液相色谱-串联质谱法检测药食两用杂交天麻及其亲本品种中的氨基酸、核苷类成分

王广政; 巩晴晴; 俞年军; 高翰博; 邢丽花; 杜方平

doi:10.13386/j.issn1002-0306.2022030299

液相色谱-串联质谱法检测药食两用杂交天麻及其亲本品种中的氨基酸、核苷类成分

doi: 10.13386/j.issn1002-0306.2022030299

王广政^{1, 2,},
巩晴晴^{1, 2},
俞年军^{1, 2, 3, 4, 5, ,},
高翰博^{1, 2},
邢丽花^{1, 3},
杜方平⁶

1.
安徽中医药大学药学院，安徽合肥 230012
2.
安徽省中医药科学院中药资源保护与开发研究所，安徽合肥 230012
3.
安徽道地中药材品质提升协同创新中心，安徽合肥 230012
4.
中药研究与开发安徽省重点实验室，安徽合肥 230012
5.
新安医学教育部重点实验室，安徽合肥 230012
6.
金寨县药用菌种植专业合作社，安徽六安 237000

基金项目: 国家自然科学基金联合基金项目（U19A2009）；安徽高校协同创新项目（GXXT-2019-043）；安徽道地中药材品质提升协同创新中心项目（皖教科〔2013〕2号）；安徽省教育厅项目（皖教秘科〔2014〕44 号）；2019年中医药公共卫生服务补助专项“全国中药资源普查项目”（财社〔2019〕43号）。

详细信息

作者简介:
王广政（1998−），男，硕士研究生，研究方向：中药资源与中药质量，E-mail：2054332851@qq.com

通讯作者:
俞年军（1965−），男，硕士，教授，研究方向：中药质量评价，E-mail：ynj2005288@sina.com

中图分类号: TS201.1
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出版历程
- 收稿日期: 2022-03-25
- 网络出版日期: 2022-12-16
- 刊出日期: 2023-01-17

Determination of Amino Acids and Nucleosides in Medicinal and Edible Hybrid Gastrodia elata and Its Parent Varieties Based on Liquid Chromatography-Tandem Mass Spectrometry

WANG Guangzheng^{1, 2
,},
GONG Qingqing^{1, 2},
YU Nianjun^{1, 2, 3, 4, 5
, ,},
GAO Hanbo^{1, 2},
XING Lihua^{1, 3},
DU Fangping⁶

1.
School of Pharmacy, Anhui University of Traditional Chinese Medicine, Hefei 230012, China
2.
Institute of Traditional Chinese Medicine Resources Protection and Development, Anhui Academy of Traditional Chinese Medicine, Hefei 230012, China
3.
Collaborative Innovation Center for Quality Improvement of Daodi Chinese Medicinal Materials, Hefei 230012, China
4.
Anhui Key Laboratory of Traditional Chinese Medicine Research and Development, Hefei 230012, China
5.
Xin'an Medical and Education Ministry Key Laboratory, Hefei 230012, China
6.
Jinzhai County Medicine Mushroom Planting Professional Cooperative, Liuan 237000, China

