Development, validation and application of an HPLC method for reduced vitamin C qualification in HBOCs solution

Figures & data

Table I. The series of pretreated mixture solutions for the hemoglobin precipitation.

X (ml)	1.0	0.8	0.6	0.4	0.2
Y (ml)	1.0	1.2	1.4	1.6	1.8

Figure 1. A₆₅₂ values of the supernatant with a different volume ratio between mobile phase solution (V_mp) and sample solution (V_s).

Figure 2. The HPLC chromatograms of the standard Vc with a concentration of 5 ppm.

Figure 3. The standard curve of Vc testing.

Figure 4. The values of peak area (Sp) at different time after the sample solution prepared.

Table II. Results of the repeatability experiment.

CVc (ppm)	Sp-1	Sp-2	Sp-3	Sp-4	Sp-5	Sp-6	RSD (%)
0.5	47,808	47,908	47,936	47,707	47,601	47,450	0.91
5	480,037	480,093	479,326	479,437	477,536	477,406	0.23
50	4,798,746	4,798,753	4,802,789	4,798,263	4,798,055	4,801,305	0.07

Table III. Results of the precision experiment.

CVc (ppm)	Sp-1	Sp-2	Sp-3	Sp-4	Sp-5	Sp-6	RSD (%)
0.5	46,699	46,683	47,119	46,861	47,009	46,670	0.41
5	472,854	474,664	474,588	473,947	473,804	473,017	0.16
50	4,749,944	4,793,040	4,770,552	4,796,801	4,787,574	4,807,878	0.44

Table IV. Results of the recovery experiment.

Labeled CVc (ppm)	Detected CVc-1 (ppm)	Detected CVc-2 (ppm)	Detected CVc-3 (ppm)	Detected CVc average (ppm)	Recovery (%)
0.5000	0.5394	0.5474	0.5430	0.5434	108.68
5.0000	5.2991	5.2869	5.2928	5.2930	105.86
50.0000	51.7840	51.7714	51.7742	51.7777	103.56

Figure 5. HPLC chromatograms of reduced Vc in HBOCs samples.

Figure 6. Concentration-time profiles of Vc concentrations during the reaction process.

Figure 7. The variation of MetHb concentrations during the reaction process.

Table V. Kinetic parameters for the reaction of Vc in HBOCs solutions at different pH values.

pH	k (mol^−1·h^−1)	T1/2 (h)
7.05	−0.1935	2.07
8.05	−0.2069	1.24
9.03	−0.2199	1.12