Zorbex rxc8

HPLC analyses of crude extract of L. corymbulosum and selected plant fractions were accomplished employing HPLC–DAD (Agilent 1200, Waldbronn, Germany) equipped with a Zorbex RXC8 (Agilent, Santa Clara, CA, USA) analytical column with 5 μm particle size and 25 mL capacity utilizing an earlier described method [47 (link)]. The mobile phase comprised eluent A (acetonitrile–methanol–water-acetic acid/5:010:85:01) and eluent B (acetonitrile–methanol–acetic acid/40:060:61). The gradient (A:B) employed was the following: 0–20 min (0 to 50% B), 21–25 min (50 to 100% B), 26–30 min (100% B) and 31–40 (100 to 0% B) at a flow rate of 1 mL/min. The samples and standards were prepared in HPLC-grade methanol (1 mg/mL), filtered using a 0.456 μm membrane filter and 20 μL was inoculated for examination. Amongst the standards rutin was assessed at 257 nm, gallic acid and catechin at 279 nm, apigenin and caffeic acid at0325 nm, while myricetin, kaempferol and quercetin were investigated at 368 nm [48 (link)]. The analysis was executed in triplicate for 10 min each, and the column was cleaned after every run. By peak integration using an external standard method, quantification was performed.

HPLC analysis of ANM and selected plant fractions (ANE and ANA) was performed using HPLC-DAD (Agilent 1200, Germany) equipped with Zorbex RXC8 (Agilent, USA) analytical column with 5 μm particle size and 25 ml capacity using previously reported method [28 (link)]. Each sample was diluted with HPLC grade methanol. Mobile phase was consisted of eluent A, (acetonitrile- methanol--water- acetic acid-/5: 10: 85: 1)-and eluent B (acetonitrile-methanol-acetic acid/40: 60:-1). The gradient (A: B) utilized was the following: 0–20 min (0 to 50 % B), 21–25 min (50 to 100 % B), 26–30 min (100 % B) and 31–40 (100 to 0 % B) at flow rate ofL1 ml/min. The standards and samples were prepared in HPLC grade methanol (1 mg/ml), filtered through 0.45 μm-membrane filter andL20 μl was injected for the analysis. Among the standards rutin was investigated at 257 nm, catechin and gallic acid at 279 nm, caffeic acid and apigenin at 325 nm while quercetin, myricetin and kampferol were analyzed atL368 nm [29 (link)]. The analysis was performed in triplicate and the column was reconditioned for 10 min after each run. Quantification was done by the integration of the peak by using the external standard method.