摘要

摘要: 实验采用超高效液相色谱与质谱联用技术精确检测金寨红乌杂交天麻及其亲本云南乌天麻、金寨红天麻中的氨基酸、核苷类成分，结合统计学方法阐明三个品种天麻之间的氨基酸、核苷类成分组成与含量差异并综合评价，为后续金寨杂交天麻的资源利用与食品开发提供科学依据。C₁₈柱（2.1 mm×100 mm，1.8 μm），流动相用0.2%甲酸水（A）-乙腈（B），体积流量为0.2 mL/min，柱温30 ℃，梯度洗脱；电喷雾离子源以多反应监测（MRM）模式测定，喷雾电压5500 V；离子化温度550 ℃；离子源气压（Gas）为241.3 kPa，雾化气（GS1）为379.2 kPa，辅助气（GS2）为379.2 kPa。实验结果运用主成分分析（PCA）和优劣解距离法（TOPSIS）进行分析并综合评价。结果显示，31种成分在0~144.58 μg/mL范围内线性关系良好，决定系数R²均大于0.999，精密度、稳定性、重复性峰面积RSD均不大于3.0%，平均回收率92.69%~99.5%，回收率RSD 1.3%~2.9%。天麻内含有丰富的氨基酸与核苷类成分，其中谷氨酸与天门冬氨酸为主要成分，杂交天麻总氨基酸、总核苷含量较亲代天麻更高，且总氨基酸呈显著性差异（P<0.05），PCA与TOPSIS法评价结果也以S1、S2杂交天麻品质最优。该实验为天麻的质量评价建立了新方法，并为金寨杂交品种的推广提供了科学依据。
- 天麻 /
- 液相色谱-串联质谱法 /
- 氨基酸 /
- 核苷 /
- 主成分分析 /
- TOPSIS法 /
- 药食两用 /
- 质量评价
Abstract: To provide a scientific basis for the follow-up resource utilization and food development of Jinzhai hybrid G. elata, the ultra-high performance liquid chromatography coupled with mass spectrometry was utilized to accurately determine the contents of amino acids and nucleosides in Jinzhai Hongwu hybrid Gastrodia elata Bl. and its parents Yunnan G. elata Bl. f. glauca S. Chow and Jinzhai G. elata Bl. f. elata. Statistical methods were also applied to clarify and comprehensively evaluate the composition and content differences in amino acids and nucleosides among the three varieties of G. elata. C₁₈ column (2.1 mm×100 mm, 1.8 μm) was used with 0.2% formic acid water (A)-acetonitrile (B) as the mobile phase at a flow rate of 0.2 mL/min. The column temperature was 30 ℃ with gradient elution. Moreover, electrospray ion source was measured in multiple reaction monitoring (MRM) mode, which was set as spray voltage at 5500 V; ionization temperature at 550 ℃; ion source gas pressure (Gas) at 241.3 kPa, atomizing gas (GS1) at 379.2 kPa, and auxiliary gas (GS2) at 379.2 kPa. The results revealed that there was a good linear relationship in the range of 0~144.58 μg/mL of the 31 components. The coefficient of determination R² was greater than 0.999, and the RSD of the precision, stability, and repeatability of the peak area was not greater than 3.0%. The average recovery rate was between 92.69% and 99.5%, while recovery rate RSD ranged from 1.3% to 2.9%. Based on all above, G. elata was rich in amino acids and nucleotides, of which glutamic acid and aspartic acid were the main components. The contents of total amino acids and nucleosides in hybrid G. elata were higher than those of its parents G. elata. And there was a significant difference in the amino acids contents of different varieties (P<0.05). In the evaluation results of PCA and TOPSIS methods, the qualities of S1 and S2 hybrid G. elata were the best. This paper established a new method for evaluating the quality of G. elata and provided a scientific basis for the development and exploitation of Jinzhai hybrid varieties.
- Gastrodia elata /
- liquid chromatography-tandem mass spectrometry /
- amino acid /
- nucleoside /
- principal component analysis /
- TOPSIS /
- homology of medicine and food /
- quality evaluation

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图 1 31种标准品的MRM图

Figure 1. MRM diagrams of 31 standard components

下载: 全尺寸图片幻灯片

图 2 不同品种天麻总氨基酸比较

Figure 2. Comparison of total amino acids of different Gastrodia elata varieties

注：不同字母标注表示呈显著性差异（P<0.05），图3同。

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图 3 不同品种天麻总核苷比较

Figure 3. Comparison of total nucleosides in different Gastrodia elata varieties

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表 1 天麻样品采集信息

Table 1. Collection information of Gastrodia elata samples

样品编号	采集地点	品种	海拔（m）	栽培方式
S1	安徽金寨班竹园	红乌天麻	581.2	林下
S2	安徽金寨汤家汇	红乌天麻	593.7	林下
S3	安徽金寨双河镇	红乌天麻	554.9	大棚
S4	安徽金寨果子园	红天麻	348.7	大棚
S5	安徽金寨吴家店	红天麻	366.0	大棚
S6	安徽金寨沙河乡	红天麻	273.1	大棚
S7	云南昭通小草坝	乌天麻	1934.2	大棚
S8	云南昭通小草坝	乌天麻	1883.7	大棚
S9	云南昭通小草坝	乌天麻	1745.4	大棚

