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2.5. Analysis of the composition of SFP The HPLC method was used to identify the monosaccharides in SFP [11]. The sample (10 mg) of SFP was hydrolyzed with 2 mol/l H2SO4 (2 ml) at 100 ◦C for 5 h, and the excess acid was neutralized with barium carbonate. After standing overnight, the neutral products were separated by centrifugation at 3000 rpm for 10 min, 100 l supernatant was collected and reacted with 1- phenyl-3-methyl-5-pyrazolone, then subsequently separated by HPLC (Shimadzu LC-10AT VP, Japan) using a Packed column (4.6 ¡Á 250 mm) with detection at 250 nm. The mobile phase were consisted of 78% phosphate buffer solution (0.05 M, KH2PO4-NaOH, pH 6.9) and 22% acetonitrile (v/v), and the samples were eluted at a flow rate of 1.0 ml/min at 28 ◦C. In this study, l-Rha, d-Gal, d-GalA, d-Glu and Ara were used as references. ÎÒ¾õµÃ¿ÉÒÔÕâÑù·Ò룺2.5.SFP×é·ÖµÄ·ÖÎö ¸ßЧҺÏàÉ«Æ×·¨³£±»ÓÃÓÚ¼ø¶¨SFPÖеĵ¥ÌÇ£¬10mlµÄSFPÑùÆ·Óë2mlŨ¶ÈΪ2mol/mlµÄ H2SO4ÔÚ100◦CµÄζÈÏÂË®½â5¸öСʱ£¬¹ýÁ¿µÄËáÔòÓÃ̼Ëá±µÖк͵ô¡£ÕâÑù·ÅÖÃÒ»¸öÍíÉϺó£¬ÒÔÿ·ÖÖÓ3000תµÄתËÙÀëÐÄ10min¿É³ýÈ¥ÖкͲúÎÊÕ¼¯100mlµÄÉÏÇåÒºÓë1-±½»ù-3-¼×»ù-5-ßÁßòͪ·´Ó¦£¬È»ºóÓøßЧҺÏàÉ«Æ×ÒÇ(ÐͺţºShimadzu LC-10AT VP, Japan)½øÐзÖÀ룬Ìî³äÖù¹æ¸ñΪ4.6 ¡Á 250 mm£¬ÔÚ250nmµÄ²¨³¤Ìõ¼þϲⶨ¡£Á÷¶¯Ïà×é³ÉΪ78%µÄÁ×ËáÑλº³åÒº (0.05 M, KH2PO4-NaOH,pH 6.9)ºÍ22%µÄÒÒëæÈÜÒº(v/v)£¬Î¶ÈΪ28 ◦C¡¢Ï´ÍÑÒºÁ÷ËÙ1.0 ml/minÏ´ÍÑÑùÆ·¡£´Ë´ÎÑо¿ÖУ¬±ê׼ƷΪL-ÊóÀîÌÇ£¬D-°±»ù°ëÈéÌÇ£¬D-°±»ù°ëÈéÌǰ·£¬»¨ÉúËÄÏ©Ëá¡£ |
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