{"id":61920,"date":"2024-12-30T10:44:17","date_gmt":"2024-12-30T10:44:17","guid":{"rendered":"https:\/\/biomedpharmajournal.org\/?p=61920"},"modified":"2025-01-06T19:00:09","modified_gmt":"2025-01-06T19:00:09","slug":"innovative-rp-hplc-technique-for-method-development-and-validation-of-propylthiouracil-tablets","status":"publish","type":"post","link":"https:\/\/biomedpharmajournal.org\/staging\/vol17no4\/innovative-rp-hplc-technique-for-method-development-and-validation-of-propylthiouracil-tablets\/","title":{"rendered":"Innovative RP-HPLC Technique for Method Development and Validation of Propylthiouracil Tablets"},"content":{"rendered":"\n

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

The\npharmaceutical industry represents companies that manufacture morale and\nmedication over the counter. Pharmaceutical research focuses on the discovery\nand development of pharmaceutical agents for administration\u23af whether\nself-administered by patients\nor provided by healthcare professionals\u23af for\ntreatment, protection, or prevention. As the field advances, innovative and\nskilled formulations are being introduced to the market. Although some dosage\nforms may be susceptible to contamination, many are highly effective. Such\ndevelopments require precise, easy-to-use, and flexible chemical analysis\ntechniques due to the significance of quality control in pharmaceuticals. Unlike\nother consumer goods, medicines must adhere to strict standards because they\ndirectly affect human health. Ensuring quality requires a dedicated department\nfor quality control and assurance [1]<\/sup>. The medication does not\nreduce the efficiency of oral or injectable thyroid Propylthiouracil is useful\nin treating hyperthyroidism because it prevents the production of thyroid\nhormone treatments, nor does it neutralize the thyroxine or triiodothyronine\npresent in the bloodstream. Propylthiouracil works by inhibiting the conversion\nof thyroxine to triiodothyronine in peripheral tissues, making it a possible\noption for managing thyroid storm. The drug is rapidly absorbed and extensively\nmetabolized. Within 24 hours, approximately 35% of the medication is excreted\nin the urine, both in its intact and conjugated forms [2]<\/sup>. The\nprimary goal of analytical chromatography is to obtain target analytes with a\nsufficient resolution in the shortest possible time. The solutes diffuse slowly\nin and out of the tiny pores in the nonporous particles, which eliminates the\nstagnant mobile phase in the intra-particle void volume [3]<\/sup>. Validation\nrefers to assessing validity and efficiency. The validation group consists of\nindividuals from various regulations. Validation is the process of providing\nwritten proof that ensures the product meets the standards for analysis\napplications [4]<\/sup>.<\/p>\n\n\n\n

This\narticle presents the systematic approach taken in the quantitative analysis for\nthe validation of propylthiouracil tablets by using reverse phase HPLC. The\nstudy elaborates on the criteria applied for selecting methods, optimization\nperformed for chromatographic conditions, parameters checked during the validation\nprocess, and analysis of real-world samples. This research will furnish\ndetailed information at each step involved in development and validation to act\nas a comprehensive guide for analysts and researchers working in the\npharmaceutical field.<\/p>\n\n\n\n

Materials and Methods<\/strong><\/p>\n\n\n\n

Chemicals and Reagents Used<\/strong><\/p>\n\n\n\n

All\nchemicals used in the analysis should be either AR grade or an equivalent\ngrade; solvents should be HPLC grade or equivalent. Monobasic potassium phosphate,\nsodium hydroxide, orthophosphoric acid, acetonitrile, methanol, and HPLC-grade\nmilliQ water.<\/p>\n\n\n\n

Working Standard and Sample Used<\/strong><\/p>\n\n\n\n

Propylthiouracil\nstandard, Propylthiouracil 50 mg tablets, and Placebo for propylthiouracil 50 mg\ntablets were given by Bio Plus Life Science Pvt Ltd., Hosur, Tamil Nadu, India.<\/p>\n\n\n\n

Instruments Used<\/strong><\/p>\n\n\n\n

Analytical\nand precision balance from Denver Instrument, HPLC with openlab software from\nAgilent 1200 and Agilent 1260 Infinity II, ultrasonicator from PCI Analytics,\nand pH meter from Weibfr.<\/p>\n\n\n\n

Preparation of Solution 5, 6<\/sup>:<\/strong><\/p>\n\n\n\n

Preparation of Buffer<\/strong><\/p>\n\n\n\n

In a 1000 mL beaker, 3.4 g of monobasic potassium phosphate was added with 500 mL of water and then sonicated to dissolve. After adjusting to a pH of 4.6\u00b1 0.05. A 0.1 N of sodium hydroxide or diluted phosphoric acid was added with 500 mL of water. A pH of 4.6\u00b1 0.05 was achieved by adding 0.1 N of sodium hydroxide or diluted phosphoric acid to the resultant solution. A 0.45\u00b5 membrane filter was used to filter this mixture, then degassed.<\/p>\n\n\n\n

