反相高效液相色谱法测定硬糖和橙汁中固黄、坚牢绿、丽春红2R、丽春红SX和丽春红3R

Determination of 5 kinds of synthetic pigments in hard candies and orange juice by reversed-phase high-performance liquid chromatography

目的建立硬糖果和橙汁中合成色素固黄、坚牢绿、丽春红2R、丽春红SX和丽春红3R的高效液相色谱检测方法。方法样品中的合成色素用聚酰胺小柱进行吸附,用乙醇-氨水-水(7:2:1,v/v/v)的解析液解吸附、提取,在吹氮浓缩仪上吹至近干,用蒸馏水溶解定容,经0.45μm滤膜过滤,取10μL注入高效液相色谱仪,以甲醇与0.02 mol/L乙酸铵溶液为流动相,色谱柱C18柱(250 mm×4.6 mm i.d.,5μm)梯度洗脱15 min分离,流速为1.0 mL/min,利用二极管阵列检测器在检测波长分别为420、630、520 nm处定量分析。结果固黄、坚牢绿分别在5.0~50.0 mg/L,0.2~50.0 mg/L范围内,丽春红2R、丽春红SX、丽春红3R在1.0~50.0 mg/L范围内峰面积与进样量呈良好的线性关系,线性相关系数大于0.999,回收率在82.5%~96.8%,相对标准偏差在2.8%~8.9%。固黄、坚牢绿、丽春红2R、丽春红SX和丽春红3R的检出限分别达到0.42、0.017、0.098、0.12mg/kg和0.082 mg/kg。结论该方法准确度高,分离效能好,结果稳定可靠。

Objective A new method for determination of fast yellow, fast green, ponceau 2R, ponceau SX and ponceau 3R in hard candies and orange juice by HPLC was developed. Methods Samples were extracted by polyamide and eluted with ethanol-ammonia solution-water (7:2:1, v/v/v). The eluate was then dried by pressure blowing concentrator and dissolved with distilled water. The above solution was filtered by 0.45μm filter before detected by HPLC. The filtered solution 10μL was separated on C18 (250 mm×4.6 mm i.d., 5μm) column using methanol-ammonium acetate aqueous solution (0.02 mol/L) as the mobile phase, and the flow rate was 1.0 mL/min. A photodiode array detector was applied to detect the compositions at wavelengths of 420, 630 nm, and 520 nm. Results This method showed a good linearity over the range of 5.0~50.0 mg/L, 0.2~50.0 mg/L for fast yellow, fast green, respectively, 1.0~50.0 mg/L for ponceau 2R, ponceau SX and ponceau 3R, respectively, with correlation coefficients (r) all above 0.999. The RSDs were in the range of 2.8%~9.8%, and average recoveries were 82.5%~96.7%. The detection limits were 0.42, 0.017, 0.098, 0.12 mg/kg and 0.082 mg/kg for fast yellow, fast green, ponceau 2R, ponceau SX and ponceau 3R, respectively. Conclusion The method is convenient, precise, and reliable, and it has a good separation ability.