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Table 4 Overview of the applied analytical methods

From: Pharmaceutical compounding of orphan active ingredients in Belgium: how community and hospital pharmacists can address the needs of patients with rare diseases

  Column HPLC Mobile phase Flow rate (ml/min) UV detection wavelength (nm) Column oven temperature (°C) Injection volume (μl)
L-arginine Nucleodur NH2-RP, 5 μm, 4.6 mm × 150 mm Isocratic: phosphate buffer pH 7.0/acetonitrile 47/53 0.8 210 30 20
L-carnitine Nucleodur NH2-RP, 5 μm, 4.6 mm × 150 mm Isocratic: phosphate buffer pH 4.7/acetonitrile 40/60 0.8 205 30 20
Primaquine phosphate Symmetry C8, 3.5 μm, 3 mm × 150 mm Isocratic: acetonitrile/tetrahydrofuran/trifluoro acetic acid/water 9/1/0.1/90 0.75 265 30 10
Pyridoxal phosphate Prevail C18, 5 μm, 4.6 mm × 250 mm Isocratic: 25 mM KH2PO4 pH 3/acetonitrile 97/3 1 212 25 20
Sodium benzoate Prevail Organic acid, 5 μm, 4.6 mm × 250 mm Isocratic: Water/acetonitrile 75/25 + 0.1% (m/V) formic acid 1.0 225 25 20
Sodium perchlorate Primesep D column, 5 μm, 3.2 mm × 150 mm equipped with suitable guard column Gradient with A: 10/90 acetonitrile/water (V/V) + 30 mM ammonium formate, B: 40/60 acetonitrile/water + 80 mM ammonium formate 0.6 -a 30 1
Sodium phenylbutyrate C18, end-capped, base-deactivated, 5 μm Isocratic: glacial acetic acid/methanol/water 1/49/50 1.1 245 35 20
MBA, PBA C18, 5 μm, 4.6 mm × 150 mm Gradient with A: 95/5 0.62% (m/V) KH2PO4 in water/methanol (V/V), B: 5/95 0.62% (m/V) KH2PO4 in water/methanol (V/V) 1.2 245 25 10
  1. Overview of the column type, mobile phase, flow rate, UV detection wavelength, column oven temperature and injection volume applied in the HPLC methods. A gradient was used for analysis the sodium perchlorate formulation (0 min: 100% A, 0% B; 12 min: 0% A, 100% B, 12.5 min: 100% A, 0% B) and for assay of MBA, PBA in the L-arginine formulation (0 min: 90% A, 10% B; 17 min: 36% A, 64% B; 22 min: 90% A, 10% B). aDetection of sodium perchlorate was performed by mass spectrometry