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

In a ratio of 20:80 (v\/v), a suitable amount of degassed acetonitrile and buffer were prepared.<\/p>\n\n\n\n

Preparation of Diluent<\/strong><\/p>\n\n\n\n

Use 1% methanol in water as diluent.<\/p>\n\n\n\n

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

About 50 mg of the working standard for propylthiouracil was accurately weighed and transferred into a 50 mLdry volumetric flask. Then, 10 mL of methanol was added, sonicated to dissolve it, and water was added to fill the flask to the recommended volume. A 5 mL of this solution was diluted to 100 mL with water and mixed.<\/p>\n\n\n\n

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

A minimum of 20 tablets was weighed to determine the averageweight. The tablets should be crushed into a fine powder. The powder weight was taken and added to a 100 mL clean, dry flask. Next, 20 mL of methanol was added and sonicated for 5 minutes. After that, 50 mL of water was added and sonicated for 15 minutes with intermittent shaking. The mixture was cooled and made up the volume with water. Using a 4.5\u00b5 membrane filter (PVDF), the mixture was filtered. The first 4 mL of the filtrate was discarded. Further, 5 mL of the solution was diluted to 1000 mL with water and mixed.<\/p>\n\n\n\n

Preparation of Placebo<\/strong><\/p>\n\n\n\n

A 100 mg equivalent of propylthiouracil placebo powder was accurately weighted and transferred into a 100 mL clean, dry volumetric flask, methanol (20 mL) was added and sonicated for 5 minutes, and then 50 mL of water was added, sonicated for 15 minutes, cooled, and made up to the flask water and mixed. Using a 0.45\u00b5 membrane filter (PVDF)for filtration. Further, 5 mL of the solution was diluted to 100 mL with water and mixed.<\/p>\n\n\n\n

Calculation for % Assay<\/strong><\/p>\n\n\n\n

Calculation of the percentage assay for propylthiouracil by using the following formula.<\/p>\n\n\n\n\n\n\n
\n

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

\n

=<\/p>\n<\/td>\n

\n

AT<\/sub><\/p>\n<\/td>\n

\n

X<\/p>\n<\/td>\n

\n

Ws<\/p>\n<\/td>\n

\n

X<\/p>\n<\/td>\n

\n

5<\/p>\n<\/td>\n

\n

X<\/p>\n<\/td>\n

\n

100<\/p>\n<\/td>\n

\n

X<\/p>\n<\/td>\n

\n

100<\/p>\n<\/td>\n

\n

X<\/p>\n<\/td>\n

\n

P<\/p>\n<\/td>\n

\n

X<\/p>\n<\/td>\n

\n

Aw<\/sub><\/p>\n<\/td>\n

\n

X<\/p>\n<\/td>\n

\n

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

\n

As<\/p>\n<\/td>\n

\n

50<\/p>\n<\/td>\n

\n

100<\/p>\n<\/td>\n

\n

WT<\/sub><\/p>\n<\/td>\n

\n

5<\/p>\n<\/td>\n

\n

100<\/p>\n<\/td>\n

\n

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

Where,\n<\/p>\n\n\n\n

AT <\/sub>= Average area of propylthiouracil peak in the chromatogram of sample solution<\/p>\n\n\n\n

As = Average area of propylthiouracil peak in the chromatogram of standard solution, as obtain under system suitability.<\/p>\n\n\n\n

Ws = Weight of working standard taken in mg<\/p>\n\n\n\n

P = Percent potency of working standard used (as is basis)<\/p>\n\n\n\n

WT <\/sub>= Weight of sample in mg<\/p>\n\n\n\n

AW <\/sub>= Average weight in mg<\/p>\n\n\n\n

L = Label claim in mg<\/p>\n\n\n\n

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

Method Validation 7-13<\/sup><\/strong><\/p>\n\n\n\n

Specificity <\/strong><\/p>\n\n\n\n

Blank, placebo, standard,\ncontrol test solution, spiked test solution, and individual impurity solution\nrelated to propylthiouracil were prepared and analysed as per the method. The\nchromatograms show there is no interference from the blank, placebo and the\nknown impurity with a retention time of peak due to propylthiouracil. Results\nare shown in Tables 1-4.<\/p>\n\n\n\n

Table 1: results obtained from standard, control and spiked test solution <\/strong><\/p>\n\n\n\n\n\n\n\n\n
\n

