柱前衍生化RP-HPLC法鉴别及测定9种补气药中的生物胺

目的：建立柱前衍生化RP-HPLC法测定白术、人参、红参、西洋参、党参、炒扁豆、刺五加、绞股蓝、红景天中11个生物胺的方法，并探讨与补气的相关性。方法：药材样品经1 mol·L^-1盐酸提取，正丁醇-二氯甲烷（1：1）萃取富集，以丹磺酰氯为柱前衍生试剂进行衍生化，RP-HPLC法测定。采用C₁₈色谱柱（200 mm×4.6 mm，5 μm），以乙腈（A）-水（B）为流动相，梯度洗脱（0~7 min，45%A→50%A；7~25 min，50%A→90%A；25~35 min，90%A；35~40 min，90%A→45%A），流速为0.8 mL·min^-1，检测波长为254 nm，柱温为30 ℃。结果：甲胺、尸胺、组胺、亚精胺与精胺的线性范围为0.10~10.00 mg·L^-1（r >0.999），色胺、腐胺的线性范围为0.10~50.00 mg·L^-1（r>0.999），2-苯乙胺、5-羟色胺、酪胺与多巴胺的线性范围为0.25~10.00 mg·L^-1（r>0.999），平均回收率在70.4%~96.3%之间。白术、党参、炒扁豆中均含有甲胺、腐胺、尸胺、亚精胺与精胺，人参、刺五加中均含有腐胺、尸胺、亚精胺与精胺，红参中含有腐胺、亚精胺与精胺，西洋参中含有腐胺、尸胺、组胺、亚精胺与精胺，绞股蓝中含有甲胺、腐胺、尸胺、酪胺、亚精胺与精胺，红景天中含有甲胺、腐胺与亚精胺。结论：在白术、人参、红参、西洋参、党参、炒扁豆、刺五加、绞股蓝、红景天中含有生物胺类成分。亚精胺是这些补气药中的共有成分，考虑到它的生物活性，这可能是中药补气和抗衰老延年益寿作用的关键成分之一。

Objective:To develop a method for the determination of eleven biogenic amines in nine kinds of Chinese Tonic-Qi herbs by RP-HPLC with pre-column derivatization,and investigate the relationship between Tonic-Qi in traditional Chinese medicine and biogenic amines.Methods:Samples were extracted with 1 mol·L^-1 hydrochloric acid,concentrated and purified by n-butyl alcohol-dichloromethane(1:1,v/v)extraction,derivatized with dansyl chloride,and separated by RP-HPLC.The separation was performed on a C₁₈ column (200 mm×4.6 mm,5 μm)with gradient elution(0-7 min,45%A→50%A;7-25 min,50%A→90%A;25-35 min,90%A;35-40 min,90%A→45%A)using acetonitrile(A)and water(B)as the mobile phase at a flow rate of 0.8 mL·min^-1.The detection wavelength was 254 nm.The column temperature was 30 ℃.Results:The calibration curves were linear in the ranges of 0.10-10.00 mg·L^-1(r >0.999)for methylamine,cadaverine,histamine,spermidine and spermine,0.10-50.00 mg·L^-1(r >0.999)for tryptamine and putrescine,0.25-10.00 mg·L^-1(r >0.999)for 2-phenylethylamine,serotonin,tyramine and dopamine,respectively.The average recoveries varied from 70.4% to 96.3%.Methylamine,putrescine,cadaverine,spermidine and spermine were found in Atractylodis Macrocephalae Rhizoma,Codonopsis Radix and Stir-baked Lablab Semen Album.Putrescine,cadaverine,spermidine and spermine were found in Ginseng Radix et Rhizoma and Acanthopanacis Senticosi Radix et Rhizoma Seu Caulis.Methylamine,putrescine,cadaverine,tyramine,spermidine and spermine were found in Ginseng Radix et Rhizoma Rubra.Putrescine,cadaverine,histamine,spermidine and spermine were found in Panacis Quinquefolii Radix.Methylamine,putrescine,cadaverine,tyramine,spermidine and spermine were found in Rhizoma Seu Herba Gynostemmatis.Methylamine,putrescine and spermidine were found in Rhodiolae Crenulatae Radix et Rhizoma.Conclusion:The biogenic amines existing in the above nine kinds of Chinese Tonic-Qi herbs.The spermidine was the biogenic amine existed in all nine kinds of Chinese Tonic-Qi herbs.In consideration of its biological activity,spermidine could possibly be one of the key constituents with Tonic-Qi and anti-aging effects of Chinese Tonic-Qi herbs.