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表 2 31种成分测定中优化的质谱条件参数

Table 2. Optimized MS parameters for the determination of 31 components

化合物	保留时间（min）	母离子	子离子	去簇电压（V）	碰撞电压（V）
甘氨酸	5.83	76.04	30.00^*、49.00	73	6
丙氨酸	6.36	90.06	44.02^*、46.78	79	10
丝氨酸	6.18	106.05	59.99^*、74.30	67	8
天门冬氨酸	6.51	134.05	87.96^*、73.80	59	10
天门冬酰胺	4.12	132.80	115.70^*、73.90	46	13
缬氨酸	6.41	118.09	72.06^*、55.06	67	8
谷氨酸	3.46	147.08	83.92^*、130.58	83	14
异亮氨酸	2.98	132.00	86.00^*、69.00	66	15
甲硫氨酸	7.01	150.06	104.03^*、133.00	91	10
精氨酸	6.88	175.12	70.20^*、116.00	88	18
组氨酸	3.52	156.08	110.03^*、93.17	95	16
苏氨酸	3.54	120.30	76.80^*、104.70	54	11
苯丙氨酸	3.55	166.10	120.05^*、103.00	56	14
亮氨酸	6.92	132.10	86.05^*、42.30	98	10
胱氨酸	3.65	240.80	151.90^*、120.00	71	18
羟脯氨酸	4.43	133.80	71.80^*、69.60	52	25
酪氨酸	3.56	182.16	136.08^*、165.10	46	17
色氨酸	5.31	205.00	188.00^*、72.2	202	15
脯氨酸	7.01	116.07	70.02^*、68.03	68	10
赖氨酸	4.29	147.00	84.00^*、130.00	52	24
γ-氨基丁酸	5.68	103.70	86.90^*、87.00	32	14
鸟嘌呤	4.20	151.80	135.00^*、135.00	62	24
腺嘌呤	6.57	136.06	119.00^*、92.00	51	24
尿嘧啶	1.78	113.04	70.00^*、95.00	111	21
次黄嘌呤	6.96	137.05	110.00^*、118.70	46	15
胸苷	5.68	243.10	127.07^*、99.00	61	13
鸟苷	4.21	284.30	152.00^*、135.00	62	15
肌苷	3.34	269.00	137.05^*、137.05	46	15
尿苷	4.21	244.90	113.00^*、70.05	103	13
腺苷	3.47	267.9	118.70^*、135.00	86	23
胞苷	1.54	244.09	94.65^*、111.80	61	10
注：表中“*”标注离子为定量离子。