Sample ID<\/strong><\/p>\n<\/td>\n

\n

Retention Time of Propylthiouracil peak ( in minutes)<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Standard solution<\/p>\n<\/td>\n

\n

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

\n

Test solution (Control)<\/p>\n<\/td>\n

\n

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

\n

Spiked test solution<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

Table 2: Results obtained from individual impurities solution<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\n

Impurity Name<\/strong><\/p>\n<\/td>\n

\n

Retention time (minutes)<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

Thiourea Impurity<\/p>\n<\/td>\n

\n

Not Detected in 272 nm<\/p>\n<\/td>\n<\/tr>\n

\n

Thiourea Impurity<\/p>\n<\/td>\n

\n

238 nm RT-1.18<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Table 3: Results obtained from control and spiked test solution<\/strong><\/p>\n\n\n\n\n\n\n\n
\n

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

\n

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

\n

Absolute difference in % Assay values<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Test solution (Control)<\/p>\n<\/td>\n

\n

98.2<\/p>\n<\/td>\n

\n

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

\n

Spiked test solution<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

Table 4: Results obtained for peak purity of Propylthiouracil<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n
\n

Sample ID<\/strong><\/p>\n<\/td>\n

\n

Purity factor<\/strong><\/p>\n<\/td>\n

\n

Result<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

Blank solution<\/p>\n<\/td>\n

\n

within the threshold limit<\/p>\n<\/td>\n

\n

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

\n

Placebo solution<\/p>\n<\/td>\n

\n

within the threshold limit<\/p>\n<\/td>\n

\n

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

\n

Standard solution<\/p>\n<\/td>\n

\n

within the threshold limit<\/p>\n<\/td>\n

\n

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

\n

Test solution (Control)<\/p>\n<\/td>\n

\n

within the threshold limit<\/p>\n<\/td>\n

\n

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

\n

Spiked test solution<\/p>\n<\/td>\n

\n

within the threshold limit<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 1: Chromatogram of specificity for standard solution with peak purity<\/strong><\/p>\n<\/p>\n

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

Figure 2: Chromatogram of specificity for sample solution with peak purity<\/strong><\/p>\n<\/p>\n

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

Forced degradation study<\/strong><\/p>\n\n\n\n

Forced\ndegradation study in propylthiouracil 50 mg tablets and its placebo. No peak\nwas detected at the retention time of propylthiouracil in the chromatogram of\ndiluent and placebo. Forced degradation studies are shown in Table 5.<\/p>\n\n\n\n

Table 5: Forced degradation of Propylthiouracil 50mg tablets results below<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Mode of degradation<\/strong><\/p>\n<\/td>\n

\n

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

\n

% Assay<\/strong><\/p>\n<\/td>\n

\n

% Degradation<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

Control sample<\/p>\n<\/td>\n

\n

Not applicable<\/p>\n<\/td>\n

\n

100.6<\/p>\n<\/td>\n

\n

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

\n

Acid degradation<\/p>\n<\/td>\n

\n

5mL of 5N HCl, at 80\u00b0C for 60 minutes<\/p>\n<\/td>\n

\n

97.7<\/p>\n<\/td>\n

\n

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

\n

Base degradation<\/p>\n<\/td>\n

\n

5mL of 5N NaOH, at 80\u00b0C for 60 minutes<\/p>\n<\/td>\n

\n

91.6<\/p>\n<\/td>\n

\n

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

\n

Photolytic degradation<\/p>\n<\/td>\n

\n

1.2million lux hours\/200watt hours square meter<\/p>\n<\/td>\n

\n

98.4<\/p>\n<\/td>\n

\n

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

\n

Thermal degradation<\/p>\n<\/td>\n

\n

3 days at 105 \u00b0C<\/p>\n<\/td>\n

\n

100.9<\/p>\n<\/td>\n

\n

No degradation<\/p>\n<\/td>\n<\/tr>\n

\n

Control sample<\/p>\n<\/td>\n

\n

Not applicable<\/p>\n<\/td>\n

\n

100.2<\/p>\n<\/td>\n

\n

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

\n

Oxidation degradation<\/p>\n<\/td>\n

\n

5 mL of 5% v\/v H2<\/sub>O2,<\/sub>bench top for 30 minutes<\/p>\n<\/td>\n

\n

93.1<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

Filter Study <\/strong><\/p>\n\n\n\n

Using the analysis method as an outline, standard and sample solutions are prepared. 0.45\u00b5m Nylon and 0.45\u00b5m PVDF filters were used to filter the standard solution as well as the sample solution. By discarding 2 mL, 4 mL, and 6 mL of the solution after it has passed through the previous filter, three sub-fractions will be gathered. Unfiltered standard solution is used as the standard. Results obtained by using different syringe filters and unfiltered standard solutions are summarised in Tables 6 and 7.<\/p>\n\n\n\n

