3. Result and discussion
3.1. Chromatography
Rf values of norethindrone acetate and dydrogesterone were obtained by dividing the distance travelled by these drugs by the solvent front (10.0 cm). The results are given in Table 1, which indicate greater Rf value of norethindrone acetate than dydrogesterone with 0.40 and 0.10 ng as limits of detection (LOD) of these pharmaceuticals, respectively. To optimize the chromatographic condition, various combinations of n-hexane and n-butanol were tried but the best separation was achieved by using n-hexane:n-butanol (90:10, v/v) solvent system on plain TLC plates. The variation of mobile phase composition was carried out on plain TLC plates by increasing amount of n-butanol. The effect of mobile phase compositions on Rf values is given in Figure 2, which indicates that Rf values increased sharply with 1.0–10.0% of n-butanol but became almost constant after this composition. This may be due to rapid increase in polarity from 1.0% to 10.0% n-butanol while the further increase of n-butanol does not increase polarity; as the concentration of low polarity n-hexane becomes negligible. Furthermore, this figure depicts that Rf values of norethindrone acetate are slightly higher than dydrogesterone at all compositions of the mobile phase, which are the characteristic features of these drugs. Furthermore, attempts have been made to improve more separation and, therefore, some TLC experiments on metal ions impregnated plates were also carried out. The selected metal ions for impregnation were Fe(II), Co(II), Ni(II), Cu(II) and Zn(II) of 0.1% concentration each separately. The results on impregnated plates were also recorded in Table 1. A comparison of the separation on plain and impregnated TLC plates from Table 1 clearly shows better results on copper impregnated TLC plates with maximum difference of Rf values and compact spot. On the other hand, the diffused spots were observed on all other metal ions impregnated TLC plates. TLC chromatograms of these two drugs on plain and copper impregnated plates are shown in Figure 3 which indicate better separation of these drugs on copper metal ion impregnated plates. It is very interesting to observe that the color of both the spots was brown on copper impregnated plates while it was yellow on plain plates. This behaviour is due to the complex formation of these drugs with copper metal ion.
3. Result and discussion3.1. ChromatographyRf values of norethindrone acetate and dydrogesterone were obtained by dividing the distance travelled by these drugs by the solvent front (10.0 cm). The results are given in Table 1, which indicate greater Rf value of norethindrone acetate than dydrogesterone with 0.40 and 0.10 ng as limits of detection (LOD) of these pharmaceuticals, respectively. To optimize the chromatographic condition, various combinations of n-hexane and n-butanol were tried but the best separation was achieved by using n-hexane:n-butanol (90:10, v/v) solvent system on plain TLC plates. The variation of mobile phase composition was carried out on plain TLC plates by increasing amount of n-butanol. The effect of mobile phase compositions on Rf values is given in Figure 2, which indicates that Rf values increased sharply with 1.0–10.0% of n-butanol but became almost constant after this composition. This may be due to rapid increase in polarity from 1.0% to 10.0% n-butanol while the further increase of n-butanol does not increase polarity; as the concentration of low polarity n-hexane becomes negligible. Furthermore, this figure depicts that Rf values of norethindrone acetate are slightly higher than dydrogesterone at all compositions of the mobile phase, which are the characteristic features of these drugs. Furthermore, attempts have been made to improve more separation and, therefore, some TLC experiments on metal ions impregnated plates were also carried out. The selected metal ions for impregnation were Fe(II), Co(II), Ni(II), Cu(II) and Zn(II) of 0.1% concentration each separately. The results on impregnated plates were also recorded in Table 1. A comparison of the separation on plain and impregnated TLC plates from Table 1 clearly shows better results on copper impregnated TLC plates with maximum difference of Rf values and compact spot. On the other hand, the diffused spots were observed on all other metal ions impregnated TLC plates. TLC chromatograms of these two drugs on plain and copper impregnated plates are shown in Figure 3 which indicate better separation of these drugs on copper metal ion impregnated plates. It is very interesting to observe that the color of both the spots was brown on copper impregnated plates while it was yellow on plain plates. This behaviour is due to the complex formation of these drugs with copper metal ion.
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