高效液相色谱法测定不同采收期余甘子叶中5种酚酸类成分含量变化

杨春兰; 麦琬婷; 陈圣杰; 叶勇; 王捷

doi:10.13386/j.issn1002-0306.2022100021

高效液相色谱法测定不同采收期余甘子叶中5种酚酸类成分含量变化

doi: 10.13386/j.issn1002-0306.2022100021

杨春兰^{1, 2,},
麦琬婷¹,
陈圣杰¹,
叶勇^1, ,,
王捷^1, ,

1.
广西医科大学药学院，广西南宁 530021
2.
玉林市第一人民医院，广西玉林 537400

基金项目: 广西艾滋病防治研究重点实验室项目（gklapt201504）；南宁市青秀区科学研究与技术开发计划项目科技惠民专项（2016FA01）；南宁市青秀区科学研究与技术开发计划项目重大专项（2015Z01）。

详细信息

作者简介:
杨春兰（1994−），女，硕士研究生，初级药师，研究方向：中药新药与保健食品的研究开发，E-mail：1021402725@qq.com

通讯作者:
叶勇（1979−），男，博士，教授，研究方向：中药新药与保健食品的研究开发，E-mail：65493278@qq.com

王捷（1969−），男，硕士，教授，研究方向：民族药保健品的研发，E-mail：2218250952@qq.com。

中图分类号: R917
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出版历程
- 收稿日期: 2022-10-08
- 网络出版日期: 2023-06-19

Changes of Contents of Five Phenolic Acids Ingredients in Phyllanthus emblica L. during Different Harvesting Times by HPLC

YANG Chunlan^{1, 2
,},
MAI Wanting¹,
CHEN Shengjie¹,
YE Yong^{1
, ,},
WANG Jie^{1
, ,}

1.
School of Pharmacy, Guangxi Medical University, Nanning 530021, China
2.
The First People's Hospital of Yulin City, Yulin 537400, China

摘要

摘要: 目的：建立同时测定不同采收期余甘子叶药材中5种酚酸类成分含量的方法，研究其变化规律，确定最佳采收期。方法：利用高效液相色谱法测定不同采收期余甘子叶药材中表没食子儿茶素、柯里拉京、诃子联苯酸、鞣花酸和诃子酸的含量并计算5种酚酸类成分的积累总量。色谱柱为Inertsil ODS-3 C₁₈色谱柱，流动相为乙腈−0.1%磷酸水溶液，梯度洗脱，使用二极管阵列检测器全波长扫描，流速为1.0 mL·min⁻¹，进样量为10 μL。并用SPSS 24.0软件进行主成分分析，选取了3个主成分，计算综合得分。结果：上述5种酚酸类成分线性关系良好，相关系数均大于0.998。平均加样回收率介于98.30%~101.22%，RSD为0.43%~2.66%。不同采收期余甘子叶中5种酚酸类成分含量存在一定差异。含量分别为0.2317~1.5074、1.9517~4.6593、2.5105~6.3325、2.2440~5.3386、13.5570~39.1897 mg·g⁻¹。不同采收期5种酚酸类成分总含量变化为7月>5月>8月>6月>1月>9月>4月>2月>10月>12月>3月>11月，以7月份总含量最高，含量达到（55.158±0.83）mg/g，呈现先降后升的新宝登录入口(中国)有限公司变化趋势。主成分分析结果表明综合得分最高的月份为7月。结论：建立的方法可用于同时测定不同采收期余甘子叶中5个酚酸类成分的含量。最适宜采收期为7月。
- 余甘子叶 /
- 采收期 /
- 高效液相色谱法 /
- 含量测定 /
- 变化规律
Abstract: Objective: To establish the method for content determination of the five phenolic acids ingredients of Phyllanthus emblica L. in different harvest time, and to investigate its variation rules so as to determine the best harvest time. Methods: The contents of Epigallocatechin, Corilagin, Scorpion Biphenyl acid, Ellagic acid and Scorpion acid of Phyllanthus emblica L. in different harvest time were determined by HPLC, and the total amount of accumulation of the five active ingredients was calculated. The chromatographic column was Inertsil ODS-3 C₁₈, acetonitrile−0.1% phosphoric acid aqueous solution as mobile phase for gradient elution. A diode array detector was used with full-wavelength scanning. Flow rate was 1.0 mL·min⁻¹. Injection was 10 μL. Used SPSS 24.0 software for principal component analysis, three principal components were selected and the comprehensive score was calculated. Results: The above five ingredients had a good linear relationship, and the correlation coefficients was more than 0.998, while the average recovery rates were 98.30%~101.22% and RSD were 0.43%~2.66%. There were some differences in the contents of the five phenolic acids ingredients in the leaves of Phyllanthus emblica L. during different harvesting periods. The contents of above five ingredients were 0.2317~1.5074, 1.9517~4.6593, 2.5105~6.3325, 2.2440~5.3386, 13.5570~39.1897 mg·g⁻¹, respectively. The change in content of the five phenolic acids ingredients in different harvest time was: July>May>August>June>January>September>April>February>October>December>March>November. The total content was the highest in July, which was (55.158±0.83) mg/g and showed a dynamic trend of decreasing first and then increasing. Principal component analysis results showed that the month with the highest comprehensive score was July. Conclusion: Established method can be used to determine the contents of the five phenolic acids ingredients of Phyllanthus emblica L. in different harvest time. July was the best harvest time for Phyllanthus emblica L.
- Phyllanthus emblica L. /
- harvesting period /
- high performance liquid chromatography (HPLC) /
- content determination /
- variation rule