Table 6: Results obtained from filter saturation study for standard<\/strong> solution<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Filter size and type<\/strong><\/p>\n<\/td>\n

\n

Quantity Discarded in ml<\/strong><\/p>\n<\/td>\n

\n

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

\n

Absolute  % difference<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

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

\n

Unfiltered standard solution<\/p>\n<\/td>\n

\n

NA<\/p>\n<\/td>\n

\n

5733363<\/p>\n<\/td>\n

\n

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

\n

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

\n

0.45 \u00b5m Nylon<\/p>\n<\/td>\n

\n

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

\n

5723530<\/p>\n<\/td>\n

\n

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

\n

4 ml<\/p>\n<\/td>\n

\n

5719118<\/p>\n<\/td>\n

\n

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

\n

6 ml<\/p>\n<\/td>\n

\n

5720648<\/p>\n<\/td>\n

\n

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

\n

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

\n

0.45 \u00b5m PVDF<\/p>\n<\/td>\n

\n

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

\n

5729777<\/p>\n<\/td>\n

\n

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

\n

4 ml<\/p>\n<\/td>\n

\n

5734088<\/p>\n<\/td>\n

\n

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

\n

6 ml<\/p>\n<\/td>\n

\n

5731705<\/p>\n<\/td>\n

\n

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

Table 7: Results obtained from filter saturation study for sample solution<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Filter size and type<\/strong><\/p>\n<\/td>\n

\n

Quantity Discarded in ml<\/strong><\/p>\n<\/td>\n

\n

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

\n

Assay %<\/strong><\/p>\n<\/td>\n

\n

Absolute % difference<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

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

\n

Unfiltered sample solution<\/p>\n<\/td>\n

\n

NA<\/p>\n<\/td>\n

\n

5651723<\/p>\n<\/td>\n

\n

99.0<\/p>\n<\/td>\n

\n

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

\n

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

\n

0.45 \u00b5m<\/p>\n

Nylon<\/p>\n<\/td>\n

\n

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

\n

5658626<\/p>\n<\/td>\n

\n

99.1<\/p>\n<\/td>\n

\n

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

\n

4 ml<\/p>\n<\/td>\n

\n

5652407<\/p>\n<\/td>\n

\n

99.0<\/p>\n<\/td>\n

\n

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

\n

6 ml<\/p>\n<\/td>\n

\n

5682430<\/p>\n<\/td>\n

\n

99.6<\/p>\n<\/td>\n

\n

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

\n

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

\n

0.45 \u00b5m<\/p>\n

PVDF<\/p>\n<\/td>\n

\n

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

\n

5660532<\/p>\n<\/td>\n

\n

99.2<\/p>\n<\/td>\n

\n

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

\n

4 ml<\/p>\n<\/td>\n

\n

5667759<\/p>\n<\/td>\n

\n

99.3<\/p>\n<\/td>\n

\n

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

\n

6 ml<\/p>\n<\/td>\n

\n

5661367<\/p>\n<\/td>\n

\n

99.2<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

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

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

According\nto the test protocol, a standard solution was prepared and stored at room\ntemperature (23\u00b0<\/strong>C- 27\u00b0C). The study\nof the stability of the solution on various days when compared to freshly made\nstandard solution is necessary. The results are shown in Table 8.<\/p>\n\n\n\n

Table 8: Results of standard solution stability at 23\u00b0C-27\u00b0C<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Standard solution stability at 23\u00b0C-27\u00b0C<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Similarity factor<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Initial<\/p>\n<\/td>\n

\n

5728333<\/p>\n<\/td>\n

\n

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

\n

24 hours<\/p>\n<\/td>\n

\n

5719319<\/p>\n<\/td>\n

\n

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

\n

48 hours<\/p>\n<\/td>\n

\n

5714535<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

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

According\nto the test method, the sample solution was prepared and stored at room\ntemperature (23\u00b0C- 27\u00b0C). The study of the stability of the solution at various\ndays when compared to a freshly made standard solution is necessary. The\nresults are shown in Table 9.<\/p>\n\n\n\n

Table 9: Results of sample<\/strong> solution stability at (23- 27) \u00b0C<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Sample solution stability at 23\u00b0C-27\u00b0C<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

% Assay<\/strong><\/p>\n<\/td>\n

\n

Absolute % difference<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Initial<\/p>\n<\/td>\n