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表 3 21种氨基酸和10种核苷的线性关系考察

Table 3. Linear relationship of 21 amino acids and 10 nucleosides

化合物	回归方程	线性范围（μg/mL）	R²	LOD（ng/mL）	LOQ（ng/mL）
甘氨酸	Y=3×10⁶X+147.2	0.034~5.68	0.9992	6.8	22.76
丙氨酸	Y=5×10⁶X−10⁶	0.15~61.43	0.9994	3.03	11.04
丝氨酸	Y=4×10⁸X−4250.3	0.11~10.28	0.9991	0.82	3.26
天门冬氨酸	Y=2×10⁸X+12493	0.024~38.29	0.9991	2.74	9.63
天门冬酰胺	Y=6×10⁸X−2×10⁵	0.13~89.45	0.9996	4.09	13.88
缬氨酸	Y=4×10⁶X−48932	0.097~14.53	0.9993	1.68	5.47
谷氨酸	Y=3×10⁸X+28292	0.19~144.58	0.9991	0.15	0.65
异亮氨酸	Y=2×10⁷X−192213	0.093~10.02	0.9992	4.2	13.74
甲硫氨酸	Y=5×10⁷X−273.7	0.027~1.805	0.9997	3.25	10.75
精氨酸	Y=3×10⁶X−1×10⁵	0.31~59.34	0.9998	3.47	12.31
组氨酸	Y=82104X−29.7	0.40~6.86	0.9995	0.76	2.39
苏氨酸	Y=8×10⁸X+10682.9	0.32~10.59	0.9996	2.11	7.03
苯丙氨酸	Y=4×10⁸X−4564.6	0.068~2.83	0.9998	4.45	14.61
亮氨酸	Y=1×10⁶X−614.9	0.51~34.30	0.9990	6.87	19.54
胱氨酸	Y=32921X−342.4	0.75~13.23	0.9992	0.16	0.57
羟脯氨酸	Y=4×10⁷X−28312.4	0.29~7.41	0.9997	0.23	0.72
酪氨酸	Y=6×10⁷X−72791	0.092~4.24	0.9994	2.49	8.63
色氨酸	Y=4×10⁷X−1×10⁶	0.058~3.92	0.9999	5.68	16.82
脯氨酸	Y=8×10⁷X−34258.3	0.11~3.08	0.9992	2.6	8.97
赖氨酸	Y=74453X−435.6	0.015~10.12	0.9991	3.64	12.06
γ-氨基丁酸	Y=2×10⁸X−432350	0.062~3.66	0.9997	1.74	6.08
鸟嘌呤	Y=4×10⁸X+2123.9	0.032~3.14	0.9998	3.91	12.97
腺嘌呤	Y=5×10⁸X−234321	0.026~2.98	0.9992	1.68	5.78
尿嘧啶	Y=1×10⁹X−33241	0.067~4.02	0.9995	3.32	10.42
次黄嘌呤	Y=1×10⁹X−9283.7	0.031~3.29	0.9991	2.41	8.73
胸苷	Y=7×10⁷X−3124.9	0.056~3.92	0.9991	5.75	18.14
鸟苷	Y=8×10⁷X−15467	0.23~5.82	0.9993	6.1	18.56
肌苷	Y=1×10⁷X−146520	0.14~4.12	0.9996	4.72	16.07
尿苷	Y=4×10⁷X+129202	0.096~4.43	0.9992	4.14	15.83
腺苷	Y=5×10⁶X−146342	0.029~3.87	0.9998	3.31	10.75
胞苷	Y=4×10⁹X+547650	0.024~8.46	0.9996	4.52	14.84

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表 4 天麻中31种测定成分的精密度、重复性、稳定性及加样回收率（n=6，%）

Table 4. Precision, repeatability, stability and sample recovery of 31 components in Gastrodia elata (n=6, %)

化合物	精密度RSD		重复性 RSD	稳定性 RSD	平均回收率	回收率 RSD
化合物	日间	日内	重复性 RSD	稳定性 RSD	平均回收率	回收率 RSD
甘氨酸	1.25	0.34	1.6	1.7	98.47	1.8
丙氨酸	0.86	0.52	2.1	1.8	97.25	1.7
丝氨酸	0.76	0.73	1.3	1.6	96.38	2.0
天门冬氨酸	0.95	1.0	1.8	2.1	98.12	2.6
天门冬酰胺	0.76	0.82	2.3	1.9	97.75	1.9
缬氨酸	0.35	0.29	1.6	2.1	96.51	2.4
谷氨酸	0.71	0.67	1.5	2.4	98.68	2.1
异亮氨酸	0.93	1.1	1.9	1.7	97.37	2.0
甲硫氨酸	1.38	1.4	2.4	2.9	95.54	1.8
精氨酸	0.87	0.72	2.6	2.5	98.38	2.3
组氨酸	0.92	0.91	2.0	1.1	95.69	2.2
苏氨酸	0.47	0.35	1.9	2.1	97.43	2.4
苯丙氨酸	1.0	0.92	1.7	1.9	96.89	2.8
亮氨酸	0.77	0.74	1.4	1.6	95.01	1.6
胱氨酸	0.48	0.39	2.3	1.2	99.50	2.7
羟脯氨酸	0.72	0.62	2.1	1.6	94.86	2.1
酪氨酸	0.57	0.69	1.7	1.1	95.33	1.9
色氨酸	0.46	0.44	2.2	1.9	97.37	2.9
脯氨酸	0.38	0.41	1.5	1.7	98.13	2.4
赖氨酸	0.31	0.28	1.4	2.1	95.34	1.3
γ-氨基丁酸	0.64	0.76	1.5	2.4	96.08	2.1
鸟嘌呤	0.86	1.2	1.2	0.93	97.21	2.6
腺嘌呤	1.12	0.91	1.9	1.2	98.48	2.9
尿嘧啶	0.92	0.87	2.3	2.0	92.69	2.8
次黄嘌呤	0.64	0.79	1.9	1.2	98.23	2.6
胸苷	0.67	0.71	1.8	1.8	99.02	2.3
鸟苷	0.93	0.82	2.0	1.4	93.31	1.4
肌苷	0.69	0.57	1.7	2.1	95.47	1.5
尿苷	0.41	0.32	0.98	2.9	96.81	2.7
腺苷	0.37	0.48	1.8	1.7	96.42	2.5
胞苷	0.64	0.56	1.4	2.4	97.40	2.6

