Sharma Rohit, Iyer Jitesh P, Chakraborti Asit K, Banerjee Uttam Chand
Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, SAS Nagar, Punjab 160062, India.
Biotechnol Appl Biochem. 2004 Feb;39(Pt 1):83-8. doi: 10.1042/BA20030137.
A method for the detection of gibberellins produced by Fusarium verticilliodes is described using HPLCMS/MS (HPLC tandem MS). A Hypersil (5 microm) octadecylsilane column with methanol/water as eluent in the ratio 3:1 at a flow rate of 0.5 ml/min was used. In the HPLCMS, GA(3) (gibberellic acid; m / z 346.3) eluted at retention time tr=3.08 min, with the corresponding mass chromatogram having peaks at m / z 346.7 and 328.8 corresponding to the M+ and M+-H2O ions respectively. The ethyl acetate extract from the broth, subjected to HPLCMS analysis under similar conditions, showed a constituent with tr=2.13 min, the mass chromatogram of which exhibited peaks at m / z 348.9 and 331.9 corresponding to the MH+ and MH+-H2O ions respectively. Comparison of the MS and MS/MS results (direct infusion) of an authentic sample of GA3 and the ethyl acetate extract from the broth revealed the formation of reduced GA3 in the broth. The present study, utilizing HPLCMS/MS, describes an improved methodology for the unambiguous determination and estimation of gibberellins from fermentation broth.
本文描述了一种使用高效液相色谱-串联质谱法（HPLC-MS/MS）检测轮枝镰孢菌产生的赤霉素的方法。使用Hypersil（5微米）十八烷基硅烷柱，以甲醇/水为洗脱剂，比例为3:1，流速为0.5毫升/分钟。在HPLC-MS中，赤霉酸（GA(3)；m/z 346.3）在保留时间tr=3.08分钟时洗脱，相应的质量色谱图在m/z 346.7和328.8处有峰，分别对应M+和M+-H2O离子。在相似条件下对发酵液的乙酸乙酯提取物进行HPLC-MS分析，结果显示有一个成分的保留时间tr=2.13分钟，其质量色谱图在m/z 348.9和331.9处有峰，分别对应MH+和MH+-H2O离子。对GA3标准品和发酵液乙酸乙酯提取物的质谱和串联质谱结果（直接进样）进行比较，发现发酵液中形成了还原型GA3。本研究利用HPLC-MS/MS描述了一种从发酵液中明确测定和估算赤霉素的改进方法。
