Identification of Illicium difengpi and Its Fake I. jiadifengpi Using DNA Barcoding / 中国实验方剂学杂志

Objective:To evaluate and compare the identification of several DNA barcoding candidate sequences on Illicium difengpi and its fake I. jiadifengpi. Method:Samples from different origins of I. difengpi and I. jiadifengpi, were collect extraction of total DNA,nuclear gene ITS2 sequence,chloroplast rbcL,matK gene sequence were selected for PCR amplification,product purification and sequencing,and CondonCode Aligner V3.7.1 was used to proofread stitching. Result:PCR amplification and sequencing of rbcL sequences of I. difengpi and I. jiadifengpi were not satisfactory. It is assumed that their rbcL sequences were too long with slow evolution,which was unsuitable for interbreeding. The success rate of matK sequencing of I. difengpi and I. jiadifengpi was 0 and 76.8%,which may be because primer standards were different for matK sequences of different groups. The results of PCR amplification and sequencing of ITS2 on I. difengpi and I. jiadifengpi were successful,with the success rate of sequencing was 89.3% and 91.2%. Analysis sequencing results, the total length of ITS2 sequences was 268 bases,and there were 2 variation sites of I. difengpi. The total length of ITS2 sequences was 430 bases,and there were 4 or 3 variation sites of I. jiadifengpi. It shows that ITS2 sequences of I. difengpi and I. jiadifengpi were short and has obvious variability and can be amplify,that ITS2 sequence was better than rbcL and matK sequence in molecular identification of I. difengpi and I. jiadifengpi. Conclusion:DNA barcoding based on ITS2 sequence was a powerful and efficient tool for identification of I. difengpi and its fake I. jiadifengpi.

Studies on chemical constituents of Clinopodium chinense / 中国中药杂志

Twenty-eight compounds were isolated and purified from Clinopodium chinense by Sephedax LH-20, ODS, MCI and preparative HPLC. Their structures were identified as apigenin (1), apigenin-7-O-β-D-glucopyranoside (2), apigenin-7-O-β-D-glucuronopyranoside (3), thellungianol (4), apigenin-7-O-β-D-rutinoside (5), luteolin (6), luteolin-4'-O-β-D-glucopyranoside (7), apigenin-7-O-β-D-pyranglycuronate butyl ester (8), luteolin-7-O-β-D-rutinoside (9), luteolin-7-O-β-D-noehesperidoside (10), acacetin (11), acacetin-7-O-β-D-glucuronopyranoside (12), buddleoside (13), naringenin (14), pruning (15), nairutin (16), isosakuranetin (17), isosakuranin (18), didymin (19), hesperidin (20), kaempferol (21), quercetin (22), kaempferol-3-O-α-L-rahmnoside (23), p-hydroxycinnamic acid (24), caffeic acid (25), cis-3-[2-[1-(3,4-dihydroxy-phenyl)-1 -hydroxymethyl]-1,3-ben-zodioxol-5-yl]-(E)-2-propenoic acid (26), mesaconic acid (27), gentisic acid 5-O-β-D-(6'-salicylyl)-glucopyranoside (28). Among them, compounds 7, 9-10, 12, 23, 26-28 were isolated from the Clinopodium for the first time. The protective effects of compounds 1-6, 8-17 and 19 against H2O2-induced H9c2 cardiomyocyte injury were tested, compounds 15 exhibited significantly protective effects. Compared with the cell viability of (62.12±6.18)% in the model, pruning exhibited viabilities of (84.25±7.36)% at 25.0 mg•L⁻¹, respectively, using quercetin as a positive control [cell viability of (84.55±8.26)%, 20 mg•L⁻¹].

Enantiomeric separation of ketoprofen by HPLC using chirobiotic V CSP and vancomycin as chiral mobile phase additives / 药学学报

<p><b>AIM</b>To establish HPLC chiral separation method for ketoprofen enantiomers by using Chirobiotic V chiral seperation phase (CSP) (A) and vancomycin as chiral mobile phase additives (B).</p><p><b>METHODS</b>The separation was first performed on Chirobiotic V CSP with the mobile phase of terahydrofran (THF)-0.5% triethylanine acetate(TEAA) buffer (15:85) at the flow rate of 0.7 mL.min-1. When using vancomycin as chiral mobile phase additive, the separation was carried out on C8 column (150 mm x 4.6 mm), the mobile phase was methanol-0.25% TEAA buffer (50:50), the flow rate was 0.7 mL.min-1. The effects of the concentration of vancomycin, organic modifier and the pH of the buffer on the resolution of ketoprofen enantiomers were investigated. Also, the feasibility of these two methods to be used as quantitative method was studied.</p><p><b>RESULTS</b>Ketoprofen enantiomers were separated at a baseline level under the chromatographic condition of both methods A and B, the resolution was 2.28 and 2.22, respectively. In method A the linearity of enantiomer was obtained from 0.5 mg.L-1 to 100 mg.L-1, the detectionlimit was 1 microgram.L-1. When using vancomycin as mobile phase additive the system was shown to have a high efficiency. In this system, the assay of enantiomer is linear from 2.5 mg.L-1 to 250 mg.L-1. The detection limit was 14.5 micrograms.L-1.</p><p><b>CONCLUSION</b>Both methods can be used to detect optical purity of S-(+)-ketoprofen.</p>