高效液相色谱法同时测定维生素D_3和维生素K_2的含量
|
摘要
目的建立高效液相色谱法同时测定维生素D_3和维生素K_2的含量的分析方法。方法以95%甲醇水:异丙醇=(98:2, V:V)为流动相,梯度洗脱,样品经C_(18)色谱柱(250 mm×4.6 mm, 5μm)分离,并于264 nm波长检测。结果维生素D3在浓度0.1282～1.2818μg/mL之间呈现良好的线性关系,r~2=0.9999,平均回收率为95.7%～100.7%,相对标准偏差为1.7%;维生素K_2在浓度0.51397～5.1397μg/mL之间呈现良好的线性关系,r=0.9999,平均回收率在97.3%～100.7%之间,相对标准偏差为1.3%。结论该方法操作简便、准确、重现性好,能同时测定维生素D_3和维生素K_2含量,可在实验室推广。
Objective To establish a method for simultaneous determination of vitamin D_3 and vitamin K_2 by high performance liquid chromatography(HPLC). Methods Using 95% methanol water: isopropanol(98:2, V:V) as mobile phase, gradient elution was performed. The sample was separated by C_(18) column(250 mm×4.6 mm, 5 μm)and detected at 264 nm wavelength. Results Vitamin D3 had a good linear relationship in range of 0.1282-1.2818μg/mL(r=0.9999), the average recovery were 95.7%-100.7%, with the RSD of 1.7%. Vitamin K_2 had a good linear relationship in range of 0.51397-5.1397 μg/mL(r~2=0.9999), the average recovery were 97.3%-100.7%, with the RSD of 1.3%. Conclusion This method is simple, accurate, and has a good reproducibility, which can be used for the simultaneous determination of vitamin D3 and vitamin K_2 in health foods, and can be popularized in the laboratories.
Objective To establish a method for simultaneous determination of vitamin D_3 and vitamin K_2 by high performance liquid chromatography(HPLC). Methods Using 95% methanol water: isopropanol(98:2, V:V) as mobile phase, gradient elution was performed. The sample was separated by C_(18) column(250 mm×4.6 mm, 5 μm)and detected at 264 nm wavelength. Results Vitamin D3 had a good linear relationship in range of 0.1282-1.2818μg/mL(r=0.9999), the average recovery were 95.7%-100.7%, with the RSD of 1.7%. Vitamin K_2 had a good linear relationship in range of 0.51397-5.1397 μg/mL(r~2=0.9999), the average recovery were 97.3%-100.7%, with the RSD of 1.3%. Conclusion This method is simple, accurate, and has a good reproducibility, which can be used for the simultaneous determination of vitamin D3 and vitamin K_2 in health foods, and can be popularized in the laboratories.
引文
[1]徐军,张慧芬,邵裕坤,等.高效液相色谱法测定人血中脂溶性维生素的含量[J].中国药学杂志,2006,(41):147-149.Xu J,Zhang HF,Shao YK,et al.Simultaneous determination of fat-soluble vitamins A,D3,25-OH-D3,E andβ-carotene in human serum by HPLC[J].Chin Pharm J,2006,(41):147-149.
[2]谭青松,付洁,何瑞国.高效液相色谱法测定鱼样中的维生素A、维生素D3和维生素E[J].动物营养学报,2007,19(5):636-640.Tan QS,Fu J,He RG.Simultaneous measurement of vitamin A,vitamin D3 and vitamin E in fish tissue by HLPC[J].Chin J Anim Nutr,2007,19(5):636-640.
[3]蒋晔,刘红菊,郝晓花.4种脂溶性维生素的非水反相HPLC测定[J].中国医药工业杂志,2005,36(2):93-94.Jiang Y,Liu HJ,Hao XH.Determination of four fat-soluble vitamins by nonaqueous RP-HPLC[J].Chin J Pharm,2005,36(2):93-94.
[4]凌龙,胡海澜,曹燕明,等.维生素K2联合维生素D3治疗青壮年早期膝骨关节炎的研究[J/CD].中华关节外科杂志(电子版),2018,12(2):197-202.Ling L,Hu HL,Cao YM,et al.Effect of vitamin K2 combined with vitamin D3 on young patients of early osteoarthritis of knee[J/CD].Chin JJoint Surg(Elect Ed),2018,12(2):197-202.
[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 review of the role of vitamin K2 in regulating on bone metabolism[J].Chin J Osteoporos 2016,22(12):1597-1600.
