2种液相色谱法测定保健品软胶囊中的维生素K_2
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摘要
目的建立正相和反相液相色谱法测定保健品种维生素K_2的含量的分析方法。方法正相法采用硅胶正相色谱柱,流动相为乙酸乙酯:正己烷(0.5:99.5, V:V);流速为1.8 mL/min;反相法采用C18反相色谱柱,流动相为甲醇;流速为1.0 mL/min。紫外检测波长均为270 nm。结果 2种方法维生素K_2浓度在5～100μg/g的范围内线性关系良好,回归系数均大于0.999。维生素K2在5.00、25.00和50.00μg/g添加水平的回收率为93.20%～99.28%,相对标准偏差小于1.8%(n=5)。正相法和反相法的检出限均为5μg/g。结论 2种方法均具备操作过程简单、回收率较高的特点,但反相色谱法具有更好的检测稳定性,更适合保健品软胶囊中维生素K2的分析检测。
Objective To establish two methods for the determination of vitamin K_2 in health foods by normal phase and reverse phase chromatography. Methods Silica gel column was used for the normal-phase method, the mobile phase was ethyl acetate: N-hexane(0.5:99.5, V:V), and the flow rate was 1.8 mL/min. In the reverse-phase method, the mobile phase was methanol, the flow rate was 1.0 mL/min. The ultraviolet detection wavelength was 270 nm. Results There was a good linear relationship between the 2 methods in the range of 5-100 μg/g, and the correlation coefficients were greater than 0.999. The recoveries of vitamin K_2 at 5.00, 25.00 and 50.00 μg/g levels ranged from 93.20% to 99.28%, and the relative standard deviation was less than 1.8%(n=5). The detection limits of both normal and reverse-phase methods were 5 μg/g. Conclusion Both methods have the characteristics of simple operation process and high recovery rate, but reversed phase chromatography has better detection stability and is more suitable for the analysis and detection of vitamin K2 in soft capsules for health care products.
Objective To establish two methods for the determination of vitamin K_2 in health foods by normal phase and reverse phase chromatography. Methods Silica gel column was used for the normal-phase method, the mobile phase was ethyl acetate: N-hexane(0.5:99.5, V:V), and the flow rate was 1.8 mL/min. In the reverse-phase method, the mobile phase was methanol, the flow rate was 1.0 mL/min. The ultraviolet detection wavelength was 270 nm. Results There was a good linear relationship between the 2 methods in the range of 5-100 μg/g, and the correlation coefficients were greater than 0.999. The recoveries of vitamin K_2 at 5.00, 25.00 and 50.00 μg/g levels ranged from 93.20% to 99.28%, and the relative standard deviation was less than 1.8%(n=5). The detection limits of both normal and reverse-phase methods were 5 μg/g. Conclusion Both methods have the characteristics of simple operation process and high recovery rate, but reversed phase chromatography has better detection stability and is more suitable for the analysis and detection of vitamin K2 in soft capsules for health care products.
引文
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[8]李向荣,方晓,陈青俊,等.反相液相色谱法同时测定食物中维生素K1和K2[J].营养学报,1998,20(1):72-75.Li XR,Fang X,Chen QJ,et al.The simultaneous determination of vitamin K1 and vitamin K2 in food by RPLC[J].Chin Clin Oncol,2007,12(2):152-155.
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[10]GB 5009.158-2016食品安全国家标准食品中维生素K1的测定[S].GB 5009.158-2016 National food safety standard-Determination of vitamin K1 in food[S].
[11]杨灵莉.维生素K2的合成方法研究[D].重庆:重庆大学,2012.Yang LL.Study on the synthesis methods of vitamin K2[D].Chongqing:Chongqing University,2012.
[12]许薇.维生素K2的合成研究[D].杭州:浙江大学,2006.Xu W.Study on the synthesis of vitamin K2[D].Hangzhou:Zhejiang University,2006.
[13]庄焕雄,陈东峰,徐孟凡,等.维生素K2对绝经后骨质疏松症的防治作用及血清组织蛋白酶K影响[J].中国骨质疏松杂志,2017,23(5):627-630.Zhuang HX,Chen DF,Xu MF,et al.The effect of vitamin K2 on the prevention and treatment of postmenopausal osteoporosis and serum cathepsin K[J].Chin J Osteoporos,2017,23(5):627-630.
[14]尉鸿飞.黄杆菌胞内维生素K2的分离纯化[D].合肥:中国科学技术大学,2018.Yu HF.Isolation,purification and physicochemical characterization of vitamin K2 from flavobacterium[D].Hefei:University of Science and Technology of China,2018.
[15]罗苗苗,胡学超,邱宏伟,等.微生物发酵法制备维生素K2研究进展[J].食品与发酵工业,2015,41(10):221-225.Luo MM,Hu XC,Qiu HW,et al.Research progress on vitamin K2 by microorganism fermentation[J].Food Ferment Ind,2015,41(10):221-225.