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图 1 对照品（A）及样品（B）HPLC色谱图（254 nm）

Figure 1. HPLC chromatograms of reference substances (A) and sample (B)(254 nm)

注：1.表没食子儿茶素；2.柯里拉京；3.诃子联苯酸；4.鞣花酸；5.诃子酸。

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图 2 不同采收期余甘子叶中表没食子儿茶素、柯里拉京、诃子联苯酸、鞣花酸的含量新宝登录入口(中国)有限公司变化

Figure 2. Dynamic change of the contents of Epigall ocatechin, Corilagin, Scorpion Biphenyl acid and Ellagic acid in the leaves of Phyllanthus emblica L. during different harvesting periods

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图 3 不同采收期余甘子叶中诃子酸的含量新宝登录入口(中国)有限公司变化

Figure 3. Dynamic change of the contents of Scorpion acid in the leaves of Phyllanthus emblica L. during different harvesting periods

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图 4 不同采收期余甘子叶各成分总量变化

Figure 4. Changes of the total components in the leaves of Phyllanthus emblica L. during different harvesting periods

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表 1 样品信息

Table 1. Information of samples

批号	产地	采集时间
S1	广西贵港市平南县	2020年1月
S2	广西贵港市平南县	2020年2月
S3	广西贵港市平南县	2020年3月
S4	广西贵港市平南县	2020年4月
S5	广西贵港市平南县	2020年5月
S6	广西贵港市平南县	2020年6月
S7	广西贵港市平南县	2020年7月
S8	广西贵港市平南县	2020年8月
S9	广西贵港市平南县	2020年9月
S10	广西贵港市平南县	2020年10月
S11	广西贵港市平南县	2020年11月
S12	广西贵港市平南县	2020年12月

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表 2 余甘子叶中5种成分标准曲线

Table 2. Calibration curves of five constituents of Phyllanthus emblica L.

成分	回归方程	线性范围（μg·mL⁻¹）	r
表没食子儿茶素	y=10⁸x−511857	22.040~110.200	0.9992
柯里拉京	y=4×10⁷x−530043	24.900~124.500	0.9989
诃子联苯酸	y=6×10⁷x−2×10⁶	28.600~143.000	0.9991
鞣花酸	y=2×10⁷x−359549	20.800~104.000	0.9991
诃子酸	y=6×10⁶x−92236	20.32~101.600	0.9993

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表 3 余甘子叶加样回收率试验结果（n=6）

Table 3. Results of recovery tests of Phyllanthus emblica L. (n＝6)

