色谱2025，Vol.43Issue(12)：1389-1396,8.DOI:10.3724/SP.J.1123.2025.06011
QuEChERS-高效液相色谱法同时测定化妆品中的维生素K1和K2及高效液相色谱-串联质谱确证
Simultaneous determination of vitamins K1 and K2 in cosmetics by QuEChERS-high performance liquid chromatography and verification by high performance liquid chromatography-tandem mass spectrometry
摘要
Abstract
Vitamins K1 and K2 are both essential fat-soluble vitamins for the human body.K1 was once used in cosmetics for its efficacy in improving dark circles by promoting periorbital circulation.However,due to its potential to cause severe allergic reactions,it is now banned in cosmetics in China and the European Union.K2,particularly the MK-7 form,has a similar structure to K1 but features a different side chain.It has gained attention for its skin benefits,including soothing,antioxidant,and anti-aging properties.Although there are no reports of its use in cosmetics in China yet,it has been registered three times as a new cosmetic ingredient,indicating significant market potential.Most existing methods for detecting vitamins K1 and K2 focus on pharmaceuticals and dietary supplements,with few studies addressing their analysis in cosmetics,especially vitamin K2.Given the structural similarity between the two compounds,a reliable method for their simultaneous determination in cosmetic products is needed.However,cosmetic matrices are complex and often interfere with analysis.Conventional sample preparation techniques,such as liquid-liquid extraction and solid-phase extraction,are time-consuming and labor-intensive.The QuEChERS method offers a faster,simpler,and more cost-effective alternative.In this study,we selected six common types of cosmetics—water-based liquids,emulsions,creams,gels,powders,and oils,to develop and validate a simultaneous quantification method for vitamins K1 and K2.A QuEChERS-based sample preparation method coupled with high performance liquid chromatography(HPLC)was developed for the simultaneous quantification of vitamins K1 and K2 in cosmetics.To enhance specificity and confirmation capability,a complementary method using high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)was also established.Samples were first pre-dispersed with saturated sodium chloride solution,extracted by n-hexane under ultrasonication,and purified using QuEChERS pretreatment technique(containing 150 mg MgSO4,50 mg PSA,and 25 mg C18).For HPLC analysis,separation was performed on a CAPCELL PAK C18 AQ column(250 mm×4.6 mm,5 µm)using methanol-isopropanol(80∶20,volume ratito)as the mobile phase,with detection at 270 nm.For HPLC-MS/MS confirmation,an ACQUITY UPLC BEH C18 column(50 mm×2.1 mm,1.7 µm)was employed,with methanol containing 0.05%(volume fraction)formic acid and 5 mmol/L ammonium formate as the mobile phase.Electrospray ionization in positive mode(ESI+)and multiple reaction monitoring(MRM)were used for detection.Both HPLC and HPLC-MS/MS methods demonstrated excellent performance for the determination of vitamins K1 and K2.In the HPLC method,both analytes showed good linearity over the range of 0.1‒50 μg/mL(r>0.999),with limits of detection(LOD)and quantification(LOQ)of 0.3 μg/g and 1.0 μg/g,respectively.The spiked recoveries ranged from 93.2%-104.5%with RSDs below 5%.For the HPLC-MS/MS method,linearity was observed in the range of 0.005‒0.5 μg/mL(r>0.999),with LOD and LOQ values of 0.02 μg/g and 0.05 μg/g,respectively.Recoveries in this case fell within 89.4%‒108.2%,accompanied by RSDs of less than 10%.The method was successfully applied to analyze 30 cosmetic samples spanning six different matrix types.Neither vitamin K1 nor K2 was detected in any sample.The proposed methodology is rapid,simple,sensitive,and accurate,making it suitable for routine determination of vitamins K1 and K2 in diverse cosmetic products.It offers reliable technical support for quality control and regulatory compliance and demonstrates the utility of QuEChERS sample preparation for the analysis of other cosmetic ingredients.关键词
QuEChERS/高效液相色谱法/高效液相色谱-串联质谱法/化妆品/维生素K1/维生素K2Key words
QuEChERS/high performance liquid chromatography(HPLC)/high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)/cosmetic/vitamin K1/vitamin K2分类
化学化工引用本文复制引用
孙倩,陈春晓..QuEChERS-高效液相色谱法同时测定化妆品中的维生素K1和K2及高效液相色谱-串联质谱确证[J].色谱,2025,43(12):1389-1396,8.基金项目
福建省药品监督管理局科技项目(2022016).Science and Technology Project of Fujian Provincial Food and Drug Administration(No.2022016). （2022016）
