Tissue-Specific Analysis of Secondary Metabolites Creates a Reliable Morphological Criterion for Quality Grading of Polygoni Multiflori Radix
Abstract
:1. Introduction
2. Results and Discussion
2.1. Sample Extraction
2.2. Optimization of Analytical Conditions
2.3. Method Validation
2.4. Quantitative Results and Discussion
2.4.1. Raw Materials
2.4.2. Micro-Dissected Tissues
3. Materials and Methods
3.1. Plant Materials, Chemicals and Reagents
3.2. Sample Preparation
3.2.1. Raw Material Extraction
3.2.2. Tissue Extraction
Laser Microdissection
Micro-Dissected Tissue Extraction
3.3. Ultra-Performance Liquid Chromatography Triple-Quadrupole Mass Spectrometry (UPLC-QqQ-MS/MS Analysis)
3.3.1. Conditions
3.3.2. Quantitative Method Validation
3.4. Data Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Acknowledgments
Conflicts of Interest
References
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Sample Availability: Samples of the trans-2,3,5,4′-tetrahydroxystilbene-2-O-β-D-glucopyranoside (trans-THSG), cis-THSG, emodin, physcion, emodin-8-O-β-D-glucosides and physcion-8-O-β-D-glucosides, gallic acid, proanthocyanidin B1, proanthocyanidin B2, epicatechin, catechin and epicatechin-3-gallate are available from the authors. |
Analyte | MRM | Collision Voltage (eV) | Calibration Curve | Sensitivity (ng/mL) | |||
---|---|---|---|---|---|---|---|
Range (ng/mL) | Equation | R2 | LODs | LOQs | |||
Gallic acid | 169.0→125.0 | 11 | 80–4000 | y = 95.80x − 605.88 | 0.9972 | 4.12 | 14.85 |
Proanthocyanidin B1 | 577.1→407.0 | 23 | 40–2000 | y = 28.09x − 105.54 | 0.9994 | 2.51 | 7.59 |
Catechin | 289.1→245.1 | 7 | 20–1000 | y = 25.33x + 256.56 | 0.9974 | 17.95 | 48.25 |
Proanthocyanidin B2 | 577.1→407.0 | 23 | 40–2000 | y = 25.93x − 48.90 | 0.9991 | 3.17 | 15.92 |
Epicatechin | 289.1→245.1 | 7 | 10–1000 | y = 31.33x − 78.60 | 0.9939 | 15.52 | 40.66 |
cis-THSG | 405.0→243.0 | 15 | 5–1000 | y = 213.54x − 348.14 | 0.9948 | 1.04 | 1.62 |
Epcatechini-3-gallate | 441.1→169.0 | 15 | 5–1000 | y = 55.85x − 82.32 | 0.9936 | 0.99 | 6.62 |
trans-THSG | 405.0→243.1 | 15 | 40–8000 | y = 196.23x − 357.59 | 0.9992 | 9.64 | 11.02 |
Emodin-8-O-β-D-glucoside | 431.1→269.1 | 27 | 40–2000 | y = 801.51x + 29.72 | 0.9911 | 4.36 | 4.58 |
Physcion-8-O-β-D-glucoside | 445.1→283.1 | 7 | 40–2000 | y = 58.92x − 103.32 | 0.9929 | 4.84 | 7.14 |
Emodin | 269.0→225.0 | 25 | 80–800 | y = 2422.72 + 16798.78 | 0.9907 | 0.22 | 1.15 |
Physcion | 283.0→240.0 | 20 | 40–2000 | y = 117.20x − 1082.86 | 0.9945 | 1.73 | 4.03 |
Repeatability (n = 6, RSD, %) | Precision (n = 6) | Spike Recovery (n = 3, Mean (RSD), %) | Stability (48 h, n = 7) | |||||||
---|---|---|---|---|---|---|---|---|---|---|
Intra-day | Inter-Day | Low | Middle | High | ||||||
