RP-HPLC法测定人工虫草中5种核苷类成分的含量

Determination of 5 nucleosides in cultured Cordyceps by RP-HPLC

为了评价虫草产品的质量,运用反相高效液相色谱法测定人工虫草中核苷类成分(尿嘧啶、尿苷、腺嘌呤、腺苷、虫草素)的含量。采用Agilent 1200高效液相色谱仪,Kromasil 100-5C18色谱柱(4.6 mm×250 mm,5μm),柱温30℃,流速1.0 mL/min,以甲醇-水梯度洗脱的方法同时测定5种核苷类成分的含量。5个核苷类成分的线性范围分别为尿嘧啶41.12—2 056μg/L(r=1.0000),尿苷80.96—4 048μg/L(r=0.9995),腺嘌呤82.56—4 128μg/L(r=0.9995),腺苷82.08—4 104μg/L(r=1.0000),虫草素81.12—4 056μg/L(r=1.0000)。平均加样回收率分别为尿嘧啶99.7%(RSD=3.1%),尿苷95.6%(RSD=6.7%),腺嘌呤93.4%(RSD=2.8%),腺苷104.1%(RSD=2.3%),虫草素99.9%(RSD=1.4%)。该方法灵敏度高、操作简便、重现性好、效率高。

In order to evaluate qualities of Cordyceps products, the contents of cultured Corclyceps＇ 5 nucleo- sides （uracil, uridine, adenine, adenosine and cordycepin ）were simultaneously determined by reverse-phase high-performance liquid chromatography （RP-HPLC） ,in which the Agilent 1200 high-performance liquid chro- matograph and the Kromasil 100-5C18 chromatographic column （4.6 mm×250 mm,5μm）were used, and the column temperature, the flow rate and the elution were respectively 30℃, 1.0 mL/min and methanol-water gradient elution. The results showed that the linearity ranges of determining the above 5 nucleosides were respec- tively 41.12--2 056μg/L （ r = 1. 0000 ）, 80.96---4 048μg/L （ r = 0. 9995 ）, 82.56--4 128μg/L （ r = 0. 9995 ） , 82.08--4 104 μg/L （ r = 1. 0000） and 81.12--4 056μg/L （ r = 1. 0000） ; And the average recovery rates of their spiked samples were respectively 99.7% （ RSD = 3. 1% ）, 95.6% （ RSD = 6.7% ）, 93.4% （RSD = 2.8% ）, 104.1% （ RSD = 2.3% ） and 99.9% （ RSD = 1.4% ）. This method was high in sensitiveness, simple in operation, good in repeatability and efficient.