HPLC-QAMS法同时测定何首乌防脱育发精华液中9种成分含量

doi:10.3969/j.issn.2097-2806.2023.07.015

摘要/Abstract

摘要：

建立高效液相色谱一测多评法（HPLC-QAMS法）同时检测何首乌防脱育发精华液中9种成分的含量，并验证所建立方法的可行性和准确性。以槲皮苷为含量参照物，甲醇-0.1%磷酸为流动相，分别采用外标法和HPLC-QAMS法同时测定12批何首乌防脱育发精华液中大黄素、大黄素甲醚、杨梅苷、槲皮苷、杨梅素、槲皮素、山柰酚、穗花杉双黄酮、扁柏双黄酮的含量，对比两种方法的结果差异，验证HPLC-QAMS法的适用性。结果显示，9种成分各线性范围良好，相关系数均大于0.999，检出限为0.027~0.151 µg/mL，定量限为0.076~0.449 µg/mL。以槲皮苷为内参比物质，大黄素、大黄素甲醚、杨梅苷、杨梅素、槲皮素、山柰酚、穗花杉双黄酮、扁柏双黄酮的相对校正因子分别为0.753 7，0.836 9，1.019 2，2.991 3，1.687 5，2.236 9，1.096 3和1.249 3。两种方法测得的何首乌防脱育发精华液中9种成分的含量差异均无统计学意义（P>0.05）。以槲皮苷为含量参照物建立的相对校正因子重现性好，HPLC-QAMS法可用于何首乌防脱育发精华液中9种成分的质量评价。

关键词: 何首乌防脱育发精华液, 高效液相色谱一测多评法, 含量测定, 相对校正因子

Abstract:

The study aimed to establish an HPLC-QAMS method for simultaneous quantitation of nine effective components in Polygonum Multiflorum Fangtuoyufa Jinghuaye, and to verify the feasibility and accuracy of the method. Quercitrin was used as the reference substance and methanol-0.1% phosphoric acid was used as the mobile phase. The contents of emodin, physcion, myricitrin, quercitrin, myricetin, quercetin, kaempferol, amentoflavone and hinokiflavone in 12 batches of Polygonum Multiflorum Fangtuoyufa Jinghuaye were determined by external standard method and HPLC-QAMS method, respectively, and the difference between the results of the two methods was compared to verify the serviceability of HPLC-QAMS method. The results show that the linear ranges of the nine components are good, the correlation coefficients are greater than 0.999, the limits of detection （LOD） are 0.027-0.151 µg/mL, and the limits of quantification are 0.076-0.449 µg/mL. The relative correlation factors are 0.753 7, 0.836 9, 1.019 2, 2.991 3, 1.687 5, 2.236 9, 1.096 3 and 1.249 3 for emodin, physcion, myricitrin, myricetin, quercetin, kaempferol, amentoflavone and hinokiflayone, respectively. There is no significant difference in the content of 9 components in Polygonum Multiflorum Fangtuoyufa Jinghuaye determined by the two methods （P>0.05）. The relative correlation factors established with quercitrin as the internal reference substance have good reproducibility. The HPLC-QAMS method can be used for the quality evaluation of nine effective components in Polygonum Multiflorum Fangtuoyufa Jinghuaye.

Key words: Polygonum Multiflorum Fangtuoyufa Jinghuaye, HPLC-QAMS, content determination, relative correlation factor

中图分类号:

TQ658

刘英, 林敏, 戴思芮. HPLC-QAMS法同时测定何首乌防脱育发精华液中9种成分含量[J]. 日用化学工业（中英文）, 2023, 53(7): 841-848.

Liu Ying, Lin Min, Dai Sirui. Simultaneous determination of nine effective components in Polygonum Multif lorum Fangtuoyufa Jinghuaye by HPLC-QAMS[J]. China Surfactant Detergent & Cosmetics, 2023, 53(7): 841-848.

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参考文献 18

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成分	回归方程	线性范围/（μg/mL）	r	检出限/（μg/mL）	定量限/（μg/mL）
大黄素	Y=2.776 2×10⁶X-806.3	0.96~192.00	0.999 5	0.107	0.304
大黄素甲醚	Y=2.485 6×10⁶X+1 017.5	0.68~136.00	0.999 2	0.084	0.257
杨梅苷	Y=2.041 9×10⁶X+663.1	1.35~270.00	0.999 1	0.151	0.449
槲皮苷	Y=2.093 2×10⁶X-1 018.0	1.57~314.00	0.999 6	0.132	0.375
杨梅素	Y=6.971 5×10⁵X-471.2	0.26~52.00	0.999 4	0.027	0.076
槲皮素	Y=1.236 5×10⁶X+753.8	0.39~78.00	0.999 1	0.043	0.127
山柰酚	Y=9.364 4×10⁵X-621.3	0.32~64.00	0.999 5	0.051	0.148
穗花杉双黄酮	Y=1.913 7×10⁶X+1 005.6	0.59~118.00	0.999 2	0.065	0.201
扁柏双黄酮	Y=1.671 5×10⁶X+1 022.9	0.46~92.00	0.999 7	0.039	0.112

