高分辨采样二维液相色谱法测定维生素D滴剂中维生素D<sub>3</sub>含量研究<sup>*</sup>

吴珏, 郭伟斌, 邱晓峰, 郑淑凤

doi:10.16155/j.0254-1793.2023-0741

药物分析杂志 >

2024 , Vol. 44 >Issue 7: 1169 - 1175

DOI: https://doi.org/10.16155/j.0254-1793.2023-0741

成分分析

高分辨采样二维液相色谱法测定维生素D滴剂中维生素D₃含量研究^*

展开

1.厦门市食品药品质量检验研究院,厦门 361012;
2.国药控股星鲨制药厦门有限公司,厦门 361026

第一作者 Tel:(0592)5619836;E-mail:jue459@126.com

收稿日期: 2023-11-20

网络出版日期: 2024-08-05

基金资助

^*厦门市市场监督管理局科技计划项目(XMSJ202305)

收起

Determination of vitamin D₃ in vitamin D drops by high resolution sampling two-dimensional chromatography^*

Expand

1. Xiamen Institute for Food and Drug Quality Control, Xiamen 361012, China;
2. Sinopharm Xingsha PharmaceuticalsXiamen Co., Ltd., Xiamen 361026, China

Received date: 2023-11-20

Online published: 2024-08-05

Fold

摘要

目的:建立高分辨采样二维液相色谱法(HiRes 2D-LC)对维生素D滴剂中维生素D₃进行准确定量分析。方法:第一维液相色谱采用Thermo HYPERSIL Gold Silica(100 mm×2.1 mm,1.9 μm)色谱柱,以正己烷-正戊醇(996∶4)为流动相,流速0.2 mL·min^-1,检测波长265 nm,柱温40 ℃;第二维液相色谱采用ShimPack Velox Hilic(50 mm×2.1 mm,2.7 μm)色谱柱,以正己烷-正戊醇-异丙醇(98∶1∶1)为流动相,流速0.5 mL·min^-1,检测波长265 nm,柱温40 ℃。二维接口采用六位十四通阀,并配置2个多中心切割阀,对前维生素D₃峰和维生素D₃峰进行多次连续切割。结果:维生素D₃质量浓度在1.018 4~5.092 mg·mL^-1范围内线性关系良好(r≥0.999 8),前维生素D₃和维生素D₃精密度试验RSD分别为0.95%和0.40%,重复性试验RSD为0.41%,平均加标回收率为101.4%。供试品溶液在4 ℃和10 ℃放置12 h稳定,RSD分别为0.58%、0.66%。采用本法测得6批维生素D滴剂样品中维生素D₃的含量分别为100.4%、101.6%、100.9%、101.6%、102.7%、101.6%,与采用2020年版《中华人民共和国药典》四部通则0722第四法所测结果基本一致。结论:本方法具有优异的灵敏度、重复性和定量准确性,且省去烦琐的样品前处理步骤,缩短分析时间,改善长样品环带来的峰展宽等问题,显著提高了维生素D₃的含量测定效率,降低分析成本,为维生素D滴剂等主成分与辅料难分离制剂的准确定量提供新方法。

关键词： 高分辨采样; 二维液相色谱法; 正相色谱法; 维生素D滴剂; 前维生素D₃; 维生素D₃; 含量测定; 峰纯度确证

本文引用格式

吴珏, 郭伟斌, 邱晓峰, 郑淑凤 . 高分辨采样二维液相色谱法测定维生素D滴剂中维生素D₃含量研究^*[J]. 药物分析杂志, 2024 , 44(7) : 1169 -1175 . DOI: 10.16155/j.0254-1793.2023-0741

