Owder of M. veneriformis were mixed with one hundred mL solvent inside a glass tube with stir bar, accurately weighted and kept at space temperature (25 ) for 60 min, two instances. The extract was cooled down for the area temperature, and created up the lost weight with solvent, then centrifuged at 1.five ?104 rpm for ten min. The supernatant was filtered by way of a 0.45- Econofilter.Figure two: Effect of various column temperatures, I: column temperature 20 ; II: column temperature 30 ; III: column temperature 40Pharmacognosy Magazine | April-June 2013 | Vol 9 | IssueJi, et al.: Determination of nucleosides and nucleobases in Mactra veneriformispartial superimposed peaks. In the end, a temperature of 30 was deemed optimal. HPLC chromatogram of a mixed operating common resolution detected having a UV set at 254 nm is shown in Figure 3-a along with the chromatogram of sample are shown in Figure 3-b.Validation with the methodand 1.01 L-1, respectively [Table 1] along with the overall recoveries have been in between 95.15 and 101.07 with RSD much less than 3.03 . The all round intra- and interday variations (RSDs) on the eight analytes have been much less than 1.21 and 1.32 [Table 2], respectively. The developed process also had good repeatability (RSD 0.4 ).Optimization of extraction procedure Optimization of extraction solventsThe linearity, regression and linear ranges of 8 analytes were determined making use of the created HPLC process. The overall LODs and LOQs had been significantly less than 0.32 L-During the preliminary investigation on the resolutionab Figure three: HPLC chromatograms of remedy of standards (a) and samples (b). Peaks: 1, uridine (102.26 L-1); 2, xanthine (113.48 L-1); 3,thymine (46.ten L-1); four,hypoxanthine (9.80 L-1); five, inosine (59.07 L-1); 6, guanine (40.98 L-1); 7, thymidine (45.62 L-1); 8, adenosine (5.55 L-1)Table 1: Linear regression information, LOD and LOQ with the investigated compoundsAnalytes Uridine Xanthine Thymine Hypoxanthine Inosine Guanosine Thymidine AdenosineaLinear regression date Regressive equation y = 268520x-38064 y = 33272x-4270.six y = 100508x-12702 y = 79302x-11290 y = 46443x-3268.7 y = 115869x-17223 y = 24972x-1595.6 y=57693x-7251.aLOQ b ( L-1)-LOD b ( L-1) 0.Buy145100-51-2 101 0.024 0.121 0.032 0.079 0.053 0.049 0.Test variety ( L ) 15.61-156.13 6.01-120.25 0.51-50.14 0.63-63.57 8.02-80.21 1.04-20.83 six.92-69.24 0.47-4.r0.9999 0.9998 0.9997 0.9996 0.9997 0.9998 0.9998 0.0.348 0.073 0.399 0.115 0.265 0.179 0.171 0.y is the worth of peak region, and x may be the value from the reference compound’s concentration ( L-1). b LOD and LOQ had been determined at S/N of about three and 10, respectively.1403850-00-9 Formula Table two: Precision, recovery, stability and repeatability of eight analytesAnalytes Uridine Xanthine Thymine Hypoxanthine Inosine Guanosine Thymidine Adenosine Precision (RSD, , n = six) Intra-day 0.PMID:23776646 23 1.21 0.03 0.83 0.39 0.66 0.11 0.63 Inter-day 1.22 1.01 1.09 0.93 0.32 0.24 1.32 0.99 Recovery ( , n = 3) Mean 99.98 98.94 99.98 100.51 101.07 99.41 one hundred.95 100.62 RSD ( ) 2.08 two.18 2.92 three.03 2.68 1.48 1.12 1.82 Stability (RSD, , n = 6) 0.09 0.99 1.64 1.75 0.34 3.41 2.76 4.26 Repeatability (RSD, , n = three) 0.22 0.36 0.07 0.09 0.23 0.17 0.21 0.Pharmacognosy Magazine | April-June 2013 | Vol 9 | IssueJi, et al.: Determination of nucleosides and nucleobases in Mactra veneriformisof separation, the impact of eight solvent, i.e., methanol: water (1:4, v/v); methanol: water (1:1, v/v); methanol; water; ethanol: water (1:four, v/v); ethanol: water (1:1, v/v); ethanol and butarol was compared. Four grams of powder of M. veneriformis.