摘要
目的建立蒜氨酸工作对照品。方法用反相高效制备液相色谱和冷冻干燥技术制备蒜氨酸精制品;用紫外、红外、质谱及核磁共振等光谱手段确证结构,用微量旋光仪测定比旋度,借助文献数据确定立体构型;分别采用反相离子对高效液相色谱紫外检测法、反相高效液相色谱紫外/蒸发光散射/质谱检测法、正相高效液相色谱紫外/蒸发光闪射/质谱检测法、阳离子交换高效液相色谱紫外检测法和AQC柱前衍生化反相HPLC-UV/MS测定其纯度;用TGA法测定干燥失重,依据WHO化学对照品建立的指导原则,确定该精制品的含量;并用NMR定量法测定其含量,验证HPLC测定结果。结果精制品的化学结构为S-烯丙基-L-半胱氨酸亚砜,HPLC纯度平均结果为99.50%(RSD为0.2%,n=7),干燥失重为0.54%,含量确定为99.0%;NMR定量法测定的平均含量98.6%(RSD为0.2%,n=5),2种方法测定结果一致。结论建立的蒜氨酸工作对照品所采用的方法和思路科学合理,测定结果准确可靠。
OBJECTIVE To establish a preparation method for working reference substance of alliin. METHODS Alliin was purified by a preparative HPLC with a preparative C18 column and dried by vacuum dried technology. The chemical structure of alliin was identified by UV, IR, MS, NMR. The stero configuration of alliin was elucidated by determinating specific rotation and comparing with literature data. The purity of purified alliin was determined by IP-RP-HPLC-UV, RP-HPLC-UV/ELSD/MS, NP-HPLC-UV/ELSD/MS, HPIC-UV and pre-column derivatization RP-HPLC-UV/ELSD/MS. The dry loss of purified alliin was determined by TGA, and the conte.nt of purified alIiin was calculated according to the WHO guideline about how to establish a chemical reference substance. In order to validate the results of HPLC, the NMR qualification analysis was used too. RESULTS The chemical structure of purified sample was S-（＋）-ally-L-cysteine sulfoxide, the average purity determined by HPLC methods was 99.50% （RSD%=0.2%, n=7） , the dry loss was 0.54%, the HPLC content of purified sample was 99.0%. the average content of purified sample determined by NMR qualification analysis was 98.6% （RSD%=0.2%, n=5） , the results by two methods was consistent. CONCLUSION The methods and scheme used in the procedure of the establishment is reasonable and applicable.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2009年第15期1177-1180,共4页
Chinese Pharmaceutical Journal