Abstract

Objective: To establish an innovative analytical method based on high resolution sampling two-dimensional chromatography (HiRes 2D-LC) for determination of the content of vitamin D₃ in vitamin D drops. Methods: Two-dimensional liquid chromatography was used. Thermo HYPERSIL Gold Silica (100 mm×2.1 mm, 1.9 μm) column was used in the first dimension with n-hexane-n-amyl alcohol (996∶4) as mobile phase. The flow rate was 0.2 mL·min^-1. The samples were injected and tested at the wavelength of 265 nm. The column temperature was 40 ℃. In the second dimension liquid chromatography, ShimPack Velox Hilic (50 mm×2.1 mm, 2.7 μm) was used as the column with n-hexane-n-pentanol-isopropanol (98∶1∶1) as the mobile phase. The flow rate was 0.5 mL·min^-1. The samples were injected and tested at the wavelength of 265 nm. The column temperature was 40 ℃. A six-position 14-way valve and was equipped with 2 multi-center cutting valves was equipped to make multiple consecutive cuts of the pre-vitamin D₃ peak and vitamin D₃ peak. Results: The calibration curves showed a good linearity at the range of 1.018 4-5.092 mg·mL^-1(r≥0.999 8). The precision test showed the RSDs of the peak area of pre-vitamin D₃ and vitamin D₃ were 0.95% and 0.40%, respectively. The repeatability test showed the RSD of vitamin D₃ content was 0.41%. The average recovery rate (n=9) was 101.4%. The test solution was stable at 4 ℃ and 10 ℃ for 12 h, and the RSDs were 0.58% and 0.66%, respectively. The contents of vitamin D₃ in the samples of vitamin D drops measured by this method were 100.4%, 101.6%, 100.9%, 101.6%, 102.7% and 101.6%, which was basically consistent with the results measured by the fourth method in General Chapter 0722 of ChP 2020 Vol Ⅳ. Conclusion: This method has a good specificity and high sensitivity to accurately determine the content of vitamin D₃ in vitamin D drops.

Key words： high resolution sampling; two-dimensional chromatography; normal plase chromatography; vitamin D drops; pre-vitamin D₃; vitamin D₃; normal-phase chromatography; peak purity confirmation

