서울特別市 保健環境硏究院報 第41號	(2005)

Report of S.I.H.E., 41:205～210   (2005)

화장품 중에 함유된 보존제 성분 조사

-페녹시에탄올의 분석

독성약품팀

윤용태․김유경․박주성․김수언․정희정․조남준

Study on the Preservative Ingredients

in Cosmetics by HPLC-The Analylsis

of Phenoxy Ethanol

Toxicopharmaceutical Team

Yong-tae Yoon, Yoo-kyung Kim, Ju-sung Bak, Su-un Kim,

Hee-jeong Jeong and Nam-joon Chough

요  약

대부분의 화장품에는 유효성분으로서 각종 영양분이 함유되어 있어 사용기간 중 화장품에 미생물의 번식 등으로 인한 변질, 부패가 없도록 방부제를 사용하게 되는데 이미다졸리디닐우레아, 파라옥시안식향산프로필, 파라옥시안식향산메칠와 더불어 페녹시에탄올이 많이 사용되고 있다. 이러한 방부제는 배합량이 많으면 접촉성피부염 같은 피부이상을 일으킬 수 있다. 본 연구에서는 48종의 수입화장품의 페녹시에탄올함량을 조사하여 이 성분이 적정하게 함유되어 있는지를 조사하였다. 하이퍼실골드칼럼(4.6×250 mm), 0.75 mM KH2PO4을 0.85% 황산에 녹인 액과 메탄올을 7대 3으로 한 액을 이동상으로 하여 214 nm에서 고속액체크로마토그래프법을 이용하여 페녹시에탄올의 정량분석을 하였다. 검량선은 9.244 ㎍/㎖～92.44 ㎍/㎖범위에서 r2은 0.9995로 양호한 직선성을 보였다. 회수율은 클린저, 크림, 스킨, 에센스, 린스, 로션, 스크럽, 팩, 샤워젤, 샴푸, 오일에 대해 실험하였으며, 평균(RSD)은 95.3%(RSD 1.3%)～105.6%(RSD 2.1%)이었다. 분석결과 화장품 48건 중 1건에서는 phenoxy ethanol이 확인되지 않았으며 47건에서는 phenoxy ethanol이 확인되었고 2건에서는 표시량의 20% 이하로 정량되었고, 2건에서는 표시량의 170%～220%, 3건에서는 460% 이상이 정량되었으나 배합허용한도 1%를 초과하지는 않았다.

Key words : phenoxy ethanol, HPLC

INTODUCTION

Cosmetic products have been used to meet women's desire to be beautiful and protect the skin from outer hazards including UV ray. Nowadays they became the necessaries of life. If they are not properly dealt with, however, they may have unexpected side effects(1).

Cosmetics contain various chemicals which are effective materials such as preservative, pigment, fragrance, etc. Some chemicals that cosmetics include are likely to be contaminated by external pollutants. If cosmetics are contaminated during the application, they may break the balance of body homeostasis, resulting in a skin trouble like dermatitis(2～5).

Generally cosmetics are applied to human skin repeatedly and continuously for a given long time. As people use them every day, the safety of cosmetics is getting important. During the app-lication period they have to be stable in quality. But it's not easy to store cosmetics as they were produced. Cosmetics have many nutrients that are vulnerable to microbes which can easily proliferate and thus contaminate the aesthetic products. That's why cosmetics contain preservative, which can prevent those products from being polluted. But excessive amount of preservative also can be a cause of skin trouble(1, 6～7).

Those mainly used in cosmetics are methyl paraben, propyl paraben, imidazolidinyl urea and phenoxy ethanol. In particular, phenoxy ethanol is being used in most cosmetics. According to KFDA notification, phenoxy ethanol should be added to a cosmetic product by no more than one percent(8).

This study selected randomly 48 imported cosmetic products with various forms such as lotion and cream, and examined whether those products were suitable to KFDA notification about phenoxy ethanol.

