Hplc clc ods c18 column

The HPLC was carried out to quantify phenolic constituents of C. sativum and C. limon (Shimadzu, Japan detector: SPD-10 AV, pump LC-10 AT). Shim pack HPLC CLC-ODS C₁₈ column of 25 cm × 4.6 mm length and width was used to perform this test. Two types of mobile phase were used named A and B. Mobile phase named as A was acetonitrile, which was run as 15% from 0 to 15 min, as 45% from 15 to 30 min and as 100% from 30 to 45 min. Second mobile phase named as B contained distilled water: acetic acid (96: 4) and its pH was adjusted at 2.27. Ultraviolet visible detector (Model: SPD- 10 AV) was used to detect sample at room temperature at a wavelength 280 nm. To draw a chromatogram, voltage units were plotted along x-axis and time in minutes plotted along y-axis. Compounds were identified by comparing previously obtained peaks of standards with samples. Compounds were quantified by external standardization method [17 (link)].

By using HPLC the total phenolic and flavonoids content of the test drug was determined. The Shim pack HPLC CLC-ODS (C-18) column with 25 cm × 4.6 mm length and width, 5 μm diameter was used. In this method, two types of mobile phases were used. Mobile phase A was acetonitrile, run as 15% for 0–15 min, 45% for 15–30 min, 100% for 30–45 min, whereas mobile phase B contained distilled water:acetic acid (94:6) and the pH was adjusted to 2.27. A UV visible (UV) detector with a wavelength of 280 nm was employed to detect the sample at room temperature. The chromatogram was generated by plotting voltage on x-axis and time on y-axis. By the comparison of reference standards and sample peaks, the compounds were identified. Instrument of HPLC was made of Shimadzu, Kyoto, Japan (detector SPD-10 AV; pump. LC-10 AT) [25 (link)].