Hplc grade methanol

HPLC-grade methanol and acetonitrile (ACN) were purchased from Merk (Darmstadt, Germany). Formic acid was obtained from Fluka (Buchs, Switzerland). LTB4-DMA purchased from Abcam (Cambridge, UK). Sphingosine, decanamide, and oleamide were purchased from Sigma-Aldrich (St. Louis, MO). Hypoxanthine, phenylalanine and L-2-chlorophenylalanine (internal standard) were obtained from Shanghai Jingchun Reagent Co. Ultrapure water was prepared with a Milli-Q water purification system (Millipore, Bedford, MA, USA).

HPLC-grade Methanol and acetonitrile (ACN) were purchased from Merk (Darmstadt, Germany). Formic acid was obtained from Fluka (Buchs, Switzerland). 5-Methylcytidine, octanoylcarnitine, glycocholic acid and decanoylcarnitine were purchased from Sigma-Aldrich (St. Louis, MO). Phytosphingosine, sphinganine and palmitoylcarnitine were purchased from Acros Organics (NewJersey, USA). Lysophosphatidylcholine (20:0) and lysophosphatidylcholine (18:2) were purchased from Larodan AB (Malmo, Sweden). Hypoxanthine, inosine, kynurenine, hippuric acid, citric acid, indoleacetic acid and L-2-chlorophenylalanine (internal standard) were obtained from Shanghai Jingchun Reagent Co. Acetyl-N-formyl-5-methoxykynurenamine (AFMK) and indolelactic acid were purchased from Haorui Chemicals and Seebio Biotech (shanghai) Co.,Ltd, respectively. Ultrapure water was prepared with a Milli-Q water purification system (Millipore, Bedford, MA, USA).

HPLC-grade methanol and acetonitrile (ACN) were purchased from Merk (Darmstadt, Germany). Formic acid was obtained from Fluka (Buchs, Switzerland). Ultrapure water was prepared with a Milli-Q water purification system (Millipore, Bedford, MA, United States).

The assay kits for superoxide dismutase (SOD), malondialdehyde (MDA) and lactic acid were purchased from Jiancheng Bioengineering Institute (Nanjing, China). Formic acid was obtained from Fluka (Buchs, Switzerland). Acetone, citrate, and ammonium formate were obtained from Sigma-Aldrich (St. Louis, MO, United States). HPLC-grade methanol and acetonitrile (ACN) were purchased from Merk (Darmstadt, Germany). Valine, sodium succinate, phenylalanine, methionine, uric acid, arachidonic acid, linoleic acid, oleic acid, and palmitic acid were purchased from Shanghai Jingchun Reagent Co. Ltd. (Shanghai, China).

Methanol extract of S. cumini seeds and 10 standards, namely, ellagic acid, gallic acid, p-coumaric acid, quercetin, protocatechuic acid, sinapic acid, caffeic acid, kaempferol, ferulic acid, and p-hydroxybenzoic acid (Sigma-Aldrich, St. Louis, MO, USA), was subjected to HPLC analysis by using Shimadzu LC instrument equipped with Hypersil C-18 column and PDA detector (Shimadzu Corporation, Kyoto, Japan). Mobile phase containing methanol/acetonitrile (98/2 v/v) was used to perform HPLC analysis [30 (link)–32 (link)]. The flow rate was set at 0.5 ml/min, and the absorbance used for this study was 254 nm. 100 μl MSE, taken from 150 ml stock solution obtained from 10 g S. cumini seed powder, was diluted with 900 μl HPLC-grade methanol and filtered through 0.2 μ filter (Millipore, MA, USA). From the diluted stock, 20 μl was used for HPLC analysis. For HPLC analysis of the standards, 1 mg/ml stocks were prepared in methanol, and 20 μl of each standard was subjected to HPLC analysis. The concentration of each component of the extract was calculated by using the area of different standards individually. The results are represented as the mean ± SD, and each experiment is performed in triplicate.