\n

5703944<\/p>\n<\/td>\n

\n

99.9<\/p>\n<\/td>\n

\n

Not Applicable<\/p>\n<\/td>\n<\/tr>\n

\n

24 hours<\/p>\n<\/td>\n

\n

5648200<\/p>\n<\/td>\n

\n

98.4<\/p>\n<\/td>\n

\n

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

\n

48 hours<\/p>\n<\/td>\n

\n

5648391<\/p>\n<\/td>\n

\n

98.2<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

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

A\nserious number of solutions containing propylthiouracil at concentrations\nlisted below were analysed to determine the linearity of the method. The\nresponse of linearity for propylthiouracil is determined from the range of\n24.916 \u00b5g\/ml to 74.748 \u00b5g\/ml. Results are illustrated in Table 10 and the\ncorresponding Figure 3- 8.<\/p>\n\n\n\n

Table 10: Results obtained for linearity<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Linearity Solution No<\/strong><\/p>\n<\/td>\n

\n

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

\n

Concentration (\u00b5g\/mL)<\/strong><\/p>\n<\/td>\n

\n

Average Area of Propylthiouracil<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

50%<\/p>\n<\/td>\n

\n

24.916<\/p>\n<\/td>\n

\n

2765964<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

75%<\/p>\n<\/td>\n

\n

37.374<\/p>\n<\/td>\n

\n

4110765<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

100%<\/p>\n<\/td>\n

\n

49.832<\/p>\n<\/td>\n

\n

5441493<\/p>\n<\/td>\n<\/tr>\n

\n

4<\/p>\n<\/td>\n

\n

125%<\/p>\n<\/td>\n

\n

62.290<\/p>\n<\/td>\n

\n

6828044<\/p>\n<\/td>\n<\/tr>\n

\n

5<\/p>\n<\/td>\n

\n

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

\n

74.748<\/p>\n<\/td>\n

\n

8171130<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

108585.7361<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

52434.80000<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Correlation  coefficient (r)<\/strong><\/p>\n<\/td>\n

\n

0.99998<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

% Deviation of Y-Intercept<\/strong><\/p>\n<\/td>\n

\n

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

\n

Regression co-efficient (r2<\/sup>)<\/strong><\/p>\n<\/td>\n

\n

0.99996<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 3: Linearity plot of Propylthiouracil<\/strong><\/p>\n<\/p>\n

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

Figure 4: linearity 50%<\/strong><\/p>\n<\/p>\n

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

Figure 5: Linearity 75%<\/strong><\/p>\n<\/p>\n

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

Figure 6: Linearity 100%<\/strong><\/p>\n<\/p>\n

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

Figure 7: Linearity 125%<\/strong><\/p>\n<\/p>\n

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

Figure 8: Linearity 150%<\/strong><\/p>\n<\/p>\n

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

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

The\naccuracy is determined by examining the solutions of placebo spiked with\npropylthiouracil active pharmaceutical ingredient (API) at three different\nlevels of concentrations in triplicate. The solutions were prepared and analysed\nas per the method. % recovery results are shown on Table 11 and corresponding\nFigure 9-11.<\/p>\n\n\n\n

Table 11: Results obtained for Propylthiouracil 50mg tablets (% Recovery)<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

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

\n

Amount of Propylthiouracil added (mg)<\/strong><\/p>\n<\/td>\n

\n

Amount of Propylthiouracil found (mg)<\/strong><\/p>\n<\/td>\n

\n

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

\n

% Mean  Recovery<\/strong><\/p>\n<\/td>\n

\n

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

\n

50 %<\/p>\n<\/td>\n

\n

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

\n

50.86<\/p>\n<\/td>\n

\n

50.813<\/p>\n<\/td>\n

\n

99.9<\/p>\n<\/td>\n

\n

100.2<\/p>\n<\/td>\n

\n

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

\n

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

\n

50.09<\/p>\n<\/td>\n

\n

50.353<\/p>\n<\/td>\n

\n

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

\n

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

\n

50.24<\/p>\n<\/td>\n

\n

50.356<\/p>\n<\/td>\n

\n

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

\n

100%<\/p>\n<\/td>\n

\n

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

\n

100.55<\/p>\n<\/td>\n

\n

100.558<\/p>\n<\/td>\n

\n

100.0<\/p>\n<\/td>\n

\n

100.1<\/p>\n<\/td>\n

\n

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

\n

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

\n

100.75<\/p>\n<\/td>\n

\n

100.617<\/p>\n<\/td>\n

\n

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

\n

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

\n

100.49<\/p>\n<\/td>\n

\n

100.811<\/p>\n<\/td>\n

\n

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

\n

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

\n

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

\n

150.30<\/p>\n<\/td>\n

\n

149.693<\/p>\n<\/td>\n

\n

99.6<\/p>\n<\/td>\n

\n

99.6<\/p>\n<\/td>\n

\n

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

\n

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

\n

150.12<\/p>\n<\/td>\n

\n

149.242<\/p>\n<\/td>\n

\n

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

\n

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

\n

150.49<\/p>\n<\/td>\n

\n

150.216<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n\n
\"\"<\/td>\n\n

Figure 9: Accuracy 50% level-1<\/strong><\/p>\n<\/p>\n

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

Figure 10: Accuracy 100% level-1<\/strong><\/p>\n<\/p>\n

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

Figure 11: Accuracy 150% level-1<\/strong><\/p>\n<\/p>\n

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

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

Repeatability\nof standard solution- System precision  <\/strong><\/p>\n\n\n\n

Repeatability solution -1 was\ninjected into the chromatographic system five times in replicates. Results are\nsummarised in Table 12.<\/p>\n\n\n\n

