Main components from cultivated and wild Nardostachyos Radix et Rhizoma by LC-MS and GC-MS / 中国中药杂志

In this study, ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and gas chromatography-mass spectrometry(GC-MS) were combined with non-targeted metabonomic analysis based on multivariate statistics analysis, and the content of five indicative components in nardosinone was determined and compared by UPLC. The main chemical components of Nardostachyos Radix et Rhizoma with imitative wild cultivation and wild Nardostachyos Radix et Rhizoma were comprehensively analyzed. The results of multivariate statistical analysis based on liquid chromatography-mass spectrometry(LC-MS) and GC-MS were consistent. G1 and G2 of the imitative wild cultivation group and G8-G19 of the wild group were clustered into category 1, while G7 of the wild group and G3-G6 of the imitative wild cultivation group were clustered into category 2. After removing the outlier data of G1, G2, and G7, G3-G6 of the imitative wild cultivation group were clustered into one category, and G8-G19 of the wild group were clustered into the other category. Twenty-six chemical components were identified according to the positive and negative ion modes detected by LC-MS. The content of five indicative components(VIP>1.5) was determined using UPLC, revealing that chlorogenic acid, isochlorogenic acid A, isochlorogenic acid C, linarin, nardosinone, and total content in the imitative wild cultivation group were 1.85, 1.52, 1.26, 0.90, 2.93, and 2.56 times those in the wild group, respectively. OPLS-DA based on GC-MS obtained 10 diffe-rential peaks. Among them, the relative content of α-humulene and aristolene in the imitative wild cultivation group were extremely significantly(P<0.01) and significantly(P<0.05) higher than that in the wild group, while the relative content of 7 components such as 5,6-epoxy-3-hydroxy-7-megastigmen-9-one, γ-eudesmol, and juniper camphor and 12-isopropyl-1,5,9-trimethyl-4,8,13-cyclotetrade-catriene-1,3-diol was extremely significantly(P<0.01) and significantly(P<0.05) lower than that in the wild group, respectively. Therefore, the main chemical components of the imitative wild cultivation group and wild group were basically the same. However, the content of non-volatile components in the imitative wild cultivation group was higher than that in the wild group, and the content of some volatile components was opposite. This study provides scientific data for the comprehensive evaluation of the quality of Nardostachyos Radix et Rhizoma with imitative wild cultivation and wild Nardostachyos Radix et Rhizoma.

Selection and validation of reference genes for quantitative real-time PCR analysis in Paeonia veitchii / 中国中药杂志

Paeonia veitchii and P. lactiflora are both original plants of the famous Chinese medicinal drug Paeoniae Radix Rubra in the Chinese Pharmacopoeia. They have important medicinal value and great potential in the flower market. The selection of stable and reliable reference genes is a necessary prerequisite for molecular research on P. veitchii. In this study, two reference genes, Actin and GAPDH, were selected as candidate genes from the transcriptome data of P. veitchii. The expression levels of the two candidate genes in different tissues(phloem, xylem, stem, leaf, petiole, and ovary) and different growth stages(bud stage, flowering stage, and dormant stage) of P. veitchii were detected using real-time fluorescence quantitative technology(qRT-PCR). Then, the stability of the expression of the two reference genes was comprehensively analyzed using geNorm, NormFinder, BestKeeper, ΔCT, and RefFinder. The results showed that the expression patterns of Actin and GAPDH were stable in different tissues and growth stages of P. veitchii. Furthermore, the expression levels of eight genes(Pv-TPS01, Pv-TPS02, Pv-CYP01, Pv-CYP02, Pv-CYP03, Pv-BAHD01, Pv-UGT01, and Pv-UGT02) in different tissues were further detected based on the transcriptome data of P. veitchii. The results showed that when Actin and GAPDH were used as reference genes, the expression trends of the eight genes in different tissues of P. veitchii were consistent, validating the reliability of Actin and GAPDH as reference genes for P. veitchii. In conclusion, this study finds that Actin and GAPDH can be used as reference genes for studying gene expression levels in different tissues and growth stages of P. veitchii.

