高效液相色谱法同时测定化妆品中17种美白成分

Simultaneous determination of seventeen kinds of whitening ingredients in cosmetics by high performance liquid chromatography

目的:建立高效液相色谱法(HPLC)同时测定化妆品中17种美白成分。方法:样品经50%甲醇溶液提取、过滤后,进行HPLC分析。采用Agilent ZORBAX Eclipse XDB-C18色谱柱(4.6 mm×250 mm,5μm),流动相为甲醇-0.02 mol·L-1磷酸二氢钾溶液,梯度洗脱,二极管阵列检测器多波长分析,熊果苷、氢醌、烟酰胺、间苯二酚、杜鹃醇、4-丁基间苯二酚、苯乙基间苯二酚和四氢木兰醇的检测波长为230 nm,抗坏血酸磷酸酯镁、抗坏血酸葡糖苷、烟酸、3-O-乙基抗坏血酸、4-甲氧基水杨酸钾、鞣花酸和甘草酸二钾的检测波长为254 nm,曲酸和苯酚的检测波长为270 nm。结果:17种成分分离度良好,峰面积与质量浓度在2~50μg·m L-1范围内呈良好的线性关系(r>0.999),霜类、乳液类、水剂类3种化妆品基质中17种美白成分的回收率良好,3个添加水平分别为0.025%、0.05%和0.25%,平均回收率(n=6)为86.9%~104.3%,RSD为0.51%~3.7%;稳定性良好,17种成分在24 h内保持稳定,RSD在0.12%~0.73%之间,检测下限在0.000 5%~0.005%之间。50批样品中检出7种美白成分,抗坏血酸磷酸酯镁、抗坏血酸葡糖苷、烟酸、熊果苷、烟酰胺、3-O-乙基抗坏血酸、甘草酸二钾的含量分别为0.021%~0.095%、0.018%~0.025%、0.019%~0.024%、0.028%~3.5%、0.027%~3.9%、0.034%~2.8%、0.023%~1.6%。结论:该方法经方法学验证,可同时测定化妆品中17种美白成分的含量。

Objective:To establish a quantitative method for simultaneous determination of 17 kinds of whitening ingredient in cosmetics by high performance liquid chromatography(HPLC).Methods:Samples were extracted by 50%methanol solution and filtered,then separated on a Agilent ZORBAX Eclipse XDB-C18 column(4.6 mm×250 mm,5μm)and detected at different wavelength.The mobile phase was methanol and 0.02 mol·L-1 monopotassium phosphate solution with gradient elution.Diode array detector(DAD)was used with the wavelength of 230 nm for arbutin,hydroquinone,nicotinamide,resorcinol,rhododendrol,4-butylresorcinol,phenylethyl resorcinol and tetrahydromagnolitol,and 254 nm for magnesium ascorbyl phosphate,ascorbyl glucoside,nicotinic acid,3-O-ethyl ascorbic acid,potassium methoxysalicylate,ellagic acid and dipotassium glycyrrhizate,and 270 nm for kojic acid and phenol.Results:The seventeen constituents were well separated.The linear range was 2-50μg·m L-1(r>0.999).The recoveries(spiked at 0.025%,0.05%and 0.25%,respectively)in 3 different types of cosmetic ranged from 86.9%to 104.3%,and the RSD range was 0.51%to 3.7%.The results of the stability tests were satisfactory and the seventeen constituents remained stable in 24 hours with all RSDs were between 0.12%and 0.73%.The lower limit of detection of the 17 whitening ingredients were in the range of 0.000 5%-0.005%.The contents of magnesium ascorbyl phosphate、ascorbyl glucoside、nicotinic acid、arbutin、nicotinamide、3-O-ethyl ascorbic acid and dipotassium glycyrrhizate in 50 batches of samples were 0.021%-0.095%,0.018%-0.025%,0.019%-0.024%,0.028%-3.5%,0.027%-3.9%,0.034%-2.8%,0.023%-1.6%,respectively.Conclusion:It is proved by methodology validation that the established method can be utilized for simultaneous determination of seventeen kinds of whitening ingredients in cosmetics.