Table 12: Results obtained for five replicate injections of standard solution<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Area of  Propylthiouracil<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

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

\n

5762988<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

5763640<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

5761356<\/p>\n<\/td>\n<\/tr>\n

\n

4<\/p>\n<\/td>\n

\n

5764861<\/p>\n<\/td>\n<\/tr>\n

\n

5<\/p>\n<\/td>\n

\n

5781881<\/p>\n<\/td>\n<\/tr>\n

\n

Mean<\/p>\n<\/td>\n

\n

5766945<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

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

 <\/p>\n\n\n

Reproducibility of sample\nsolution- Method precision <\/strong><\/p>\n\n\n\n

Six test solutions of a single batch\nwere analysed as per the proposed methodology and the results as given in Table\n13.<\/p>\n\n\n\n

Table 13: Results obtained from six sample preparation from Method precision -Assay: Analyst 1<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

Propylthiouracil % assay<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

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

\n

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

\n

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

\n

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

\n

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

\n

4<\/p>\n<\/td>\n

\n

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

\n

5<\/p>\n<\/td>\n

\n

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

\n

6<\/p>\n<\/td>\n

\n

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

\n

Mean<\/p>\n<\/td>\n

\n

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

\n

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

\n

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

 <\/p>\n\n\n

Intermediate precision \u2013\nRuggedness<\/strong><\/p>\n\n\n\n

By comparing the analysis of the\npropylthiouracil 50 mg tablet samples conducted on different days by different\nanalysts using different instruments and columns, the robustness of the method\nwas verified. The outcomes of Analyst 2\u2019s examination of the sample solution are\ndisplayed in Table 14. Table 15 shows the mean, standard deviation, and\npercentage RSD for the two sets of data.<\/p>\n\n\n\n

Table 14: Results obtained from six sample preparation of intermediate precision -Assay: Analyst 2<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

% Assay<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

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

\n

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

\n

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

\n

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

\n

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

\n

4<\/p>\n<\/td>\n

\n

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

\n

5<\/p>\n<\/td>\n

\n

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

\n

6<\/p>\n<\/td>\n

\n

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

\n

Mean<\/p>\n<\/td>\n

\n

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

\n

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

\n

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

 <\/p>\n\n\n

Table 15: Overall compilations of method precision and intermediate precision: Assay<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Sample<\/strong> Number<\/strong><\/p>\n<\/td>\n

\n

% Assay<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Intermediate precision<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

99.8<\/p>\n<\/td>\n

\n

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

\n

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

\n

99.9<\/p>\n<\/td>\n

\n

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

\n

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

\n

100.0<\/p>\n<\/td>\n

\n

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

\n

4<\/p>\n<\/td>\n

\n

99.1<\/p>\n<\/td>\n

\n

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

\n

5<\/p>\n<\/td>\n

\n

99.0<\/p>\n<\/td>\n

\n

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

\n

6<\/p>\n<\/td>\n

\n

98.6<\/p>\n<\/td>\n

\n

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

\n

Mean<\/p>\n<\/td>\n

\n

99.4<\/p>\n<\/td>\n

\n

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

\n

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

\n

0.6<\/p>\n<\/td>\n

\n

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

\n

Overall mean<\/p>\n<\/td>\n

\n

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

\n

Overall RSD (%)<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

Robustness<\/strong><\/p>\n\n\n\n

By\nverifying that the system suitability parameters were met and deliberately\nchanging variables such as flow rate, organic content of the mobile phase\ncomposition, pH variation and detection wavelength, the procedure\u2019s robustness\nwas tested. Every condition was examined for a sample solution.\nPropylthiouracil assay percentage and system suitability were determined. Altered\nparameters from an optimised method for the robustness of the assay and results\nare tabulated from Tables 16 and 18.<\/p>\n\n\n\n

Table 16: Altered parameters from optimized method for robustness of assay<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n
\n

