Tcc 3000rs column compartment

Samples from the purification process were lyophilized and then dissolved in the mixture of water, acetonitrile, and formic acid (94.9, 5, 0.1% v/v/v). Immediately before HPLC, samples were filtered through a 0.22 μm filter and analyzed on apparatus Dionex 3000 RS-HPLC equipped with a DGP-3600 pump, a WPS-3000 TLS TRS autosampler, a TCC-3000 RS column compartment and a DAD-3000 RS diode array detector variable (Dionex Corporation, United States). The chromatography column was a 50 × 3.1 (i.d)-millimeter Thermo Scientific Hypersil GOLD with 1.9-micron particles (Part no. 25002-052130). The following chromatographic conditions were applied: the mobile phase: solvent A—0.1% formic acid in water; solvent B—0.1% formic acid in acetonitrile. The program was as flows: 0 min—5% solvent B; 1 min—5% solvent B; 21 min—95% solvent B; 21.5 min—95% solvent B; 21.6 min—5% solvent B; 30 min—5% solvent B. The inject sample volume was 4 μl. The flow rate was 0.4 ml/min and the eluent was monitored at 254 nm at room temperature. The peptide was analyzed in MALDI-TOF-MS using α-cyano-4-hydroxycinnamic acid as a matrix. Five microliters of the peptide solution (1 mg/ml) was mixed with 5 μl of the matrix (1 mg/ml in 80% acetonitrile containing 1% TFA).

Water extracts (1% w/v) were prepared from the A’, B’, and F’ bulk samples as described in Section 4.2 to determine the glycoalkaloid composition. To calculate the percentage of glycoalkaloids recovered, α-solamarine was added as an internal standard to each potato leaf extract sample (final concentration of 10 ng μL⁻¹). The control sample consisted of α-solamarine at the same concentration as that dissolved in water. A total of 750 μL of acetonitrile acidified with 1% formic acid was added to the same amount of potato leaf extract sample and passed through a sterilizing filter (0.2 μm, Nalgene™). Then, the glycoalkaloid fraction was isolated using the solid phase associated with the QuEChERS (UTC) technique. The supernatant was diluted 10-fold with methanol. HPLC–MS analysis was performed on a Dionex 3000 RS-HPLC equipped with a DGP−3600 pump, a WPS-3000 TLS TRS autosampler, a TCC-3000 RS column compartment (Dionex Corporation, CA, USA), and a Bruker micrOTOF-QII mass spectrometer (Bruker Daltonics, Bremen, Germany). The chromatographic column was a 50 × 3.1 (i.d.)-millimeter Thermo Scientific Hypersil GOLDc column with 1.9 μm particles (Part No. 25002–052130, Serial No. 0110796A6, Lot No. 10922).
The results are expressed as the frequency of each compound in the glycoalkaloid fraction found in the sample.

All of the HPLC analyses were performed with a Dionex U-3000 series equipped with a SR-3000 Solvent Rack, a LPG-3400SDN Quaternary Pump, a WPS-3000SL Auto sampler, a TCC-3000RS Column compartment, a DAD-3000RS detector, and a Chromeleon 7 chromatography workstation (Thermo Fisher Scientific, Waltham, MA, USA). An Agilent ZORBAX Extend-C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase consisted of acetonitrile (A) and water (B). The gradient program was developed as follows: 15–28% A for 0–22 min, 28–37% A for 22–35 min, 37–50% A for 35–45 min, and 50–75% A for 45–60 min. The flow rate was maintained at 1.0 mL/min and the column temperature at 30 °C. The injection volume was 10 μL and the detective wavelength was selected at 205 nm.