Mean (ng/mL) | RSD, % | Mean (ng/mL) | RSD, % | Mean (ng/mL) | RSD, % | |||||
Gallic acid | 10.81 | 394.15 | 3.01 | 471.86 | 7.69 | 96.71 (4.58) | 97.23 (4.15) | 94.94 (1.53) | 445.58 | 8.73 |
Proanthocyanidin B1 | 7.43 | 959.60 | 2.47 | 999.00 | 4.08 | 90.73 (6.29) | 91.72 (5.46) | 90.85 (4.85) | 1042.45 | 7.81 |
Catechin | 9.35 | 293.40 | 4.43 | 329.87 | 9.01 | 101.89(7.75) | 95.14(6.51) | 98.52(1.86) | 315.646 | 7.38 |
Proanthocyanidin B2 | 8.31 | 341.51 | 2.21 | 357.12 | 4.16 | 96.17(4.73) | 93.72(1.52) | 96.08(3.70) | 369.96 | 5.54 |
Epicatechin | 3.48 | 205.35 | 5.58 | 221.47 | 4.09 | 102.69(4.13) | 107.37(1.31) | 101.70(4.44) | 203.26 | 5.68 |
cis-THSG | 4.22 | 2012.47 | 1.21 | 2198.07 | 1.79 | 82.51(10.18) | 102.87(9.21) | 87.96(10.49) | 2107.79 | 4.76 |
Epcatechini-3-gallate | 7.27 | 440.67 | 7.25 | 448.75 | 5.69 | 103.50(5.13) | 95.52(8.18) | 100.89(2.06) | 456.16 | 4.23 |
trans-THSG | 2.42 | 7396.96 | 2.28 | 7932.62 | 6.72 | 95.15(6.80) | 92.82(5.34) | 85.98(8.56) | 7865.36 | 5.95 |
Emodin-8-O-β-D-glucoside | 3.19 | 2124.69 | 0.48 | 2365.74 | 0.78 | 94.72(1.47) | 100.49(1.99) | 97.93(1.58) | 2272.80 | 4.48 |
Physcion-8-O-β-D-glucoside | 3.58 | 503.18 | 1.54 | 558.48 | 1.59 | 94.11(2.09) | 95.36(4.40) | 85.81(0.38) | 529.89 | 4.66 |
Emodin | 2.02 | 719.23 | 8.32 | 669.05 | 9.95 | 99.05(2.92) | 106.78(2.56) | 101.18(2.33) | 682.54 | 5.77 |
Physcion | 3.40 | 605.38 | 4.49 | 569.70 | 6.19 | 97.94(9.05) | 92.87(6.61) | 99.27(8.78) | 578.33 | 3.97 |
Repeatability (n = 6, RSD, %) | Precision (n = 6) | Spike Recovery (n = 3, mean (RSD), %) | Stability (48 h, n = 7) | |||||||
---|---|---|---|---|---|---|---|---|---|---|
Intra-Day | Inter-day | Low | Middle | High | ||||||
Mean (ng/mL) | RSD, % | Mean (ng/mL) | RSD, % | Mean (ng/mL) | RSD, % | |||||
Gallic acid | 2.85 | 144.60 | 3.08 | 147.10 | 8.14 | 108.78(5.00) | 101.82(15.80) | 120.95(10.18) | 142.90 | 3.82 |
Proanthocyanidin B1 | 9.55 | 54.92 | 5.65 | 60.05 | 4.27 | 111.34(11.31) | 109.33(10.52) | 111.09(4.81) | 51.82 | 5.19 |
Catechin | 6.11 | 113.16 | 7.96 | 141.29 | 6.68 | 109.47(5.94) | 102.63(11.29) | 98.44(5.67) | 135.61 | 5.60 |
Proanthocyanidin B2 | 6.25 | 96.75 | 6.14 | 103.44 | 6.50 | 100.39(11.61) | 99.32(3.13) | 100.73(0.33) | 100.70 | 7.46 |
Epicatechin | 3.68 | 63.25 | 7.48 | 72.78 | 4.76 | 100.98(7.78) | 100.80(9.17) | 109.27(9.68) | 78.55 | 8.64 |
cis-THSG | 4.25 | 3040.38 | 2.09 | 3682.49 | 3.91 | 113.93(9.18) | 103.54(6.08) | 99.46(5.65) | 3378.57 | 5.82 |
Epcatechini-3-gallate | 6.33 | 162.27 | 6.03 | 178.00 | 4.79 | 93.66(8.48) | 97.62(6.72) | 90.98(10.54) | 131.54 | 4.90 |
trans-THSG | 5.91 | 4296.71 | 0.56 | 4921.06 | 5.25 | 109.77(6.74) | 112.86(4.00) | 102.28(9.76) | 7249.63 | 7.37 |
