290 JOURNAL OF COSMETIC SCIENCE
Topical application of glucose may promote skin homeostasis by maintaining the
skin microbiome i.e., functioning as a prebiotic.3 It has not been clearly established,
though, a topical glucose effect on the microflora involved in skin diseases such as
Propionibacterium acnes (acne), or Malassezia spp. (folliculitis, dandruff and pityriasis)
etc. Other prebiotics have been demonstrated to inhibit the inflammation-causing
bacterium Propionibacterium acnes, in favor of beneficial species of the skin.4,5 It has
been manifested that excess glucose in hyperglycemic abscesses can significantly
enhance S. aureus virulence potential, thus worsening infection.6 Since glucose often
enters cosmetic formulations through added extracts, it is not always mentioned in the
product’s list of ingredients. In such specific applications, the existence of an analytical
method for glucose quantification in the frame of quality control may potentially be
important.
In the area of topical pharmaceuticals, the availability of a direct method to inform on
glucose content may be important for specific applications/patient groups: Although
systemic hyperglycemia prevents wound healing, topical preparations of sugar-based
compounds and honey have provided, in specific studies, promising results as a means
toward wound repair.7,8 No contribution to systemic hyperglycemia was observed.8 Another
study indicated that high glucose inhibits keratinocyte and fibroblast migration as well
as wound healing in vivo, in a concentration-dependent manner.9 Last, in the domain of
nutrition, there is a need to quantify glucose in emulsions used for the purpose of parenteral
nutrition.10
The glucose oxidase-based photometric method is commonly used for determining glucose
in solution11, either in biological fluids or food extracts, and presents the advantages of
increased specificity and ease of analysis, since several commercial kits relying on this
principle are available. A large number of alternative methods have been published
including spectrophotometric, electrophoretic and chromatographic methods.12 In general,
chromatographic methods (usually HPLC or GC) and mass spectrometry are the gold
standard methods of analysis of organic analytes in cosmetics and personal care products.13
Although specific and sensitive, they present the drawback of requiring strong analytical
expertise, expensive equipment, and typically extensive sample preparation.14 A number
of spectrophotometric methods are also commonly used such as Raman, IR, UV-Vis13,
which although simpler, still require sample preparation prior to analysis, which may
often involve toxic waste generation. Additionally, immunological analytical techniques
have been proposed for quantification of a number of analytes in cosmetics and personal
care products15, which are highly specific and sensitive, however they suffer from high
analysis cost.
We here describe a low-cost and low-waste, extraction-free, smartphone-based
methodology for in situ analysis of glucose in cosmetic formulations. The methodology
relies on the glucose oxidase (GOD)-catalyzed specific glucose conversion to a colored
product through peroxidase (POD)11 inside the cosmetic preparation. Smartphone-
based digital image colorimetry is then employed to capture, calibrate, and quantify
responses. Smartphone-based image analysis is emerging as an increasingly popular
technique, with applications over a range of domains.16 The greatest advantage of the
proposed method over existing analysis setups is the lack of a pretreatment step. The
format’s simplicity and the lack of need for analytical equipment also make the method
very attractive.
291 IN SITU ANALYSIS OF GLUCOSE IN COSMETIC FORMULATIONS
MATERIALS AND METHODS
CHEMICALS AND REAGENTS
Glucose monohydrate puriss was provided by Riedel-de Haën. The GOD/POD commercial
kit was from Biosis (Athens, Greece). The working reagent (WR) provided therein contained
a 0.2 Μ buffer pH 7.1, GOD 18 IU/ml, POD 2 U/ml, 0.25 mM 4-amino phenazone,
10 mM phenol derivative, and relevant cofactors. All cosmetics tested were commercial
products available in the local market by Cosmetic S.A. (Athens, Greece).
APPARATUS
Pictures were taken with a 48-Megapixel camera of a Samsung Galaxy S10, in a portable
white photo studio (PULUZ (China), cube of an edge of 23 cm) equipped with a single
row of LED light (at 3 cm from the edge and parallel to it). An opening exists at the
top center of the cube, where the smartphone camera is positioned. A Brookfield DV-E
digital viscometer was used for viscosity measurements. Conductivity measurements were
performed using a Cond 50 Violab conductivity meter equipped with a conductivity cell
2301T.
PREPARATION OF STANDARDS AND QUALITY CONTROL SAMPLES
Stocks and quality controls (QCs) of glucose in an O/W emulsion, a shampoo, or a gel,
were freshly prepared by dissolving a suitable amount of glucose monohydrate powder or
of a 1.0% weight per volume (w/v) glucose aqueous solution. Standards and QCs were then
processed as described below.
PROCEDURE FOR QUANTITATIVE DETERMINATION OF GLUCOSE IN AN O/W EMULSION OR GEL
The color-generating reaction was run at 25°C. For this purpose, 1.0 g of a cosmetic
product, containing varying concentrations of glucose, and 642 µL of the WR of the
GOD/POD commercial kit were added into a vial and were thoroughly mixed, manually.
Depending on the emulsion, the addition may have to be performed in aliquots to avoid
phase separation. The mixture was loaded into the wells of a microstrip, and its surface was
flattened with the flat end of a spatula. Around 5 minutes after reaction initiation and no
longer than 20 minutes after WR addition, a picture of the colored formulation was taken
with a smartphone camera. When dilution was required to lower emulsion glucose levels
down within the linear range, emulsion B (composition in Table I) was used as the diluent.
PROCEDURE FOR PICTURE CAPTURING AND ANALYSIS
A 48-megapixel mobile phone camera (lens aperture: F2.0, magnification factor, focal
length (35mm): 25.9 mm, automatic adjustment of brightness) and a white photo studio,
equipped with a single row of LEDs was used for capturing pictures of the cosmetic
samples. The camera was positioned at the opening at the center of the top side of the
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