[16]覃佳强.维生素K治疗儿童继发性骨质疏松的机理及疗效观察[D].重庆:重庆医科大学,2004.Qin JQ.Observation on the effect and mechanism of secondary osteoporosis in children treated with vitamin K[D].Chongqing:Chongqing Medical University,2004.
[2]方瑞斌,雷泽,刘忠厚.维生素K2与骨健康[J].中国骨质疏松杂志,2013,19(2):191-198.Fang RB,Lei Z,Liu ZH.Vitamin K2 and bone health[J].Chin JOsteoporos,2013,19(2):191-198.
[3]康丽娟,李宝强,徐传伟.维生素K对营养性维生素D缺乏性佝偻病患儿骨钙素羧化率的影响[J].儿科药学杂志,2018,24(10):19-21.Kang LJ,Li BQ,Xu CW.Effect of vitamin K on osteocalcin carboxylation rate in children with nutritional vitamin D deficiency rickets[J].J Pediat Pharm,2018,24(10):19-21.
[4]周小芸.维生素K2抗肝癌作用的研究进展[J].临床肿瘤学杂志,2007,12(2):152-155.Zhou XY.Research progress of anti-hepatoma effect of vitamin K2[J].Chin Clin Oncol,2007,12(2):152-155.
[5]张萌萌.维生素K2调节骨代谢的生物学研究回顾[J].中国骨质疏松杂志,2016,22(12):1597-1600.Zhang MM.A biological study re vie w of the role of vitamin K2 in re gulating on bone me tabolism[J].Chin J Osteoporos,2016,22(12):1597-1600.
[6]GB 14880-2012食品安全国家标准食品营养强化剂使用标准[S].GB 14880-2012 National food safety standard-Standards for the use of food nutrition enhancers[S].
[7]木晓云,董跃伟,温晓江,等.反相高效液相色谱法测定维生素K2(20)[J].云南大学学报(自然科学版),2008,30(4):405-407.Mu XY,Dong YW,Wen XJ,et al.Determination of menatetrenone by RP-HPLC[J].J Yunnan Univ(Nat Sci Ed),2008,30(4):405-407.
[8]李向荣,方晓,陈青俊,等.反相液相色谱法同时测定食物中维生素K1和K2[J].营养学报,1998,20(1):72-75.Li XR,Fang X,Chen QJ,et al.The simultaneous determination of vitamin K1 and vitamin K2 in food by RPLC[J].Chin Clin Oncol,2007,12(2):152-155.
[9]孙家慧.高效液相色谱法快速测定维生素K2[J].天津化工,2011,25(6):55-56.Sun JH.Rapid determination of vitamin K2 by high performance liquid chromatography[J].Tianjin Chem Ind,2011,25(6):55-56.
[10]GB 5009.158-2016食品安全国家标准食品中维生素K1的测定[S].GB 5009.158-2016 National food safety standard-Determination of vitamin K1 in food[S].
[11]杨灵莉.维生素K2的合成方法研究[D].重庆:重庆大学,2012.Yang LL.Study on the synthesis methods of vitamin K2[D].Chongqing:Chongqing University,2012.
[12]许薇.维生素K2的合成研究[D].杭州:浙江大学,2006.Xu W.Study on the synthesis of vitamin K2[D].Hangzhou:Zhejiang University,2006.
[13]庄焕雄,陈东峰,徐孟凡,等.维生素K2对绝经后骨质疏松症的防治作用及血清组织蛋白酶K影响[J].中国骨质疏松杂志,2017,23(5):627-630.Zhuang HX,Chen DF,Xu MF,et al.The effect of vitamin K2 on the prevention and treatment of postmenopausal osteoporosis and serum cathepsin K[J].Chin J Osteoporos,2017,23(5):627-630.
[14]尉鸿飞.黄杆菌胞内维生素K2的分离纯化[D].合肥:中国科学技术大学,2018.Yu HF.Isolation,purification and physicochemical characterization of vitamin K2 from flavobacterium[D].Hefei:University of Science and Technology of China,2018.
[15]罗苗苗,胡学超,邱宏伟,等.微生物发酵法制备维生素K2研究进展[J].食品与发酵工业,2015,41(10):221-225.Luo MM,Hu XC,Qiu HW,et al.Research progress on vitamin K2 by microorganism fermentation[J].Food Ferment Ind,2015,41(10):221-225.
[16]覃佳强.维生素K治疗儿童继发性骨质疏松的机理及疗效观察[D].重庆:重庆医科大学,2004.Qin JQ.Observation on the effect and mechanism of secondary osteoporosis in children treated with vitamin K[D].Chongqing:Chongqing Medical University,2004.