HPLC测定鹿胎软胶囊中4种嘌呤的含量

Determination of Four Kinds of Purines in Deer Fetus Soft Capsule by HPLC

目的：建立HPLC法测定鹿胎软胶囊中4种嘌呤的含量。方法：色谱条件：流动相为0.02 mol/L KH2PO4缓冲液含有1 mmol/L庚烷磺酸钠（pH=3.8）∶甲醇=97∶3,检测波长254 nm,流速1.5 mL/min。4种嘌呤的线性关系：次黄嘌呤：Y1=83695X1＋355（r1=0.9998）,线性范围：0.040~0.667 mg/mL;黄嘌呤：Y2=50638X2＋39（r2=0.9989）,线性范围：0.008~0.119 mg/mL;鸟嘌呤：Y3=30269X3-9562（r3=0.9924）,线性范围：0.018~0.279 mg/mL;腺嘌呤：Y4=38975 X4-8671（r4=0.9989）,线性范围：0.027~0.399 mg/mL。4种嘌呤的平均加样回收率分别为98.1%、98.6%、98.0%和97.5%,RSD分别为1.0%、0.4%、0.8%和0.6%。结果：3批鹿胎软胶囊中次黄嘌呤、黄嘌呤、鸟嘌呤和腺嘌呤的含量为116.5~132.0μg/粒、21.2~23.0μg/粒、48.6~54.3μg/粒和68.9~75.2μg/粒。结论：该方法操作简便、准确、重复性好,为鹿胎软胶囊中嘌呤的分离和定量分析提供了依据。

Objective：To establish an HPLC method for simultaneous determination of four kinds of purines in deer fetus soft capsule.Methods：Four kinds of purines were detected and determined by HPLC.The mobile phase was 0.02 mol/L KH 2 PO 4（ containing 1 mmol/L heptane sulfonic acid sodium,pH=3.8）-methanol=97 ∶ 3.Detection wavelength was 254 nm and flow rate was 1.5 mL/min.The linear relationship of four kinds of purines was as follows：hypoxanthine：Y 1=83695X 1＋355（ r 1=0.9998）,with the linear range 0.040~0.667 mg/mL;xanthine：Y 2=50638X 2＋39（ r 2=0.9989）,with the linear range 0.008~0.119 mg/mL;guanine：Y 3=30269X 3- 9562（ r 3=0.9924）,with the linear range 0.018~0.279 mg/mL;adenine：Y 4=38975X 4- 8671（ r 4=0.9989）,with the linear range 0.027~0.399 mg/mL.The average sample recovery rate of hypoxanthine,xanthine,guanine and adenine were 98.1%,98.6%,98.0% and 97.5%,with RSD 1.0%,0.4%,0.8% and 0.6%,respectively.Results：The content of hypoxanthine,xanthine,guanine and adenine in 3 lots of deer fetus soft capsule were 116.5~132.0 μg/capsule,21.2~23.0 μg/capsule,48.6~54.3 μg/capsule and 68.9~75.2 μg/capsule,respectively.Conclusions：This method is simple,accurate and reproducible,which provides a basis for quality control of purines in deer fetus soft capsule.