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表 5 不同品种天麻中21种氨基酸和10种核苷的含量测定结果（$\bar x $±s，μg/g，n=3）

Table 5. Contents of 21 amino acids and 10 nucleosides in Gastrodia elata of different varieties ($\bar x $±s, μg/g, n=3)

No.	红乌天麻			红天麻			乌天麻
No.	S1	S2	S3	S4	S5	S6	S7	S8	S9
甘氨酸	790.62±38.24^a	790.12±27.24^a	786.69±31.41^a	592.64±21.48^c	585.13±22.95^c	523.82±17.38^d	559.03±24.16^cd	611.55±27.47^c	683.15±28.58^b
丙氨酸	733.34±26.52^a	669.02±35.12^b	709.33±37.15^ab	386.05±27.67^ef	349.83±12.77^f	414.84±10.56^e	530.33±27.17^cd	502.53±28.67^d	564.23±12.76^c
丝氨酸	81.81±1.35^b	74.93±2.22^bc	110.86±7.34^a	62.97±4.74^cd	52.99±4.96^d	66.35±5.93^c	75.49±2.36^bc	55.37±6.74^cd	71.71±7.62^bc
天门冬氨酸	1181.16±49.42^a	1091.34±41.56^ab	1084.25±37.54^ab	1024.61±36.06^b	1036.74±42.77^b	1006.24±46.36^b	1124.83±54.56^ab	1012.54±27.74^b	1045.71±48.46^b
天门冬酰胺	19.13±0.27^c	30.18±0.51^bc	31.26±1.74^bc	24.01±1.52^c	32.07±1.04^b	28.97±1.54^bc	24.54±0.17^c	88.87±7.73^a	30.13±1.77^bc
缬氨酸	585.76±36.64^a	511.72±29.22^b	578.56±28.33^a	562.86±41.86^ab	369.33±14.16^c	427.55±26.43^c	430.81±36.62^c	408.12±19.47^c	496.54±32.87^b
谷氨酸	1357.44±78.28^a	1304.14±77.39^a	1218.85±69.59^a	1267.18±84.96^a	1183.00±65.77^a	1245.37±92.43^a	1196.75±79.26^a	1252.34±103.21^a	1161.75±87.329^a
异亮氨酸^*	410.93±43.48^ab	459.28±39.26^ab	380.36±31.49^bc	288.73±24.78^c	367.16±41.79^c	291.09±17.84^c	286.38±36.35^c	376.28±32.85^bc	472.78±51.85^a
甲硫氨酸^*	327.84±14.56^a	347.04±9.66^a	148.46±10.74^c	200.87±18.94^b	209.06±12.52^b	94.48±8.74^d	188.56±20.57^b	138.32±15.44^c	87.52±4.76^d
精氨酸^*	481.59±39.54^c	542.41±16.66^bc	682.42±33.73^a	667.66±28.23^a	593.56±32.83^b	650.46±33.32^a	518.91±21.56^c	556.12±28.36^bc	583.54±30.36^bc
组氨酸	263.42±11.31^a	252.90±9.25^ab	244.64±11.21^ab	169.61±11.66^e	194.71±9.93^d	190.35±8.52^de	236.43±12.76^b	181.13±8.85^de	214.93±10.45^c
苏氨酸^*	483.24±39.45^a	265.81±29.63^c	357.21±32.77^b	170.92±15.71^d	207.86±19.62^d	180.26±10.43^d	166.03±9.44^d	165.71±14.89^d	178.21±16.63^d
苯丙氨酸^*	37.72±1.82^a	26.56±0.94^b	24.82±1.76^b	17.97±1.01^c	16.84±0.72^c	15.13±1.89^c	14.03±2.74^c	16.25±1.92^c	10.88±0.86^d
亮氨酸^*	780.69±19.31^a	783.5±21.54^a	782.43±32.86^a	782.27±65.21^a	794.57±24.74^a	776.03±53.9^a	780.19±32.6^a	778.2±35.2^a	790.2±22.1^a
胱氨酸	1102.35±42.71^a	1052.11±76.2^ab	998.41±64.76^abc	933.15±24.96^bc	950.11±51.66^bc	897.63±64.96^c	793.22±63.96^c	859.31±54.43^c	1088.16±46.81^a
