目的 采用高效液相色谱串联二极管阵列检测(reverse phase-high performance liquid chromatography-diode array detector,RP-HPLC-DAD)测定市售骨化三醇胶丸中主要成分与其异构体。方法 采用Symmetry C18色谱柱(4.6 mm×250 mm,5 μm),以水-乙腈-甲醇混合溶液为流动相,梯度洗脱,采用二极管阵列检测器。应用Chem Station色谱工作站进行分析。结果 骨化三醇与其异构体线性范围分别为0.171 4~1.36和0.161 3~1.28 μg·mL-1,最低检测限分别为39.75和40.90 ng·mL-1,最低定量限分别为141.6和136.4 ng·mL-1,方法专属性良好,精密度与稳定性的RSD值均小于1.2%,加样回收率大于98%,对于3个厂家的骨化三醇药品进行测定,平均含量为72.71%、80.35%和78.18%。结论 本方法简便、快速、准确、易于推广,适合于骨化三醇胶丸中骨化三醇和反式骨化三醇的测定。
Abstract
OBJECTIVE To establish a new method for determination of the main active ingredient and its isomer in commercially available calcitriol capsules by reverse phase-high performance liquid chromatography-diode array detector (RP-HPLC-DAD). METHODS The chromatographic separation was performed on a Symmetry C18 column (4.6 mm×250 mm, 5 μm) with gradient elution of water-acetonitrile-methanol. The mass spectrometer was equipped with diode array detector. Using Chem Station chromatography workstation, the main ingredient and its isomer in commercially available calcitriol capsules were analyzed. RESULTS The linear ranges of calcitriol and its isomer were 0.171 4-1.36 and 0.161 3-1.28 μg·mL-1, the detection limits were 39.75 and 40.90 ng·mL-1 and the limits of quantification were 141.6 and 136.4 ng·mL-1, respectively. The specificity of the method was good, the RSD values of the precision and stability tests were less than 1.2%, and the recoveries were greater than 98%. The average contents of calcitriol capsules from three manufacturers were 72.71%, 80.35%, and 78.18%, respectively. CONCLUSION The method is simple, rapid, accurate, and easy to be popularized, which is suitable for the determination of calcitriol and trans-calcitriol in calcitriol capsules.
关键词
骨化三醇 /
胶丸 /
高效液相色谱串联二极管阵列检测
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Key words
calcitriol /
softgel capsules /
RP-HPLC-DAD
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中图分类号：
R944
R917
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参考文献
[1] BAO A Y, LI Y, TONG Y Q, et al. 1,25-Dihydroxyvitamin D and cisplatin synergistically induce apoptosis and cell cycle arrest in gastric cancer cells [J]. Int J Mol Med, 2014, 33(5):1177-1184.
[2] PILON C, URBANET R, WILLIAMS T A, et al. 1α,25-Dihydroxyvitamin D3 inhibits the human H295R cell proliferation by cell cycle arrest: a model for a protectiverole of vitamin D receptor against adrenocortical cancer [J]. J Steroid Biochem Mol Biol, 2014, 140:26-33.
[3] BROWN J M, SECINARO K, WILLIAMS J S, et al. Evaluating hormonal mechanisms of vitamin D receptor agonist therapy in diabetic kidney disease: the VALIDATE-D study [J]. BMC Endocr Disord, 2012,3(1):1-9.
[4] WOOTTON A M. Improving the measurement of 25-hydroxyvitamin D [J]. Clin Biochem Rev, 2005, 26(1):33-36.
[5] BASIT S. Vitamin D in health and disease: a literature review [J]. Br J Biomed Sci, 2013, 70(4):161-172.
[6] BP 2014[S]. 2014.
[7] CARTER G D, CARTER C R, GUNTER E, et al. Measurement of vitamin D metabolites: an international perspective on methodology and clinical interpretation [J]. J Steroid Biochem Mol Biol, 2004, 89-90(1-5):467-471.
[8] DING S, SCHOENMAKERS I, JONES K, et al. Quantitative determination of vitamin D metabolites in plasma using UHPLC-MS/MS [J]. Anal Bioanal Chem, 2010, 398(2):779-789.
[9] STRATHMANN F G, LAHA T J, HOOFNAGLE A N. Quantification of 1α,25-Dihydroxy vitamin D by immunoextraction and liquid chromatography-tandem nass spectrometry [J]. Clin Chem, 2011, 57(9):1279-1285.
[10] FARRELL C J, MARTIN S, MC WHINNEY B, et al. State-of-the-art vitamin D assays: a comparison of automated immunoassays with liquid chromatography-tandem mass spectrometry methods [J]. Clin Chem, 2012, 58(3):531-542.
[11] CHEN Y S, XU W J, LV D M. HPLC assay of calcitriol pills [J]. Chin J Pharm Anal(中国药物分析杂志), 2005,25(2):140-141.
[12] LI J, YANG C. Study on the stability of calcitriol soft capsules[J]. Chin J Pharm Anal (中国药物分析杂志), 2009,18(2):22.
[13] HEDMAN C J, WIEBE D A, DEY S, et al. Development of a sensitive LC-MS/MS method for vitamin D metabolites: 1,25 Dihydroxyvitamin D2&3 measurement using a novel derivatization agent [J]. J Chromatogr B Analyt Technol Biomed Life Sci, 2014, 953-954(1):62-67.
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脚注
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基金
北京市职业院校教师素质提高工程“特聘专家项目”
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