<\/p>\n

3452.9<\/p>\n

3444.8<\/p>\n

26.38<\/p>\n

0.7657<\/p>\n<\/td>\n

\n

314268<\/p>\n

314394<\/p>\n

314942<\/p>\n

314727<\/p>\n

314904<\/p>\n

314749<\/p>\n

314664<\/p>\n

274.15<\/p>\n

0.0871<\/p>\n<\/td>\n

\n

0.346<\/p>\n

0.348<\/p>\n

0.353<\/p>\n

0.333<\/p>\n

0.340<\/p>\n

0.354<\/p>\n

0.347<\/p>\n

0.0055<\/p>\n

1.585<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Accuracy<\/strong><\/p>\n\n\n\n

The formulation solution previously analyzed was diluted with pure drugs to evaluate the method’s accuracy. To analyze the mixture, a method was proposed. A calculation was made to determine the amount of drug recovered. Our study found that the percentage recovery for HPTLC, RP-HPLC, and UV methods was 1000.13% to 100.63%. RSDs were found to be less than 2%. Table 4 shows the results.<\/p>\n\n\n\n

Table 4: Data for Accuracy study for the proposed methods<\/strong>.<\/p>\n\n\n\n\n\n\n\n\n
\n

Various Methods<\/strong><\/p>\n

 <\/strong><\/p>\n<\/td>\n

\n

Concentration (%)<\/strong><\/p>\n<\/td>\n

\n

Added Amount<\/strong><\/p>\n

\u00b5g\/ml<\/strong><\/p>\n<\/td>\n

\n

Amount Found<\/strong><\/p>\n

\u00b5g\/ml<\/strong><\/p>\n<\/td>\n

\n

Recovery<\/strong><\/p>\n

Mean (%)<\/strong><\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

SD<\/strong><\/p>\n<\/td>\n

\n

RSD (%)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

HPTLC<\/p>\n<\/td>\n

\n

50<\/p>\n

100<\/p>\n

150<\/p>\n<\/td>\n

\n

100.00<\/p>\n

200.00<\/p>\n

300.00<\/p>\n<\/td>\n

\n

90.10<\/p>\n

180.21<\/p>\n

270.45<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

100.13<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

0.0307<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

0.0305<\/p>\n<\/td>\n<\/tr>\n

\n

HPLC<\/p>\n<\/td>\n

\n

50<\/p>\n

100<\/p>\n

150<\/p>\n<\/td>\n

\n

4.00<\/p>\n

8.00<\/p>\n

12.00<\/p>\n<\/td>\n

\n

3.98<\/p>\n

8.12<\/p>\n

11.95<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

100.16<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

1.1547<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

1.1527<\/p>\n<\/td>\n<\/tr>\n

\n

U0056<\/p>\n<\/td>\n

\n

50<\/p>\n

100<\/p>\n

150<\/p>\n<\/td>\n

\n

15.00<\/p>\n

30.00<\/p>\n

45.00<\/p>\n<\/td>\n

\n

15.09<\/p>\n

30.32<\/p>\n

45.11<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

100.63<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

0.4110<\/p>\n<\/td>\n

\n

 <\/strong><\/p>\n

0.4082<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Ruggedness<\/strong><\/p>\n\n\n\n

A ruggedness validation was performed on the developed method. An independent lab is rugged if it can maintain high precision over multiple days while using multiple analysts, instruments, and reagents from different sources. The results are shown in Table 5. The % of RSD was 1.283, 0.643, and 0.994% for HPTLC, HPLC, and UV, respectively.<\/p>\n\n\n\n

Table 5: Data for Ruggedness study<\/strong>.<\/p>\n\n\n\n\n\n\n\n\n
\n

Method<\/strong><\/p>\n<\/td>\n

\n

Different conditions<\/strong><\/p>\n<\/td>\n

\n

Average Percentage<\/strong><\/p>\n<\/td>\n

\n

SD<\/strong><\/p>\n<\/td>\n

\n

(%) RSD<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

 <\/strong>HPTLC<\/strong><\/p>\n<\/td>\n

\n

Analyst-I Analyst-II<\/p>\n<\/td>\n

\n

101.10<\/p>\n

99.28<\/p>\n<\/td>\n

\n

1.286<\/p>\n<\/td>\n

\n

1.283<\/p>\n<\/td>\n<\/tr>\n

\n

HPLC<\/strong><\/p>\n<\/td>\n

\n

Analyst-I Analyst-II<\/p>\n<\/td>\n

\n

98.42<\/p>\n

99.32<\/p>\n<\/td>\n

\n

0.636<\/p>\n<\/td>\n

\n

0.643<\/p>\n<\/td>\n<\/tr>\n

\n

UV<\/strong><\/p>\n<\/td>\n

\n

Analyst-I Analyst-II<\/p>\n<\/td>\n

\n

99.43<\/p>\n

98.04<\/p>\n<\/td>\n

\n

0.982<\/p>\n<\/td>\n

\n

0.994<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Discussion<\/strong><\/p>\n\n\n\n