MATERIALS AND METHODS

1. Collection of samples

Forty eight imported cosmetics were analyzed. The list of collected samples is at the Table 1.

Table 1. The classification of sample
Sample No.	Manufacturing company	Type	Number of samples
1	A	cleansing lotion	1
2	A	cream	3
3	A	essence	3
4	A	cleansing cream	1
5	A	pack	1
6	A	skin lotion	1
7	B	essence	1
8	B	hair rinse	1
9	B	hair spray	1
10	B	hair tonic	1
11	B	hair cream	2
12	B	lotion	1
13	B	cream	2
14	C	cream	2
15	C	lotion	3
16	C	scrub	1
17	C	pack	1
18	C	skin lotion	1
19	C	shower gel	1
20	C	shampoo	1
21	D	cream	1
22	E	cream	1
23	F	skin lotion	1
24	F	lotion	1
25	F	shampoo	1
26	G	essence	3
27	G	skin lotion	1
28	G	lotion	1
29	G	pack	1
30	H	oil	1
31	H	skin lotion	1
32	H	pack	2
33	H	essence	1
34	H	cream	3

2. Reagents and instrumentation

1) Reagents

Phenoxy ethanol(Sigma Aldrich Co., USA)

Potassium phosphate, monobasic(Junsei chemical Co., Japan)

Methanol(J.T.Baker , USA, HPLC solvent)

Sulfuric acid(Matsunoen chemicals Ltd.. Japan, harmful metal analysis)

2) Instrumentation

HPLC(Injector：Waters 717 plus autosampler,

Pump : Waters 510,

Detector : Waters 486,

Data program : Autochrome 2000)

Sonicator(Fisher Scientific FS7652H)

3. Condition of analysis

1) Standard solution and Calibration

The phenoxy ethanol was used as a stan-dard for quantitative analysis. The 23.11 ㎎ of phenoxy ethanol was dissolved in 25.0 ㎖ volumetric flask with methanol. This solution was used as stock solution. The volume 1, 2, 3, 4, 5, 10 ㎖ of the stock solution were taken and added with methanol to make 50 ㎖ in total. By using these six standard solutions and external standard method, the calibration curve was made.

Fig. 1. Chromatogram of phenoxy ethanol in standard.

2) Sample solution

About 0.4～4.0 g of each sample of cosmetics was weighed accurately, sonicated with methanol for 30 minutes, added with methanol to make 50 ㎖ and filtered through a membrane filter with pore size of 0.45 ㎛.

3) Recovery test

The known amounts of phenoxy ethanol standard were added to the known concentration of sample solution. Suitable amounts of these samples were weighed accurately, sonicated with methanol for 30 minutes, added with methanol to make 50 ㎖ and filtered through a membrane filter with pore size of 0.45 ㎛.

4) HPLC assay

The analysis of standard solutions and sample solutions were carried out at the room temperature with the conditions of the Table 2(9).

Table 2. Analytical conditions of HPLC
Column	Hypersil Gold 4.6 × 250 mm
Mobile phase	0.75 mM KH2PO4 in 0.85% sulfuric acid：methanol(7：3)
Wavelength	214 nm
Flow rate	0.7 ㎖/min
Injection volumn	10 ㎕
Detector	Waters 486

RESULTS AND DISCUSSION

Six solutions containing phenoxy ethanol at concentrations ranging from 9.244 ㎍/㎖ to 92.44 ㎍/㎖ were analyzed in a bid to make the calibration curve. The regression equation of phenoxy ethanol calculated with peak area(y, mV) and concentration(x, ㎍/㎖) was y=36.788x＋0.4701(r2=0.9995) over con-centration range 9.244 ㎍/㎖ to 92.44 ㎍/㎖. The retention time of phenoxy ethanol in standard solutions was 25.9400 mins and the retention factor was 4.6(Fig. 1～3).