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

\n

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

\n

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

\n

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

\n

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

\n

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

\n

Flow rate \u00b1 0.1 mL\/min<\/p>\n<\/td>\n

\n

1.0 mL\/min<\/p>\n<\/td>\n

\n

0.9 mL\/min<\/p>\n<\/td>\n

\n

1.1 mL\/min<\/p>\n<\/td>\n<\/tr>\n

\n

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

\n

Wavelength \u00b1 2 nm<\/p>\n<\/td>\n

\n

272 nm<\/p>\n<\/td>\n

\n

270 nm<\/p>\n<\/td>\n

\n

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

\n

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

\n

Buffer  pH \u00b1 0.1<\/p>\n<\/td>\n

\n

4.6<\/p>\n<\/td>\n

\n

4.5<\/p>\n<\/td>\n

\n

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

\n

4<\/p>\n<\/td>\n

\n

Mobile phase organic \u00b1 2%<\/p>\n<\/td>\n

\n

ACN: buffer (200:800)<\/p>\n<\/td>\n

\n

ACN: buffer (196:804)<\/p>\n<\/td>\n

\n

ACN: buffer (204:796)<\/p>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n

 <\/p>\n\n\n

Effect\nof variation in flow rate, organic variation and <\/strong>wavelength variation:<\/strong><\/p>\n\n\n\n

Table 17: System suitability for flow rate variation, organic variation and wavelength variation<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

Flow Variations<\/strong><\/p>\n<\/td>\n

\n

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

\n

Column Efficiency<\/strong><\/p>\n<\/td>\n

\n

Tailing factor<\/strong><\/p>\n<\/td>\n

\n

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

\n

% Assay<\/strong><\/p>\n<\/td>\n

\n

Absolute % Difference<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Unchanged (1.0 mL\/min)<\/p>\n<\/td>\n

\n

0.2<\/p>\n<\/td>\n

\n

73186<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.2<\/p>\n<\/td>\n

\n

98.8<\/p>\n<\/td>\n

\n

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

\n

High Flow rate 1.1 mL\/min<\/p>\n<\/td>\n

\n

0.2<\/p>\n<\/td>\n

\n

63286<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.3<\/p>\n<\/td>\n

\n

98.2<\/p>\n<\/td>\n

\n

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

\n

Low Flow rate 0.9 mL\/min<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

74076<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.2<\/p>\n<\/td>\n

\n

98.3<\/p>\n<\/td>\n

\n

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

\n

High M.P organic (\uff0b2%)<\/p>\n<\/td>\n

\n

0.0<\/p>\n<\/td>\n

\n

71688<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.5<\/p>\n<\/td>\n

\n

98.4<\/p>\n<\/td>\n

\n

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

\n

Low M.P organic (- 2%)<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

70992<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.4<\/p>\n<\/td>\n

\n

98.2<\/p>\n<\/td>\n

\n

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

\n

High Wavelength (274nm)<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

73350<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.2<\/p>\n<\/td>\n

\n

98.3<\/p>\n<\/td>\n

\n

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

\n

Low Wavelength (270nm)<\/p>\n<\/td>\n

\n

0.2<\/p>\n<\/td>\n

\n

73029<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.2<\/p>\n<\/td>\n

\n

98.2<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

Effect of variation in buffer pH<\/strong><\/strong><\/p>\n\n\n\n

Table 18: System suitability from variation in buffer pH<\/strong><\/p>\n\n\n\n\n\n\n\n\n
\n

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

\n

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

\n

Column Efficiency<\/strong><\/p>\n<\/td>\n

\n

Tailing factor<\/strong><\/p>\n<\/td>\n

\n

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

\n

% Assay<\/strong><\/p>\n<\/td>\n

\n

Absolute % Difference<\/strong><\/p>\n<\/td>\n<\/tr>\n

\n

Unchanged (pH 4.6)<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

67533<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.6<\/p>\n<\/td>\n

\n

98.4<\/p>\n<\/td>\n

\n

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

\n

High pH (pH 4.7)<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

73033<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.5<\/p>\n<\/td>\n

\n

98.4<\/p>\n<\/td>\n

\n

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

\n

Low pH (pH 4.5)<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

66959<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

100.5<\/p>\n<\/td>\n

\n

98.3<\/p>\n<\/td>\n

\n

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

 <\/p>\n\n\n

System Suitability<\/strong><\/p>\n\n\n\n

During\nthe validation investigation, a standard solution was injected on various days.\nThe propylthiouracil peak was calculated from a standard solution using system\nsuitability software. Using standard solution-1, the percentage RSD of the area\nratio of the five replicate injections for the propylthiouracil peak was\ndetermined. % similarity factor calculated from standard solution-2 against\nstandard solution-1. Data is shown in Table 18. A typical set of system\nsuitability chromatograms is shown in the appendix.<\/p>\n\n\n\n

