Simple and sensitive high performance liquid chromatographic (HPLC) method for the determination of the selegiline in human plasma

Figures & data

Figure 1. Chemical structure of selegiline.

Figure 2. Typical chromatograms of the HPLC method developed for selegiline assay in human plasma: (A) the HPLC chromatogram for a blank plasma; (B) chromatogram of a human plasma spiked to a 100 ng/ml drug concentration; (C) chromatogram of LOQ (10 ng/ml) of the method.

Table 1. Within-run variations and accuracy of the HPLC method for quantitation of selegiline (n = 3)

Nominal added concentration (ng/ml)	Sample number	Measured concentration (ng/ml)	Mean (SD)	CV %	Accuracy	Mean (SD)
10	1	10.12	10.04 (0.07)	0.7	95.23	95.54 (1.37)
	2	9.98			94.36
	3	10.03			97.05
50	1	49.51	50.22 (1.7)	3.39	99.24	97.12 (1.84)
	2	52.17			95.89
	3	48.99			96.23
100	1	99.46	100.33 (0.93)	0.93	100.02	99.23 (0.70)
	2	101.32			99.02
	3	100.23			98.65

Table 2. Between-run variations and accuracy of the HPLC method for quantitation of selegiline (n = 3)

Nominal added concentration (ng/ml)	Run number	Measured concentration (ng/ml)	Mean (SD)	CV %	Accuracy	Mean (SD)
10	1	9.85	10.06 (0.19)	1.94	97.90	99.23 (1.62)
	2	10.23			98.78
	3	10.12			101.02
50	1	50.32	49.68 (0.75)	1.51	96.55	98.79 (2.34)
	2	49.87			101.23
	3	48.85			98.59
100	1	100.16	99.86 (0.3)	0.3	101.02	99.63 (1.25)
	2	99.87			99.32
	3	99.56			98.56

Table 3. Repeatability of the test results for spiked plasma containing 50 ng/ml selegiline

Sample	Peak area	Mean (SD)	CV %	Retention time (min)	Mean (SD)	CV %
1	133546	133188.2 (326.98)	0.26	4.98	4.99 (0.014)	0.28
2	133054			5.01
3	132987			4.99
4	133654			4.97
5	132867			5.00
6	133021			4.99

Table 4. Relative recovery of selegiline by the HPLC method (n = 3)

Nominal added concentration (ng/ml)	Sample number	Recovery (%)	Mean (SD)
10	1	95.21	98.77 (3.41)
	2	102.01
	3	99.11
50	1	96.18	96.62 (4.30)
	2	101.14
	3	92.56
100	1	101.25	99.21 (4.22)
	2	102.04
	3	94.36

Table 5. Intermediate precision of the test results for spiked plasma containing 50 ng/ml selegiline

Sample	Peak area	Mean (SD)	CV %	Retention time (min)	Mean (SD)	CV %
1	132956	133190.8 (351.77)	0.28	4.97	4.99 (0.014)	0.28
2	132863			4.96
3	133065			5.01
4	133620			4.96
5	132987			4.99
6	133654			4.98

Table 6. Data showing stability of selegiline in human plasma at different QC levels (n = 5)

Stability	15 (ng/ml)	50 (ng/ml)	100 (ng/ml)
Short-term stability	95.45	94.32	91.01
Freeze and thaw stability	97.85	96.54	98.12
Long-term stability	96.32	98.21	96.32
Post-preparative stability	98.12	92.30	94.12