Fig. 2. Chromatogram of phenoxy ethanol in sample.

The recovery test was carried out 3 times over 11 imported cosmetics by HPLC.

Table 3. Recovery rate of phenoxy ethanol in spiked samples
Order	Type	1	2	3	Mean(%)	RSD(%)
1	Cleansing lotion	96.5	95.2	94.1	95.3	1.3
2	Cream	97.0	94.8	97.3	96.4	1.4
3	Skin lotion	98.7	99.8	99.3	99.3	0.6
4	Essence	99.5	97.3	98.8	98.5	1.1
5	Hair rinse	98.5	99.0	100.7	99.4	1.2
6	Lotion	97.6	97.2	99.2	98.0	1.1
7	Scrub	107.9	103.4	105.4	105.6	2.1
8	Pack	95.9	97.6	93.4	95.6	2.2
9	Shower gel	96.8	96.6	98.9	97.4	1.3
10	Shampoo	94.4	96.0	98.8	96.4	2.3
11	Oil	106.2	105.7	103.1	105.0	1.6

The means(RSD) of recovery rates in clean-sing lotion, cream, skin lotion, essence, hair rinse, lotion, scrub, pack, shower gel, shampoo and oil were 95.3%(1.3%), 96.4% (1.4%), 99.3%(0.6%), 98.5%(1.1%), 99.4% (1.2%), 98.0%(1.1%), 105.6% (2.1%), 95.6%(2.2%), 97.4%(1.3%), 96.4%(2.3%) and 105.0%(1.6%)(Table 3).

Fig. 3. Calibration curve of phenoxy ethanol standard.

The result of analysis of 48 imported cosmetics showed one cosmetic did not contain phenoxy ethanol, 47 cosmetics included phenoxy ethanol, among which two cosmetics contained less than 20 percent of indicated

