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枸杞子中甜菜碱含量测定方法的建立和提取方法的优化 Δ

黄钰馨 ，马 玲 ，李 苗 ，郑国保 ，马小荣（1.宁夏回族自治区药品检验研究院，银川 750002；2.宁夏农林
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科学院农业生物技术研究中心，银川 750002）
中图分类号 R927.2 文献标志码 A 文章编号 1001-0408（2020）14-1700-04
DOI 10.6039/j.issn.1001-0408.2020.14.07

摘 要 目的：建立枸杞子中甜菜碱的含量测定方法，并优化其提取方法。方法：采用高效液相色谱法测定枸杞子中甜菜碱的含
量，色谱柱为Waters Spherisorb NH2，流动相为乙腈-0.01 mol/L磷酸二氢钾水溶液（75∶25，V/V），流速为0.7 mL/min，检测波长为195
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nm，柱温为30 ℃，进样量为10 μL。以甜菜碱含量为指标，在单因素试验的基础上，采用L9 （3）正交试验设计对枸杞子中甜菜碱超
声提取的甲醇体积分数、提取时间、料液比进行筛选并进行验证。测定10批枸杞子中甜菜碱的含量，并与2015年版《中国药典》收
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录的薄层色谱法测定结果进行比较。结果：甜菜碱检测质量浓度的线性范围为2.035～2 035.04 μg/mL（R ＝0.999 3）；检测限、定
量限分别为0.410、2.051 μg/mL，平均加样回收率为97.41％～98.86％（RSD为0.8％～1.4％，n＝3），精密度、重复性、稳定性（24 h）
试验的RSD均不大于1.2％。最优提取方法为料液比1∶30（g/mL）投料、甲醇超声提取45 min。3次验证试验所得提取液中甜菜碱
的平均含量为2.30％（RSD＝0.43％，n＝3）。10批枸杞子中甜菜碱的含量为1.91％～2.55％，与薄层色谱法的测定结果（1.88％～
2.60％）无明显差异（相对误差为－1.92％～2.7％）。结论：成功建立了枸杞子中甜菜碱的含量测定方法，并优化了其提取工艺。
关键词 枸杞子；甜菜碱；高效液相色谱法；含量测定；提取方法

Establishment of the Content Determination Method of Betaine in Lycium barbarum and Optimization of
the Extraction Method
HUANG Yuxin ，MA Ling ，LI Miao ，ZHENG Guobao ，MA Xiaorong（1. Ningxia Hui Autonomous Region
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Institute of Drug Control，Yinchuan 750002，China；2. Agricultural Biotechnology Research Center of Ningxia
Academy of Agricultural and Forestry Sciences，Yinchuan 750002，China）
ABSTRACT OBJECTIVE：To establish the content determination method of betaine in Lycium barbarum，and to optimize the
extraction method. METHODS：HPLC method was used to determine the content of betaine in L. barbarum. The determination was
performed on Waters Spherisorb NH2 column with mobile phase consisted of acetonitrile-0.01 mol/L monopotassiun phosphate
aqueous solution（75 ∶ 25，V/V）at the flow rate was 0.7 mL/min. The detection wavelength was 195 nm，and column temperature
was 30 ℃. The sample size was 10 μL. Using the content of betaine as index，on the basis of single factor tests，L9 （3）orthogonal
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test design were used to select the methanol volume fraction，extraction time and solid-liquid ratio of betaine in L. barbarum by
ultrasonic extraction. The contents of betaine in 10 batches L. barbarum were determined，and compared with the results of TLC
included in 2015 edition of Chinese Pharmacopeia. RESULTS：The linear range of betaine was 2.035-2 035.04 μg/mL（R ＝0.999 3）.
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The limits of detection and quantification were 0.410 μ g/mL and 2.051 μ g/mL，respectively. The average recovery were
97.41％-98.86％（RSDs were 0.8％-1.4％，n＝3）. RSDs of precision，reproducibility and stability（24 h）tests were not higher
than 1.2％. The optimal extraction method included solid-liquid ration of 1 ∶ 30（g/mL），ultrasonic extraction with methanol for 45
min. The average content of betaine in the extract from the three validation tests was 2.30％（RSD＝0.43％，n＝3）. The contents
of betaine in 10 batches L. barbarum were 1.91％ -2.55％ ，which was no significantly different from the results of TLC
（1.88％-2.60％）（RE were －1.92％-2.79％）. CONCLUSIONS：The content determination method of betaine in L. barbarum was
established successfully，and the extraction process was optinized.
KEYWORDS Lycium barbarum；Betaine；HPLC；Content determination；Extraction method

枸杞子为茄科植物宁夏枸杞（Lycium barbarum L.） 实、叶、柄中主要的生物碱类成分，在人体内起甲基供体
[5]
的干燥成熟果实 ，是我国传统中药材，具有治疗糖尿 的作用 ，具有促进脂肪代谢、调节渗透压、抗应激等功
[1]
[9]
[3]
[4]
[2]
病 、保护视网膜 、抗氧化 等作用。甜菜碱是枸杞果 效 [6-8] ，被认为是枸杞子的主要活性物质之一 。目前，
2015 年版《中国药典》（一部）记载的枸杞子中甜菜碱含
Δ 基金项目：国家自然科学基金资助项目（No.41661108）
量测定方法为薄层色谱法 ，该方法操作繁琐、人为因素
[1]
＊主管药师，硕士。研究方向：药物分析。电话：0951-4104813。
影响较大、灵敏度低、分离效果和重现性较差。高效液
E-mail：coco_901125@163.com
# 通信作者：副主任药师。研究方向：药物分析。电话：0951- 相色谱法（HPLC）作为一种现代分离分析技术，已被广
4104813。E-mail：763746007@qq.com 泛应用于中药化学成分的分离分析和含量测定中 [10-12] 。
·1700 · China Pharmacy 2020 Vol. 31 No. 14 中国药房 2020年第31卷第14期