高效液相色谱法同时测定漱口水中多种抑菌成分含量

doi:10.3969/j.issn.2097-2806.2025.11.016

Abstract

Abstract:

This study developed a high-performance liquid chromatography （HPLC） method for the simultaneous determination of five antibacterial components （chlorhexidine, cetylpyridinium chloride, ethylparaben, sodium methylparaben, and domiphen bromide） in mouthwash. Mouthwash samples were extracted using acetonitrile-0.05 mol/L phosphate buffer （pH 3.8） at a volume ratio of 20∶80. Ultrasonic extraction parameters were set as follows: power of 300 W, extraction time of 20 minutes, temperature of 40 ℃, and solid-to-liquid ratio of 1∶4. An Agilent 1260 HPLC system, equipped with a diode array detector （DAD）, was employed using an Agilent ZORBAX Eclipse Plus C18 reversed-phase column （250 mm×4.6 mm, 5 μm）. The column temperature was maintained at 30 ℃, with an injection volume of 10 μL, flow rate of 1.0 mL/min, detection wavelength of 260 nm, and total run time of 30 minutes. Methodological validation demonstrates that for intraday precision, the relative standard deviations （RSDs） of the five components range from 2.33% to 4.72%, while interday precision RSDs range from 1.01% to 4.21%. Linear regression analysis reveals that all five components exhibit excellent linearity with the correlation coefficients （r）>0.999 5, the limits of detection （LOD） ranging from 0.09 to 0.18 μg/mL and the limits of quantification （LOQ） ranging from 0.27 to 0.54 μg/mL. Spike recovery experiments indicate that the average recovery rates of the five components range from 96.02% to 104.65%, with the corresponding RSDs of 1.74% to 4.53%. Detection of 20 commercial mouthwash samples show that chlorhexidine is detected only in samples S15 and S19 at the concentrations of 1.25 and 1.27 mg/mL, respectively. Cetylpyridinium chloride is detected in 7 samples with the concentrations ranging from 0.13 to 0.75 mg/mL. Ethylparaben is detected solely in sample S4 at 0.15 mg/mL. Sodium methylparaben is found only in sample S18 at 1.19 mg/mL. Domiphen bromide is detected exclusively in sample S20 at 0.32 mg/mL, and the remaining 9 samples show no detection of the target components. In conclusion, this method enables simultaneous and rapid detection of the five target antibacterial components in commercial mouthwashes.

Key words: mouthwash, high-performance liquid chromatography, antibacterial components, simultaneous detection, quality control

CLC Number:

TQ658

Lin Cao, Mingwei Liu. Simultaneous determination of multiple antibacterial components in mouthwash by high performance liquid chromatography[J].China Surfactant Detergent & Cosmetics, 2025, 55(11): 1493-1500.

Figures/Tables 9

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References 10

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抑菌成分	回收率/%
抑菌成分	甲醇-水（50∶50）	甲醇-水（20∶80）	乙腈-水（50∶50）	乙腈-水（20∶80）	乙腈-磷酸盐缓冲液（pH 3.8）（20∶80）
氯己定	85.32±2.10	78.45±3.21	89.67±1.98	82.14±2.67	98.76±1.23
西吡氯铵	88.91±1.78	83.22±2.54	93.34±0.85	91.05±1.92	95.12±1.45
尼泊金乙酯	92.57±1.25	86.43±2.11	87.62±1.67	80.33±2.88	84.21±2.09
对羟基苯甲酸甲酯钠	91.24±1.67	88.76±1.98	88.95±1.54	83.47±2.34	86.54±1.87
度米芬	89.63±1.89	84.12±2.45	93.78±1.32	87.29±2.01	90.45±1.66

流动相体系	平均分离度（Rs）	目标物拖尾因子	保留时间RSD/%	柱压/MPa
甲醇-水	1.21±0.18	1.38±0.09	2.15±0.30	18.3±1.22
乙腈-水	1.63±0.11	1.15±0.05	1.24±0.18	13.8±0.91
乙腈-0.05 mol/L磷酸盐（pH 3.8）	1.85±0.12	1.08±0.03	0.48±0.05	12.5±0.85

抑菌成分	回归方程	r	线性范围/（μg/mL）	LOD/（μg/mL）	LOQ/（μg/mL）
氯己定	y=8.02x-0.18	0.999 9	0.5~200	0.11	0.33
西吡氯铵	y=3.19x-0.15	0.999 5	0.5~200	0.09	0.27
尼泊金乙酯	y=6.38x+0.32	0.999 7	0.5~200	0.15	0.45
对羟基苯甲酸甲酯钠	y=7.98x+0.21	0.999 8	0.5~200	0.13	0.39
度米芬	y=4.79x+0.25	0.999 6	1.0~200	0.18	0.54

