丹参注射剂中丹参素、原儿茶酸、原儿茶醛和丹酚酸B的同时测定

Simultaneous Determination of Danshensul, Protocatechuic Acid Protocatechuic Aldehyde and Salviamolic Acid B in Salvia miltiorrhiza Injection

目的:建立高效液相色谱法测定丹参注射剂中丹参素、原儿茶酸、原儿茶醛和丹酚酸B等4种有效成分的含量。方法:色谱柱为Hypersil C18(4.6 mm ×250 mm,5μm);流动相由甲醇(A)和5%冰醋酸(B)组成,进行梯度洗脱,在0～5 min,B体积分数为90%,5～10 min,B体积分数由90%线性改变至65%,10～20 min,维持B体积分数为65%,流速为1.0 mL·min-1;柱温30℃,检测波长为281 nm;对羟基苯甲酸作为内标物。结果:丹参素、原儿茶酸、原儿茶醛和丹酚酸B浓度分别在3.95～47.4μg·mL-1(r=0.9996)、4.47～53.6μg·mL-1(r=0.9996)、5.14～61.7μg·mL-1(r=0.9999)和8.40～117μg·mL-1(r=0.9994)范围内呈良好线性关系,丹参素、原儿茶酸、原儿茶醛、丹酚酸B的平均回收率分别为99.45%(RSD=3.6%,n=6),97.57%(RSD=2.8%,n=6),100.2%(RSD=3.9%,n=6)和100.2%(RSD=3.2%,n=6)。测定了6批丹参注射液样品。结论:方法简便,分离效果好,能同时测定丹参注射剂中丹酚酸B、丹参素、原儿茶醛、原儿茶酸等4种水溶性成分的含量,结果准确可靠。

Objective: To develop a HPLC method for simultaneous determination of four water soluble components in Salvia miltiorrhiza injection. Methods:The analysis was carried on a Hypersil C18(4. 6 mm ×250 mm,5 μm)col-umn. The mobile phase was composed of methanol(A) and 5% acetic acid(B)with gradient elution(0 -5 min, 90% B,5 -10 min,90% -65% B,10 -20 min,65% B,volumn fraction)and the flow rate was 1.0 mL·min-1. The detection wavelength was at 281 nm and column temperature was 30℃. The 4 - hydroxy - benzoic acid was used as the internal substance. Results: The linear range was 3. 95 - 47. 4 μg·mL-1 (r = 0. 9996 ) for Danshensu, 4.47-53.6μg· mL-1(r=0. 9996)for protocatechuic acid, 5. 14 - 61. 7μg· mL -1(r = 0. 9999 ) for protocate-chuic aldehyde and 8.40-117μg· mL-1 (r = 0. 9994) for salviamolic acid B. The average recoveries were 99. 45% (RSD = 3. 6% , n = 6 ) for Danshensu, 97. 57% ( RSD = 2. 8% , re = 6) for protocatechuic acid, 100. 2% (RSD =3. 9% ,n =6)for protocatechuic aldehyde and 100. 2% (RSD =3. 2% ,n =6) for salviamolic acid B. Conclusion: This method is rapid,simple and accurate.