[6]张殿元.维主素K缺乏所致颅内出血33例综合报告[J].中华儿科杂志,1982,20(4):221.Zhang DY.A comprehensive report of 33 cases of intracranial hemorrhage caused by vitamin K deficiency[J].Chin J Pediat,1982,20(4):221.
[7]王艳红,刘银坤,叶胜龙,等.维生素K2对人肝癌细胞的抗黏附和抗侵袭作用[J].中华肿瘤杂志,2004,26(12):220.Wang YH,Liu YK,Ye SL,et al.The anti-adhesion and anti-invasion effect of vitamin K2 on human liver cancer cells[J].Chin J Oncol,2004,26(12):220.
[8]邹志强.维生素K2的研究进展[J].中国骨质疏松杂志,2005,11(3):389-392.Zou ZQ.Research advance on vitamin K2[J].Chin J Osteoporos,2005,11(3):389-392.
[9]吴元锋,郑裕国.微生物法生产维生素K2(MK)[J].科技通报,2004,20(5):428-433.Wu YF,Zheng YG.Microbial production of vitamin K2(MK)[J].Bull Sci Technol,2004,20(5):428-433.
[10]刘倩,阳延松,莫薇薇,等.维生素K2及Gas6与维持性血透患者血管钙化相关性研究[J].海南医学,2017,28(5):696-697.Liu Q,Yang YS,Mo WW,et al.Correlation between vitamin K2,Gas6and vascular calcification in maintenance hemodialysis patients[J].Hainan Med J 2017,28(5):696-697.
[11]赵明杰,黄静静,王伟伟,等.不同血液透析方式对维生素K2及腹主动脉钙化的影响[J].实用医学杂志,2018,33(8):1323-1326.Zhao MJ,Huang JJ,Wang WW,et al.Effects of different hemodialysis methods on vitamin K2 and abdominal aortic calcification[J].J Pract Med,2018,33(8):1323-1326.
[12]王萍,陈丽艳,孙银玲,等.基于维生素K2的纳豆发酵条件优化[J].食品工业,2018,39(7):54-57.Wang P,Chen LY,Sun YL,et al.Optimization of natto fermentation conditions based on vitamin K2[J].Food Ind,2018,39(7):54-57.
[13]曾红燕,邹晓莉,黎源倩,等.HPLC同时测定保健食品中维生素A、D3、E和β-胡萝卜素[J].分析试验室,2008,2(27):15-18.Zeng HY,Zhou XL,Li YQ,et al.Simultaneous determination of retinol,vitamin D3,tecopherol andβ-carotene in health food by HPLC[J].Chin JAnal Lab,2008,2(27):15-18.
[14]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软胶囊中K2(20)含量[J].分析试验室,2009,5(28):33-35.Mu XY,Dong YW,Wen XJ et al.Determination of the content of K2(20)in vitamin K2 soft capsule by RP-HPLC[J].Chin J Anal Lab,2009,5(28):33-35.
[15]胡代花,张嘉昕,韩豪,等.UPLC法快速测定婴儿维生素D3滴剂、维生素D3片、维生素AD滴剂及维生素D2片中维生素D3和维生素D2[J].药物分析杂志,2016,36(8):1409-1414.Hu DH,Zhang JX,Han H,et al.Rapid determination of vitamin D3,D2 in baby vitamin D3 drops,vitamin D3 tablets,vitamin AD drops and vitamin D2 tablets by UPLC[J].Chin J Pharm Anal,2016,36(8):1409-1414.
[16]徐建中,王颖妤,严为留,等.维生素K2合成途径中主要酶对MK-7产量的影响[J].生物技术通报,2016,32(11):248-254.Xu JZ,Wang YY,Yan WL,et al.Effects of major enzymes in the biosynthetic pathway of vitamin K2 on MK-7 production[J].Biotechnol Bull,2016,32(11):248-254.
[2]谭青松,付洁,何瑞国.高效液相色谱法测定鱼样中的维生素A、维生素D3和维生素E[J].动物营养学报,2007,19(5):636-640.Tan QS,Fu J,He RG.Simultaneous measurement of vitamin A,vitamin D3 and vitamin E in fish tissue by HLPC[J].Chin J Anim Nutr,2007,19(5):636-640.
[3]蒋晔,刘红菊,郝晓花.4种脂溶性维生素的非水反相HPLC测定[J].中国医药工业杂志,2005,36(2):93-94.Jiang Y,Liu HJ,Hao XH.Determination of four fat-soluble vitamins by nonaqueous RP-HPLC[J].Chin J Pharm,2005,36(2):93-94.