Phenolic acid from the fruits of forsythia suspensa (thunb.) vahl / 中国药学杂志

OBJECTIVE: To study the chemical constituents from the fruits of Forsythia suspensa (Thunb.) Vahl.. METHODS: Compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, Sephadex LH-20 and reversed-phase HPLC. Their structures were elucidated by physiochemical properties and spectral analysis. RESULTS: Sixteen compounds were isolated and identified as 3, 4, α-trihydroxy-methyl phenylpropionate (1), protocatechaldehyde (2), vanillic acid (3), p-hydroxybenzoic acid (4), p-hydroxylbenzylalcohol (5), p-hydroxyphenylethyl alcohol (6), 2-(4-methoxyphenyl) acetaldehyde (7), 2-(4-hydroxyphenyl) acetic acid (8), 4-hydroxyphenethyl 2-(4-hydroxyphenyl) acetate (9), p-coumatic acid (10), methyl 2-(4-hydroxyphenyl) acetate (11), 3, 4-dihydroxyphenylethanol (12), 1, 2, 4-benzentriol (13), 1-(4- hydroxyphenyl)-2, 3 -dihydroxypropan-1-one(14), 4-hydroxybenzaldehyde (15), and isovanillic acid (16). CONCLUSION: Compounds 1-3, 5, 7, 9, 11, and 14 are isolated from this plant for the first time.

Lignan Constituents of Patrinia villosa (Thunb.) Juss / 中国药学杂志

OBJECTIVE: To study the chemical constituents of Patrinia villosa (Thunb.) Juss. METHODS: The compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by physiochemical property and spectral analysis. RESULTS: Eleven compounds were isolated and identified as(7R, 8S)-3, 3', 5-trimethoxy-4', 7-epoxy-8, 5'-neolignan-4, 9, 9'-triol-9-O-β-D-glucopyranoside(1), massonianoside D(2), (7R, 8S)-dihydroxydehydrodiconiferyl alcohol-4-O-β-D-glucopyranoside(3), (7S, 8R)-dihydroxydehydrodiconiferyl alcohol-4-O-β-D-glucopyranoside(4), 7R, 8S-glochidioboside(5), lariciresinol-4-O-β-D-glucopyranoside(6), lariciresinol-9-O-β-D-glucopyranoside(7), lariciresinol-4'-O-β-D-glucopyranoside(8), tortoside B(9), tanegool(10), and tanegool-7'-methyl ether(11). CONCLUSION: All compounds are isolated from Patrinia genus for the first time.

Chemical constituents of Patrinia villosa / 中草药

Objective: To study the chemical constituents of Patrinia villosa. Methods: Compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by physiochemical property and spectral analysis. Results: Fourteen compounds were isolated and identified as isololiolide (1), citroside A (2), grasshopper kectone (3), (E)-4-Hydroxy-3,3,5-trimethy1-4- (3-oxobut-1-en-1-yl)-cyclohexan-1-one (4), bluemenol A (5), pubinernoid A (6), robinin (7), puerarin (8), 5-(1'-hydroxyethyl)-methyl nicotinate (9), 2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]-propance-1,3-diol (10), 1-O-(β-D-glucosyl)-2-[2-methoxy-4-(3-hydroxypropyl)- phenoxy]-propan-3-ol (11), dihydrosinapyl alcohol (12), 3,5-dimethoxyl-4-hydroxyl-phenylpropanol-9-O-β-D-glucopyranoside (13), and 2-phenylethy-α-L-arabinopyranosyl-(1″→6')-β-D-glucopyranoside (14). Conclusion: All compounds are isolated from the plants of Patrinia Juss. for the first time.

Chemical constituents from fruits of Forsythia suspense / 中草药

Objective: To study the chemical constituents from the fruits of Forsythia suspensa. Methods: Compounds were isolated by a combination of various chromatographic techniques including column chromatography over macroporous resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by physiochemical property and spectral analysis. Results: Thirteen compounds were isolated and identified as salidroside (1), forsythoside E (2), quinolacetic acid (3), rengyolone (4), senecio lactone (5), cleroindicin C (6), 4-hydroxy-4-isopropylcyclohex-1-enecarboxylic acid (7), oleuropeic acid (8), rel-(1S,2S,4S)-trihydroxy-p-menthane (9), azelaic acid (10), 2-hydroxy-succinic acid 4-methylester (11), glochidioboside (12), and (+)-isolariciresino-9-O-β-D-glucopyranoside (13). Conclusion: Compounds 5-13 are isolated from genus Forsythia Vahl. for the first time.

Phenylethanoid glycosides from fruits of Forsythia Suspensa / 中草药

Objective: To study the chemical constituents from the fruits of Forsythia suspensa. Methods: Compounds were isolated by combination of various chromatographic techniques including column chromatography over macroporous resin, silica gel, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by physicochemical property and spectroscopic analysis. Results: Two compounds were isolated from the 50% ethanol extract of the fruits of F. suspensa and identified as 7R-suspensaside methyl ether (1) and 7S-suspensaside methyl ether (2). Conclusion: Compound 1 is a new compound named forsythoside K.