待测成分	取样量（g）	已知含量（mg）	加入量（mg）	测得量（mg）	加样回收率（%）	平均加样回收率（%）	RSD（%）
表没食子儿茶素	0.2503	0.7537	0.7541	1.5074	99.95	98.30	2.66
	0.2506	0.7534	0.7541	1.4981	98.75
	0.2504	0.7530	0.7541	1.4572	93.38
	0.2507	0.7531	0.7541	1.4884	97.51
	0.2509	0.7533	0.7541	1.5093	100.25
	0.2501	0.7534	0.7541	1.5072	99.96
柯里拉京	0.2503	0.9759	0.9763	1.9508	99.86	101.22	1.61
	0.2506	0.9750	0.9763	1.9716	102.08
	0.2504	0.9749	0.9763	1.9529	100.17
	0.2507	0.9755	0.9763	1.9918	104.10
	0.2509	0.9753	0.9763	1.9526	100.10
	0.2501	0.9755	0.9763	1.9618	101.02
诃子联苯酸	0.2503	1.2553	1.2552	2.5103	99.98	99.59	0.43
	0.2506	1.2551	1.2552	2.5004	99.21
	0.2504	1.2553	1.2552	2.5102	99.98
	0.2507	1.2554	1.2552	2.5003	99.18
	0.2509	1.2556	1.2552	2.5104	99.97
	0.2501	1.2550	1.2552	2.5003	99.21
鞣花酸	0.2503	1.2646	1.2644	2.5091	98.43	98.73	1.59
	0.2506	1.2641	1.2644	2.5090	98.46
	0.2504	1.2642	1.2644	2.4893	96.89
	0.2507	1.2641	1.2644	2.5098	98.52
	0.2509	1.2644	1.2644	2.5091	98.44
	0.2501	1.2642	1.2644	2.5496	101.66
诃子酸	0.2503	25.0499	25.0498	50.0895	99.96	99.76	0.60
	0.2506	25.0498	25.0498	50.0947	99.98
	0.2504	25.0497	25.0498	50.1995	100.40
	0.2507	25.0490	25.0498	50.0983	100.00
	0.2509	25.0493	25.0498	49.7689	98.68
	0.2501	25.0495	25.0498	49.9803	99.52

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表 4 不同采收期余甘子叶主成分分析结果

Table 4. Analysis result of principal component of Phyllanthus emblica L. in different harvest periods

成分	初始特征值			提取载荷平方和
成分	总计	方差百分比	累积（%）	总计	方差百分比	累积（%）
1	2.939	58.779	58.779	2.939	58.779	58.779
2	1.006	20.127	78.906	1.006	20.127	78.906
3	0.704	14.071	92.976	0.704	14.071	92.976
4	0.314	6.289	99.265
5	0.037	0.735	100.000

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表 5 不同采收期余甘子叶成分得分系数矩阵

Table 5. Component score coefficient matrix of Phyllanthus emblica L. in different harvest periods

成分	成分1	成分2	成分3
表没食子儿茶素	−0.151	0.831	0.330
柯里拉京	0.303	0.252	0.370
诃子联苯酸	0.322	0.171	0.305
鞣花酸	0.288	−0.275	0.149
诃子酸	0.198	0.368	−1.030

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表 6 不同采收期余甘子叶药材主成分因子得分

Table 6. Principal component factor scores of cotyledons of Phyllanthus emblica L. in different harvest periods

采收期	FAC1_1	FAC2_1	FAC3_1	F1	F2	F3	综合得分	排序
1月	−0.0965	−0.1108	−1.2927	−0.1655	−0.1111	−1.0847	−0.2928	6
2月	0.4601	−1.4049	−0.0756	0.7888	−1.4091	−0.0634	0.1840	4
3月	−1.7355	0.4689	2.0786	−2.9752	0.4703	1.7440	−1.5152	12
4月	−1.2300	1.5178	−1.0001	−2.1086	1.5223	−0.8392	−1.1305	11
5月	1.1485	0.1173	0.2495	1.9689	0.1176	0.2093	1.3019	3
6月	1.1862	0.2021	0.4505	2.0336	0.2027	0.3780	1.3867	2
7月	1.6468	1.2298	1.0025	2.8231	1.2335	0.8412	2.1791	1
8月	0.2132	0.2473	−0.9695	0.3656	0.2480	−0.8135	0.1617	5
9月	−0.4574	0.4817	−1.0873	−0.7841	0.4831	−0.9123	−0.5292	9
10月	−0.0900	−1.0962	−0.3398	−0.1543	−1.0994	−0.2851	−0.3787	7
11月	−0.5825	−1.8420	0.4206	−0.9987	−1.8475	0.3529	−0.9779	10
12月	−0.4629	0.1890	0.5634	−0.7935	0.1895	0.4727	−0.3891	8

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