[4]凌龙,胡海澜,曹燕明,等.维生素K2联合维生素D3治疗青壮年早期膝骨关节炎的研究[J/CD].中华关节外科杂志(电子版),2018,12(2):197-202.Ling L,Hu HL,Cao YM,et al.Effect of vitamin K2 combined with vitamin D3 on young patients of early osteoarthritis of knee[J/CD].Chin JJoint Surg(Elect Ed),2018,12(2):197-202.
[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 review of the role of vitamin K2 in regulating on bone metabolism[J].Chin J Osteoporos 2016,22(12):1597-1600.
[6]张殿元.维主素K缺乏所致颅内出血33例综合报告[J].中华儿科杂志,1982,20(4):221.Zhang DY.A comprehensive report of 33 cases of intracranial hemorrhage caused by vitamin K deficiency[J].Chin J Pediat,1982,20(4):221.
[7]王艳红,刘银坤,叶胜龙,等.维生素K2对人肝癌细胞的抗黏附和抗侵袭作用[J].中华肿瘤杂志,2004,26(12):220.Wang YH,Liu YK,Ye SL,et al.The anti-adhesion and anti-invasion effect of vitamin K2 on human liver cancer cells[J].Chin J Oncol,2004,26(12):220.
[8]邹志强.维生素K2的研究进展[J].中国骨质疏松杂志,2005,11(3):389-392.Zou ZQ.Research advance on vitamin K2[J].Chin J Osteoporos,2005,11(3):389-392.
[9]吴元锋,郑裕国.微生物法生产维生素K2(MK)[J].科技通报,2004,20(5):428-433.Wu YF,Zheng YG.Microbial production of vitamin K2(MK)[J].Bull Sci Technol,2004,20(5):428-433.
[10]刘倩,阳延松,莫薇薇,等.维生素K2及Gas6与维持性血透患者血管钙化相关性研究[J].海南医学,2017,28(5):696-697.Liu Q,Yang YS,Mo WW,et al.Correlation between vitamin K2,Gas6and vascular calcification in maintenance hemodialysis patients[J].Hainan Med J 2017,28(5):696-697.
[11]赵明杰,黄静静,王伟伟,等.不同血液透析方式对维生素K2及腹主动脉钙化的影响[J].实用医学杂志,2018,33(8):1323-1326.Zhao MJ,Huang JJ,Wang WW,et al.Effects of different hemodialysis methods on vitamin K2 and abdominal aortic calcification[J].J Pract Med,2018,33(8):1323-1326.
[12]王萍,陈丽艳,孙银玲,等.基于维生素K2的纳豆发酵条件优化[J].食品工业,2018,39(7):54-57.Wang P,Chen LY,Sun YL,et al.Optimization of natto fermentation conditions based on vitamin K2[J].Food Ind,2018,39(7):54-57.
[13]曾红燕,邹晓莉,黎源倩,等.HPLC同时测定保健食品中维生素A、D3、E和β-胡萝卜素[J].分析试验室,2008,2(27):15-18.Zeng HY,Zhou XL,Li YQ,et al.Simultaneous determination of retinol,vitamin D3,tecopherol andβ-carotene in health food by HPLC[J].Chin JAnal Lab,2008,2(27):15-18.
[14]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软胶囊中K2(20)含量[J].分析试验室,2009,5(28):33-35.Mu XY,Dong YW,Wen XJ et al.Determination of the content of K2(20)in vitamin K2 soft capsule by RP-HPLC[J].Chin J Anal Lab,2009,5(28):33-35.
[15]胡代花,张嘉昕,韩豪,等.UPLC法快速测定婴儿维生素D3滴剂、维生素D3片、维生素AD滴剂及维生素D2片中维生素D3和维生素D2[J].药物分析杂志,2016,36(8):1409-1414.Hu DH,Zhang JX,Han H,et al.Rapid determination of vitamin D3,D2 in baby vitamin D3 drops,vitamin D3 tablets,vitamin AD drops and vitamin D2 tablets by UPLC[J].Chin J Pharm Anal,2016,36(8):1409-1414.
[16]徐建中,王颖妤,严为留,等.维生素K2合成途径中主要酶对MK-7产量的影响[J].生物技术通报,2016,32(11):248-254.Xu JZ,Wang YY,Yan WL,et al.Effects of major enzymes in the biosynthetic pathway of vitamin K2 on MK-7 production[J].Biotechnol Bull,2016,32(11):248-254.