The HPLC system (,,) Shimadzu Kyoto Japan used in this study included two LC-10ADvp pumps an SIL-10ADvp, autosampler and. An M10Avp photo - diode array detector. The analytical column.Was a 250mmL4.6mm I.D. Stainless steel tube packed with, Ascentis RP-Amide 5 mm particle size.(Supelco Inc.,,, Bellefonte PA USA). Solution A of the mobile phase mixture was, acetonitrile / water 40 / 60 V / V and solution. B was 75 / 25 V / v. HPLC elution was carried out with 100% A for 8 min followed by, a linear gradient from 100% A to 100% B. For 22 min and then held for 15 min. The flow rate of the mobile phase was 1.5 ml Min1 the column, temperature was 40 1C. And the injection volume was 20 ml. The temperature and humidity during the entire sampling period were monitored by using. A data logger (TR-72U, Thermo Recorder T&R Co.Ltd, Nagano Japan), to record at the 10 min interval.
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