羟脯氨酸	505.74±18.53^a	443.75±27.86^b	411.13±22.51^bc	393.43±12.96^bc	335.73±16.68^cd	376.13±13.96^c	442.11±27.63^b	313.67±42.36^d	483.72±28.33^ab
酪氨酸	50.96±2.41^b	29.84±4.52^c	22.66±1.63^cd	30.78±2.56^c	17.62±1.33^d	27.48±3.76^cd	24.45±3.41^cd	71.02±10.43^a	23.46±1.69^cd
色氨酸^*	8.18±0.21^a	7.67±0.12^b	4.43±0.28^e	5.74±0.02^d	6.62±0.11^c	6.58±0.25^c	4.37±0.36^e	4.16±0.24^e	8.02±0.33^ab
脯氨酸	534.23±28.26^a	403.13±16.75^b	385.61±18.87^b	376.92±9.45^b	309.65±8.57^cd	286.14±13.77^d	328.46±12.82^c	290.44±15.34^d	269.96±20.48^d
赖氨酸^*	3.74±0.09^c	3.98±0.13^c	6.53±0.52^a	3.63±0.08^c	2.84±0.16^d	3.49±0.08^c	5.2±0.06^b	4.91±0.25^b	5.1±0.22^b
γ-氨基丁酸	77.25±7.63^b	95.30±5.78^a	32.08±4.64^ef	44.41±5.89^d	26.73±1.13^f	41.34±2.52^de	40.9±3.65^de	81.2±4.28^b	53.81±7.32c
鸟嘌呤	7.34±0.53^a	8.07±0.35^a	5.61±0.52^c	3.32±0.06^e	4.61±0.38^d	2.37±0.08^f	2.06±0.07^f	0.55±0.03^g	3.07±0.24^e
腺嘌呤	5.11±0.42^a	7.65±0.62^a	3.67±0.21^b	3.54±0.05^b	0.88±0.04^d	2.75±0.12^c	3.09±0.25^b	1.12±0.09^d	2.61±0.21^c
尿嘧啶	172.41±15.3^a	11.83±0.82^f	35.31±2.91^e	90.97±4.12^bc	79.45±8.34^c	21.04±1.41^f	89.02±4.85^bc	97.15±4.23^b	54.00±1.90^d
次黄嘌呤	53.73±3.24^bc	42.74±2.49^c	61.61±7.42^b	40.66±4.95^c	34.81±1.77^c	31.61±8.53^c	56.68±4.85^b	38.11±2.98^c	117.1±12.6^a
胸苷	18.64±2.52^ab	20.82±1.95^a	6.31±0.53^d	8.82±1.06^cd	16.77±2.74^b	8.12±0.72^cd	8.15±0.93^cd	5.06±0.12^d	10.92±1.05^c
鸟苷	142.26±4.73^b	179.86±12.64^a	94.19±10.82^c	42.54±0.84^e	130.08±7.84^b	39.55±2.74^e	57.47±4.95^d	132.38±10.6^b	87.46±9.21^c
肌苷	95.91±3.49^a	90.01±2.31^a	72.36±4.62^b	77.41±3.27^b	71.69±5.28^b	74.08±7.32^b	74.35±4.12^b	89.49±5.22^a	75.93±2.51^b
尿苷	113.12±6.56^c	193.44±11.86^a	55.09±6.57^e	54.72±4.56^e	59.39±6.96^e	59.24±7.24^e	74.27±4.91^e	161.5±13.5^b	96.39±7.22^d
腺苷	131.97±7.49^a	132.24±12.63^a	131.91±9.25^a	132.07±10.98^a	132.12±6.33^a	132.78±9.98^a	131.28±8.36^a	132.07±11.3^a	131.98±16.52^a
胞苷	6.34±0.24^b	7.47±0.23^a	5.76±0.38^b	4.16±0.13^c	4.61±0.11^c	5.78±0.18^b	6.46±0.72^b	4.21±0.21^c	4.02±0.15^c
氨基酸总量	9816.83±210.56^a	9108.51±254.68^b	9000.67±195.36^b	8005.34±206.95^cd	7641.36±276.29^d	7549.43±158.35^d	7771.55±269.63^cd	7767.61±384.51^cd	8322.87±295.57^c
核苷总量	746.67±21.56^a	693.97±29.26^b	471.81±17.67^e	458.07±29.34^e	534.31±27.94^d	376.49±16.63^f	503.33±19.26^de	661.76±26.26^b	583.33±18.43^c
必需氨基酸总量	2533.93±95.46^a	2436.25±73.39^a	2386.66±83.03^a	2137.79±63.20^bc	2198.51±72.13^b	2017.52±83.48^bc	1963.67±62.15^c	2039.95±92.09^bc	2136.25±69.25^bc
注：不同字母标注表示同一行数据具有显著性差异（P<0.05）；“*”标记表示人体必需氨基酸。