混合对照品溶液	?
混合对照品溶液	大黄素	大黄素甲醚	杨梅苷	杨梅素	槲皮素	山柰酚	穗花杉双黄酮	扁柏双黄酮
1	0.758 2	0.827 4	1.003 5	2.970 1	1.690 4	2.250 2	1.120 9	1.247 1
2	0.747 5	0.816 8	1.001 9	2.976 1	1.675 6	2.238 2	1.078 1	1.249 9
3	0.752 0	0.855 1	1.046 8	2.964 9	1.695 0	2.234 3	1.104 9	1.246 3
4	0.756 1	0.833 6	1.016 7	3.005 4	1.671 9	2.247 4	1.088 7	1.257 9
5	0.757 8	0.856 9	1.022 8	3.074 1	1.714 9	2.203 8	1.088 7	1.241 4
6	0.750 8	0.831 8	1.023 4	2.957 5	1.677 4	2.247 7	1.096 6	1.253 4
平均值	0.753 7	0.836 9	1.019 2	2.991 3	1.687 5	2.23	1.096 3	1.249 3
RSD/%	0.57	1.90	1.61	1.46	0.95	0.78	1.37	0.46

仪器	色谱柱	?
仪器	色谱柱	大黄素	大黄素甲醚	杨梅苷	杨梅素	槲皮素	山柰酚	穗花杉双黄酮	扁柏双黄酮
LC-20AC	Agilent ZORBAX ODS C₁₈	0.750 2	0.822 1	1.001 6	2.956 3	1.671 6	2.214 3	1.079 2	1.236 7
	Welch UltimateLP-C₁₈	0.753 7	0.835 4	1.019 7	2.990 6	1.685 2	2.239 2	1.095 1	1.248 2
	Waters ODS C₁₈	0.759 6	0.857 9	1.037 9	3.035 2	1.699 7	2.258 7	1.114 9	1.253 6
Waters e2695	Agilent ZORBAX ODS C₁₈	0.748 2	0.821 5	1.000 4	2.954 3	1.670 1	2.212 4	1.078 7	1.234 6
	Welch UltimateLP-C₁₈	0.752 4	0.832 3	1.018 2	2.988 7	1.684 4	2.238 1	1.094 4	1.246 4
	Waters ODS C₁₈	0.758 8	0.854 1	1.036 5	3.034 5	1.698 9	2.257 6	1.113 2	1.251 8
平均值		0.753 8	0.837 2	1.019 1	2.993 3	1.685 0	2.236 4	1.095 9	1.245 2
RSD/%		0.61	1.86	1.59	1.19	0.76	0.88	1.44	0.63

流速/ （mL/min）	?
流速/ （mL/min）	大黄素	大黄素甲醚	杨梅苷	杨梅素	槲皮素	山柰酚	穗花杉双黄酮	扁柏双黄酮
0.8	0.750 6	0.820 3	1.001 6	2.950 1	1.671 3	2.216 3	1.080 2	1.241 6
1.0	0.753 2	0.830 7	1.014 3	2.992 6	1.688 5	2.227 6	1.096 1	1.249 7
1.2	0.759 7	0.851 2	1.036 1	3.027 9	1.699 7	2.249 8	1.107 8	1.256 2
平均值	0.754 5	0.834 1	1.017 3	2.990 2	1.686 5	2.231 2	1.094 7	1.249 2
RSD/%	0.62	1.89	1.72	1.30	0.85	0.76	1.27	0.59

柱温/℃	?
柱温/℃	大黄素	大黄素甲醚	杨梅苷	杨梅素	槲皮素	山柰酚	穗花杉双黄酮	扁柏双黄酮
25	0.748 1	0.820 0	1.006 2	2.952 4	1.670 3	2.214 6	1.078 3	1.241 3
30	0.754 3	0.837 1	1.019 6	2.992 7	1.685 9	2.237 8	1.097 3	1.249 6
35	0.758 4	0.849 8	1.037 9	3.029 6	1.699 5	2.258 1	1.111 5	1.254 7
平均值	0.753 6	0.835 6	1.021 2	2.991 6	1.685 2	2.236 8	1.095 7	1.248 5
RSD/%	0.69	1.79	1.56	1.29	0.87	0.97	1.52	0.54