抑菌成分	加标质量浓度/（μg/mL）	实测质量浓度/（μg/mL）	平均回收率/%	RSD/%
氯己定	10	9.65, 10.14, 9.84, 9.65, 10.02, 10.28	99.30	2.62
	50	48.81, 51.62, 54.73, 53.91, 52.44, 50.91	104.14	4.10
	100	97.64, 98.82, 94.58, 89.31, 96.15, 99.60	96.02	3.91
西吡氯铵	10	9.43, 10.00, 9.97, 10.01, 10.01, 9.67	98.48	2.47
	50	47.23, 47.76, 48.75, 48.45, 48.75, 51.69	97.54	3.18
	100	107.29, 105.85, 101.91, 104.60, 104.20, 104.02	104.65	1.74
尼泊金乙酯	10	10.45, 9.40, 9.61, 9.94, 9.42, 9.67	97.48	4.06
	50	48.96, 48.50, 50.39, 50.59, 47.40, 52.51	99.45	3.65
	100	104.19, 101.59, 97.93, 103.04, 98.62, 102.90	101.38	2.52
对羟基苯甲酸甲酯钠	10	10.35, 10.50, 9.74, 9.50, 9.80, 9.68	99.28	4.03
	50	51.29, 48.87, 48.17, 44.57, 49.13, 49.07	97.03	4.53
	100	103.77, 96.74, 99.25, 96.27, 101.28, 98.80	99.35	2.84
度米芬	10	10.84, 9.91, 9.95, 10.48, 10.46, 9.72	102.27	4.21
	50	54.52, 51.41, 49.62, 53.37, 51.07, 49.65	103.21	3.85
	100	98.99, 107.56, 103.28, 97.97, 98.81, 99.32	100.99	3.68

质量浓度/（μg/mL）	重复	实测质量浓度/（μg/mL）
质量浓度/（μg/mL）	重复	A	B	C	D	E
5	1	5.04	4.68	5.36	5.06	4.91
	2	4.67	5.05	5.22	5.19	5.02
	3	5.30	5.06	4.95	4.99	5.15
	4	4.95	4.99	5.32	5.14	5.16
	5	4.93	4.90	4.98	5.24	5.04
	6	5.28	4.98	5.16	4.83	4.88
	RSD/%	4.72	2.86	3.30	2.95	2.33
50	1	46.70	48.06	50.52	48.20	48.04
	2	50.00	47.83	52.63	46.36	51.60
	3	50.05	49.72	51.17	50.71	49.86
	4	49.68	50.93	50.94	51.21	46.83
	5	49.95	51.04	47.78	48.26	51.63
	6	53.49	50.56	49.82	46.32	49.21
	RSD/%	4.31	2.88	3.20	4.30	3.87
150	1	149.28	150.20	145.33	152.82	152.52
	2	146.05	142.59	144.58	154.52	147.22
	3	158.91	149.84	150.46	146.64	150.56
	4	144.64	143.07	151.83	151.16	158.58
	5	157.46	153.24	148.22	145.78	141.98
	6	150.88	150.45	152.70	155.95	156.18
	RSD/%	3.88	2.94	2.27	2.75	3.99

Simultaneous determination of multiple antibacterial components in mouthwash by high performance liquid chromatography

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实验号	A 功率/W	B 时间/min	C 温度/℃	D 料液比	总回收率/%
1	200	10	30	1∶2	82.64±2.32
2	200	20	40	1∶4	89.12±2.54
3	200	30	50	1∶8	85.43±2.85
4	300	10	40	1∶8	91.37±2.12
5	300	20	50	1∶2	92.14±2.31
6	300	30	30	1∶4	90.89±2.63
7	400	10	50	1∶4	88.70±2.97
8	400	20	30	1∶8	86.24±3.03
9	400	30	40	1∶2	87.55±2.72

质量浓度/（μg/mL）	重复	实测质量浓度/（μg/mL）
质量浓度/（μg/mL）	重复	A	B	C	D	E
5	D1-1	5.10	5.05	5.05	4.80	5.06
	D1-2	5.25	5.15	5.04	5.21	5.15
	D2-1	4.86	5.30	5.16	4.67	5.15
	D2-2	4.98	4.80	5.07	5.09	5.07
	D3-1	4.78	5.25	4.95	4.99	4.68
	D3-2	4.97	4.96	4.95	4.78	4.96
	RSD/%	3.37	3.69	1.58	4.21	3.53
50	D1-1	48.02	49.40	51.68	49.09	49.75
	D1-2	50.47	49.28	49.46	49.53	47.08
	D2-1	47.91	52.78	47.52	49.78	48.30
	D2-2	49.53	49.49	50.45	49.66	49.53
	D3-1	47.37	46.98	48.17	50.34	50.65
	D3-2	50.63	49.79	47.63	47.96	46.64
	RSD/%	2.87	3.73	3.42	1.64	3.26
150	D1-1	152.11	149.46	152.90	151.27	157.43
	D1-2	155.82	139.71	152.85	148.94	150.42
	D2-1	159.37	152.84	152.00	151.75	144.76
	D2-2	155.86	149.73	150.83	157.71	147.98
	D3-1	148.06	148.78	153.94	143.17	149.75
	D3-2	149.01	151.04	149.71	147.54	141.78
	RSD/%	2.87	3.08	1.01	3.24	3.61

样品	ρ（氯己定）/（mg/mL）	ρ（西吡氯铵）/ （mg/mL）	ρ（尼泊金乙酯）/（mg/mL）	ρ（对羟基苯甲酸甲酯钠）/（mg/mL）	ρ（度米芬）/ （mg/mL）
S1	—	—	—	—	—
S2	—	—	—	—	—
S3	—	—	—	—	—
S4	—	0.22	0.15	—	—
S5	—	0.75	—	—	—
S6	—	0.25	—	—	—
S7	—	0.45	—	—	—
S8	—	—	—	—	—
S9	—	—	—	—	—
S10	—	—	—	—	—
S11	—	0.75	—	—	—
S12	—	0.75	—	—	—
S13	—	—	—	—	—
S14	—	—	—	—	—
S15	1.25	—	—	—	—
S16	—	0.13	—	—	—
S17	—	—	—	—	—
S18	—	—	—	1.19	—
S19	1.27	—	—	—	—
S20	—	—	—	—	0.32

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