Table 19: System suitability observed on different days<\/strong><\/p>\n\n\n\n\n\n\n\n\n\n\n\n\n\n
\n

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

\n

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

\n

Asymmetry factor<\/strong><\/p>\n<\/td>\n

\n

Plate count<\/strong><\/p>\n<\/td>\n

\n

% Recovery<\/strong><\/p>\n<\/td>\n<\/tr>\n<\/thead>\n

\n

Specificity<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

68693<\/p>\n<\/td>\n

\n

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

\n

Forced degradation<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

69139<\/p>\n<\/td>\n

\n

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

\n

Method precision and Filter study<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

1.1<\/p>\n<\/td>\n

\n

65511<\/p>\n<\/td>\n

\n

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

\n

Linearity<\/p>\n<\/td>\n

\n

0.2<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

67151<\/p>\n<\/td>\n

\n

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

\n

Accuracy<\/p>\n<\/td>\n

\n

0.2<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

65918<\/p>\n<\/td>\n

\n

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

\n

Intermediate Precision<\/p>\n<\/td>\n

\n

0.1<\/p>\n<\/td>\n

\n

1.1<\/p>\n<\/td>\n

\n

61400<\/p>\n<\/td>\n

\n

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

\n

Robustness<\/p>\n<\/td>\n

\n

0.2<\/p>\n<\/td>\n

\n

1.0<\/p>\n<\/td>\n

\n

73186<\/p>\n<\/td>\n

\n

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

\u00a0<\/p>\n\n\n

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

The\nabsolute difference between the % assay of spiked and un-spiked samples meets\nthe acceptance criteria, and the peak purity plot of propylthiouracil peak from\nstandard, control and spiked test solutions shows that the peak is homogenous\nand no co-eluting peaks are seen in specificity. The peak purity plot of the propylthiouracil\npeak from degradation sample solutions indicates that the results are within\nthe threshold limits. Based on the above data, a 0.45\u00b5m PVDF filter is suitable\nfor standard and sample preparation, and discard the 4 ml of filtrate. The\nstandard and sample solutions are stable up to 48 hours at ambient temperature\n(23\u00b0C-27\u00b0C). The data response in\nlinear throughout the working range. The method\u2019s acceptable recovery\nlevel is between 50% and 150% of the sample concentration. The method\u2019s precision\nis of acceptance level. Robustness indicates that the method is robust. All\nvalidation parameters were met in terms of system suitability, meeting the\npredetermined acceptance criteria.<\/p>\n\n\n\n

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

 A novel simple and sensitive reversed-phase\nHPLC isocratic method has been developed and validated for the assay of\npropylthiouracil tablets. The method was found to be linear (R2) is 0.999\nwithin the analytical range of 24.916 \u00b5g\/mL to 4.748 \u00b5g\/mL. The results\ndemonstrated that the method was both accurate and reproducible. Therefore, the\ndeveloped chromatographic method can be used for estimation of propylthiouracil\ntablets. <\/p>\n\n\n\n

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

   <\/strong>The analytical R&D department of BioPlus\nLife Sciences Pvt Ltd, Hosur, Tamil Nadu, India, provided support for the\nconducted experiment. The author expresses gratitude to Vinayaka Mission’s\nCollege of Pharmacy Principal, Prof. Dr. Kumar M., at VMRF(DU), Salem.\nFurthermore, I would like to express my gratitude to everyone who assisted and\nencouraged the research, particularly for the authorship.<\/p>\n\n\n\n

Funding Sources<\/strong><\/p>\n\n\n\n

The author(s) received no financial support for the\nresearch, authorship, and\/or publication of this article<\/p>\n\n\n\n

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

The\nauthor(s) do not have any conflict of interest<\/strong><\/p>\n\n\n\n

Data Availability Statement<\/strong><\/p>\n\n\n\n

This statement does not\napply to this article<\/p>\n\n\n\n

Ethics Statement<\/strong><\/p>\n\n\n\n

This research did not involve human participants, animal subjects, or any material that requires ethical approval.<\/p>\n\n\n\n

Informed Consent Statement<\/strong><\/p>\n\n\n\n

This study did not involve human participants, and therefore, informed consent was not required.<\/p>\n\n\n\n

Clinical Trial Registration<\/strong><\/p>\n\n\n\n

This\nresearch does not involve any clinical trials<\/p>\n\n\n\n

Author\u2019s Contribution<\/strong><\/p>\n\n\n\n

The\nstudy was designed by Umamaheshwari D, who also headed the study’s analyses and\nliterature searches, wrote the protocol, carried out the statistical analysis,\nand wrote the first draft of the manuscript. The final manuscript was read and\napproved by all authors.<\/p>\n\n\n\n

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

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