Emodin-8-O-β-D-glucoside | 4.92 | 193.50 | 2.44 | 241.92 | 3.34 | 113.83(1.50) | 109.74(15.73) | 101.83(2.13) | 359.83 | 6.05 |
Physcion-8-O-β-D-glucoside | 11.17 | 25.00 | 6.44 | 31.79 | 9.84 | 109.61(6.19) | 103.38(8.95) | 115.32(8.42) | 28.27 | 9.00 |
Emodin | 6.23 | 183.42 | 1.66 | 192.12 | 4.33 | 102.25(8.27) | 107.03(11.56) | 109.24(1.84) | 218.22 | 8.14 |
Physcion | 11.22 | 90.94 | 1.92 | 85.36 | 5.69 | 94.96(4.87) | 90.12(9.48) | 92.84(6.01) | 68.27 | 3.54 |
Sample No. | Grade | Type (a) | Cultivated Location | Collection Date |
---|---|---|---|---|
PMR-RMA1 | 1 (b) | RM | Guizhou Province, Shibing Country | 2017-1-18 |
PMR-RMA2 | 1 | RM | Guizhou Province, Shibing Country | 2017-1-16 |
PMR-RMA3 | 1 | RM | Guizhou Province, Shibing Country | 2017-2-18 |
PMR-RMA4 | 2 | RM | Guizhou Province, Shibing Country | 2017-1-18 |
PMR-RMA5 | 2 | RM | Guizhou Province, Shibing Country | 2017-2-16 |
PMR-RMA6 | 2 | RM | Guizhou Province, Shibing Country | 2017-2-18 |
PMR-RMA7 | 3 | RM | Guizhou Province, Shibing Country | 2017-1-18 |
PMR-RMA8 | 3 | RM | Guizhou Province, Shibing Country | 2017-2-16 |
PMR-RMA9 | 3 | RM | Guizhou Province, Shibing Country | 2017-2-18 |
PMR-TA1 | 1 | T | Guizhou Province, Shibing Country | 2017-2-16 |
PMR-TA2 | 2 | T | Guizhou Province, Shibing Country | 2017-2-16 |
PMR-TA3 | 3 | T | Guizhou Province, Shibing Country | 2017-2-16 |
PMR-TB1 | 1 | T | Guangdong Province, Xinxing Country | 2017-2-8 |
PMR-TB2 | 2 | T | Guangdong Province, Xinxing Country | 2017-2-8 |
PMR-TB3 | 3 | T | Guangdong Province, Xinxing Country | 2017-2-8 |
PMR-TC1 | 1 | T | Hubei Province, Lizhou City | 2017-1-16 |
PMR-TC2 | 2 | T | Hubei Province, Lizhou City | 2017-1-16 |
PMR-TC3 | 3 | T | Hubei Province, Lizhou City | 2017-1-16 |
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Liang, L.; Xu, J.; Liang, Z.-T.; Dong, X.-P.; Chen, H.-B.; Zhao, Z.-Z. Tissue-Specific Analysis of Secondary Metabolites Creates a Reliable Morphological Criterion for Quality Grading of Polygoni Multiflori Radix. Molecules 2018, 23, 1115. https://doi.org/10.3390/molecules23051115
Liang L, Xu J, Liang Z-T, Dong X-P, Chen H-B, Zhao Z-Z. Tissue-Specific Analysis of Secondary Metabolites Creates a Reliable Morphological Criterion for Quality Grading of Polygoni Multiflori Radix. Molecules. 2018; 23(5):1115. https://doi.org/10.3390/molecules23051115
Chicago/Turabian StyleLiang, Li, Jun Xu, Zhi-Tao Liang, Xiao-Ping Dong, Hu-Biao Chen, and Zhong-Zhen Zhao. 2018. "Tissue-Specific Analysis of Secondary Metabolites Creates a Reliable Morphological Criterion for Quality Grading of Polygoni Multiflori Radix" Molecules 23, no. 5: 1115. https://doi.org/10.3390/molecules23051115