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

Table 4 Overview of the applied analytical methods

	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

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

ISSN: 1750-1172