The\nquantitation and detection limits are shallow, indicating that the method is\nsensitive. Compared with the developed method, the reported Reverse Phase HPLC11<\/sup>\nand Ultra Violet13<\/sup> methods had higher LOD and LOQ values. Therefore,\nthe developed method was more sensitive than the reported method (Table -6).<\/p>\n\n\n\n

% RSD value below 1.3, considered acceptable, validated the robustness and repeatability of the method. (Acceptance criteria \u2013 not more than 2 %). Precision and repeatability were good for the developed method. <\/strong>For the developed RP-HPTLC, HPLC, and UV spectroscopic methods, the RSD values were less than 2%. RSD values were not affected by excipients used in formulations. As a result, it can be concluded that the method is accurate. This resulted in the methods being more accurate as a result of this. With the three methods, the estimation of drugs was accurate, precise, and rapid. HPLC was more sensitive than the other methods when comparing the sensitivity of the three methods since the LOD and LOQ values in HPTLC and UV methods were lower than those of HPLC and UV. Therefore, it is concluded that all three methods can be successfully applied for the routine analysis of the quality <\/strong>control of the BDQ in bulk and as tablets for dosage forms as part of the routine quality control process.<\/p>\n\n\n\n

Table 6: Comparison between the reported method and advanced method<\/strong>.<\/p>\n\n\n\n\n\n\n\n
\n

HPLC and UV<\/strong><\/p>\n<\/td>\n

\n

Reported method (11&13)<\/strong><\/p>\n<\/td>\n

\n

Developed method<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Linearity<\/strong><\/p>\n

LOD LOQ<\/strong><\/p>\n<\/td>\n

\n

10-100 \u00b5g\/ml<\/p>\n

0.3272\u00b5g\/ml 0.9525\u00b5g\/ml<\/p>\n<\/td>\n

\n

5-25 \u00b5g\/ml<\/p>\n

0.0070<\/p>\n

0.0212<\/p>\n<\/td>\n<\/tr>\n

\n

Linearity<\/strong><\/p>\n

LOD LOQ<\/strong><\/p>\n<\/td>\n

\n

15-75 \u00b5g\/ml<\/p>\n

2.72 \u00b5g\/ml<\/p>\n

8.23 \u00b5g\/ml<\/p>\n<\/td>\n

\n

8-40 \u00b5g\/ml<\/p>\n

0.1791<\/p>\n

0.5428<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Conclusion<\/strong><\/p>\n\n\n\n

A simple, accurate, and\nreproducible quantitative analysis of BDQ in tablet form was developed and\nvalidated by HPTLC, reversed-phase HPLC, and UV visible spectrophotometry.\nAccording to the analysis results, the concentration range was linear, and the\nvalue of the correlation coefficient was r2<\/sup><0.999 for the three\nmethods. Compared to the developed method, the reported RP-HPLC and UV methods\nhad higher linearity ranges. Thus, by utilizing the developed method, it is\npossible to estimate BDQ when the least amount of drug needs to be\nadministered. <\/p>\n\n\n\n

Acknowledgment<\/strong><\/p>\n\n\n\n

We acknowledge the management of Vels Institute of Science, Technology and Advanced Studies (VISTAS), Pallavaram, Chennai-600 117, Tamil Nadu, India, for providing a research facility.<\/p>\n\n\n\n

Conflict of Interest<\/strong><\/p>\n\n\n\n

We declared that there are no Conflicts of Interest.<\/p>\n\n\n\n

Funding Support<\/p>\n\n\n\n

There is no funding sources.<\/p>\n\n\n\n

References<\/strong><\/p>\n\n\n\n

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    Introduction BDQ Chemically known as (1R, 2S) 1(6Bromo-2methoxy-3quinolyl) 4-dimethylamino-2(1- naphthyl) […]<\/p>\n","protected":false},"author":15,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[113],"tags":[],"class_list":["post-55302","post","type-post","status-publish","format-standard","hentry","category-vol17no1"],"_links":{"self":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/55302","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/users\/15"}],"replies":[{"embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/comments?post=55302"}],"version-history":[{"count":5,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/55302\/revisions"}],"predecessor-version":[{"id":57448,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/posts\/55302\/revisions\/57448"}],"wp:attachment":[{"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/media?parent=55302"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/categories?post=55302"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/biomedpharmajournal.org\/staging\/wp-json\/wp\/v2\/tags?post=55302"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}