参考文献

[1] 廖祥鹏, 张增利, 张红红, 等. 维生素D与成年人骨骼健康应用指南(2014年简化版)[J]. 中国骨质疏松杂志, 2014, 20(6): 1011
LIAO XP, ZHANG ZL, ZHANG HH, et al. Application guideline for vitamin D and bone health in adult Chinese (2014 Starter Edition)[J]. Chin J Osteoporosis, 2014, 20(6): 1011
[2] 中华人民共和国药典2020年版. 四部[S]. 2020: 105
ChP 2020. Vol Ⅳ[S]. 2020: 105
[3] 范世德, 廖晓君, 邱正霞, 等. RP-HPLC法测定钙佳锌口服液中维生素D₂含量[J]. 中国医药导报, 2010, 7(13): 59
FAN SD, LIAO XJ, QIU ZX, et al. Content determination of vitamin D₂ in Gaijiaxin oral liquid by RP-HPLC[J]. China Med Her, 2010, 7(13): 59
[4] 冯国. 反相高效液相色谱法测定维D₂磷酸氢钙片中维生素D₂含量[J]. 中国药业, 2013, 22(12): 91
FENG G. Determination of vitamin D₂ content in vitamin D₂ and calcium hydrogen phosphate tablets by RP-HPLC[J]. China Pharm, 2013, 22(12): 91
[5] 许强, 刘金生, 王轶鹏, 等. 反相高效液相色谱法测定维生素D₂和维生素D₃[J]. 理化检验(化学分册), 2011, 47(10): 1168
XU Q, LIU JS, WANG YP, et al. RP-HPLC determination of vitamin D₂ and vitamin D₃ [J]. Phys Test Chem Anal(Part B: Chem Anal), 2011, 47(10): 1168
[6] 李秀明, 徐建. UPLC-MS/MS测定维生素AD滴剂的含量[J]. 甘肃医药, 2021, 40(1): 59
LI XM, XU J. Determination of the content of vitamin AD drops by UPLC-MS/MS[J]. Gansu Med J, 2021, 40(1): 59
[7] KO DH, JUN SH, NAM Y, et al. Multiplex LC-MS/MS for simultaneous determination of 25-hydroxyvitamin D, 24, 25-dihydroxyvitamin D₃, albumin, and vitamin D-binding protein with its isoforms: one-step estimation of bioavailable vitamin D and vitamin D metabolite ratio[J]. J Steroid Biochem Mol Biol, 2021, 206:105796
[8] 赵雅清, 付冬梅, 刘艳芳, 等. 离线二维液相色谱法分离巴天酸模根的化学成分[J]. 色谱, 2018, 36(1): 37
ZHAO YQ, FU DM, LIU YF, et al. Separation of chemical compositions in root of Rumex patientia L. with off-line two-dimensional liquid chromatography[J]. Chin J Chromatogr, 2018, 36(1): 37
[9] 张艳海, 张大伟, 孟兆青, 等. 在线二维液相色谱法快速测定桂枝茯苓胶囊中芍药苷、丹皮酚、苦杏仁苷和肉桂酸的含量[J]. 中国中药杂志, 2013, 38(23): 4088
ZHANG YH, ZHANG DW, MENG ZQ, et al. Rapid determination of four components in Guizhi Fuling capsule with online two-dimensional liquid chromatography[J]. China J Chin Mater Med, 2013, 38(23): 4088
[10] 沈灿杰, 马阳, 陈夏霖, 等. 多中心切割二维液相色谱法同时测定九味熄风颗粒中10种成分[J]. 中草药, 2022, 53(14): 4333
SHEN CJ, MA Y, CHEN XL, et al. Simultaneous quantitation of 10 components in Jiuwei Xifeng granules by multiple heart-cutting two dimensional liquid chromatography[J]. Chin Tradit Herb Drugs, 2022, 53(14): 4333
[11] 孔滢, 刘红, 彭洪薇, 等. 二维液相色谱法同时测定人血浆中伊马替尼及去甲伊马替尼药物浓度及临床应用[J]. 中国现代应用药学, 2022, 39(17): 2229
KONG Y, LIU H, PENG HW, et al. Simultaneous determination of imatinib and N-desmethyl imatinib in human plasma by two-dimensional liquid chromatography and its clinical application[J]. Chin J Mod Appl Pharm, 2022, 39(17): 2229
[12] 王智聪, 傅荣杰, 吉建国, 等. 高分辨采样二维液相色谱法同时测定金银花中绿原酸和木犀草苷含量[J]. 色谱, 2019, 37(2): 201
WANG ZC, FU RJ, JI JG, et al. Simultaneous determination of chlorogenic acid and cynaroside contents in Lonicerae Japonicae Flos by high resolution sampling two-dimensional liquid chromatography[J]. Chin J Chromatogr, 2019, 37(2): 201
[13] 林思荣, 王勇, 林晨, 等. 柱切换高效液相色谱法测定维生素D滴剂、维生素AD滴剂中维生素D的含量[J]. 药物分析杂志, 2021, 41(3): 491
LIN SR, WANG Y, LIN C, et al. Determination of vitamin D content in vitamin D drops and vitamin AD drops by column-switching high-performance liquid chromatography [J]. Chin J Pharm Anal, 2021, 41(3): 491
[14] 徐雄飞, 谭灵芝. 正相HPLC法测定碳酸钙D₃片中VD₃的含量[J]. 广东化工, 2015, 42(22):178
XU XF, TAN LZ. Determination of vitamin D₃ content in calcium carbonate D₃ tablets by positive phase HPLC[J]. Guangdong Chem Ind, 2021, 41(3): 49
[15] 王英新, 王凤英, 吴学萍, 等. HPLC法测定碳酸钙D₃咀嚼片(Ⅱ)维生素D₃含量[J]. 齐鲁工业大学学报, 2017, 31(1): 16
WANG YX, WANG FY, WU XP, et al. Determination of vitamin D₃ in calcium carbonate and vitamin D₃ chew table by HPLC[J]. J Qilu Univ Technol, 2017, 31(1): 16
[16] 徐硕, 徐文峰, 金鹏飞, 等. HPLC快速测定维生素D₃软胶囊中的维生素D₃[J]. 华西药学杂志, 2018, 33(1):84
XU S, XU WF, JIN PF, et al. Rapid determination of vitamin D₃ in vitamin D₃ soft capsule by HPLC [J]. West China J Pharm Sci, 2018, 33(1): 84
[17] 王珵, 施洁明, 张立雯, 等. HPLC法测定维生素D含量结果计算方法的研究[J]. 中国药品标准, 2020, 21(2): 107
WANG C, SHI JM, ZHANG LW, et al. Investigation on the calculation methods of vitamin D assay by HPLC[J]. Drug Stand China, 2020, 21(2): 107

Options

文章导航

模态框（Modal）标题

摘要

本文引用格式

Abstract

参考文献