Table 4. Concentration of phenoxy ethanol in cosmetics
No.	Type	Labelled (g/100g)	1	2	3	Mean(g/100g)	RSD(%)	Content(%)
1	Cleansing lotion	0.2	0.225	0.212	0.210	0.216	3.8	107.9
2	Cream	0.5	0.477	0.467	0.463	0.469	1.6	93.8
3	Cream	0.5	0.500	0.487	0.502	0.496	1.7	99.3
4	Essence	0.2	0.000	0.000	0.000	0.000	0.0	0.0
5	Cleansing cream	0.2	0.205	0.205	0.204	0.205	0.4	102.4
6	Cream	0.3	0.351	0.349	0.345	0.349	0.9	116.2
7	Pack	0.2	0.340	0.345	0.341	0.342	0.8	171.0
8	Skin lotion	0.3	0.318	0.314	0.320	0.317	0.9	105.8
9	Essence	0.2	0.219	0.208	0.205	0.211	3.5	105.4
10	Essence	0.4	0.439	0.425	0.432	0.432	1.6	108.0
11	Essence	0.5	0.605	0.606	0.603	0.605	0.3	120.9
12	Hair rinse	0.3	0.292	0.298	0.291	0.294	1.2	97.9
13	Hair spray	0.4	0.444	0.464	0.485	0.464	4.4	116.1
14	Hair tonic	0.3	0.306	0.305	0.309	0.307	0.7	102.2
15	Hair cream	0.2	0.206	0.210	0.208	0.208	1.1	104.1
16	Hair cream	0.4	0.422	0.426	0.431	0.426	1.1	106.5
17	Lotion	0.3	0.286	0.285	0.288	0.286	0.5	95.5
18	Cream	0.3	0.330	0.341	0.337	0.336	1.7	112.1
19	Cream	0.3	0.281	0.282	0.285	0.283	0.6	94.3
20	Cream	0.3841	0.345	0.356	0.369	0.357	3.4	92.8
21	Cream	0.3675	0.324	0.363	0.353	0.347	5.8	94.3
22	Lotion	0.666	0.671	0.689	0.684	0.681	1.4	102.3
23	Scrub	0.5	0.459	0.470	0.456	0.462	1.7	92.4
24	Lotion	0.23575	0.230	0.230	0.232	0.231	0.5	97.8
25	Pack	0.5	0.486	0.500	0.499	0.495	1.6	99.0
26	Skin lotion	0.02175	0.021	0.017	0.020	0.019	11.7	87.8
27	Shower gel	0.36	0.350	0.353	0.357	0.353	1.0	98.1
28	Lotion	0.5076	0.498	0.505	0.510	0.504	1.2	99.4
29	Shampoo	0.5076	0.499	0.520	0.509	0.509	2.0	100.3
30	Cream	0.25	0.251	0.239	0.244	0.245	2.4	97.8
31	Cream	0.4638	0.369	0.375	0.371	0.372	0.7	80.1
32	Skin lotion	0.14	0.160	0.167	0.165	0.164	2.2	117.1
33	Lotion	0.14	0.147	0.144	0.138	0.143	3.2	102.0
34	Shampoo	0.14	0.139	0.146	0.143	0.143	2.5	101.9
35	Essence	0.2	0.154	0.169	0.172	0.165	5.7	82.6
36	Essence	0.2	0.189	0.203	0.201	0.198	3.8	98.8
37	Skin lotion	0.5	0.091	0.090	0.088	0.090	1.7	17.9
38	Lotion	0.1	0.467	0.464	0.461	0.464	0.6	464.0
39	Pack	0.9	0.082	0.080	0.077	0.080	3.2	8.9
40	Essence	0.1	0.091	0.089	0.083	0.088	4.7	87.7
41	Oil	0.36	0.348	0.334	0.328	0.337	3.1	93.5
42	Skin lotion	0.06	0.133	0.133	0.126	0.131	3.0	217.7
43	Pack	0.1216	0.113	0.112	0.117	0.114	2.4	93.7
44	Essence	0.8	0.796	0.782	0.791	0.790	0.9	98.7
45	Cream	0.0539	0.048	0.042	0.045	0.045	6.7	83.5
46	Cream	0.3633	0.319	0.306	0.297	0.307	3.6	84.6
47	Cream	0.054	0.298	0.290	0.288	0.292	1.8	540.6
48	Pack	0.1	0.469	0.493	0.462	0.475	3.5	474.8

amounts and two cosmetic contained 170 percent to 220 percent of indicated amount, three cosmetics contained more than 460 percent of indicated amount. However all of them did not include phe-noxy ethanol more than one percent of their weights.

CONCLUSION

Using high-performance liquid chromato-graphic method, 48 imported cosmetics were analyzed. The result of analyzing the content of phenoxy ethanol is as follows.

1.  The analysis was carried out using Hypersil Gold 4.6×250 ㎜ at room temperature. The wavelength was 214 ㎚. The potassium phosphate mono-basic, 0.75 mM in 0.85% sulfuric acid : methanol (7:3) were served as mobile phase at a flow rate 0.7 ㎖/min.

2.  The regression equation of phenoxy ethanol calculated with peak area(y, mV) and con-cen-tration(x, ㎍/㎖) was y=36.788x+0.4701(r2＝0.9995) over con---cen-tration range 9.244 ㎍/㎖ to 92.44 ㎍/㎖. The retention time of phenoxy ethanol in standard solutions was 25.9400min.

3.  Recovery rates in cleansing lotion, cream, skin lotion, essence, hair rinse, lotion, scrub, pack, shower gel, sham-poo and oil were 95.3%(RSD 1.3%)～105.6%(RSD 2.1%)

4.  One cosmetic did not contain phenoxy ethanol, forty seven cosmetics included phenoxy ethanol, among which two cosmetics contained less than 20 percent of indicated amounts, two cosmetics contained 170 percent to 220 percent of indicated amount and three cosmetics contained more than 460 percent of indicated amount.

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