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表 6 氨基酸主成分分析及综合评价

Table 6. Principal component analysis and comprehensive evaluation of amino acids

样品	主成分					F	排名
样品	F1	F2	F3	F4	F5	F	排名
S1	6.68	−0.38	−1.65	−0.48	0.09	2.80	1
S2	2.90	0.78	−0.44	0.94	−0.39	1.50	2
S3	1.35	−1.07	3.63	−1.06	0.29	0.86	3
S4	−1.53	−1.40	−0.64	−0.82	1.72	−1.03	7
S5	−2.74	−1.41	−1.67	1.82	0.04	−1.56	8
S6	−2.88	−1.33	−0.70	−1.36	−0.17	−1.81	9
S7	−1.26	−0.77	0.41	−0.18	−2.26	−0.80	6
S8	−1.99	4.22	−0.59	−1.45	0.08	−0.46	5
S9	−0.53	1.36	1.64	2.60	0.59	0.49	4

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表 7 核苷主成分分析及综合评价

Table 7. Principal component analysis and comprehensive evaluation of nucleosides

样品	主成分		F	排名
样品	F1	F2	F	排名
S1	2.53	−0.41	1.13	2
S2	3.38	0.77	1.68	1
S3	−0.42	0.83	−0.08	3
S4	−0.68	−1.16	−0.48	7
S5	0.35	−1.92	−0.10	4
S6	−0.95	−0.54	−0.52	8
S7	−1.29	1.08	−0.46	5
S8	−1.44	−0.36	−0.72	9
S9	−1.48	1.69	−0.46	6

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表 8 样品中31种成分TOPSIS分析结果

Table 8. TOPSIS analysis results of 31 components in the samples

项	总成分			排序结果
项	正理想解距离D⁺	负理想解距离D⁻	相对贴近度C_i	排序结果
S1	0.916	1.789	0.661	1
S2	1.305	1.433	0.523	2
S3	1.634	0.897	0.355	4
S4	1.690	0.700	0.293	6
S5	1.784	0.693	0.28	7
S6	1.923	0.366	0.16	9
S7	1.864	0.499	0.211	8
S8	1.731	1.049	0.377	3
S9	1.703	0.894	0.344	5

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